World Cotton Research Conference-5

WORLD COTTON RESEARCH CONFERENCE-5
Renaissance Convent ion Cent re, Mumbai 7-11 November 2011

Theme: Technologies for Prosperity
www.excelpublish.com
WORLD COTTON RESEARCH CONFERENCE-5
Renaissance Convent ion Cent re, Mumbai 7-11 November 2011
Theme: Technologies for Prosperity

(Oral Presentations of WCRC-5)

Editors
Dr. K.R. Kranthi
Dr. M.V. Venugopalan
Dr. R.H. Balasubramanya
Dr. Sandhya Kranthi
Dr. Sumanbala Singh
Dr. Blaise

Organized by

International Cotton Advisory Committee,
Washington, DC, USA
Indian Society for Cotton Improvement,
Mumbai, India
Indian Council of Agriclutural Research,
New Delhi, India

EXCEL INDIA PUBLISHERS
New Delhi
Book of Papers

First Impression: 2011
Indian Society for Cotton Improvement, Mumbai
World Cotton Research Conference on Technologies for Prosperity
ISBN: 978-93-81361-51-1
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Preface
Every advance in civilization has been denounced as unnatural while it was recent
Betrand Russell
Indeed, it has now become more common than ever before, to debate, denounce but adopt changes.
And, when it happens, adoption of new technologies happens in a manner like never before. The recent
biotech advances in cotton epitomized the saga of technologies that became game-changers to emerge
victorious, unscathed, but wading through adversities.
Cotton is natures gift to mankind. The gentle fibers pass through tough mechanical rigor to become
soft fabric that clothes and makes civilizations. For more than seven thousand years, cotton has been the
best possible natures fabric and the world reveres it now. Cotton in many countries means livelihood,
employment and food security. With a current share of 36.0% in fabrics, cotton continues to remain as
the most skin friendly of all apparel available to mankind. The global cotton production in 2010 was 25.1
M tons from 34.0 M hectares. With population at 6.8 billion in 2010, the average per-capita utilization of
fibers is estimated to be 10.4 kg. The global production of fibers increased from 52.0 M tons in 2000 to
72.5 M tons in 2010 at an average growth rate of 3.3% and is expected to reach 138 M tons by 2030 with
an estimated 48.0 M tons of cotton production. The world population is estimated to increase at an annual
growth rate of 1.4% to reach 8.2 billion by 2030. It is being speculated that the declining supply of raw
materials and oil reserves in the future will constrain man-made fiber production and the demand for
cotton will increase. Even today the demand for cotton would increase if the entire production and
processing system is made more cost effective by improving yield per unit input. The cotton demand may
increase, if the petroleum reserves become a limiting factor for the production of man-made fibers.
Yields have to increase without any further area spread. Science and technologies alone will show the
way. As we move on, to cater to the needs of burgeoning population, it becomes our collective
responsibility to tread a path that is in consonance with nature. For several decades, cotton cultivation
and post-harvest processing had become input intensive and chemical dependent. Together we will
succeed as a global family only when we shall be able to discover methods of agriculture and industrial
processes that will be least disruptive to the environment and profitable for all concerned in production
and processing.
W.R. Whitney, chemist and founder of General Electric Company, said
In the advance of civilization, it is new knowledge that paves the way, and the pavement is eternal
The papers presented at the conference represent New Knowledge and reflect the trend towards
progress in science and technologies for a better future. We earnestly hope that the book of papers will be
treasured by the cotton fraternity.
Editors

Contents
Preface v
COTTON IMPROVEMENT AND BIOTECHNOLOGY
1. Genetic Diversity Analysis in Cotton Germplasm
Prafulla Naphade, Pandurang Kulkarni, Rahul Ramekar,
Ashok Jaybhaye Chandrashekhar Chaporkar, Bharat Char
and Venugopal Mikkilineni 3

2. Creating Novel Diversity and using Comprehensive Methods for
Their Further Use in Hybrid ResearchAn Exercise
in Gossypium hirsutum L.
Rajesh S. Patil, Bharathkumar, Kasu Pawar, Sudheendra Ashtaputre,
Ishwarappa Katageri, Basavaraj Khadi, Bhuvaneshwaragouda Patil,
Shreekanth Patil and Shekhar L. 8
3. New Cotton Germplasm as an Intermediate Cycle Called
SP 8 Development by the National Institute
of Agricultural TechnologyINTA
A. Tcach Mauricio, A.F. Poisson, Ivan Bonacic, Silvia Ibalo,
Alex Montenegro Daniel Ojeda

and Mariano Cracogna 13

4. Introgression of High Fibre Strength Trait
to Upland Cotton using Marker-Assisted Selection
Nallathambi Kannan, P. Selvakumar, R. Krishnamoorthy, D. Raja,
M. Bhuvaneshwari, V. Subramanian and M. Ramasami 17
5. Estimation of Genetic Parameters for Yield
and Fibre Quality Traits in Inter-Specific Crosses
of Cotton (Gossypium spp.)
Gunasekaran Mahalingam, Krishnasamy Thiyagu
and Nagasamy Nadarajan 28
6. Introgression of Desirable Characters
for Growing Cotton in Pakistan
Abid Mahmood, Jehanzeb Farooq and Noor-Ul-Islam 35
7. Temporal Changes in Metabolically Important Enzymes
and Solutes act as Trigger for Epidermal Cell Conversion
to Fibre Initials in Cotton
Gopalakrishnan N., A.H. Prakash and Y.L. Balachandran 43

8. Study of Interspecific Hybrids (Gossypium hirsutum x G. barbadense)
for Heterosis and Combining Ability
K.P.M. Dhamayanthi 51
9. Predicting F

Performance from Their Parental Charectaristics
in Upland Cotton (Gossypium hirsutum L.)
R.K. Gumber, Pankaj Rathore and J.S. Gill 56
10. Thermosensitive Genetic Male Sterility System
in Cotton (G. arboreum L.)
S.M. Palve, V. Santhy, S.R. Bhat, S. Laxman, Rajesh Patil, B.M. Khadi,
Sonali Virkhede and Priyanka Bihariya 62
11. Heterosis for Yield and Yield Attributing Traits
in Arboreum Cotton (Gossypium arboreum L.)
S.B. Lalage, N.D. Deshmukh, I.S. Halakude and J.C. Rajput 69

viii Contents
12. Multi-Level Determination for Heat Tolerance of Cotton Cultivars
Nicola S. Cottee, Michael P. Bange, Daniel K.Y. Tan, J. Tom Cothren 72
13. Genetic Variability in Single Plant Selections
for Improving Drought Tolerance in Upland Cotton
Suman B. Singh, A.H. Prakash and Amol A. Karpe 80
14. Genetic Parameters of Physiological Traits for Salinity Tolerance
in Diverse Genotypes of Cotton (Gossypium hirsutum L.
and Gossypium arbadense L.)
Hosseini Gholamhossein

and Behdarvand Pejman 85
15. Marker-Assisted Selection
for Improving Drought Resistance in Cotton
Yehoshua Saranga, Avishag Levi and Andrew H. Paterson 89
16. Tak FA8: New Jassid Tolerant Cotton Variety
P. Seburuang,W. Sirichumpan,P. Nakapan,S. Thaitad,S. Lapbunjob
A. Traisiri,N.T. Khumla,S. Areerak,S.A. Juttupornpong,N. Panlai,
A. Kasivivat,P. Sangsoda,R. Chuekittisak, B. Kumseub,
P. Pulcha and K. Khockakang 92
17. High Boll Weight and High Ginning Outturn
The Major Tools for Breaking Yield Barriers
in Gossypium arboreum
Punit Mohan, S. Manickam, S.K. Verma, D. Pathak,
A.S. Singh and Tarun Kumar Das 95

18. Development of Naturally Coloured Gossypium hirsutum
Cotton Genotypes Suitable for Textile Industry
through Genetic Improvement
Manjula S. Maralappanavar, Vikas V. Kulkarni, Somshekhar,
C. Madhura, S.S. Patil, K. Narayanan, K.J. Sanapapamma
and Jyoti V. Vastrad 100
19. Divergent Selection for Yield and Earliness
in Cotton (Gossypium hirsutum L)
P. Michalakopoulos, C. Goulas, A. Katsiotis and S. Rangasamy 104
20. Development of Recombinant Inbred Lines
for Fibre Quality Traits in Gossypium hirsutum L
Jagmail Singh, Babita Chaudhary, Preeti Srivastva,
Sapna Tiwari and Mukesh Kumar Sharma 111
21. Elite Cotton Varieties in the Zimbawean Private
Sector Research Programme
Mandiveyi Jeremiah Kudzayi 115
22. Development of Biotic Stress Resistance Transgenic
Diploid Cotton Utilizing Agrobacterium
and Shoot Apical Meristem Cells
Sukhadeo Nandeshwar, Pranjib Chakraborty, Kanchan Singh,
Mithila Meshram and Bipinchandra Kalbande 120
23. Cloning and Characterization of Cellulose Synthase Genes
from Arabidopsis thaliana
Balasubramani G., Amudha J., Sahare S. and Kranthi K.R. 129
24. Cotton Transgenic with DRE-binding Transcription
Factor Gene (DREBA) and Zinc Finger Gene (ZF) Confers
Enhanced Tolerance to Drought
Amudha J., G. Balasubramani, A.H. Prakash, Shweta C.,
K.C. Bansal and K.R. Kranthi 136
Contents ix
25. Study of Heterosis in Inter Varietal Crosses
of Asiatic Cotton (Gossypium herbaceum L)
N.N. Patel, D.U. Patel, D.H. Patel, K.G. Patel, S.K. Chandran and V. Kumar 149
26. Assessment of Genetic Diversity for Improved Fibre Quality
Traits in G. barbadense Accessions to Widen Cotton Gene Pool
Amala Balu P., D. Kavithamani and S. Rajarathinam 153
COTTON PROTECTION
27. Survival of Helicoverpa armigera on Bt Cotton Hybrids
in IndiaCan We Buy the Interpretations
A. Prabhuraj, Y.B. Srinivasa and K. Muralimohan 161
28. Field Performance of F
,
-F

and

non-Bt of BG-II
(MRC-707 Bt) and JKCH-97 Bt Against Bollworms of Cotton
G.T. Gujar, G.K. Bunker, B.P. Singh and V. Kalia 165
29. Intrinsic Rate of Increase and Life Parameters
of Cotton Leaf Eating Caterpillar Spodoptera litura
on Bollgard II Hybrids
Golla M.V. Prasada Rao, T. Sujatha, G.A.D. Grace,
N.V.V.S.D. Prasad and V. Chenga Reddy 174
30. Field Efficacy of Widestrike Bt Cotton, Expressing CryAc
and CryF Proteins, Against Lepidopteran Pests in India
Moudgal R.K., Chetan Chawda, Gajendra Baktavachalam
Sundara Rajan and Gary D. Thompson 184

31. Influence of Weather Parameters on Population
of Mealybug, Phenacoccus solenopsis and
its Natural Enemies on Bt Cotton
B.V. Patil, S.G. Hanchinal, M. Bheemanna and A.C. Hosamani 193
32. Insecticide Induced Resurgence of Mealybug,
Phenacoccus solenopsis Tinsley in Cotton
Rishi Kumar, Dinesh Swami, Vijender Pal and K.R. Kranthi 198
33. Species Diversity, Pestiferous Nature, Bionomics
and Management of Mirid Bugs and Flower Bud
Maggots: the New Key Pests of Bt Cottons
S. Udikeri, S. Kranthi, K.R. Kranthi, N. Vandal, A. Hallad
S.B. Patil and B.M. Khadi 203
34. Influence of Spatial Cropping Patterns of Cotton Cultivation
on Population Dynamics of Mirid Bug,
Creontiades biseratense (Distant)
B. Dhara Jothi, T. Sonai Rajan, V.S. Nagrare, M. Amutha,
Rishi Kumar and T. Surulivelu 210
35. Determination of Economic Injury Level for Defoliator
Spodoptera litura (Fab.) on Bt Cotton
M. Bheemanna, S. Hanchinal, A.K. Hosamani and R. Chowdary 216
36. Development of Metapopulation Approach
for Landscape-level Lygus hesperus
Management in Texas
M.N. Parajulee, R.B. Shrestha, W.O. Mcspadden and S.C. Carroll 220

x Contents
37. Survival of Pink Bollworm, Pectinophora gossypiella
(Saunders) in Bt and Non Bt Cotton In Normal
and Late Sowing with A Special Emphasize
to Avoid Population Pressure
S. Mohan and S. Nandini 229
38. Dynamics of Biotypes b and q of Bemisia tabaci
in Cotton Fields and Their Relevance
to Insecticide Resistance
A.R. Horowitz, H. Breslauer, M. Rippa, S. Kontsedalov, M. Ghanim,
P. Weintraub and I. Ishaaya 232

39. Gambit of IPM for Insect Resistant Transgenic Cotton
N.V.V.S. Durga Prasad, G.M.V. Prasad Rao and V. Chenga Reddy 239
40. Cotton Pest Management Programmes using Threshold-Based
Interventions Developed by CIRAD and its Partners
in SubSaharan African Countries
Silvie P.J., Adegnika M.A., Akantetou K.P., Ayeva B., Bonni G.,
Brevault T., Gautier C., Hma O., Houndete T.A., Ochou G., Prudent P.
Renou A. and Togola M. 244

41. Can Natural Refuges Delay Insect Resistance to Bt Cotton
Brvault Thierry
,
, Nibouche Samuel, Achaleke Joseph and Carrire Yves 255
42. Can Tomato be a Potential Host Plant for Pink Bollworm
N. Ariela, S. Harpaz Liora, R. Mario, S.

Roee and H.A. Rami 258
43. Impact of IRM Strategies on Bt Cotton in Andhra Pradesh
T.V.K. Singh, N.V.V.S.D. Prasad, S. Sharma and S. Dayakar 261
44. Efforts to Mitigate Stickiness Problem in Sudan
A. Abdelatif and E. Babiker 265

45. Present Status of Mealy Bug Phenacoccus solenopsis (Tinsley)
on Cotton and Other Plants in Sindh (Pakistan)
Khuhro S.N., A.M. Kalroo and R. Mahmood 268
46. Changing Scenario of Cotton Diseases in India
The Challenge Ahead
D. Monga, K.R. Kranthi, N. Gopalakrishnan and C.D. Mayee 272
47. Emerging and Key Insect Pests on Bt Cotton
Their Identification, Taxonomy,
Genetic Diversity and Management
S. Kranthi, K.R. Kranthi, Rishi Kumar, Dharajothi, S.S. Udikeri,
G.M.V. Prasad Rao, P.R. Zanwar, V.N. Nagrare, C.B. Naik, V. Singh
V.V. Ramamurthy and D. Monga 281
48. Efficacy of Triazoles in Management
of Major Fungal Foliar Diseases of Cotton
A.S. Ashtaputre, N.S. Chattannavar, S. Patil, Rajesh
N.K. Pawar and G.N. Hosagoudar 287
49. Damage Caused in Cotton by Different Levels of Ramulosis in Brazil
Alderi Emdio De Arajo, Alexandre Cunha De Barcellos Ferreira

and Camilo De Lelis Morello 290
50. Insecticidal Toxin Genes from Bacterial Symbiont
of Thermotolerant Isolate of Heterorhabditis indica,
Entomopathogenic Nematode
Nandini Gokte-Narkhedkar, Kanchan Bhanare, Prachi Nawkarkar,
Prashanth Chiliveri and K.R. Kranthi 293
Contents xi
51. Identification and Characterization of a Novel Source
of Resistance to Root-Knot Nematode in Cotton
Mota C. Fabiane, Giband Marc, Carneiro, D.G. Marina, Silva,
H. Esdras, Furlanetto Cleber, Nicole Michel, Barroso,
A.V. Paulo and M.D.G. Regina 298
52. Predominance of Resistance Breaking Cotton Leaf
Curl Burewala Virus (ClCuBuv) in Northwestern India
Prem A. Rajagopalan, Amruta Naik, Prashanth Katturi, Meera Kurulekar
Ravi S. Kankanallu

and Radhamani Anandalakshmi 304
COTTON PRODUCTION, PHYSIOLOGY AND ECONOMICS
53. Cotton Genotypes Performance under Rainfed
and Irrigated Conditions in two Regions of Northern Argentina
Marcelo Paytas and Jose Tarrago 309

54. The Adaptation of Irrigated Cotton to the Tropical Dry Season
S.J. Yeates 312

55. Which Carbon Footprint Tool for the Cotton Supply Chain
F. Visser, P. Dargush and C. Smith 323
56. Studies on the Seed Cotton Yield, Growth and Yield
Contributing Characters of New Bt Cotton Hybrids
under Varied Agronomic Manipulations
Kulvir Singh, Harmandeep Singh, R.K. Gumber and Pankaj Rathore 338
57. Evaluation of Cotton Genotypes for High Density Planting
Systems on Rainfed Vertisols of Central India
M.V. Venugopalan, A.H. Prakash, K.R. Kranthi, Rachana Deshmukh,
M.S. Yadav and N.R. Tandulkar 341
58. Pruning and Detopping Studies in Bt-Cotton
S.S. Hallikeri 347
59. Input use Efficiency, Productivity, Profitability and Sustainability
of Bt Cotton Based Multi Tier System with Nutrient Levels
K. Sankaranarayanan, P. Nalayini, C.S. Praharaj and N. Gopalakrishnan 350

60. Effects of Prolonged and Integrated Use
of Organics and Inorganics on the Performance of Cotton
S.N. Upperi and V.B. Kuligoud 359
61. Response of Cotton to Bio Boron and its Use Efficiency
in Vertic Ustropept Soil of Tamil Nadu, India
P. Janaki and S.

Meena 364

62. A Thermal Optimum Approach to Irrigation Scheduling
in Australian Drip Irrigated Cotton
W.C. Conaty, J.E. Neilsen, J.R. Mahan, B.G. Sutton and D.K.Y. Tan 369
63. Efficient Water Management Technology
for Sustainable Cotton Production in Central India
V. Kumar, R.G. Patil and J.G. Patel 376
64. Biodegradable Polyethylene MulchingA New Approach
for Moisture Conservation, Weed Control
and Enhanced Productivity of Winter
Irrigated Cotton-Maize System
P. Nalayini, K. Sankaranarayanan, K. Velmourougane and M. Suveetha 386

xii Contents
65. Comparative Study of Different Weeding Methods
on Cotton Crop under Drip Irrigation System
Dil Baugh Muhammad, M.N. Afzal, I. Raza and P.L. Dupont 392
66. Comparative Efficiency and Economic Viability
of Herbicides for Controlling Weeds in
Bt Cotton (Gossypium hirsutum L.)
J.G. Patel, V.C. Raj, V.P. Usadadiya, R.R. Parmar, C.M. Sutaria,
R.L. Leva and V. Kumar 396
67. Agronomic Management and Benefits
of Glyphosate Tolerant Transgenic Cotton Hybrids
C. Chinnusamy, C. Nithya, P. Muthukrishnan and S. Jeyaraman 399
68. Evaluation of Pyrithiobac Alone and in Combination
with Grassy Herbicides on Weed Control in Cotton
A.S. Rao 406
69. Defining Optimal Application Rate and Timing
of Mepiquat Chloride for Cotton Grown
in Conditions that Promote Excessive Vegetative Growth
G.D. Collins, R. Wells, R. Riar and K.L. Edmisten 410

70. Effect of Cool Conditions on Cotton Seedlings
D.K.Y. Tan, S. Ormiston, M.P. Bange and J.S. Amthor 416

71. Increased Nutrient Uptake and Salinity Tolerance
in AhCMO Tansgenic Cotton
Huijun Zhang, Jianlong Dai and Hezhong Dong 420

72. Improvement of Partial Root-Zone Soil Environment
Increases Salinity Tolerance of Cotton
Hezhong Dong, L.I. Weijiang and L.I. Zhenhuai 432
73. Do Female-Led Farms Perform Less Well
in Cotton Production? Insight from Hebei Province (China)
Michel Fok and Guiyan Wang 435

74. Debunking the Myths
J. Reed, E. Barnes and P. O'Leary 442
75. Analysis of Growth and Instability of Cotton Production in India
Anuradha Narala and A.R. Reddy 449

76. Total Factor Productivity of Cotton in Gujarat (India)
A.R. Reddy, S.M. Yelekar, R.B. Petkar and N. Anuradha 454

77. Transfer of Technology Initiatives for Profitable
and Sustainable Cotton Farming in India
An Empirical Analysis
S. Usha Rani and S.M. Wasnik 461
POST HARVEST PROCESSING
78. Enzyme/ Zinc Chloride Pretreatment of Short-Staple Cotton Fibres
for Energy Reductionduring Nano-Fibrillation
by Refining Process
N. Vigneshwaran, Vilas Karande, G.B. Hadge,
S.T. Mhaske and A.K. Bharimalla 471
79. Optimal Cotton Covered Jute, Nylon
and Metal Core Spun Yarns for Functional Textiles
Production and Characterization
S.K. Chattopadhyay, A. Yadav, V.V. Kadam, Bindu V., D.L. Upadhye,
V.D. Gotmare and A.K. Jeengar 477
Contents xiii
80. The Cotton Length Analysis using the Lengthcontrol
Iwona Frydrych, Anna Pabich and Jerzy Andrysiak 487

81. An Innovative Bio-chemical Approach
for Low Energy and Less Polluting Scouring of Cotton Textiles
P.V. Varadarajan, R.H. Balsubramanya, Nayana D. Nachane,
Sheela Raj and R.R. Mahangade 493
82. The Within Bale Repeatability of Standardized InstrumentS
for Testing Cotton Fiber Produced in Africa
E. Lukonge,

M. Aboe, Gourlot, J.P. Goz

and E. Hubl 500
83. A Vision for Technical Textiles in this Decade
A. Subramaniam 508
84. Cotton Stalk: An Additional Raw Material to Board Industry
R.M. Gurjar, P.G. Patil, A.J. Shaikh and R.H. Balasubramanya 510
85. Differential Speed Setting Facility for Roller
and Beater in Gins for Higher Ginning Rates
S.B. Jadhav and K.R.K. Iyer 524
86. Influence of Quality Attributes of Individual Bales
on Yarn Quality
R.P. Nachane 531
87. Development of an Automatic Roller Grooving Machine
for Making Helical Grooves on Rollers Used
in Roller Ginning Machines
T.S. Manoj Kumar, V.G. Arude

and S.K. Shukla 537
88. The Effect of Quarantine Treatments on the Physical Properties
of Cotton Fibresand Their Subsequent Textile
Processing Performance
M.H.J. Van, Der Sluijs, F. Berthold and V. Bulone 543

89. The Impact of Cotton Fibre Maturity on Dye Uptake & Low
Stress Mechanical Properties of the Fabric
S. Venkatakrishnan and R.P. Nachane 553
90. Exploration of Residual Hazardous Compounds on Cotton Fibers
Syed Zameer Ul Hassan and Jiri Militky 561
91. Studies on Composition of Oil and Fatty Acid
in Bt and Non Bt Cotton (G. hirsutum)
Harijan Nagappa and Khadi B.M. 568
92. The Properties of the Naturally-Pigmented Cotton Cultivated
in Nakornsawan Field Crop Research Center Thailand
Piyanut Jingjit

and Parinya Seebunruang 572
AUTHOR INDEX 577
Cotton Improvement and Biotechnology
Genetic Diversity Analysis in Cotton Germplasm

Prafulla Naphade
1
, Pandurang Kulkarni
1
, Rahul Ramekar
2
, Ashok Jaybhaye
2

Chandrashekhar Chaporkar
1
, Bharat Char
2
and Venugopal Mikkilineni
2
1
Research & Development (Cotton Breeding)
2
Research & Development (Molecular Breeding and Applied Genomics),
Maharashtra Hybrids Seeds Co. Ltd., Jalna, India
AbstractCrop germplasm diversity contributes significantly to the development of improved crop cultivars aimed at
increasing crop productivity. In this study, we have selected 192 proprietary inbred lines of Gossypium hirsutum that
show variable phenotype for traits such as leaf hair density, leaf texture, boll size, plant architecture (type), fibre
quality parameters, maturity group, and response to biotic and abiotic stresses. This germplasm pool was screened
with 54 polymorphic Microsatellite markers. It was found that 47 loci out of the 54 loci show polymorphism between
any two lines. The similarity index values ranged between 41% to 98%. Three major dendrogram clusters and twelve
minor dendrogram clusters were observed. These results suggested that there is a high degree of genetic diversity in the
cotton germplasm which was screened.
INTRODUCTION
Allelic diversity naturally present in the germplasm pool and characterization of the allelic diversity
determines the genetic diversity present in the germplasm pool. This forms the basis for continuous
evolution. Genetic diversity and the knowledge on relationship between genotypes are of great
importance for crop breeding. It creates a resource pool of alleles and enables pooling of novel alleles
and helps in creating new allelic combinations which result in creation of novel genotypes. From a
practical crop breeding perspective, understanding the genetic variability will serve as a guide to
choosing the parents from a larger pool of germplasm. Crossing individuals that are genetically distant
can result in developing superior hybrids with higher heterotic potential and hence higher yields.
Molecular level study of the genetic diversity will also help in situations where quantitative traits are
desirable and in field conditions it is difficult to evaluate the lines due to the effect of the environment on
the phenotype (Weir, 1990).
Cotton productivity and the future of cotton breeding efforts, as in many other agronomic crops, also
depend on genetic diversity of cotton gene pools. Worldwide cotton breeders and producers have
expressed concern over the narrow genetic basis of cultivated cotton germplasm that has caused a decline
in yield and quality. Globally cotton breeding programme are working with a narrow germplasm pool
thus resulting in genetic bottleneck through historic domestication events and selection
(Iqbal et al., 1997).
Assessment of the genetic diversity of cotton cultivars is essential to breeding strategies, such as the
characterization of individuals, accessions, and for the choice of parental genotypes in breeding
programs. For any meaningful plant-breeding programme, accurate determination of genetic diversity is
an essential step for an effective utilization of germplasm resources. An accurate estimation of genetic
diversity can be invaluable in the selection of diverse parental combinations to generate progenies with
maximum genetic variability and heterosis. In addition, introgression of desirable traits from diverse/wild
germplasm into the elite cultivars to broaden the genetic base is possible (Ulloa et al., 2007). Estimation
of genetic diversity based on the morphological and biochemical markers has its limitations due to
environmental variations. Molecular marker techniques on the other hand have evolved as powerful tools
for genetic diversity analysis and in establishing relationships between cultivars. Molecular genetic
techniques using DNA markers have been increasingly used to characterize and identify novel
germplasm for use in the crop breeding process (Zhang et al., 2003). A systematic assessment of genetic
resources will also help to identify the specific crosses to be made and hence decrease the number of
1
4 World Cotton Research Conference on Technologies for Prosperity
crosses to be designed in a breeding program. This will enable better utilization and management of
germplasm resources and also help enlarge the germplasm base hence removing the bottle necks in
breeding (Karp, 2002). Classification of germplasm based on the geographic regions would also be
valuable in understanding the structure of the cotton germplasm gene pools.
The development of abundant cotton SSR markers has stimulated more effort in molecular
characterization of cotton germplasm around the world (Blenda et al., 2006; Zhang et al., 2008). DNA-
based markers, microsatellite or simple sequence repeats (SSR) are co-dominant markers to assess
genome level diversity. SSR markers have been used as tools in genotype identification and variety
protection, seed purity evaluation, germplasm characterization, diversity studies, gene and quantitative
trait locus (QTL) analysis, pedigree analysis and marker assisted breeding. SSR markers have played an
important role in the dramatic progress of cotton genetics and genomics. Being both co-dominant and
multi-allelic, microsatellites are highly reproducible and informative genetic markers (Morgante et al.,
2002; Turkoglu et al., 2010). Another advantage of SSR markers is that they are highly transferable
across species especially within a genus (Saha et al., 2004). The objectives of this study are 1) to evaluate
the genetic diversity among selected cotton cultivars, and 2) to provide essential information for future
marker-assisted breeding and to facilitate a more efficient use of germplasm in cotton breeding.
MATERIALS AND METHODS
One hundred and ninety two Gossypium hirsutum germplasm accessions were included for genetic
diversity study. This germplasm is the proprietary core cotton collections developed at Maharashtra
Hybrid Seeds Co. Ltd., Jalna, India.
Flow chart illustrating the methodology

Leaf crushing was done on a paint shaker and DNA extraction was done by Silica method
(unpublished protocol). To develop a core set of polymorphic markers, we screened 278 markers across
13 elite germplasm lines and identified 54 polymorphic markers which were eventually converted into
core set of SSR markers. This core set of 54 polymorphic markers were used to screen the 192
germplasm lines.

Polym
Taq DNA
chloride, 0
and 0.5 un
(Perkin-E
DNA at 9
72 C (Ste
Fragment R
Amplified
the ABI37
Data Analy
Analyzed
amplificat
NTSYS-p
using Unw
RESULTS A
The dendr
two lines
The cluste
and add t
genetic di
useful for
using SSR
Mahyco c
distinct ge
genetic va
further ex
merase chain
A buffer (10x
0.1% gelatin
nits of Taq p
lmer/Applied
95 C for 5 m
ep 3) for 45s
Run
d PCR produ
730 genetic a
ysis
data was c
tion =9, for
pc software w
weighted Pai
AND DISCUSSI
rogram has i
was observe
enetic differ
to the impor
iversity amo
r parental sel
R markers ge
cotton germ
enetic backg
ariability for
xploitation of
G
reactions w
x contains 10
n), 0.2mM dN
polymerase.
d Biosystem
minutes, follo
After 35 cyc
uct was dilute
analyzer and
onverted to
r all 192 ac
was used fo
ir-Group Met
ON
identified 3 m
ed at 41% w
rences betwe
rtance of hy
ong cotton c
ection of div
enerated not
mplasm, but
ground for di
r majority of
f their genetic
Fig. 1: Chromat
Genetic Diversit
were performe
00 mM Tris
NTPs, 5 pico
Temperature
ms). The am
owed by 35 c
cles, the fina
ed to 1:15 ra
d allele callin
a binary fo
ccessions. T
or cluster an
thod Arithm
major and 12
while the hig
een selected
ybridization
cultivars. Th
verse plants f
t only essent
also provide
iversifying co
f the traits ca
c potential.
togram File Gener
ty Analysis in C
ed in 15-l v
- pH 9.0, 50
o moles of fl
e cycling wa
mplification p
cycles of 94
al extension t
atio with dist
g was done b
ormat, where
The Numeric
nalysis. Simi
metic Average
2 minor clust
ghest similari
genotypes, a
as well as b
e findings o
for cotton br
ial informati
ed a useful
otton breedin
an be used in
ated by ABI3730 9
otton Germplas
volumes con
00 mM Potas
luorescent la
s conducted
profile consi
C (Step 1) f
temperature o
tilled water.
by GeneMap
e, band pres
cal Taxonom
larity matrix
e (UPGMA).
ters (Fig.2).
ity between
as they each
back-crossin
on genetic re
eeding. The
ion for unde
guide for s
ng program.
n a future cot
96 Well Capillary E
sm
ntaining 40ng
ssium chlorid
abeled forwa
on a GeneA
isted of an
for 30s, 55 C
of 72 C was
Fragment an
pper software
ent = 1, ban
my Multivar
x and dendro
.
The lowest
any two line
h form a sep
ng as an effe
elationship a
characteriza
erstanding ge
selecting spe
The promisi
tton breeding
Electrophoresis
g of template
de, 15 mM m
ard and rever
Amp PCR Sy
initial denat
C (Step 2) fo
s held for 6 m
nalysis was d
e (Figure 1).
nd absent =
riate Analys
ogram were
similarity be
es was 98%
arate cluster
ective tool i
and distinctiv
ation of cotto
enetic divers
ecific germp
ing cultivars
g to design h

5
e DNA, 1x
magnesium
rse primers
ystem 9700
turation of
or 45s, and
minutes.
done using

0 and no
sis System
generated
etween any
(Table 1).
r or group,
in creating
veness are
on cultivars
ity of elite
plasm with
s with high
hybrids for
6
1
1 1.00
2 0.81
3 0.82
4 0.75
5 0.88
6 0.83
7 0.51
8 0.89
9 0.74
10 0.74
11 0.90
12 0.82
13 0.82
14 0.80
15 0.82
16 0.84
17 0.86
18 0.82
19 0.78
CONCLUSIO
This study
that there
potential.
characters
genotypes
correlated
associatio
2 3

1.00
0.84 1.00
0.80 0.80 1
0.88 0.83 0
0.80 0.83 0
0.76 0.77 0
0.82 0.84 0
0.87 0.80 0
0.88 0.84 0
0.79 0.79 0
0.85 0.91 0
0.89 0.85 0
0.88 0.81 0
0.85 0.92 0
0.86 0.82 0
0.86 0.90 0
0.76 0.79 0
0.85 0.78 0
ON
y shows that
is considera
The genetic
s using withi
s can be use
d with phen
on studies wh
World Cot
TABL
4 5 6

.00
.75 1.00
.74 0.87 1.0
.64 0.70 0.72
.79 0.86 0.8
.79 0.84 0.8
.86 0.83 0.7
.80 0.86 0.7
.82 0.84 0.82
.85 0.84 0.8
.88 0.84 0.8
.80 0.84 0.84
.73 0.91 0.84
.80 0.90 0.8
.70 0.83 0.7
.74 0.85 0.7
t considerab
able scope fo
c diversity da
in cluster don
d for hybrid
otypic diver
hich will enab
tton Research C
E 1: SAMPLE SIMILARIT
7 8

0
2 1.00
6 0.54 1.00
5 0.68 0.83
3 0.71 0.84
9 0.53 0.90
2 0.71 0.83
1 0.62 0.86
1 0.68 0.82
4 0.73 0.85
4 0.66 0.81
7 0.71 0.83
7 0.63 0.85
7 0.58 0.80
ble variability
for developm
ata also help
nor or with m
d developmen
rsity analysi
ble us to iden
Conference on T
Fig. 2
TY MATRIX GENERATED
9 10

1.00
0.86 1.00
0.74 0.82 1
0.82 0.83 0
0.80 0.79 0
0.83 0.89 0
0.78 0.80 0
0.85 0.76 0
0.80 0.76 0
0.78 0.75 0
0.82 0.84 0
y exists in th
ment of super
ps in improv
maximum si
nt with dive
is may lead
ntify the mar
Technologies fo
BY NTYSY SPC SOF
11 12 1

1.00
0.79 1.00
0.77 0.85 1.0
0.84 0.82 0.8
0.78 0.84 0.8
0.75 0.88 0.8
0.80 0.88 0.9
0.80 0.80 0.7
0.79 0.87 0.7
he proprietar
rior cotton li
ement of sel
milarity. On
rse gene poo
d to the dev
rker-trait ass
or Prosperity
FTWARE
3 14 15

00
85 1.00
87 0.82 1.00
88 0.78 0.85
90 0.82 0.90
76 0.75 0.76
77 0.84 0.78
ry cotton ger
ines and hyb
lected lines f
n the other ha
ol. Also, gen
velopment o
ociations at t
16 17

0
5 1.00
0 0.92 1.00
6 0.73 0.80
8 0.82 0.83
rmplasm wh
brids with hi
for specific
and genetical
netic diversit
f haplotype
the germplas

18 19

1.00
0.74 1.00
hich shows
igher yield
agronomic
lly distinct
ty analysis
maps for
sm level.
Genetic Diversity Analysis in Cotton Germplasm 7

Fig. 3: Chromatogram File Generated by ABI3730 96 Well Capillary Electrophoresis
REFERENCES
[1] Blenda, A., Scheffler J., Scheffler B., Palmer M., Lacape J. M., Yu J. Z., Jesudurai C., Jung S., Muthukumar, S.,
Yellambalase, P., Ficklin, S., Staton, M., Eshelman, R., Ulloa, M., Saha, S., Burr, B, Liu, S., Zhang, T., Fang, D., Pepper,
A., Kumpatla, S., Jacobs, J., Tomkins, J., Cantrell, R., and Main, D. (2006). CMD: a Cotton Microsatellite Database
resource for Gossypium genomics. BMC Genomics 7:132
[2] Iqbal, M.J., Aziz, N., Saeed, N.A., Zafar, Y., Malik, K.A. (1997). Genetic diversity evaluation of some cotton varieties by
RAPD analysis. Theor. Appl. Genet. 94: 139-144.
[3] Karp, A. (2002). The new genetic era: will it help us in managing genetic diversity? In: Managing
[4] Plant Genetic diversity. (Eds.): J.M.M. Engels, V.R. Rao, A.H.D. Brown and M.T. Jackson.
[5] International Plant Genetic Resources Institute, Rome, Italy, 43-56.
[6] Krishnasamy Thiyagu, Narayanan Manikanda Boopathi, Nagasamy Nadarajan, Ayyanar Gopikrishnan, Pandi Selvakumar,
Santoshkumar Magadum and Rajasekar Ravikesavan. (2011) Sampling and exploitation of genetic variation exist in locally
adapted accessions using phenotypic and molecular markers for genetic improvement of cotton. Genecon. 10: 129-153.
[7] Morgante, M, Hanafey, M. and Powell, W. (2002). Microsatellites are preferentially associated with nonrepetitive DNA in
plant genomes. Nat. Genet. 30: 194-200
[8] Saha, S., Wu, J., Jenkins, J.N., McCarty, J.C. Jr, et al. (2004). Effect of chromosome substitutions from Gossypium
barbadense L.3-79 into G. hirsutum L. TM-1 on agronomic and fiber traits. J. Cotton Sci. 8: 162-169.
[9] Turkoglu, Z., Bilgener, S., Ercisli, S., Bakir, M., et al., (2010). Simple sequence repeat-based assessment of genetic
relationships among Prunus rootstocks. Genet. Mol. Res. 9: 2156-2165.
[10] Ulloa, M., Brubaker, C. and Chee, P. (2007). Cotton. In: Genome Mapping & Molecular Breeding (Kole C, ed.). Vol. 6.
Technical Crops Springer, New York.
[11] Weir, B. S. (1990). Genetic data analysis: methods for discrete population genetic data. Sinauer Associates, Inc. publishers.
Sunderland, Massachusetts. 377.
[12] Zhang, Y., Wang, X.F., Li, Z.K., Zhang G.Y. and Ma Z.Y, (2011). Assessing genetic diversity of cotton cultivars using
genomic and newly developed expressed sequence tag-derived microsatellite markers. Gen. Mol. Res. 10 (3): 1462-1470.
Coefficient
0.01 0.07 0.14 0.20 0.26
100MW
1
122
111
119
190
11
100
127
103
105
136
145
32
99
163
156
102
161
74
98
5
88
113
133
134
148
132
191
131
6
135
8
44
125
165
117
143
164
53
166
40
48
182
169
115
108
150
118
140
141
104
121
178
97
157
186
106
107
129
158
Creating Novel Diversity and using Comprehensive

Methods for Their Further Use in Hybrid Research
An Exercise in Gossypium hirsutum L.
Rajesh S. Patil, Bharathkumar, Kasu Pawar, Sudheendra Ashtaputre,
Ishwarappa Katageri, Basavaraj Khadi, Bhuvaneshwaragouda Patil,
Shreekanth Patil and Shekhar L.
UAS, Dharwad, India
AbstractCotton breeding is a continuous endeavour aiming to produce better genotypes and hybrids. The present
exercise involved choosing the F
1
hybrids, from national trials, as parents and then employing methods to assess the
diversity produced in the F
5
segregants leading to identification of elite lines which can be used in further hybrid
research. The two parts of study spanning a period of five years began in 2007-08 and was initiated with an objective
to isolate superior Gossypium hirsutum genotypes related to yield and fibre properties from double crosses whose F
1

parents were chosen for their diversity and superior traits. Segregants from a three-way cross and also the respective
single cross parents of double crosses were included in the study. In all there were 115 lines drawn from five double
cross, one three-way cross and six single cross hybrids in F
5
which were evaluated in an augmented design during
kharif of 2010-11.
Five genotypes viz., Line-632, Line-131, Line-642, Line-1151 and Line-1101 had better yields ranging from 8.90
to 21.77 per cent over best check Sahana with mean yields higher than 20.27q/ha. Line-632 had the highest seed cotton
yield of 21.70 q/ha which was 21.77 per cent better than Sahana (17.82 q/ha). It also had superior fibre length.
In second part of the study, the diversity generated was assessed through K-means clustering. Seven clusters were
formed accommodating the 115 lines. The second step was to employ a simple method called Path-of-productivity
analysis to identify the different paths the top 12 lines took towards producing higher yields. As expected, they did
have differences in their paths to higher yield attributable to their differential genetic makeup. In addition, these 12
genotypes fell in five different clusters identified in the previous step. Considering both tests, 10 genotypes were finally
identified to be included in a diallel to pave way for hybrid research. Lines- 632, 131, 642, 1151, 11101, 1081, 531,
391, 8141, and 12111 were the chosen genotypes.
INTRODUCTION
Genetic diversity is at the heart of all plant breeding activities. Crossing over and recombination among
the chromosomes of a heterozygote leads to the formation of genetically dissimilar gametes. Such
gametes of two heterozygotes can be brought together when we use F
1
hybrids as parents of a double
cross. Creating and harnessing novel genetic diversity through such conventional means is one method of
obtaining superior segregants. In the present study, the F
1
hybrids which served as parents of the double
crosses were chosen from the different cotton growing zones of India in the hope that geographical
diversity would contribute to the diverseness of the hybridization material. Greater the genetic diversity
better would be the release of variability in the segregants. In the later generations (say F
4
/F
5
), where
these desirable segregants are fairly stabilized, they can be evaluated against checks. Productive
segregants are isolated in each generation via individual plant selection. After extensive yield
performance trials, the new genotypes can be released as new varieties. Freom here starts the next
activity. The genetic variability created can be harnessed for heterosis breeding. The new genotypes can
be subjected to diversity analysis and diverse groups can be identified from which genotypes can be
picked for hybridization. A method called Path-of-Productivity has been described and now, can be
used in conjunction with diversity analysis to identify genotypes that can serve as parents of new hybrids.
The parents can be brought together in a diallel cross to identify superior hybrids. These hybrids will
again help in embarking upon a fresh cycle of recombination and creation of diversity.

2
Creating Novel Diversity and using Comprehensive Methods for Their Further Use in Hybrid 9
Six intra-hirsutum hybrids of cotton were identified from the All India Coordinated Crop Improvement
Project trials during 2005-06 and were used as parents in producing double crosses and a three-way cross
in 2006-07. From 2007-08 onwards, individual plant selections were made based on productivity and
fibre properties in each generation till 2009-10. One hundred and fifteen plants belonging to different
crosses were identified in 2009-10 for evaluation during 2010-11. These hundred and fifteen genotypes in
F
4
/F
5
were obtained through individual plant selection (IPS) from five double crosses, one three way
cross and six single cross (parents) hybrids. The details are given in Tables 1 and 2. These 115 genotypes
were evaluated in augmented design with five check varieties during kharif 2010-11 at Agricultural
Research Station, Dharwad to identify productive genotypes. Analysis of variance (ANOVA) for
augmented designII (Federer, 1977) for all characters was carried out separately. Parameters based on
the mean performance of the varieties and also parameters of genetic variability for the different traits
were obtained. GCV and PCV values were calculated as per Burton (1951) and heritability (broad sense)
was obtained as per Johnson et al., (1955). Selection efficiency reflected in genetic advance and GAM
was assessed as per Johnson et al., (1955). In the present study, a simple method called Path-of-
productivity (Rajesh Patil et al., 2007), used earlier in arboreum cotton with some degree of success, has
been outlined which helps in finding out differences in the trait contributions to the final yield of a
genotype. If two such genotypes with different paths to productivity are hybridized one can expect hybrid
vigour as there could be underlying genetic differences responsible for their differing path-of-
productivity. As an adjunct to this, conventional genetic diversity analysis can be done to decide upon the
genotypes to be chosen as parents in a hybridization program. Diversity generated was assessed through
K-means clustering using Systat software. The most productive 12 lines were considered for path-of-
productivity analysis. The 10 lines, selected after the path-of-productivity analysis, were allocated to
their respective clusters to see if they fell in diverse clusters. Together, the methods can identify parents
amenable to a hybridization program.
TABLE 1: HYBRIDS FROM AICCIP TRIALS AND THEIR PERFORMANCE FEATURES ACROSS THE THREE COTTON GROWING ZONES OF INDIA DURING 2005-06
THAT SERVED AS PARENTS OF THE DOUBLE CROSSES
Hybrid North Zone (6 Locations) Central Zone (7 Locations) South Zone (6 Locations)
S
e
e
d

C
o
t
t
o
n

Y
i
e
l
d

(
k
g
/
h
a
)

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i
b
r
e

L
e
n
g
t
h

(
m
m
)

F
i
b
r
e

S
t
r
e
n
g
t
h

(
g
/
t
e
x
)

S
:
L

R
a
t
i
o

S
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d

C
o
t
t
o
n

y
i
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l
d

(
k
g
/
h
a
)

F
i
b
r
e

L
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n
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t
h

(
m
m
)

F
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e

S
t
r
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(
g
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S
:
L

R
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C
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t
t
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y
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d

(
k
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F
i
b
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l
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(
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s
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n
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(
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S
:
L

r
a
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o

GSHH-2201 1284 26.40 20.40 0.77 2060 26.90 21.40 0.80 2127 30.20 23.00 0.76
VBCH-2312 1669 30.30 21.90 0.72 1808 30.80 24.10 0.78 1988 29.70 24.40 0.82
CHATRAPATHI 1148 33.10 25.70 0.78 1977 33.30 25.40 0.76 1882 32.80 22.90 0.70
BCHH-1232 1430 31.30 22.20 0.71 2046 29.80 22.70 0.76 2235 32.00 22.50 0.70
JKCH-2022 1228 29.60 22.10 0.75 2103 31.10 22.80 0.73 2709 32.10 22.80 0.71
RATNA 1265 29.70 20.60 0.69 1970 32.70 24.00 0.73 2056 29.50 24.50 0.83
Note: S:L ratio is the fibre strength to length ratio, a combined parameter to judge fibre property
TABLE 2: LIST OF COTTON GENOTYPES DERIVED FROM DOUBLE AND SINGLE CROSS HYBRIDS INCLUDED FOR EVALUATION AT ARS DHARWAD DURING KHARIF 2010-11
Entry No F4 progeny of Cross Progenies Entry No F5 Progeny of Cross Progenies
Double Cross Hybrids Single Cross Hybrids
DC-1. GSHH-2201 RATNA 10 DC-7 GSHH-2201 6
DC-2. VBCH-2312 RATNA 3 DC-8 VBCH-2312 13
DC-3. CHATRAPATHI RATNA 17 DC-9 CHATRAPATHI 9
DC-4. BCHH-1232 RATNA 4 DC-10 BCHH-1232 8
DC-5. JKCH-2022 RATNA 11 DC-11 JKCH-2022 14
Three-way Cross Hybrids DC-12 RATNA 12
DC-6. RCR 4 x RATNA 7
Note: Altogether, a total of 115 progenies/genotypes were evaluated.

RESULTS AND DISCUSSION
The genotypes were evaluated in augmented design for productivity traits and also for fibre properties.
The ANOVA revealed that the variability generated in the experimental material across all the traits was
larger. The various genetic parameters have been given in Table 3. The top five genotypes viz., DC-632,
DC-131, DC-642, DC-1151 and DC-1101 were superior to the zonal and local check, Sahana, in both
yield as well as fibre properties. The performance of selected superior genotypes against the two released
check varieties across seed cotton yield and fibre traits has been given in Table 6. Genotypes DC-632
(2170 kg/ha), DC-131 (2064 kg/ha) and DC-642 (1993 kg/ha) were superior to checks Sahana (1782
kg/ha) and RAH-100 (1457 kg/ha). Superiority in fibre length (23.93 % over Sahana in DC-391) and
fibre strength (20.08 % over RAH-100 in DC-11101) has been recorded. Genotype DC-632 apart from
having a yield superiority of 17.89 per cent over best check was also superior for the fibre properties.
Another genotype DC-771 had a fibre length of 31.30 mm and strength of 25.50 g/tex.
TABLE 3: VARIABILITY PARAMETERS FOR DIFFERENT MORPHOLOGICAL CHARACTERS AMONG SINGLE AND DOUBLE CROSS DERIVED LINES AT ARS DHARWAD DURING KHARIF 2010-11
V
a
r
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(
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S
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a
t

5
0
%

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p
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(
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(
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(
%
)

H
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(
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C
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(
g
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)

Mean 97.20 2.10 22.10 49.40 24.00 7.30 1.20 4.40 4.40 27.70 8.70 4.70 35.30 29.10 19.30
Maximum 140.50 4.00 37.00 70.00 39.00 10.00 1.90 13.10 7.30 35.80 10.00 6.10 39.60 37.80 39.10
Minimum 70.00 0.90 15.40 33.00 17.40 3.80 0.70 1.10 1.70 20.40 6.50 3.40 30.20 21.10 4.30
Vg 105.87 0.06 4.81 17.25 5.85 0.22 0.01 0.49 0.01 1.41 0.48 0.23 2.79 7.55 23.18
Vp 160.68 0.38 11.65 46.85 12.20 0.95 0.08 2.86 0.62 7.77 0.72 0.40 4.86 8.65 59.80
PCV 13.04 29.28 15.44 13.86 14.55 13.37 24.01 38.46 17.95 10.07 9.74 13.52 6.25 10.11 40.07
GCV 10.59 11.86 9.92 8.41 10.08 6.37 9.13 15.84 2.38 4.28 7.95 10.23 4.73 9.44 24.94
hbs(%) 65.89 16.40 41.30 36.82 47.95 22.69 14.46 16.98 1.76 18.11 66.57 57.18 57.46 87.22 38.76
GA (%) 17.20 0.21 2.90 5.19 3.45 0.46 0.09 0.59 0.03 1.04 1.16 0.75 2.61 5.29 6.17
GAM (%) 17.70 9.89 13.14 10.51 14.38 6.25 7.15 13.45 0.65 3.76 13.36 15.93 7.39 18.16 31.99
TABLE 4: PATH-OF-PRODUCTIVITY ANALYSIS IN THE 12 MOST PRODUCTIVE GENOTYPES OF THE 115 NEW GENOTYPES PRODUCED AND EVALUATED
Mean Values of 12 Most Productive Genotypes for 16 Traits
Genotypes 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
DC-632 39.07 105.10 3.00 22.80 24.40 50.00 28.80 8.80 1.50 6.30 6.20 3.60 1.50 32.60 34.84 48.26
DC-131 37.15 82.90 1.50 16.80 17.80 47.20 24.60 7.20 1.44 13.10 2.84 14.90 2.20 34.80 34.17 34.70
DC-642 35.88 106.60 1.00 18.90 21.30 54.00 27.80 7.80 1.30 7.32 4.90 4.30 1.70 32.90 34.91 60.95
DC-1151 35.47 99.50 2.20 21.00 23.00 51.00 27.00 7.20 1.00 6.20 5.72 7.40 4.20 33.80 43.69 22.26
DC-1101 34.95 101.30 2.40 19.20 20.20 50.20 24.80 8.00 1.56 6.70 5.22 14.50 2.30 34.50 37.54 51.30
DC-11101 31.65 103.50 2.10 23.40 25.40 51.00 30.00 6.60 1.10 5.70 5.55 7.60 5.20 34.80 37.71 16.26
DC-1081 31.47 92.30 2.40 19.40 21.40 47.00 28.00 7.80 1.10 5.70 5.52 1.90 1.10 35.20 37.12 34.90
DC-531 31.00 86.40 2.20 16.10 18.30 46.00 30.00 7.60 1.10 9.20 3.37 17.60 4.00 31.70 38.24 43.00
DC-391 30.95 82.40 1.80 22.00 23.00 49.20 31.00 7.30 1.78 6.80 4.55 18.70 4.00 34.80 34.79 54.00
DC-8141 30.93 95.10 3.30 19.60 22.00 49.00 29.60 7.70 1.10 6.50 4.76 3.70 1.30 34.10 38.10 114.62
DC-1131 30.39 92.80 1.80 20.00 22.00 48.00 29.00 8.10 1.00 8.20 3.71 15.10 4.10 34.70 38.70 23.78
DC-12111 29.80 108.00 2.10 23.60 25.80 48.00 29.00 6.70 0.80 5.70 5.23 9.30 5.20 38.00 36.78 44.90
Group Mean 33.23 96.33 2.15 20.23 22.05 49.22 28.30 7.57 1.23 7.29 4.80 9.88 3.07 34.33 37.22 45.74
Overall Mean 19.21 97.15 2.22 22.10 23.96 49.45 28.16 7.29 1.22 4.39 4.36 9.32 2.81 34.83 37.10 52.07
The per se performance and per cent deviation values of the top 12 genotypes from the overall mean
for all traits have been given in Table-4&5. Twelve genotypes were considered for path-of-productivity
analysis as the mean seed cotton yield of these 12 genotypes was higher than the two checks. The group
mean of the 12 genotypes was higher than the overall mean for 60 per cent of the traits. Important traits
like seed cotton yield, number of bolls, boll weight and photosynthesis had above average expression.
Negative but desirable expression was seen in plant height, number of monopodia and length of
sympodium at 50 per cent plant height. The per cent deviations across the contributing traits, showed
differences among the genotypes. These differences can safely be assumed to be arising out of genetic
Creating Novel Diversity and using Comprehensive Methods for Their Further Use in Hybrid 11
differences among the lines. All the 12 genotypes were high yielding but had different paths to
productivity owing to differential gene architecture. The differences among the path to productivity of the
12 genotypes, when 2 lines are compared against each other at a time, shows that lines DC-1101 and DC-
1131 showed less than 50 per cent difference with other lines. Both these lines can be conveniently
dropped from any hybridization programme. The other 10 lines viz., DC-391, DC-531, DC-642, DC-131,
DC-1151, DC-1081, DC-11101, DC-632, DC-12111 and DC-8141 can be used to set up a diallel
crossing set which will help ultimately to identify superior hybrids. The line DC-632 can be crossed to
any of these three lines viz., DC-131, DC-1081 or DC-531 as all the three pairs of parents showed more
than 60 per cent trait difference between the parents of the cross. Using the path-of-productivity can
thus lead to proper choice of parents for a planned production of hybrids.
TABLE 5: MEAN DEVIATIONS OF TOP GENOTYPE VALUES FROM OVERALL MEAN ACROSS ALL TRAITS
Genotypes 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
DC-632 103.38 8.18 35.14 3.17 1.84 1.11 2.27 20.71 22.95 43.51 42.24 -61.37 -46.62 -6.40 -6.09 -7.32
DC-131 93.39 -14.67 -32.43 -23.98 -25.71 -4.55 -12.64 -1.23 18.03 198.41 -34.96 59.87 -21.71 -0.09 -7.90 -33.36
DC-642 86.78 9.73 -54.95 -14.48 -11.10 9.20 -1.28 7.00 6.56 66.80 12.39 -53.86 -39.50 -5.54 -5.90 17.06
DC-1151 84.64 2.42 -0.90 -4.98 -4.01 3.13 -4.12 -1.23 -18.03 41.23 31.21 -20.60 49.47 -2.96 17.76 -57.26
DC-1101 81.94 4.27 8.11 -13.12 -15.69 1.52 -11.93 9.74 27.87 52.62 19.64 55.58 -18.15 -0.95 1.19 -1.48
DC-11101 64.76 6.54 -5.41 5.88 6.01 3.13 6.53 -9.47 -9.84 29.84 27.35 -18.45 85.05 -0.09 1.64 -68.78
DC-1081 63.82 -4.99 8.11 -12.22 -10.68 -4.95 -0.57 7.00 -9.84 29.84 26.63 -79.61 -60.85 1.06 0.05 -32.98
DC-531 61.37 -11.07 -0.90 -27.15 -23.62 -6.98 6.53 4.25 -9.84 109.57 -22.72 88.84 42.35 -8.99 3.07 -17.42
DC-391 61.11 -15.18 -18.92 -0.45 -4.01 -0.51 10.09 0.14 45.90 54.90 4.39 100.64 42.35 -0.09 -6.23 3.71
DC-8141 61.01 -2.11 48.65 -11.31 -8.18 -0.91 5.11 5.62 -9.84 48.06 9.14 -60.30 -53.74 -2.10 2.70 120.12
DC-1131 58.20 -4.48 -18.92 -9.50 -8.18 -2.93 2.98 11.11 -18.03 86.79 -15.00 62.02 45.91 -0.37 4.31 -54.33
DC-12111 55.13 11.17 -5.41 6.79 7.68 -2.93 2.98 -8.09 -34.43 29.84 19.91 -0.21 85.05 9.10 -0.86 -13.77
Group Mean 72.96 -0.85 -3.15 -8.45 -7.97 -0.47 0.50 3.80 0.96 65.95 10.02 6.04 9.13 -1.45 0.31 -12.15
Note: Group mean is of 12 genotypes and overall mean is of 115 genotypes
To make this simple test for diversity assessment more comprehensive, the conventional cluster
analysis through K-means was also performed. The cluster details are presented in Table-7. The ten
genotypes picked up on the basis of path-of-productivity analysis fell in 5 different clusters showing
their diverse genetic make-up. This analysis also proves the genetic diversity existing among the ten lines
which can be used in a hybridization set-up based on the path-of-productivity analysis. The parents of
each of the three pairs of crosses suggested above also belonged to different clusters making them ideal
parents for a heterotic cross.
INDEX FOR THE 16 DIFFERENT TRAITS
1 Seed Cotton Yield
(g/plant)
5 Number of Nodes Per
Plant
9 Stem diameter (cm) 13 Transpiartion Rate
(mol of H2OmS )
2 Plant height(cm) 6 SL at 50% plant
height(cm)
10 Number of bolls 14 Leaf temperature (0c)
3 Monopodia per
plant
7 Angle of sympodium
at 50% plant height
(deg)
11 Boll weight (g) 15 Chlorophyll
content(mg/gm fresh
weight) of leaf)
4 Sympodia per plant 8 Inter boll distance (cm) 12 Photosynthesis
(mol of CO
2
m S )
16 RWC (%)
TABLE 6: PERFORMANCE SUPERIORITY OF SELECTED GENOTYPES OVER TWO CHECKS ACROSS YIELD AND FIBRE PROPERTIES
% Improvement over Sahana % Improvement over RAH-100 Seed Cotton
Yield (kg/ha) Seed Cotton
Yield
Fibre
Length
Fibre
Strength
Seed Cotton
Yield
Fibre
Length
Fibre
Strength
For Both Seed Cotton Yield and Fibre Properties
DC-632 17.89 13.59 8.48 32.83 9.06 11.16 2170
DC-642 10.56 13.59 8.48 26.87 9.06 11.16 1993
For Fibre Properties Only
DC-11101 -1.34 10.70 17.67 17.09 6.02 20.08 --
DC-391 -3.50 23.93 11.26 15.22 19.90 13.85 --
Mean values of checks Sahana RAH-100
1782 kg/ha 26.70 mm 20.50 g/tex 1457 kg/ha 28.10 mm 19.90 g/tex --

TABLE 7: THE SEVEN CLUSTERS SHOWING THE DIVERSITY OF THE TEN GENOTYPES SELECTED ON THE BASIS OF PATH-OF-PRODUCTIVITY
Clusters Number of Genotypes Genotypes Selected on the Basis of Path-of-Productivity
I 20 DC-391, DC-531
II 20 DC-642
III 18 DC-131, DC-1151, DC-1081, DC-11101
IV 26 DC-632, DC-12111
V 5 --
VI 10 DC-8141
VII 16 --
REFERENCES
[1] Burton, G.W., (1951). Quantitative inheritance in pearlmillet (Pennisetum glaucum S. and H.). Agron., 43:404-417.
[2] Federer, W. T., (1977). Experimental design; Theory and Application. McMillan, New York.
[3] Johnson, H.W., Robinson, H.F. and Comstock, R.E. (1955). Estimates of genetic and Environmental Variability in soybean.
Agron., 47:314-318.
[4] Rajesh, S. Patil, Shreekant S. Patil, Rashmi, Bhuvaneshwargouda, R. Patil and Khadi, Basavaraj M. (2007). Path-of-
Productivity A method to handle genetic material using F1s in cotton (Gossypium arboreum L.). Proceedings of World
Cotton Research Conference 4, Lubbock, Texas, USA.

New Cotton Germplasm as an Intermediate Cycle

Called SP 48114 Development by the National
Institute of Agricultural TechnologyINTA
A. Tcach Mauricio, A.F. Poisson, Ivan Bonacic, Silvia Ibalo, Alex Montenegro,
Daniel Ojeda

and Mariano Cracogna

Estacin Experimental Agropecuaria INTA Senz Pea, Chaco, Argentina
AbstractCotton in Chaco, Argentina, is grown under rainfed conditions. The availability of water during flowering
determines the retention of fruiting structures. Better performance depends mainly on water availability during the
reproductive phase. In varieties with short cycle, the losses due to stress are the most and is becoming necessary to
develop varieties as intermediate types helps to compensate losses due to stress at flowering. The objective of this
investigation was to select cotton with intermediate habit and agronomic traits equivalent to early types. From the F2
populations of a cross between lines SP 99138 x SP 99035 in 1994/1995 season, selection was made by visual
observation and an individual plant was obtained. The selection plant was named as SP48114 after following pedigree
method of breeding. During 2004/2005 the elite population was part of a network regional comparative trials
conducted at 4 locations for 3 seasons. From the F3 generation onwards it was tested for ginning and the progenies
were artificial infected with Xanthomonas axonopodis pv malvacearum. All susceptible plants were discarded. F8
generation was tested for the blue disease caused by cotton leaf roll dwarf virus (CLRDV) through artificial infection
and only resistance lines were selected. The line SP 48114 is characterized by greater differentiation of fruiting points
on the main stem 5 % more than Guazuncho 3 INTA. This commercial variety has short cycle and high boll retentions
in first fruiting branches. The selected line SP 48114 maintains boll retentions in the inferior part of the plant similar
to Guazuncho 3 INTA and continued to flower for more days. This feature increases the flowering period for about 10
days and improves the compensation at time of water stress. The fiber parameters viz., fiber length is 29 mm, strength
31 g / tex and lint percentage about 39 to 40.
INTRODUCTION
In Argentina, the main province of cotton-growing area is Chaco, where cotton is grown only under
rainfed conditions (SAGPyA, 2009). The precipitation in this area is erratic and irregular during growing
season, which increases the risk in the production. It necessiates the research work in the Argentinean
cotton industry for improvement in water use efficiency (Payta 2010).
The cotton has xerophytic adaptation, however 53% of the area is cultivated under irrigated
conditions in the world (Hearn, 1994). When the cotton plant cross dry conditions, its vegetative grows is
terminate, being very difficult to restart vegetative growth and produce more squares and flowers
(Hearn 1994). Fryxel (1986) observed various strategies in wild species, of adaptations to arid conditions,
life cycle being one of them.
The main objectives of INTAs programme for cotton breeding is development of varieties with short
cycle (Royo et al., 2007). Sekloka et al., 2007 found that varieties with short cycle showed better
performance in dry conditions as it may run away to dry period. The problem is that typical sowing date
may not run away to dry period. The varieties with short cycle can cluster the flowering and compensate
the eventual loss when its peak phase coincides with the stress. Further, flowering can be maintained for
more time in varieties with intermediates cycle, without any losses and hence such varieties are to be
developed. The objective in this investigation was to select a cotton line with intermediate cycle and
agronomic traits equivalent to early materials.

3
The Argentine breeding programme follows the classical pedigree method (Allard 1960; Poisson 2005).
From the cross of two different genotypes, the following generations were selfed and individual plants
and progeny row selections were carried out from the F2 to successive generations. From the F4
generation onward, the seeds were not selfed and were collected to carry out replicated evaluation trials.
In the AES Saenz Pea the germplasm lines were evaluated until the F6 generation. The previous
generations were inoculated with the pathogens causing bacterial blight. The F8 were infected with
aphids which are vector for virus causing blue disease. After the F7 generation, selected lines were
evaluated at four additional localities viz., El Colorado, Reconquista, Colonia Benitez (dryland); and
Santiago del Estero (irrigated). Current commercial cultivars were used in all trials for comparison
purposes. Several trials in four locations at 3 seasons were conducted. The complete process is shown in
the picture No. 1. Trials were planted as a randomized complete block design with four replications in
plots of two rows, 10 m long. Plants were separated at a distance of 10 cm. in the row and at one m.
between rows. Bolls were hand-picked in each plot to determine the yield. Thirty selected bolls were
used to determine GOT by baby ginner lint turnout and using HVI equipped for fiber parameters.
In the last season in 1 meters of row, numbers of fruiting branch and plant height in several trials
were also studied. The dates were analysed with Infostat software and averages were separated with test
LSD Fisher.
The flow chart for the line development from the F2 populations, the plant that made the progeny row
and successive testing generations is show in Fig 1. This material with intermediate cycle finished the
process of test in the 2006-2007 season, but is not registered still.
CYCLE
The differentiations of new nodes in the main stem and successive fruit point on fruiting branch were at
regular intervals, generating the typical pyramidal shape present in cotton (Hearn 1994). This process can
be maintained for more time in SP 48114 compared with Guazuncho 3 INTA, the late variety with short
cycle. SP 48114 during 2010-2011showed that ( at 110 days after planting), the growth cycle ended with
2 to 4 more potential fruiting branch than Guazuncho 3 INTA, in addition the final height was 12 to 7 cm
more than Guazuncho 3 INTA. Both the parameters are associated with the growth cycle. This feature
could allow obtaining better performance in dry conditions. The relative performance is
shown in Table 1.
TABLE 1: AGRONOMIC PARAMETERS REGISTERED AT 110 DAYS AFTER SOWING, SEASON 2010-11. PRESIDENCIA ROQUE SENZ PEA, CHACO. DATAS
BY THE SAME LETTER ARE NOT DIFFERENT AT 5% PROBABILITY LEVEL.
Line/ Variety No. of Fruiting Branch Plant Height cm
SP 48114 14,25 a 94,25 a
SP 8461 12,75 ab 92 ab
SP 44825 12,5 ab 74,5 c
Poraite INTA 12,1 b 76,7 c
Guazuncho 3 INTA 12 b 81,5 bc
CV 9,64 10,2
SANITY
The line presented high resistance to bacterial blight caused by Xanthomonas axonopodis pv
malvacearum, because in the process of selections it was artificially infected from F3 to F12 (Fig1.). In
addition to this process it was also infected with cotton leaf roll dwarf virus (CLRDV) during F8
generations. When the plant developed the symptoms, the resistant lines were selected. The sanitary
performance was achieved by INTAs varieties (Poisson 2002).
New Cotton Germplasm as an Intermediate Cycle Called SP 48114 Development by the National Institute of Agricultural 15

Fig. 1: Process of Breeding Used for Development SP 48114 from Year 1993
YIELD AND FIBER PROPRIETIES
The line was evaluated in several trials from the season 2004-2005 which showed good performance and
achieved the first positions in the test in relation to commercial varieties (Royo et al., 2007). During the
dry and wet conditions of 2009-2010 and 2010-2011, the line SP 48114 showed better performances than
varieties with short cycle (Guazuncho 3 INTA and Poraite INTA). (Table 2 and 3). Both experiments
were grown in Presidencia Roque Saenz Pea, Chaco, with four replications each. The differential
behaviour can be explained for more possibilities to maintain the process of flowering for more days.
Sekloca (et. al., 2007) found that the varieties with intermediate cycle present better performance in
medium conditions, related to drought and humidity. The lint turnout % in both experiments for SP
48114 was better (Table 2 and 3). In dry conditions, the fiber length was 1 to 4 mm shorter in SP 48114
in comparision to Guazuncho 3 INTA (Table 2). However, in wet conditions the fiber properties was
similar than that of Guazuncho 3 INTA (Table 3).
Thus, it is possible to select lines with more differentiations fruit point at growing stations and
maintain similar agronomic parameters as varieties with short cycle.
TABLE 2: LINT YIELD AND QUALITY PARAMETERS FOR PCIA. ROQUE SENZ PEA, CHACO, 2009-10. THE DATA FROM THE TRIAL WITH 2 COMMERCIAL CULTIVARS, 3 PROMISING LINES,
INCLUDING SP 48114. SEASON WITH DRY CONDITIONS DURING FLOWERING. DATE BY THE SAME LETTER ARE NOT DIFFERENT AT 5% PROBABILITY LEVEL
Varieties/ Line Lint Yield (kg/ha) Lint Turnout (%) Length (mm) Strength (g/Tex) Micronaire Index
SP 48114 674 a 39,5 a 25,9 ab 28,7 b 4,7 a
SP 48666 639 a 38,4 a 25,1b 28,6 b 4,7 a
SP 81424 590 a 37,3a 26,05ab 29,8 ab 4,6 a
Poraite INTA 452 b 38,1 a 26,6 a 30,4 ab 4,4 a
Guazuncho 3 INTA 411 b 39,4 a 26,5 a 31,7 a 4,6 a
CV 12,1 6,6 3,3 4,6 11,6
TABLE 3: LINT YIELD AND QUALITY PARAMETERS FOR PCIA. ROQUE SENZ PEA, CHACO, IN THE YEAR 2010-11. THE DATES FROM THE TRIAL WITH 2 COMMERCIAL CULTIVARS 3 PROMISING
LINE, INCLUDING SP 48114, SEASON WITH WET CONDITIONS. DATE BY THE SAME LETTER ARE NOT DIFFERENT AT 5% PROBABILITY LEVEL
Varieties /line Lint Yield (kg/ha) Lint Turnout (%) Length (mm) Strength (g/Tex) Micronaire Index
SP 48114 962 a 41,2 ab 29,3 a 31,3 a 4,7
Poraite INTA 695 b 40,4 b 28,3 a 32,5 a 4,5
SP 48666 645 b 41,2 ab 27,9 b 31,5 a 4,6
Guazuncho 3 INTA 662 c 41,6 a 29,6 b 32,1 a 4,6
SP 6180 494 c 39,3 c 27,2 b 31,6 a 4,5
CV 14,1 1,43 1,74 2,69 4,7
Winter 1993 cross in green house between SP 99138 SP 99035
Season 1993/1994 F1 Generations
Visual Selection of individual
Plant
Progeny row
Agronomic characterization and
artificial infected with
xanthomonas axonopodis pv
malvacearum
Seasons 1996/1997 2003/2004
F4-F12 -Generations
Agronomic Testing, artificial
infected with xanthomonas
axonopodis pv malvacearum and
selecting and in F8, artificial
infected with cotton leaf rolf duarf
virus (CLRDV) and selecting
resistance lines
Seasons 2004/2005 2006/2007
Regional comparative trials in 4
locations for 3 seasons
REFERENCES
[1] Allard, R. W. (1960). Principles of plant breeding. John Wiley. N.Y. 473 p.
[2] Fryxell, P. A. (1986). Ecological adaptations of Gossypium species. pp. 1-7 In Mauney, J.R and Steward, J.McD. (Eds).
Cotton Physiology. The Cotton Foundations, Memphis, TN.
[3] Hearn, A. B. (1994). The principles of cotton water relations and their application in management. World Cotton Research
Conference 1:66-92.
[4] Paytas, M. (2010). Improving cotton yield under water limiting conditions in Argentina. Repor. ICAC Research Program.
[5] Poisson, J. A. F. (2002). Breve historia de la produccin de algodn en la Argentina. In: 1923-1 de agosto-2002. De Chacra
Oficial a Estacion Experimental. 79 anos de investigacin algodonera en el centro de la provincia del Chaco. Editorial
INTA EEA Saenz Pena, Centro Regional Chaco-Formosa.Pag.8
[6] Poisson, J. A. F., Bonacic, I., Royo, O. and Ibalo, Y. S. (2005). Mejoramiento gentico de algodn. Ano Agrcola
2004/2005. In: Proyecto Nacional de Algodon. Informe de avance No 1. 2o Reunin anual. Sosa M.A. y O. Peterlin (Ed).
Ediciones Instituto Nacional de Tecnologa Agropecuaria. Pages 9-11.
[7] Royo, O. M., Poisson Juan, A. F.; Bonacic, I., Montenegro, A., Ibalo, S. I., Mazza, S., and Gimnez, L. (2007). Direction of
Cotton Breeding in Argentina. In: Proceedings of the World Cotton Research Conference. Lubok Texas
[8] Sekloka, E. And Jacques, L. (2007). Early-compact American and late-vegetative African cotton ideotypes can address the
increasing diversity of cropping conditions in Africa. 4 Word Cotton Research.
Introgression of High Fibre Strength Trait

to Upland Cotton using Marker-Assisted Selection
Nallathambi Kannan, P. Selvakumar, R. Krishnamoorthy, D. Raja,
M. Bhuvaneshwari, V. Subramanian and M. Ramasami
Rasi R and D Centre, Rasi Seeds (P) Ltd., Attur636102, Tamil Nadu, India
AbstractCotton fibre is a basic raw material used in the textile industry. In recent years, changes in spinning
technology have resulted in the need of unique and often increased cotton fibre quality, especially fibre strength. In this
concern, an attempt was made to improve fibre strength of G. hirsutum by utilizing G. barbadense as donor through
Backcross (BC) and Modified Back Cross (MBC) pedigree breeding methods following marker-assisted selection using
Simple Sequence Repeats (SSR) markers. The Phenotypic Co-efficient of Variation (PCV), Genotypic Co-efficient of
Variation (GCV), heritability and genetic advance was studied in 475 numbers of F
2
populations. The result showed
fibre strength varied from 18.0 to 36.0 g/tex and 32 % of plants in the population fall under 27.0 to 36.0 g/tex group.
The PCV was higher than GCV which shows fibre strength is highly influenced by environment. The moderate
heritability and high genetic advance was observed for fibre strength; hence the selection is effective for this trait and
the heritability is due to additive genes effect. The identified SSR markers for fibre strength have been utilised to select
the high fibre strength plants in each generations. In BC
1
F
1
generation, fibre strength varied from 24.4 to 32.7 g/tex.
After continuous selection of high fibre strength plants using molecular markers in each generation, we obtained high
productive progenies with high fibre strength that ranged from 30.0 to 35.7 g/tex having more recurrent background
genome in BC
1
F
8
generations. High recovery of hirsutum background with high strength and different staple length
progenies were obtained in modified backcross population. Thus the high strength hirsutum lines developed will serve
as a donor for introgressing the fibre strength to improve the elite parental lines through marker-assisted background
selection.
INTRODUCTION
Cotton is the most preferred natural fibre in the world and plays a major role in the economy of
agriculture and industry. Among the four cultivated species, Gossypium hirsutum is well known for high
yield and dominates the worlds cotton fibre production followed by the Gossypium barbadense that is
known for superior fibre qualities. In cotton improvement, in addition to yield enhancement of lint, the
fibre qualities such as staple length, fibre strength, and fineness and maturity are very important. The
demand for improved fibre quality by textile industry will continue. Improvements in textile processing,
particularly advances in spinning technology, have led to increased emphasis on breeding cotton for
improved fibre characteristics, especially strength. (Rahman and Malik, 2008). The requirements in
textile spinning machinery with the adoption of rotor spinning, demands fibres with high strength to meet
out spinning productivity. Most of the presently developed cotton varieties have low fibre strength of 18
to 24 g/tex. Genetic variation for the fibre qualities are very limited in most of the currently cultivated
Gossypium hirsutum cotton. Thus there is an urgent need to introduce fibre strength characteristics from
Gossypium barbadense to upland cotton while maintaining the cotton fibre yield.
Cotton fibre strength trait is governed by several genes located in several loci of chromosomes and
are inherited quantitative way and thus influenced by quantitative trait loci (QTLs). Most traits in
breeding programs are quantitatively inherited, complicating their manipulation through phenotypic
and/or genomic approaches. Each of the QTLs has relatively small effects and is influenced by genotype
and environment showing strong GxE interaction, which leads to low genetic advance in cotton
improvement (Kohel, 1999ab).
Cotton fibre strength trait is governed by several genes located in several loci of chromosomes and
are inherited quantitative way and thus influenced by quantitative trait loci (QTLs). Most traits in
breeding programs are quantitatively inherited, complicating their manipulation through phenotypic
and/or genomic approaches. Each of the QTLs has relatively small effects and is influenced by genotype
and environment showing strong GxE interaction, which leads to low genetic advance in cotton
improvement (Kohel, 1999ab).
4
The modified backcross method followed for pyramiding of multiple traits is one of the ways by
which the inherent fibre strength trait can be transferred to an upland cotton elite line. Experiments in
cotton showed the negative linkage between yield and fibre traits and following modified backcross
(MBC) is expected to circumvent this effect. However, due to several QTLs involved for both yield and
fibre traits, the breeding cycle is expected to be longer.
The identification and utilization of molecular markers make it possible for plant breeders to find a
rapid and precise approach of marker-assisted selection (MAS) of desirable plants with target traits.
Introgressing the traits of interest can be followed using molecular markers that are mapped flanking or
tightly linked with the traits being incorporated. Following the advancement of MAS and MBC method,
it is expected to have selection for both recurrent parent background as well as genes to be introgressed
from non-recurrent parent. The use of MAS facilitates a faster introgression since plants can be sampled
and genotypes with target traits can be identified even at the early stage of development. Among the
available types of molecular markers, microsatellite markers simple sequence repeats (SSR) have shown
to be the most adequate for breeding programs due to their co-dominance and multi-allelic
characteristics, and for their ability to automate the process.
The main objective of the study has been to improve fibre strength of G. hirsutum by introgression of
QTLs associated with fibre strength from G. barbadense by means of backcross (BC) and modified
backcross (MBC) pedigree breeding methods using fibre strength QTL SSR markers. Thus a
combination of MBC with MAS for selection of desirable cotton lines with enhanced yield and high fibre
strength was followed in our breeding strategy.
The present investigation was also undertaken to study the phenotypic and genotypic coefficient of
variability, phenotypic and genotypic variances, heritability and genetic advance of the variation existed
in F
2
and F
3
population originated from the inter-specific crosses in cotton.
In the present study, the field experiments were conducted at the Rasi Seeds (P) Ltd., Research Farm,
Attur, Salem (District) Tamil Nadu state (INDIA).
The salient features of parents involved in the backcross and modified backcross are furnished in
Table 1. The breeding scheme, number of plants raised and number of plants selected in each generation
of backcross and modified backcross are shown in Figs. 1 to 4. The experiments were raised in the winter
season (August February). All the recommended cultural practices of cotton production in the area
were done periodically.
TABLE 1: SALIENT FEATURES OF PARENTS INVOLVED IN THE STUDY
Parents Species Used as Boll Weight
(g)
Ginning % Span
Length
(mm)
Lint Index Fibre
Strength
(g/tex)
Fineness
(Mic)
Uniformity

Ratio
RC 64 G. hirsutum Recurren
t
Medium
(4.0-4.8)
Medium
(33-36)
Long
(30-32)
Medium
4.5-5.5
Medium
(24-26)
Medium
(4.0-4.2)
Excellent
(47-49)
t
Medium to Big
(4.9-5.6)
Low
(31-33)
Extra Long
(33-35)
Medium
4.0-5.0
Strong
(26 28g/tex)
Fine
(3.3-3.7)
Excellent
(47-48)
t
Medium to Big
(4.8-5.8)
Low
(30-32)
Extra Long
(34-36)
Medium
4.0-5.0
Strong
(25-27)
Fine
(3.5-3.8)
Excellent
(47-48)
t
Big
(5.3-6.0)
Medium
(33-35)
Extra Long
(33-35)
Medium
4.0-5.0
Medium
(24-25)
Fine
(3.5-3.7)
Excellent
(47-49)
RC 45SB G. barbadense Donor Small
(2.8-3.7)
Low
(25 -28)
Extra Long
(37-40)
Medium
4.0-5.0
Very Strong
(33-36)
Very Fine
(2.8-3.1)
Excellent
(48-50)
Phenotypic Characters
Selected plants in each single plant progeny were observed and their biometrical and fibre quality traits
were recorded. The genetic analysis for the traits such as boll weight (g), Number of bolls/plant, ginning
percentage (GP %), lint index (LI), seed index (SI), single plant yield (g) and fibre quality parameters
Introgression of High Fibre Strength Trait to Upland Cotton using Marker-Assisted Selection 19
were done in the F
2
population along with their parents. The fibre quality traits viz., 2.5% span length
(mm), uniformity ratio (%), fibre fineness (micronaire), fibre strength (g/tex) and elongation were
estimated by High Volume Instrument USTER
HVI Spectrum in ICC mode.

Fig. 1: The Breeding Scheme, Number of Plants Raised and Number of Plants Selected Based on MAS in the Backcross Population

Fig. 2: The Breeding Scheme, Number of Plants Raised and Number of Plants Selected Based on MAS in the Modified Backcross Population (I)
First G.hirsutum(RC64) x G.barbadense (RC45SB)
Season
2002(W) Identification of polymorphic markers of both parents (658 markers were screened and 454 were pol
Second G.hirsutum (RC 64 ) x F1
Season F1 backcross with the recurrent parent
2003(W)
475 F2 individuals were rasied and genotyping were done with 158 polymorphic markers based on low and high
Third BC1F1 1. 276 plants were raised
Season 2. 15 high fibre strength plants with more recurrent background were selected
2004(W) based on phenotypic and genotypic data (MAS) and forwarded to next generation
Fourth BC1F2 1. Individual 15 plant progenies were grown. (40 plants/progeny)
Fifth BC1F3 1. Individual 8 plant progenies were grown. (21 plants/progeny)
Season 2. Homozyous progenies similar to recurrent
2006(S) parent with high fibre strength 17 plants were selected based on phenotypic and genotypic data (MAS)
3.Recurrent plant type with high fibre strength plants will be forwarded from superior progenies
Sixth BC1F4 1. Individual 17 plant progenies were grown. (20 plants/progeny)
2006(W) parent with high fibre strength 67 plants were selected based on phenotypic and genotypic data (MAS)
Seventh BC1F5 1. Individual 67 plant progenies were grown. (20 plants/progeny)
Eighth BC1F6 1. Individual 82 plant progenies were grown. (20 plants/progeny)
Ninth BC1F7 1. Individual 148 plant progenies were grown. (20 plants/progeny)
Tenth BC1F8 1. Individual 54 plant progenies were grown. (20 plants/progeny)
2010(W) parent with high fibre strength 36 plants was selected and forwarded
Season
2003(W)
Third RC 62 X BC1F1 1. 276 plants were raised
Season 2.More G.hirsutum plant type with high fibre strength plants
2004(W) to be crossed with G.hirsutum recurrent parent(RC62)
Fourth MBC1F1 1. 309 plants were raised
Season 2. High fibre strength plants with more recurrent background were selected
Fifth MBC1F2 1. Individual 305 plants were raised
Sixth MBC1F3 1. Individual 71 plant progenies were grown. (20 plants/progeny)
Seventh MBC1F4 1. Individual 44 plant progenies were grown. (20 plants/progeny)
Eighth MBC1F5 1. Individual 17 plant progenies were grown. (10 plants/progeny)
Ninth MBC1F6 1. Individual 25 plant progenies were grown. (10 plants/progeny)
2010(W) parent with high fibre strength 27 plants were selected and forwarded

Fig. 3: The Breeding Scheme, Number of Plants Raised and Number of Plants Selected in the Modified Backcross Population (II)

Fig. 4: The Breeding Scheme, Number of Plants Raised and Number of Selected Based on MAS in the Modified Backcross Population (III)
Mean values were used for different statistical analysis. Analysis of variance and genotypic and
phenotypic variation were calculated following Singh and Chaudhury (1985). Phenotypic coefficient of
variation (GCV), Genotypic coefficient of variation (PCV) were estimated using the formula suggested
by Burton (1952), while genetic advance (GA) as percent means and genetic advance as percentage of
mean (GA %) was estimated by the formula given by Lush (1949) and Johnson et al. (1955). The
estimates of broad-sense heritability were computed as suggested by Allard (1960).
Season
2003(W)
2005(W) based on phenotypic and genotypic data (MAS) and forwarded to next generation*
2006(W) based on phenotypic and genotypic data (MAS) and forwarded to next generation*
Eighth MBC1F5 1. Individual 23 plant progenies were grown. (10 plants/progeny)
Ninth MBC1F6 1. Individual 33 plant progenies were grown. (10 plants/progeny)
2010(W) parent with high fibre strength plants 20 were selected based on phenotypic and genotypic data (MAS)
Season
2003(W)
Genotyping using SSR Markers
F
2
mapping populations were developed from the interspecific cross between G.hirsutum (RC 64) and
G.barbadense (RC 45SB) for the identification of SSR markers associated with fibre strength trait.
Young leaf samples were collected from 475 F
2
individuals and DNA was extracted using modified
Davis protocol. PCR was conducted in a total volume of 10 l with 10 ng of cotton DNA, 1 x PCR buffer
(without MgCl
2
), 1.5 mM MgCl
2,
0.1 M dNTPs, 0.2 M of each primer and 0.5 units of Taq DNA
polymerase. The cycling conditions for PCR were as follows: 5 min for 94

C; 35 cycles of 94
C for 45 s,
57C for 45 s, 72C for 60 s; 72C for 5 min; 4C for preservation. Amplified DNA fragments were
resolved in 6% denatured polyacrylamide gel [(acryl amide: bisacrylamide (19:1)] and stained with silver
nitrate.
We employed 658 SSR primers including BNL, NAU, JESPR and CIR etc., for the identification of
polymorphism between the two parents. The polymorphic primers were used to screen the bulked low
and high fibre strength DNA samples and selected primers were subsequently used to genotype the F
2

individuals. Only unambiguous distinct bands were scored. QTLs for cotton fibre strength in F
2

population were identified using MAPMAKER 2.0 and QTL CARTOGRAPHER (version 1.15)
respectively. The SSR markers associated with the fibre strength QTL were used in the backcross and
modified backcross breeding program.
Genotyping the BC and MBC Samples
Marker-assisted selection was conducted for every generation of backcrossing and modified backcrossing
with the markers associated with fibre QTLs based on the F
2
population. The markers covering the fibre
strength QTLs that were used in MAS are RAS 72, RAS 158, RAS 215, RAS 223, RAS 224, RAS 230,
RAS 306 and RAS 304.The selection of plants with high fibre strength trait at every generation was
based on the markers and phenotypic data.
The first and foremost criterion to be considered in any breeding programme is the magnitude of the
genetic variability present in the base population which is prime requirement for starting a judicious
breeding programme for combining desirable characters into the elite lines. In the present investigation
the estimates of mean, range, phenotypic and genotypic coefficients of variation, heritability and genetic
advance as per cent of mean in F
2
generation are calculated and presented in Table 2. There were large
differences in the variances for most of the characters under study. The high variance (10.2) of fibre
strength character in F
2
population indicates that the presence of sufficient amount of variability which
had been generated in segregating populations (Pradeep and Sumalini, 2003). The distribution of fibre
strength in F
2
generations is given in Fig. 6. The distribution range of fibre strength in F
2
was between 18
g/tex to 36 g/ tex. The 27% of plants out of 475 plants showed moderate fibre strength (26-28 g/tex).
Furthermore, 3 plants in F
2
showed above 34 g/tex which was higher than the donor parent, suggesting
transgressive segregation for the trait. The variation and transgressive segregation observed for fibre
strength has practical implication for combining fibre strength in upland cotton.
TABLE 2: THE ESTIMATES OF MEAN, RANGE, HERITABILITY, GENETIC ADVANCE, GENETIC ADVANCE PER CENT OF MEAN, PCV AND GCV OF F
2
GENERATION (RC 64 X RC 45 SB)
Characters Mean Range Variance Heritability (h
2
%) GA GA% of Mean PCV % GCV%
Boll Weight (g) 3.3 1.9-4.6 0.4 84.5 1.3 39.1 19.0 17.5
Number of bolls/plant 67.5 32.0-127.0 445.8 46.9 43.5 64.4 31.3 21.4
Ginning percentage (%) 29.5 22.5-38.7 9.3 56.1 6.3 21.3 10.3 7.7
Lint index 3.8 1.9-6.6 0.6 8.9 1.6 41.5 20.1 6.0
Seed index 9.2 5.4-14.0 2.4 15.1 3.2 34.3 16.7 6.5
2.5% span length (mm) 32.5 26.1-38.1 6.3 83.1 5.2 15.9 7.7 7.0
Fibre strength (g/tex) 43.7 40.6-47.5 10.2 59.1 6.6 24.0 11.6 9.0
Uniformity ratio 27.4 18.4-36.1 1.7 78.4 2.7 6.1 3.0 2.6
Elongation 5.6 4.0-12.0 0.7 65.8 1.7 30.5 14.8 12.0
Micronaire 2.8 2.0-4.2 0.2 39.6 0.9 31.3 15.2 9.6
Seed cotton yield (g)/plant 152.5 82.3-266.7 2482.5 30.1 102.6 67.3 32.7 17.9
Although range can provide a preliminary idea about the variability but coefficient of variation is
reliable as it is independent of unit of measurement. The extent of variability as measured by PCV and
GCV also gives information regarding the relative amount of variation.

Fig. 5: Frequency Distribution of Fibre Strength Trait in F
2
Population (475 Plants)
The estimates of phenotypic coefficients of variation (PCV) ranged from 2.98 for fibre uniformity
ratio to 32.68 % for seed cotton yield per plant and the corresponding values for genotypic coefficients of
variation (GCV) were 2.64 % for fibre uniformity ratio and 21.83 % for number of bolls per plant (Table
2). The phenotypic coefficient of variation which measures total variation was found to be greater than
genotypic coefficient of variation for all the characters indicating some degree of environmental
influence on the traits.
It is not the magnitude of variation but the extent of heritable variation, which matters most for
achieving gains in selection programme. The coefficient of variation indicates only the extent of variation
for a character and does not discriminate the variability into heritable and non-heritable portion. The
heritability worked out in broad sense would suggest how far the variation is heritable and selection is
effective. A perusal of heritability estimates indicated that the characters such as boll weight, fibre length,
uniformity ratio and fibre elongation have high heritability (Table 2). Such high heritability estimates
have been found to be helpful in making selection of superior genotypes on the basis of phenotypic
performance for quantitative characters. The characters viz., number of bolls per plant, ginning
percentage, fibre strength, mircronaire and seed cotton yield per plant had moderate heritability. Though
the heritability estimates are the true indicators of genetic potentiality of the genotypes which can be used
as a tool for selection, changes in the values of the heritability due to fluctuations of the environmental
factors detract for total dependence on such estimates. However, heritability estimates when considered
in conjunction with the predicted genetic gain form a reliable tool for selection. They indicate the
expected genetic advance of a character in response to the certain selection pressure imposed on them
and also provide an idea about the gene action involved in the expression of various polygenic traits
involving several QTLs.
High heritability coupled with high genetic advance as per cent of mean was noticed for the
characters boll weight and elongation. This indicates that additive gene action was responsible for the
inheritance of these traits and the selection in the early generation could be fruitful in improving these
characters (Kumaresan, et. al., 2000). In contrast the characters lint index and seed index have low
heritability and high genetic advance as per cent of mean. The fibre strength character has moderate
heritability and high genetic advance as per cent of mean indicates that success through simple selection
could be expected in the early generation as this trait is having the additive gene action.
Marker-Assisted Selection (MAS) using Simple Sequence Repeats (SSR)
Based on limited DNA polymorphism in upland cotton for markers available to date, and limited
application of markers for cotton improvement, sound MAS breeding strategy is important for
incorporating QTLs associated with fibre traits are successfully used in crop improvement. We have
screened 658 SSR primers for the identification of polymorphism between the two parents. Of the 658
primer, 454 primers were polymorphic between the parents, 158 primers were polymorphic between
bulked low and high fibre strength samples (Fig. 6). Subsequently 158 polymorphic primers obtained in
0
5
10
15
20
25
30
1
8
2
0
2
0
2
2
2
2
2
4
2
4
2
6
2
6
2
8
2
8
3
0
3
0
3
2
3
2
3
4
3
4
3
6
3
6
3
8
F
r
e
q
u
e
n
c
y
i
n
%
Range
F2

bulked an
three min
Among th
and RAS
selection
phenotypi
M 1
1, 2
3 Bu
4, 5
M 1
B, H
1 to 1
17 to

M 1
B, H
1 to 1
11 to
Introgre
nalysis were
or QTLs for
hese markers
304) molec
in the BC
ic data are gi
Fig. 6: SSR
kb ladder
G.barbadens
ulked low fib
Bulked high
Fig. 7:
kb ladder
G.barbaden
6 Low fibre
46 High fib
Fig. 8: S
kb ladder
G.barbaden
0 Low fibre
25 High fib
ession of High F
used to geno
r fibre streng
, eight SSR (
cular marker
and MBC
iven in Fig. 1
R Screening Gener
se & G.hirsu
bre strength D
h fibre strengt
: SSR Profiles Gen
nse & G.hirs
e strength in
bre strength i
SSR Profiles Gener
nse & G.hirs
e strength in
bre strength i
Fibre Strength T
otype the 47
gth from the
(RAS 72, RA
rs associated
(Fig. 8).The
1 4.
rated for Bulked lo
utum
DNA of F
2
in
th DNA of F
nerated for F
2
Low
sutum
dividual sam
individual sa
rated for BC
1
F
8
low
sutum
dividual sam
individual sa
Trait to Upland C
75 number o
strain have b
AS 158, RAS
d with fiber
e number o
ow and High Fibre
ndividuals

(<
F
2
individuals
and High Fibre St
mples of F
2
(<
amples of F
2
w and high Fibre St
mples of BC
1
F
amples of BC
Cotton using Ma
f F2 individ
been identifi
S 215, RAS 2
strength QT
of selected p
Strength DNA of F
24 g/tex)
s

(< 30 g/tex)
rength Individuals
< 24 g/tex)
(> 30 g/tex)
trength Individuals
F
8
(< 27 g/te
C
1
F
8
(> 31 g/
arker-Assisted S
duals (Fig. 7)
ied and tagge
223, RAS 22
TLs were use
plants based
F
2
Samples with SS
)
s with the Primer R
s with the Primer
ex)
/tex)
Selection
). One major
ed with DNA
24, RAS 230
ed for mark
d on the ma

SR Primers

RAS 72

RAS 223
23
r QTL and
A markers.
0, RAS 306
ker-assisted
arkers and
TABLE 3: THE ESTIMATES OF MEAN, RANGE AND VARIANCE OF BACKCROSS POPULATIONS (RC 64 X (RC 64 X RC45 SB)
Generation Number of
bolls/Plant
Boll
Weight
(g)
Ginning
Percenta
ge (%)
Lint
Index
Seed
Index
2.5% Span
Length
(mm)
Fibre
Strength
(g/tex)
Uniformity
Ratio
Elongation Micronaire
BC1F1 Mean 58.0 3.8 31.8 5.0 10.7 35.1 27.2 48.9 6.1 3.5
Range 16.0-185.0 2.5-5.5 22.1-38.6 2.3-7.6 5.5-19.2 30.3-38.3 24.4-32.7 43.3-54.4 3.6-9.9 2.4-5.1
Variance 584.1 0.4 7.1 0.7 3.7 2.6 4.9 5.4 1.9 0.3
BC1F2 Mean 47.6 4.1 32.3 4.8 10.0 33.7 27.1 46.0 3.6 5.8
Range 20.0-146.0 2.9-5.2 26.8-36.0 3.3-6.4 7.2-11.5 27.1-37.1 26.0-30.7 41.1-49.4 2.5-4.7 4.1-8.7
Variance 576.6 0.2 5.8 0.6 1.1 5.0 1.1 3.9 0.3 1.0
BC1F3 Mean 42.4 3.0 31.4 4.3 9.3 30.4 30.5 46.8 4.6 4.1
Range 18.0-89.0 1.8-4.5 28.5-34.5 3.0-6.0 6.4-12.7 26.4-36.1 26.1-35.6 44.9-48.6 3.8-6.4 2.8-5.9
Variance 301.4 0.3 2.7 0.4 1.5 3.8 9.9 0.9 0.4 0.8
BC1F4 Mean 77.5 3.1 35.1 4.5 8.3 28.3 29.2 47.6 5.6 4.6
Range 36.0-142.0 2.0-4.9 27.8-39.8 3.0-6.5 6.0-11.3 24.4-31.0 26.3-34.1 44.7-49.1 4.3-7.9 2.2-5.8
Variance 625.2 0.3 4.8 0.4 1.1 1.6 3.4 0.5 0.4 0.6
BC1F5 Mean 105.7 3.0 34.2 4.2 8.1 28.7 29.4 47.0 5.5 4.3
Range 48.0-147.0 2.0-4.4 25.6-40.9 2.3-6.6 4.6-12.3 23.3-33.0 22.9-37.0 44.5-51.0 4.1-7.0 2.5-5.7
Variance 285.8 0.2 4.3 0.5 1.4 2.6 4.5 1.0 0.2 0.3
BC1F6 Mean 69.0 3.1 32.9 4.6 9.3 28.8 29.4 46.5 5.9 4.7
Range 13.0-152.0 1.8-4.7 26.9-39.8 2.8-6.5 5.2-13.8 25.7-32.3 24.5-33.6 44.2-49.4 4.7-7.6 2.5-5.9
Variance 554.4 0.3 3.9 0.5 1.7 2.0 3.5 0.9 0.2 0.4
BC1F7 Mean 88.3 3.4 33.3 5.0 10.0 28.9 30.0 48.0 5.5 4.9
Range 38.0-124.0 2.0-5.1 28.0-38.8 3.4-6.5 6.5-12.5 25.2-32.7 27.3-34.3 46.1-50.3 4.5-6.7 3.2-6.2
Variance 332.1 0.2 3.5 0.2 1.0 1.4 2.0 0.6 0.2 0.3
BC1F8 Mean 37.0 3.2 32.4 5.1 10.6 29.4 30.4 47.8 5.6 5.3
Range 12.0-78.0 2.0-4.6 22.3-38.4 2.9-6.7 7.0-13.4 26.4-41.0 26.9-35.7 42.8-50.8 4.3-6.9 3.0-6.1
Variance 134.5 0.2 5.5 0.4 1.0 2.7 3.2 0.9 0.2 0.3
RC 64
(Recurrent
Parent)
Mean 143.0 5.5 33.3 6.4 12.9 31.6 24.1 47.8 5.4 6.7
RC 45SB
(Donor
Parent)
Mean 127.0 3.6 28.4 4.7 12.0 38.0 33.1 49.7 2.8 4.8
The introgression of fibre strength character into upland cotton utilizing Gossypium barbadense as
donor through back cross and modified back cross method was done in the present investigation. Since
the fibre strength trait is controlled by several QTLs, the introgression was done by one back cross
followed by pedigree method. The mean, range and variance of observed characters for back cross
generations are given in the Table 3. The mean values of fibre strength in all the backcross generations
(BC
1
F
1
- BC
1
F
8
) indicates that the progenies are having high fibre strength (>27 g/tex). The high fibre
strength plants in each back cross generations were selected based on phenotypic selection coupled with
genotypic selection utilizing identified fibre strength linked SSR markers. The high fibre strength plants
with recurrent parent background were selected by utilizing molecular markers and phenotypic data. The
distribution range of fibre strength in backcross population (Fig. 9) reveals that in each generation the
number of plants fall under the high fibre strength group has been increased by the effective selection of
combining phenotypic and genotypic information. In BC
1
F
1
generation the frequency of plants fall under
high fibre strength (>30g/tex) is 14 per cent while in the BC
1
F
8
generation the frequency is 60 per cent.
Furthermore in BC
1
F
8
generation, the fibre strength values were ranged from 26.9 to 35.7 g/tex and seven
plants were having highest fibre strength values of above 34 g/tex. The high fibre strength plants in the
advance generations are have high phenotypic similarities to the recurrent parent type and thus the
molecular markers are effectively used in the selection of target alleles with high background of recurrent
parent.


Fig. 9: Frequency Distribution of Fibre Strength Trait in Backcross Generations
The modified backcross method has been used for pyramiding the multiple traits into upland cotton
besides the introgression of fibre strength traits. The three upland cotton lines namely RC 62, RC 67 and
RC 92 were utilized as recurrent parent and high fibre strength BC
1
F
1
plants were used as donor plant to
develop a three modified back cross population (Figs. 2- 4) The advantage of this proposed modified
backcross breeding method was to obtain the more recurrent genome background with high fibre strength
as the frequency of the undesirable genes from the donor parents was reduced similar to the reported by
Li and Pan, 1990.
TABLE 4: THE ESTIMATES OF MEAN, RANGE AND VARIANCE OF MODIFIED BACKCROSS (I) POPULATIONS
Bolls/Plant
Boll
Weight
(g)
Ginning
Percentage
(%)
Lint
Index
Seed
Index
2.5%
Span
Length
(mm)
Fibre
Strength
(g/tex)
Uniformity
Ratio
MBC1F1 Mean 90.1 4.6 31.1 5.0 11.1 35.6 26.5 44.7 4.8 3.4
Range 56.0-154.0 3.1-5.9 25.8-36.1 3.7-6.0 8.5-
14.1
32.2-38.4 25.0-28.6 43.2-46.6 3.6-6.0 2.6-4.1
Variance 501.1 0.5 5.6 0.4 1.6 2.4 0.5 0.6 0.3 0.2
MBC1F2 Mean 85.4 3.6 31.7 4.5 9.5 32.8 27.1 45.5 5.5 3.1
Range 19.0-176.0 2.1-5.6 2.2-42.4 2.9-6.9 3.55-
14.4
29.1-37.5 25.7-30.1 42.8-47.2 4.2-7.2 1.8-4.7
Variance 686.1 0.4 14.6 0.7 2.9 3.3 1.0 0.6 0.4 0.3
MBC1F3 Mean 99.4 3.8 32.5 4.8 10.0 32.4 28.1 44.4 5.4 3.5
Range 56.0-135.0 2.8-5.4 25.6-39.9 3.8-6.3 7.5-
14.4
28.3-38.1 25.9-30.8 42.0-47.2 4.1-7.1 2.6-4.4
Variance 301.7 0.3 6.4 0.3 1.5 3.4 1.2 1.0 0.3 0.2
MBC1F4 Mean 98.0 3.7 31.3 5.0 11.0 32.6 26.9 44.2 6.1 3.8
Range 45.0-142.0 2.0-6.5 23.4-37.7 2.9-7.0 6.3-
14.4
28.7-36.2 25.0-29.9 41.9-47.3 5.0-8.3 2.4-5.3
Variance 314.8 0.7 5.7 0.6 2.4 2.3 0.6 0.8 0.4 0.3
MBC1F5 Mean 89.0 4.0 33.9 5.3 10.4 33.3 28.4 45.8 5.4 3.6
Range 68.0-116.0 2-5.5 24.7-46.0 3.6-6.9 5-
14.25
30.6-35.8 25.7-31.8 44.0-47.2 4.3-6.5 2.6-4.5
Variance 111.6 0.4 5.6 0.5 1.8 1.5 1.9 0.5 0.2 0.2
MBC1F6 Mean 72.6 4.3 32.7 5.5 11.3 33.0 29.2 45.5 5.5 4.2
Range 19.0-122.0 2.9-5.9 21.4-36.9 4.1-7.4 8.35-
16.86
29.2-35.2 25.2-32.3 44.0-47.9 4.6-6.8 2.9-5.1
Variance 457.3 0.3 3.4 0.4 1.8 2.1 1.9 0.6 0.2 0.1
RC 64
(Recurrent
Parent)
Mean 143.0 5.5 33.3 6.4 12.9 31.6 24.1 47.8 6.7 5.4
RC 62
(Recurrent
Parent)
Mean 66.0 4.9 31.3 5.9 12.9 37.2 28.5 47.2 5.9 3.8
RC 45SB
(Donor
Parent)
Mean 127.0 3.6 28.4 4.7 12.0 38.0 33.1 49.7 2.8 4.8

0
10
20
30
40
50
60
70
80
90
100
1
8
2
0
2
0
2
2
2
2
2
4
2
4
2
6
2
6
2
8
2
8
3
0
3
0
3
2
3
2
3
4
3
4
3
6
3
6
3
8
F
r
e
q
u
e
n
c
y
i
n
%
Range
BC1F1
BC1F2
BC1F3
BC1F4
BC1F5
BC1F6
BC1F7
In the modified backcross generations the high fibre strength has been improved significantly. The
mean, range and variance of fibre strength for modified back cross generations are given in the Table 4 to
6. The fibre strength values of modified backcross generations ranged from 27.2 to 37.6 g/tex in MBC
1
F
4

(III), 25.3 to 32.4 g/tex in MBC
1
F
6
(I) and 24.4 to 32.8 g/tex in MBC
1
F
6
(II). The marker based selected
advanced progenies in the modified backcross generations are having uniform high fibre strength with
high similar phenotypic characters of the recurrent parents. The selected high strength progenies were
grouped into different staple length group in order to meet out the textile industry requirement of various
counts.
TABLE 5: THE ESTIMATES OF MEAN, RANGE AND VARIANCE OF MODIFIED BACKCROSS (II) POPULATIONS
Bolls/Plant
Boll
Weight
(g)
Ginning
Percentage
(%)
Lint
Index
Seed
Index
2.5% span
Length
(mm)
Fibre
Strength
(g/tex)
Uniformity
Ratio
MBC1
F1
Mean 92.7 4.6 32.2 5.3 10.9 36.3 44.5 26.9 5.2 3.6
Range 38.0-178.0 3.8-6.5 28.7-36.2 4.3-6.3 5.8-13.3 33.0-39.7 43.2-46.0 26.0-29.3 4.4-7.5 3.0-4.5
Variance 921.6 0.4 3.6 0.3 2.2 3.4 0.5 0.7 0.4 0.2
MBC1
F2
Mean 93.6 3.8 32.5 4.8 10.0 33.4 26.8 45.3 5.3 3.4
Range 46.0-124.0 2.2-6.5 25.7-39.4 2.26-7.7 5.4-14.1 29.7-36.8 26.0-29.7 43.4-47.5 4.2-7.1 2.3-6.0
Variance 312.2 0.7 8.2 0.7 2.0 2.9 0.6 0.8 0.3 0.5
MBC1
F3
Mean 105.5 3.9 31.4 4.4 9.6 32.2 29.5 44.6 5.1 3.6
Range 76.0-128.0 2.7-5.8 27.4-36.6 3.6-5.6 7.7-12.1 28.5-34.8 26.9-33.0 42.9-46.8 4.1-6.5 2.8-4.7
Variance 185.8 0.7 4.4 0.2 1.0 1.9 3.0 0.8 0.4 0.3
MBC1
F4
Mean 97.1 4.0 28.7 4.6 11.3 33.5 28.6 43.8 5.6 3.5
Range 62.0-134.0 2.4-6.0 23.0-35.8 2.9-12.3 1.3-14.0 29.8-37.2 25.1-31.8 41.9-45.6 4.5-8.1 2.4-5.02
Variance 283.2 0.6 6.6 0.8 1.9 2.2 0.6 1.6 0.7 0.3
MBC1
F5
Mean 93.1 4.3 29.6 4.5 10.7 33.3 28.4 45.7 5.0 3.3
Range 68.0-120.0 2.3-5.9 24.2-47.5 3.0-5.9 5.8-13.8 29.5-37.3 21.9-32.0 43.7-47.7 3.9-6.5 2.2-4.3
Variance 200.7 0.5 7.9 0.4 2.1 2.6 2.7 0.7 0.3 0.2
MBC1
F6
Mean 56.2 4.2 29.6 4.6 11.0 33.4 29.2 45.3 4.8 3.8
Range 18.0-118.0 2.0-5.7 24.5-34.7 3.4-5.5 8.5-14.0 28.5-35.6 24.3-32.7 43.1-47.7 4.1-6.1 2.7-4.9
Variance 239.4 0.3 3.4 0.1 0.9 1.5 2.4 0.5 0.2 0.1
RC 64
(Recur
-rent
Parent)
Mean 143.0 5.5 33.3 6.4 12.9 31.6 24.1 47.8 6.7 5.4
RC 67
(Recur
-rent
Parent)
Mean 93.0 5.0 31.8 5.9 12.6 37.0 27.3 47.7 3.3 6.1
RC
45SB
(Donor
Parent)
Mean 127.0 3.6 28.4 4.7 12.0 38.0 33.1 49.7 2.8 4.8
TABLE 6: THE ESTIMATES OF MEAN, RANGE AND VARIANCE OF MODIFIED BACKCROSS (III) POPULATIONS
Bolls/Plant
Boll
Weight
(g)
Ginning
Percentage
(%)
Lint
Index
Seed
Index
2.5% Span
Length
(mm)
Fibre
Strength
(g/tex)
Uniformity
Ratio
MBC1F1
Mean 76.5 4.9 32.4 5.8 12.2 34.9 26.9 45.4 4.3 3.7
Range 34.0-133.0 2.5-6.9 27.7-36.7 4.7-7.0 10.2-17.5 25.8-37.8 26.1-28.7 44.0-46.6 3.7-5.5 2.5-4.5
Variance 471.5 0.9 5.2 0.4 2.5 6.1 0.6 0.5 0.1 0.3
MBC1F2
Mean 97.0 4.1 34.2 5.5 10.6 33.3 27.4 45.8 4.8 4.9
Range 75.0-124.0 2.0-6.1 28.0-40.4 3.6-7.8 7.4-14.4 0.5-36.6 26.0-30.7 43.6-47.9 2.7-7.7 2.6-9.2
Variance 270.9 0.7 6.9 0.8 2.8 1.7 1.2 0.8 1.5 3.2
MBC1F3
Mean 74.3 4.2 33.7 5.6 10.9 31.0 29.1 46.9 5.4 4.0
Range 50.0-111.0 2.5-6.0 29.9-38.6 3.1-7.6 7.3-14.4 25.9-36.2 25.8-33.7 44.7-49.9 4.5-6.3 2.5-5.5
Variance 151.5 0.6 5.6 0.8 2.2 4.2 3.0 1.5 0.2 0.6
MBC1F4
Mean 51.5 4.2 33.1 5.8 11.7 31.3 31.4 46.7 5.4 4.9
Range 18.0-98.0 2.6-6.8 26.7-45.3 2.6-7.9 5.9-17.1 26.8-41.3 27.2-37.5 42.7-49.3 4.2-7.4 3.4-5.6
Variance 228.0 0.5 4.7 0.5 2.2 3.3 2.8 0.9 0.3 0.2
Table 6 (Contd.)

Table 6 Contd.
RC 64
(Recurrent
Parent)
Mean 143.0 5.5 33.3 6.4 12.9 31.6 24.1 47.8 6.7 5.4
RC 92
(Recurrent
Parent)
Mean 78.0 5.6 34.6 7.6 14.3 35.8 28.4 49.2 6.6 3.6
RC 45SB
(Donor
Parent)
Mean 127.0 3.6 28.4 4.7 12.0 38.0 33.1 49.7 2.8 4.8
The application of DNA markers in backcross breeding program is dependant by the precision of
associated markers as well as by the cost effectiveness of marker-assisted selection. Marker-assisted
selection was found useful in developing genotypes with combinations of favourable alleles. The main
reasons supporting the utilization of molecular markers in cotton breeding programs are the 100%
heritability of the markers and their lower cost. Hence, the molecular markers were used in a
backcrossing scheme to improve the fibre strength traits in upland cotton efficiently.
Further to the present investigation the selected BC
1
F
8
and MBC
1
F
6
plants will be forwarded to the
next generation. The progeny test row may be conducted to select the best uniform high yielding
progenies with high fibre strength. Thus the developed introgressed high fibre strength upland cotton line
can be utilized for introgressing the fibre strength to improve the available elite parental lines through
marker assisted background selection to develop high fibre strength hybrids.
REFERENCES
[1] Allard, R.W. 1960. Principles of Plant Breeding. New York: John Willy and Sons, Inc.
[2] Burton, G.M. 1952. Quantitative inheritance in grasses. Proc. 6
th
Int. Grassland Cong., 1: 277-283.
[3] Chen, H., N. Qian., W.Z. Guo., Q.P. Song., B.C. Li., F.J. Deng., C.G. Dong and T.Z. Zhang (2009). Using three
overlapped RILs to dissect genetically clustered QTL for fibre strength on Chro.D8 in Upland cotton. Theor. Appl. Genet.,
119: 605612.
[4] Dudley, J.W. and R.H. Moll.1969. Interpretation and use of estimates of heritability and genetic variances in plant breeding.
Crop. Sci., 9(3):257-262.
[5] Johnson, H. W., H.F. Robinson and R.E. Comstock. 1955. Genotypic and phenotypic correlation in soybean and their
implications in selection. Agron. J., 47 : 477-483.
[6] Kohel, R.J.1999a. Cotton Improvement: A Perspective. Cotton World 1: (in press).
[7] Kohel, R.J.1999b. Cotton germplasm resources and the potential for improved fibre production and quality, In: A.S. Basra
(Ed.), Cotton fibres, pp. 167182. The Haworth Press, Inc, NY.
[8] Kumaresan, D., J. Ganesan and S. Ashok. 2000. Genetic analysis of qualitative characters in cotton (Gossypium hirsutum
L.). Crop Res. Ind., 19: 481-484.
[9] Lacape, J.M., T.B. Nguyen, B. Courtois, J. L. Belot, M. Giband, J. P. Gourlot, G. Gawryziak, S. Roques and B. Hau. 2005.
QTL analysis of cotton fibre quality using multiple Gossypium hirsutum x Gossypium barbadense backcross generations.
Crop. Sci., 45: 123-140.
[10] Li, W.H and J.J. Pan. 1990. Effect of modified backcross in breeding upland cotton cultivars. J. Nanjijng. Agri. univ., 13:
232-235.
[11] Lush, J.N. 1949. Animal breeding plans. The collegiate Press. Amer. Iowa Ed. 3.
[12] Pradeep, T. and K. Sumalini. 2003, Impact of mating systems on genetic variability in segregating generation of Asiatic
cotton (Gossypium sp.). Indian J. Genet., 63 : 143-147.
[13] Rahman,

S

and T.A. Malik. 2008. Genetic analysis of fibre traits in cotton. Int. J. Agri. Biol., 10: 209212.
[14] Singh, R.K and B.D. Chawdhury. 1985. Biometrical methods in quantitative genetic analysis, Kalyani Publications, New
Delhi.
[15] Zhang, H.B., Y. Li., B. Wang and P.W. Chee.2008. Recent advances in cotton genomics. Int. J. Plant. Genomics., 2008:
742304.
Estimation of Genetic Parameters for Yield

and Fibre Quality Traits in Inter-Specific
Crosses of Cotton (Gossypium spp.)
Gunasekaran Mahalingam, Krishnasamy Thiyagu and Nagasamy Nadarajan
Department of Cotton, Centre for Plant Breeding and Genetics, Tamil Nadu Agricultural
University, Coimbatore641003 India
AbstractTo study the nature and magnitude of gene effects for seed cotton yield and fibre quality traits in cotton
(Gossypium spp), the generation mean analysis was carried out using the following four crosses of different cotton
cultivars: SVPR 2 Suvin, SVPR 2 Barbados, TCH 1218 Suvin and TCH 1218 Barbados. The P
1
, P
2
, F
1
, F
2
,
B
1
and B
2
of these generations were studied for yield and fibre quality traits. The analysis showed the presence of
additive, dominance and epistatic gene interactions for these characters. Duplicate type epistasis played a greater role
than complementary epistasis. To harness the additive gene action, simple selection procedures or pedigree method of
breeding is sufficient. Heterosis breeding procedures are effective in harnessing dominance gene action to the full
extent. When additive and dominant gene actions are pre-dominant, the bi-parental mating design or reciprocal
recurrent selection can be used for further recombination of alleles to produce desirable segregants.
INTRODUCTION
Cotton is an important natural fibre crop of global importance grown commercially in about 111
countries with a global area of 30.29 million hectares accounting for 105.06 million bales (217.724 kgs)
of production with a productivity of 755 kg/ha. In India, cotton is cultivated in an area of nearly 10.17
million hectares which is the largest in the world, with a production of 29.20 million bales (2009-10)
ranking second next to China. Cotton plays a key role in the national economy by way of its contribution
in trade, industry, employment and foreign exchange earnings. The average productivity of cotton in
India is the lowest among cotton-growing nations of the world.
Though India is a pioneer in cultivation of hybrids on a commercial scale, the productivity of cotton
has been practically stagnant during last few years. In order to increase the yield potential, it is desirable
to efficiently utilize the available genetic variability. Genetic analysis of quantitative traits further helps
to elucidate the nature and magnitude of genetic variation present in the population. The estimates of
gene effects in a plant improvement programme have a direct bearing upon the choice of breeding
procedure to be followed. Additive gene effects are useful in the development of pure lines whereas
dominance and epistatic effects can be used to exploit hybrid vigour. In tetraploid cotton, various studies
have been conducted to study the nature and magnitude of gene effects in the inheritance of different
quantitative characters and involvement of both additive and non-additive gene effects have been
reported by many workers (Nadarajan et al., 1999; Patel et al., 2007). In the present investigation,
additive dominance and epistatic gene effects were estimated by Generation mean analysis for yield and
fibre quality traits in four inter-specific crosses involving G. hirsutum and G. barbadense and suggest
suitable breeding methods for genetic improvement of cotton.
The genetic materials used in this study consisted of two each of G. hirsutum and G. barbadense
genotypes namely SVPR 2 and TCH 1218; and Suvin and Barbados respectively. Two G. hirsutum
genotypes have good combiner, good adaptability, jassid resistance, drought tolerance and higher yield
and the remaining two G. barbadense genotypes have good fibre length, strength and fineness. The
experiment was performed with a set of six generations viz., P
1
, P
2
, F
1
, F
2
, B
1
and B
2
in four cross
combinations viz., SVPR 2 Suvin (cross 1), SVPR 2 Barbados (cross 2), TCH 1218 Suvin (cross 3)
5
Estimation of Genetic Parameters for Yield and Fibre Quality Traits in Inter-Specific Crosses of Cotton (Gossypium spp.) 29
and TCH 1218 Barbados (cross 4) to study the presence of interaction effects in governing the traits of
seed cotton yield and yield attributes and fibre quality traits. The F
1
plants were randomly chosen from
each cross and used as male parent for back crossing programme, whereas the concerned parents of each
cross were used as female parents to produce B
1
(P
1
F
1
) and B
2
(P
2
F
1
) progenies. Simultaneously, F
2

seeds were produced by selfing F
1
plants. All these generations were produced during two cropping
seasons and, as such, all the six generations had to be grown together during the same cropping season.
The F
1
, F
2
, B
1
and B
2
generations of the four crosses were raised along with their parents in a
randomized block design (RBD) with two replications during winter 200809 at Department of Cotton,
Centre for Plant Breeding and Genetics, Tamil Nadu Agricultural University, Coimbatore. Four different
crosses were randomized in a block followed by different generations within each cross. The spacing was
90 60 cm. Ten plants in a row is same for all the generations of each crosses but the number of rows
varied as two rows each for non-segregating population such as P
1
, P
2
and F
1
; 40 rows for F
2
and 20 rows
for each of B
1
and B
2
. Since the non-segregating generations represent the homogeneous population
while the segregating generations represent the heterogeneous population the sample size (i.e. number of
plants analyzed) varied as 20 plants for P
1
, P
2
and F
1
, 400 plants for F
2
and 200 plants in each of B
1
and
B
2
. The recommended agronomic practices and need-based plant protection measures were followed to
obtain good crop stand. Observations were recorded on number of bolls per plant (NB), boll weight (g)
(BW), seed cotton yield per plant (SCY), ginning percentage (%) (GP), 2.5 per cent span length (mm)
(SL) and bundle strength (g/tex) (BS).
The generation means were calculated by taking the average over all the replications for each
generation. The variance and the corresponding standard errors of the means were calculated from the
deviations of the individual values from the pooled mean for each of the generation for every cross. To
test the adequacy of the additive-dominance model the individual scaling test given by Mather (1949) as
well as joint scaling test by Cavalli (1952) were applied. First, simple additive-dominance model
consisting of mean [m], additive [d] and dominance [h] gene effects was tried and the adequacy of the
model was tested statistically by examining the goodness of fit between the estimated generation means
and the observed generation means from the above three parameters for three degrees of freedom.
Wherever the data failed to fit the simple additive dominance non-epistatic-model, the analysis was
proceeded further by perfect fit estimate of the six parameters m, [d], [h], additive additive [i], additive
dominance [j] and dominance dominance [l] on the assumptions of an additive dominance model
with digenic interactions as proposed by Jinks and Jones (1958).
The mean of the six generations with four crosses for six traits are presented in Table 1, with the mean
values for the scaling, joint scaling and their interaction effects being presented in Table 2.
Among the parents, P
1
generation showing higher performance for boll weight, ginning percentage
and seed cotton yield per plant in all the crosses except TCH 1218 Barbados, where P
2
mean was
higher for seed cotton yield per plant. With respect to 2.5 per cent span length and bundle strength, P
2

was superior than P
1
in all the four crosses. Among the different generations all the F
1
s performed better
for number of bolls per plant and seed cotton yield per plant. With regard to 2.5 per cent span length and
bundle strength, F
1
mean was lower than the greater parent in all the crosses. Among the segregating
generations of F
2
, B
1
and B
2
, the B
1
generation showed higher mean performance in all the crosses for all
the traits except 2.5 per cent span length and bundle strength were B
2
showed its superiority over B
1

generations.
A simple additive dominance model was non-adequate as inferred the significance of any one of the
scales and joint scaling test. Hence, an epistatic digenic interaction model was found to be fit for all the
six traits. The additive, dominance and epistatic types of gene interaction in each cross for different trait
were found to be different from each other.

TABLE 1: MEAN VALUES OF DIFFERENT CHARACTERS OVER SIX GENERATIONS IN SIX CROSSES OF COTTON
Characters and
Generations
SVPR 2 Suvin SVPR 2 Barbados TCH 1218 Suvin TCH 1218 Barbados
Number of Bolls Per Plant
P
1
26.90 0.50 25.50 0.43 24.70 0.56 23.70 0.56

P
2
20.70 0.37 27.80 0.42 16.20 0.53 28.70 0.54

F
1
41.70 0.54 45.70 0.73 28.90 0.64 40.80 0.84

F
2
23.64 0.31 22.41 0.32 19.69 0.29 26.75 0.38

B
1
24.63 0.44 27.00 0.51 20.58 0.39 27.34 0.56

B
2
22.77 0.50 25.07 0.50 20.33 0.40 29.42 0.58

Boll Weight (g)
P
1
4.82 0.09 4.74 0.06 4.68 0.08 4.84 0.05

P
2
4.13 0.06 4.01 0.05 4.09 0.04 4.07 0.04

F
1
3.73 0.03 3.71 0.05 4.57 0.05 4.55 0.05

F
2
2.79 0.04 3.08 0.03 2.77 0.04 3.66 0.04

B
1
3.02 0.06 3.44 0.05 3.37 0.05 3.73 0.05

B
2
2.74 0.06 2.86 0.04 2.61 0.04 3.51 0.05

Seed Cotton Yield Per Plant
P
1
104.65 3.32 100.76 2.34 100.37 2.87 95.14 2.21

P
2
61.28 2.41 96.53 2.06 56.28 2.28 96.99 1.17

F
1
134.97 2.64 149.38 2.92 114.16 2.80 169.78 3.45

F
2
50.31 1.19 60.36 1.44 45.49 1.21 81.74 1.92

B
1
60.99 1.93 83.60 2.29 54.12 1.60 90.36 2.56

B
2
47.38 1.87 62.75 2.06 42.69 1.47 87.73 2.76

Ginning Percentage (%)
P
1
36.97 0.40 36.31 0.33 34.87 0.47 34.06 0.62

P
2
33.77 0.47 32.87 0.32 30.56 0.32 33.99 0.45

F
1
33.29 0.35 32.00 0.37 34.07 0.57 33.22 0.38

F
2
30.22 0.20 31.54 0.19 30.44 0.17 30.78 0.17

B
1
32.93 0.33 33.94 0.30 31.25 0.22 32.75 0.23

B
2
29.68 0.29 31.55 0.28 31.67 0.23 30.80 0.26

2.5% Span Length (mm)
P
1
25.46 0.36 25.36 0.31 30.00 0.42 30.06 0.49

P
2
37.28 0.32 34.56 0.29 37.08 0.40 34.54 0.28

F
1
34.20 0.24 33.56 0.25 36.26 0.42 35.10 0.37

F
2
30.34 0.26 29.49 0.23 31.25 0.35 31.45 0.24

B
1
28.47 0.45 28.18 0.38 31.19 0.53 31.28 0.40

B
2
34.94 0.16 31.96 0.38 33.60 0.42 32.16 0.38

Bundle Strength (g/tex)
P
1
18.36 0.21 18.22 0.10 21.18 0.33 21.18 0.33

P
2
27.12 0.15 26.94 0.29 27.20 0.15 26.86 0.27

F
1
24.26 0.26 25.00 0.35 24.36 0.41 24.70 0.41

F
2
22.49 0.26 21.90 0.20 22.91 0.51 23.44 0.23

B
1
20.83 0.39 21.06 0.32 21.19 0.66 22.23 0.30

B
2
25.48 0.39 25.39 0.33 25.18 0.54 25.50 0.41

Number of Bolls Per Plant
Among the genetic effects, the mean effect was higher in SVPR 2 Suvin and TCH 1218 Barbados.
The additive effect (d) was found to be positively significant in SVPR 2 Suvin and TCH 1218 Suvin,
while dominance effect (h) was positive in SVPR 2 Barbados and TCH 1218 Barbados. This
indicated the pre-dominance of both additive and dominance main effects. In case of interaction effects,
the dominance dominance was positively significant for all the crosses, while the additive additive
interaction effect was positive for all the crosses and significantly positive in SVPR 2 Barbados and
TCH 1218 Barbados. The additive dominance effect was negatively significant in TCH 1218 Suvin
and positively significant in SVPR 2 Barbados. The signs of (h) and (l) were opposite in SVPR 2
Suvin and TCH 1218 Suvin, while in the same direction in SVPR 2 Barbados and TCH 1218
Barbados. This indicated the presence of both complementary and duplicate epistasis for this trait. In
general, both additive and dominance main effects, predominant of dominance dominance and additive
additive interaction effects along with both complementary and duplicate dominance epistasis were
noticed for number of bolls per plant. Shanti (1998), Esmail et al. (1999) and Ramalingam and Sivasamy
(2003) reported additive, dominance dominance and additive additive interaction effects for this trait.
TABLE 2: SCALING TEST AND GENETIC EFFECTS FOR DIFFERENT CHARACTERS IN FOUR CROSSES OF COTTON
Cross A B C D JST m (d) (h) (i) (j) (l) 1 2 3 4
SCY
C1 ** ** ** * ** 67.46**
7.47
21.69*
2.05
-36.10**
19.91
15.51*
7.19
-8.08*
3.38
203.61**
13.54
-6.27 157.79 227.20 DE
C2 ** ** ** ** ** 47.36**
8.58
2.12
1.56
-50.04*
22.47
51.28**
8.44
18.74**
3.45
152.06**
15.13
-23.66 52.16 222.08 DE
C3 ** ** ** - ** 66.67**
6.77
22.04**
1.83
-132.19**
17.38
11.65
6.51
-10.62**
2.84
179.68**
12.00
-6.00 154.23 201.95 DE
C4 ** ** ** ** ** 66.83**
0.82
-0.93
1.25
-43.31
27.79
29.24**
0.75
3.56
3.97
146.27**
18.43
46.67 44.24 179.07 DE
NB
C1 ** ** ** - ** 23.57**
1.84
3.10**
0.31
-17.84**
4.82
0.23
1.82
-1.25
0.74
35.97**
3.19
-5.75 20.94 37.45 DE
C2 ** ** ** ** ** 12.14**
1.95
-1.15**
0.30
7.49
5.15
14.51**
1.92
3.08**
0.78
26.06**
3.52
-6.52 8.64 43.65 CE
C3 ** ** ** - ** 17.40**
1.67
4.25**
0.39
-2.33
4.32
3.05
1.62
-4.00**
0.68
13.84**
2.95
-0.55 6.58 20.89 DE
C4 ** ** ** ** ** 19.67**
2.26
-2.50**
0.39
7.18
5.90
6.53**
2.22
0.42
0.89
13.95**
4.02
-2.87 9.68 20.9 CE
BW
C1 ** ** ** - ** 4.12**
0.22
0.34**
0.06
-4.94**
0.58
0.36
0.21
-0.06
0.10
4.55**
0.37
-14.53 5.28 4.98 DE
C2 ** ** ** - ** 4.09**
0.19
0.36**
0.04
-3.66**
0.49
0.28
0.18
0.22**
0.08
3.28**
0.32
-10.17 4.02 3.78 DE
C3 ** ** ** ** ** 3.50**
0.20
0.29**
0.05
-4.01**
0.51
0.88**
0.20
0.47**
0.08
5.08**
0.33
-13.83 4.30 6.43 DE
C4 ** ** ** - ** 4.63**
0.21
0.39**
0.03
-3.79**
0.55
-0.18
0.21
-0.18*
0.08
3.71**
0.36
-9.72 4.18 4.07 DE
GP
C1 ** ** ** ** ** 31.03**
1.23
1.60**
0.31
-5.49
3.24
4.34**
1.19
1.65**
0.54
7.75**
2.15
-3.43 7.09 13.74 DE
C2 - * ** ** ** 29.76**
1.13
1.72**
0.23
4.86
2.97
4.83**
1.10
0.67
0.47
-2.62
1.99
2.83 6.58 8.13 DE
C3 ** - ** ** ** 28.63**
0.97
2.15**
0.28
1.79
2.54
4.08**
0.93
-2.57**
0.42
3.65**
1.92
0.83 3.94 10.3 CE
C4 * * ** ** ** 31.03**
1.23
1.60**
0.31
-5.49
3.24
4.34**
1.19
1.65**
0.54
7.75**
2.15
-3.43 7.09 13.74 DE
*Significant at 5% level, **Significant at 1% level; C1SVPR 2Suvin; C2SVPR 2Barbados; C3 TCH 1218Suvin;
C4TCH 1218Barbados; A, B, C, DScales; JSTJoint scaling test; mmid parent; (d)additive; (h)dominance; (i)additive
additive; (j)additive dominance; (l)dominance dominance; 1h/d; 2Total main effect; 3Total interaction effect; 4Type
of epistasis; CE Complementary Epistasis; DEDuplicate Epistasis
Boll Weight
Among the genetic effects estimated, the additive main effect (d) was pre-dominant and showed
positively significant values in all crosses and was greater than dominance effects (h) where negatively
significant interaction effect was noticed in all the crosses. The dominance dominance interaction
effect (l) was found to be positively significant in all crosses. The additive dominance interaction effect
(j) was positively significant in SVPR 2 Barbados and TCH 1218 Suvin, while additive additive
interaction was positively significant in TCH 1218 Suvin. Duplicate dominance epistasis was
confirmed in all the crosses due to the opposite signs of (h) and (l). On the whole, additive gene effects
and epistatic effects, especially of dominance dominance and additive dominance type were found to
govern this trait along with duplicate type of epistasis. This is in conformity with the results of Sandhu et
al. (1992). Rajendra Kumar and Raveendran (2001) reported the presence of additive effects for boll
weight. Ramalingam and Sivasamy (2003) expressed dominance dominance and additive dominance
interaction effect for this trait.
Seed Cotton Yield Per Plant
The perusal of six generations using generation mean analysis indicated that additive gene effects were
prevalent for majority of the crosses viz., SVPR 2 Suvin, SVPR 2 Barbados and TCH 1218 Suvin.
In SVPR 2 Suvin and TCH 1218 Suvin, additive effect was positively significant. The dominance
effect (h) was negatively significant in all the crosses except TCH 1218 Barbados which had negative
effect. The dominance dominance interaction effect (l) was pre-dominant in all the crosses where it was
positively significant followed by additive additive effect which was also positively significant in all
the crosses except TCH 1218 Suvin. The additive dominance interaction effect (j) was positively
significant in SVPR 2 Barbados and negatively significant in SVPR 2 Suvin and TCH 1218 Suvin.
The signs of (h) and (l) were opposite in direction in all the crosses. This indicated that seed cotton yield
per plant was found to be governed by additive gene effects and interaction effects pre-dominantly of
dominance dominance type followed by additive additive with duplicate type of epistasis. All the
three types of interaction for this trait were reported by Singh and Chahal (2004) and Singh et al. (2008).
Shanti (1998) reported additive effect while Esmail et al. (1999) reported additive additive gene
interactions but Jagtap (1995) reported both type of gene actions for this trait.
Ginning Percentage
Among the genetic effects estimated, interaction effects were witnessed in all crosses. In these crosses,
the additive genetic effect was positive in TCH 1218 Barbados and significantly positive in rest of the
crosses viz., SVPR 2 Suvin, SVPR 2 Barbados and TCH 1218 Suvin, while the dominance gene
effects (h) was found to be positive in SVPR 2 Barbados and TCH 1218 Suvin and negative in SVPR
2 Suvin and TCH 1218 Barbados. The additive additive interaction effect was found to be
positively significant in all the crosses. The additive dominance effect (j) was positively significant in
SVPR 2 Suvin and TCH 1218 Barbados and positive for SVPR 2 Barbados. The dominance
dominance effect (l) was positively significant in SVPR 2 Suvin and TCH 1218 Suvin, positive in
TCH 1218 Barbados and negative in SVPR 2 Barbados. The signs of (h) and (l) were dissimilar in all
crosses except TCH 1218 Suvin. Hence it appears that, the additive effect along with all the three types
of interaction with duplicate dominance epistasis existed pre-dominantly for ginning percentage. Panchal
et al. (1994) reported the presence of additive, additive additive, additive dominance and dominance
dominance interactions for ginning percentage. Rajendra Kumar and Raveendran (2001) and Shanti
(1998) reported additive gene effects and Sandhu et al. (1992) reported all the three types of epistasis
gene interactions.
Per Cent Span Length
The mean effect estimated by the perfect fit method for digenic interactions in all the crosses was highly
and positively significant. The interaction effects were witnessed in all crosses. In these crosses, the
dominance genetic effect was positive in SVPR 2 Barbados and TCH 1218 Suvin, positively
significant in SVPR 2 Suvin and negative in TCH 1218 Barbados, while the additive gene effect (d)
was found to be negatively significant in all the four crosses. The additive additive interaction effect
was found to be positively significant in SVPR 2 Suvin and TCH 1218 Suvin and positive for the rest
of the crosses. The additive dominance (j) and dominance dominance effect (l) were positively
significant for TCH 1218 Barbados, positive for SVPR 2 Barbados and TCH 1218 Suvin and
negative for SVPR 2 Suvin. Hence it appears that, the dominance effect along with additive additive
gene action followed by additive dominance and dominance dominance gene action with duplicate
and complementary epistasis exists for 2.5 per cent span length. Mehetre et al. (2003) reported the
presence of dominance, additive additive and dominance dominance interaction for 2.5 per cent span
length. Thangaraj et al. (2002) and Singh and Chahal (2004) reported additive, dominance and duplicate
epistasis for 2.5 per cent span length.
Bundle Strength
The mean effect was positively significant and higher than other genetic effects. The interaction effects
were witnessed by significance of scale C in all crosses. The dominance effect (h) was positively
significant in SVPR 2 Barbados and positive for SVPR 2 Suvin and TCH 1218 Barbados and
negative in TCH 1218 Suvin. The additive effect (d) was negatively significant in all crosses. The
additive additive effect (i) was positive in all the crosses except SVPR 2 Barbados, where it was
positively significant. The additive dominance effect (j) was positive in SVPR 2 Barbados and
negative in SVPR 2 Suvin, TCH 1218 Suvin and TCH 1218 Barbados. The dominance
dominance interaction effect (l) was negative in SVPR 2 Suvin and SVPR 2 Barbados and positive
for rest of the crosses. The signs of (h) and (l) were opposite in all crosses except TCH 1218 Barbados
indicating the presence of duplicate dominance epistasis. To conclude, this trait was controlled by
dominance gene effects and interaction effects mainly of additive additive with duplicate dominance
type of epistasis. Mehetre et al. (2003) reported additive, dominance and additive additive gene effects.
Hendawy et al. (1999) reported additive and additive additive with duplicate type of epistasis for this
trait.
CONCLUSION
The gene action analyzed through generation mean analysis indicated that both additive and dominance
gene effects were found to control most of the important yield contributing and fibre quality traits viz.,
number of bolls per plant, boll weight, seed cotton yield per plant and ginning percentage, while
dominance gene action was prevalent for the traits viz., 2.5 per cent span length and bundle strength. One
or more type of epistatic interaction effects was prevalent for all the characters. Duplicate dominance
type of epistasis was witnessed in all characters, while complementary epistasis was also noticed in
certain crosses for the traits number of bolls per plant and 2.5 per cent span length.
To harness additive gene action, simple selection procedures or pedigree breeding method is
sufficient. But the presence of dominance gene action in most of the characters warrants postponement of
selection to later generations after effecting crosses. Heterosis breeding procedures are effective in
harnessing dominance gene action to the full extent. Both additive and dominance gene actions play
major role in several characters. In such circumstances biparental mating design or reciprocal recurrent
selection can be followed which allows further recombination of alleles to produce desirable segregants.
These methods can also be well adopted in order to harness the epistatic interactions by way of breaking
the undesirable linkages. Diallel selective mating system could also be followed to break such
undesirable linkages between two or more genes and to produce desirable recombinants.
REFERENCES
[1] Cavalli, L.I. (1952). An analysis of linkage in quantitative inheritance. In: Quantitative inheritance. E.C.R.Reeve and
C.H.Waddington (eds.), HMSO, London. pp135-144.
[2] Esmail, R.M., Hendawy, F.A., Rady, M.S. and Abd-El-Hamid, A.M.. (1999). Genetic studies on yield and yield
components in one inter and two intraspecific crosses of cotton. Egyptian J. Agron., 21: 3751.
[3] Hendawy, F.A., Rady, M.S., Abd-El-Hamid, A.M. and Esmail, R.M. (1999). Inheritance of fibre traits in some cotton
crosses. Egyptian J. Agron., 21: 1536.
[4] Jagtap, D.R. (1995). Genetic components of heterosis and selection potential of crosses in generation means of upland
cotton. J. Indian Soc. Cotton Improv., 20(3): 142148.
[5] Jinks, J.L. and Jones , R.M. ( 1958). Estimation of the components of heterosis. Genetics, 43: 223234.
[6] Mather, K. (1949). Biometrical genetics. Dover publication in New York. p58.
[7] Mehetre, S.S., Rajput, H.J., Shinde, G.C. and Mokate, A.S. (2003). Genetics of fibre quality traits in intraspecific crosses of
G. hirsutum cotton. J. Indian Soc. Cotton Improv., 28(3): 132136.
[8] Nadarajan, N., Kumaresan, D., Ponnusamy, K. and Azhuguvel, P. (1999). Genetic analysis of fibre quality characters in
upland cotton (Gossypium hirsutum L.). In: International seminar on cotton and its utilization in the 21st century, Dec. 10-
12, CIRCOT, Mumbai. p43.
[9] Panchal, S.S., Patel, J.A., Patel, S.A. and Dahal, K.C. (1994). Genetic architecture of lint component characters in
interspecific hybrids of cotton. Gujarat Agril. Univ. Res. J., 19(2): 6264.
[10] Patel, K.G., Patel, R.B., Patel, M.I. and Kumar, V.(2007). Genetic of yield, fibre quality and their implications in breeding
of interspecific cross derivatives of cotton. J. Cotton Res. Dev., 21: 153-157.
[11] Rajendra Kumar, P. and Raveendran, T.S. (2001). Genetic evaluation of yield and yield components in upland cotton
through triple test cross analysis. Indian J. agric. Sci., 71: 6264.
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cotton (Gossypium hirsutum). Madras Agric. J., 90(79): 472477.
[13] Sandhu, B.S., Gill, M.S. and Mittal, V.P. (1992). Genetic architecture of Gossypium arboreum. Indian J. Genet., 52(3):
257260.
[14] Shanti, R.M. (1998). Investigation on genetic potential and biochemical compounds related to resistance of Helicoverpa
armigera (Hubner) in the racial and wild species derivatives of Gossypium spp. Ph.D Thesis, Tamil Nadu Agricultural
University, Coimbatore, India. (Unpublished)
[15] Singh, P. and Chahal, G.S. (2004). Simultaneous improvement of yield and fibre quality in upland cotton (Gossypium
hirsutum L.). Indian J. agric. Sci., 74(12): 643648.
[16] Singh, P., Chahal, G.S. , Mittal, V.P. and Brar, K.S. (2008). Genetic analysis of yield components and fibre quality
characters in upland cotton (Gossypium hirsutum L.). Indian J. Genet., 68(1): 3337.
[17] Thangaraj, K., Raveendran, T.S. and Jehangir, K.S. (2002). Heterosis and inbreeding depression in interracial derivatives of
Gossypium hirsutum L. for fibre quality. J. Indian Soc. Cotton Improv., 27(1): 1518.
Introgression of Desirable Characters

for Growing Cotton in Pakistan
Abid Mahmood, Jehanzeb Farooq and Noor-Ul-Islam
Cotton Research Institute, Ayub Agricultural Research Institute, Faisalabad, Pakistan
AbstractCotton crop face both biotic and abiotic stresses. Keeping in view the emerging problems of cotton in
Pakistan, a breeding programme was initiated to develop cotton varieties which can withstand under changing
environment especially at high temperature and Cotton Leaf Curl Virus (CLCuV) conducive conditions. A series of
experiments was conducted to select parental lines which can contribute for high temperature tolerance, low input
requirements, earliness, good fibre squality traits, CLCuV tolerance and plants suitable for high density planting. A
comprehensive breeding programme was initiated to combine desirable characters in resulting breeding material.
Screening of breeding material was carried out in CLCuV hot spot areas. At the same time fibre quality, earliness and
high yield were also considered during selection. For low input requirement varieties like FH-113 and FH-942 were
developed. For heat and CLCuV tolerance 24 Lines were developed. Some of the lines like MNH-886, FH-142 and
MNH-456 showed promising results in this respect. For high density planting of cotton, the line FH-114 showed good
performance at densities of 10,0000 plants per hectare. It was recommended that for late sowing of cotton, plant to
plant distance of FH-114 may be reduced to 12cm to achieve desirable yield per unit area. The varieties developed
during these studies exhibited the highest yield with good fibre quality in CLCuV hot spot areas. It is expected that in
future more improved germplasm will be available which will enhance the productivity of cotton in Pakistan.
INTRODUCTION
The importance of upland cotton, (Gossypium hirsutum L), is evident from the fact that it is the worlds
leading fibre producing species (Dutt et al., 2004; Fryxell, 1992). It contributes 60% in the total foreign
exchange through the exports of value added products (Iqbal et al. 2005). Cotton accounts for 8.6 % of
the value added in agriculture and about 1.8 % to GDP of Pakistan (Anonymous, 2007). Pakistan is the
4
th
largest cotton producing country in the world after Peoples Republic of China, USA and India, and 3
rd

largest consumer of cotton after People Republic of China and India (Akhtar, 2005).
Keeping in view the geographical situation and farming system of Pakistan breeding of cotton for the
development of earliness, heat tolerance, low input requirements, CLCuV resistance and better quality
traits are the main objectives. These breeding objectives help to protect environment, boost cotton
productivity and make cotton production a profitable venture. Most of the cotton growers in Pakistan are
poor and cannot afford heavy expenses of inputs like fertilizer, pesticides and irrigation water. Varieties
having deep root system and higher stature require less irrigation water thus can meet the needs of the
farmer.
Similarly the varieties with Bt gene provide resistance against bollworms thus reduce the cost of
pesticides to control these insects. Bt. Cotton provides an alternative by replacing insecticides with a
toxin within the plant. According to Layton et al. (1997) overall performance of Bt. Cotton was better
than conventional varieties. Transgenic Bt. cotton can effectively control specific lepidopterous species
(Arshad et al., 2009).
For the last many years one of the main causes of low per acre yield of cotton in Pakistan is CLCuV
attack. In the early nineties cotton varieties like CIM-1100, CIM-448, CIM-446, CIM-443, MNH-552,
MNH-554 were developed which were having resistance against CLCuV. Build up of this disease in the
recent years resulted in high inoculum pressure under which varieties which were initially showing some
tolerance later became susceptible (Mahmood et al., 2003; Shah et al., 2004).
In several districts of Punjab and Sindh severe summer temperatures exceeding 45
0
C cause
considerable damage to cotton crop (Rehman et al., 2004). Fruit setting in upland cotton is severely
6
effected if day temperature of >30
0
C remained for a period of 13 hrs or more (Reddy et al., 1992).
Further, it is the need of the day to improve fibre quality in the Hirsutum genotypes, to fulfill the
requirements of growing textile industry.
Cotton plant sets its bolls (fruit) over a period of about 80 days. Delayed in that period allows various
environmental factor to act and effect maturation period (Iqbal et al., 2003). Late maturating types are
ultimately affected by a later pest pressure. Cotton fiber quality is primarily influenced by late maturity of
the genotype and by environmental conditions as the secondary factors (Subhan et al., 2001). Earliness in
cotton is a complex polygenic trait influenced by a number of factors like morphology, phenology,
physiology and environmental attributes (Shah et al., 2010). Earliness allows development of crop during
period of favorable moisture and timely picking prevent the crop from unfavorable weather (Rauf et al.,
2005). The benefit of growing early maturing cotton cultivars is the provision of proper time for rotation
of other crops allowing timely sowing of wheat in cotton wheat cotton cropping system as in Pakistan
(Ali et al., 2003). The cultivation of early maturing cultivars not only minimizes the use of pesticides, but
the expenses incurring on other inputs like irrigation water and fertilizer will also be reduced.
In Pakistan under cotton wheat cotton rotation system planting of cotton become late due to
delayed harvesting of wheat crop. Delayed planting of cotton poses many problems including severe
insect pest attack, incidence of CLCuV and poor growth of the plants. To get better yield in late plantings
(high plant population) might be beneficial provided that the late sown genotypes be CLCuV tolerant and
fit to high population. Close row spacing and high plant populations in ultra-narrow rows lead to more
rapid canopy closure than in wider rows (Robinson, 1993), which leads to increased light interception
and reduced weed competition (Kreig, 1996).
The main objective of these studies was to develop germplasm having combination of characters
including Bt (Cry-1 Ac), earliness, heat tolerance, resistance to CLCuV and less fertilizer requirement. In
addition this germplasm should fit in cotton-wheat-cotton rotation system.
In order to combat the threats of cotton bollworms, CLCuV, heat stress and for the development of
varieties having less fertilizer requirement comprehensive breeding programme was initiated. For this
purpose parents having desirable characters were identified. The detail of characteristics of the parent
lines is given in Table 1.
TABLE 1: NAMES AND VARIOUS CHARACTERISTICS OF THE LINES USED AS PARENT IN VARIOUS EXPERIMENTS INCLUDED IN THESE STUDIES
Parents Distinctive Features
FH-925 Early sympodial type variety with medium boll. Having tolerance to heat and CLCuV.
Nucot-N33B
An Australian line with Bt gene and was used to transfer Bt in local germplasm possessing good fibre and
plant traits.
FH-925 Early sympodial type variety with medium boll. Having tolerance to heat and CLCuV.
Nucot-N33B
An Australian line with Bt gene and used for transfer it in local germplasm possessing good fibre and plant
traits.
FH-900(S)
Resistance against CLCuV, relatively tolerant to heat stress, good fiber quality, fitness in cotton-wheat
rotation system and adaptability to wider range of environments
CIM-125 Medium in maturity, intermediate in growth habit and possess good fibre traits.
FH-207
Tall with long sympodial branches, moderately resistant to sucking pest and bollworms. Suitable for Low
CLCV intensity areas of non-core zone.
MNH-770
Medium tall plant with tolerance to CLCuV and heat. Its distinctive feature is yellow pollen.
MNH-609 Medium tall plant with tolerance to CLCuV and heat.
A systematized crossing programme was carried out to combine the desirable characters in resulting
breeding material. For this purpose after crossing F
1
and F
2
were raised. In F
2
single plants with desirable
characters were selected. Progeny selection was carried out from F
3
to F
6
on the basis of seed cotton yield
and fibre quality. The detail of the selected genotypes included in the studies is given in Table-2.

Introgression of Desirable Characters for Growing Cotton in Pakistan 37
TABLE 2: PEDIGREE AND VARIOUS TRAITS OF LINES INCLUDED IN VARIOUS EXPERIMENTS CONDUCTED AT COTTON RESEARCH INSTITUTE, FAISALABAD
Genotypes Parentage Distinctive Features
FH-113 FH-925 NuCot-N33B
Tall growing Bt variety possesses moderate tolerance to CLCuV. It has strong
stem, prolonged flowering duration and require low inputs.
FH-114 FH-925 NuCot-N33B
A compact input intensive Bt variety with high tolerance to CLCuV and heat
stress. Best suited to high plant population in late sowing.
FH-942 FH-900(S) CIM-121
A spreading growth habit variety most suitable for water-scarce areas. Possess
good boll size and fibre quality of national standards.
MNH-886 FH-207 MNH-770
A revolutionary variety with respect to CLCuV. Intermediate to spreading in its
growth habit. Boll size of this variety is medium and can tolerate heat stress in
core areas of Punjab.
MNH-456 FH-207 x MNH-609 Highly tolerant to CLCuV
A series of experiments were carried out in targeted environments and conditions to find out various
lines of cotton. The studies were carried out at experimental area of Cotton Research Institute,
Faisalabad. The soil had pH was 8.2, EC 1.3 dSm-1, nitrogen 0.030% and available Phosphorous 7.0 mg
kg-1. The detail of experiments is as under.
Experiment 1: Selection of Lines for Low Fertilizer Requirement
These studies were conducted following Split Plot design with 4 replications. The fertilizer doses were
randomized as main plots and genotypes as sub-plot. The line to line distance was kept as 75cm and plant
to plant distance at 30cm. Each plot was consisted of 5 lines with 5 meter length. Three genotypes viz.,
FH-113, FH-942 and FH-1000 were included in these studies. Sowing of the genotypes was carried out
on May 3 in 2003-04 and on May 12 in 2004-05. The phosphorus and potash were applied basaly at the
time of sowing while the nitrogen was applied in three splits i.e., with first irrigation, at the time of start
of square formation and at blooming stage. There were three treatments, i.e full doze of recommended
fertilizer (NPK, 114:54:62), half dose of fertilizer (NPK, 57:27:31) and control, (where no fertilizer was
applied). Before the experiment the soil was exhausted by growing maize crop in the field. All other
agronomic practices and insect control measures was carried out as standard procedure. At maturity data
of seed cotton yield/ha was recorded. Analysis of variance was carried out and means of genotypes were
compared by LSD.
Experiment 2: Selection of CLCuV Tolerant Lines
In this experiment high yielding with good fibre quality character lines were included. Twenty four
advance lines of different genetic background were sown in three replications to identify lines tolerant to
CLCuV. Each genotype was planted in two rows with row length of 5 meter. Standard agronomic
practices were applied. To maximize CLCuV inoculum pressure the pesticide for whitefly was not
applied throughout the experiment. Data for CLCuV was recorded following the rating system described
by Akhtar et al, 2010. The procedure of rating CLCuV incidence is briefly mentioned in Table 3.
TABLE 3: DISEASE SCALE FOR RATING COTTON LEAF CURL VIRUS DISEASE
Symptoms Disease
Index%
Rating Disease
Response
Complete absence of symptoms 0 0 Immune
Thickening of few small scattered veins or only presence of leaf enations on one or
few leaves of a plant observed after careful observations.
0.110 1 Highly resistant
Thickening of small group of veins, no leaf curling, no reduction in leaf size and
boll setting.
10.1-20 2 Resistant
Thickening of all veins, minor leaf curling and deformity of internode with minor
reduction in leaf size but no reduction in boll setting.
20.1-30 3
Moderately
resistant
Severe vein thickening, moderate leaf curling followed by minor deformity of
internodes and minor reduction in leaf size and boll setting.
30.1-40 4
Moderately
susceptible
Severe vein thickening, moderate leaf curling and deformity of internodes with
moderate reduction in leaf size and boll setting followed by moderate stunting.
40.1-50 5 Susceptible
Severe vein thickening, leaf curling, reduction in leaf size, deformed internodes and
stunting of the plant with no or few boll setting.
>50 6
Highly
susceptible
Foliar outgrowths (enation) will be marked with E where observed.
To calculate severity index (SI) and per cent disease index (PDI) of the genotype under study,
Individual symptomatic plant ratings for each genotype was added and divided by the number of infected
plants to calculate the corresponding SI. The PDI was calculated using the following formula:
Percent Disease index=
Sum of all disease ratings of the selected plants at random

100
Total no. of plants assessed. 6
Experiment 3: High Density with Late Cotton Sowing Studies
These studies were carried out to find out suitable lines for high density planting of cotton. For this
purpose cotton lines with short sympodia (FH-114 and FH-2015) and intermediate sympodia FH-113
were selected. The cotton was sown after the harvesting of wheat. The trial was sown on June 10, 2004-
05. All agronomic practices were carried out as standard except line to line distance was kept 37cm and
plant to plant was 12cm. Plant population 43000 and 10,0000/ha was maintained. At maturity, data on
seed cotton yield and yield components were recorded. Ginning out turn was calculated after ginning of
50 gm sample of seed cotton and other fibre quality traits like staple length, maturity and fineness was
measured.
Experiment 4: Heat Tolerance Studies
For the identification of heat tolerant cotton genotypes the same 24 genotypes which were studied in
CLCuV screening experiment were included. The genotypes showed maximum boll retention at highest
temperature of the flowering period of cotton was identified as heat tolerant. The experiment was sown
on April, 15 in 2003-04 and on April, 12 in 2004-05. The data for boll retention was recorded from June
15 to July 15 each year. These studies were carried out at two locations i.e., Faisalabad and Multan. The
data of maximum temperature of experimental sites is given in Table 4. The location of Multan was
included in this study as it has the highest temperature in Punjab. Boll retention percentage was
calculated by dividing the number of retained boll by total number of flowered fruiting sites.
TABLE 4: MAXIMUM TEMPERATURE FOR THE MONTH OF JUNE AND JULY DURING 2003-04 AND 2004-05
Location Month Temperature (
0
C) 2003-04 Temperature(
0
C) 2004-05
Multan
June 43.5
0
C 43
0
C
July 44.5
0
C 44
0
C
Faisalabad
June 41.0
0
C 41
0
C
July 42.0
0
C 41.5
0
C
The results of the fertilizer trial in two year study i.e. 2003-04 and 2004-05 indicated highly significant
differences (P>0.05) for varieties, fertilizer doses and their interaction. On the basis of first year, FH-113
surpassed all varieties by producing a yield of 2860kg/ha at full dose of fertilizer followed by FH-942,
(2770kg/ha) and FH-1000 (2133kg/ha). At half dose of fertilizer FH-113 also produced maximum yield
of 2470kg/ha followed by FH-942 (2120kg/ha) and FH-1000 (1880kg/ha). At control, FH-113 produced
maximum yield of 1560kg/ha. During second year again FH-113 produced maximum yield of 2903kg/ha
at full dose of fertilizer than FH-942 (2685kg/ha) and FH-1000 (2237kg/ha). At half dose FH-113
showed its superiority by producing yield of 2575kg/ha which is significantly higher than the other two
varieties. It is evident from the results of both years that the variety FH-113 not only performed well at
full dose of fertilizer but also produced highest yield at half dose of fertilizer. Khan et al. (1994) and Latif
et al. (1994) were of the view that 100 kg N was the optimum N requirement for cotton under Faisalabad
and Sakrand conditions, respectively. But in present studies FH-113 produced promising yield at half
dose of fertilizer also. The detailed results of analysis of variance and means of the genotypes at three
levels of fertilizer are given in Table 5 and 6.

Results of the screening experiment against CLCuV revealed that three genotypes viz., MNH-609,
MNH-886 and MNH-456 showed resistance to CLCuV by showing disease severity of 1.65, 1.69 and
1.74 respectively and disease index of 17.8, 18.7 and 19.3% respectively (Table 7). These genotypes
possess Cry1 Ac gene and hence have no attack of boll worms. However, FH-114, FH-941, FH-942 and
BH-168 were among moderately susceptibility genotypes as the disease index of these genotypes was
below 50%. Remaining seventeen genotypes showed high susceptibility ranging from 58.35 to 80.12%.
The strains resistant to CLCuV sometimes again become susceptible due to the emergence of new strains
of virus which ultimately results in resistance break down (Mansoor et al., 2003, Akhtar et al., 2002). In
Pakistan, previously successful efforts have been made to develop virus resistant varieties resulted in
recovery of production that was improved from 8.04 million bales in 1993-94 to 11.17 million bales in
1999-2000, (Anonymous, 2001). In the present studies resistance to CLCuV exhibited by some lines is
encouraging though continued efforts are needed to develop resistant germplasm against CLCuV and
various new strains of virus.
For high population studies the genotypes with compact structure, FH-114, FH-2015 and
intermediate growth habit (FH-113) were evaluated. The morphological (plant height, sympodia per
plant, bolls per plant, boll weight, seed cotton yield) and fibre quality traits like ginning out turn%, fibre
length, strength and fineness were studied at population densities of 43000 and 10,0000 plants/ha. FH-
114 gave a mean plant height of 96.4cm, 12.8 sympodial per plant, 12.0 bolls per plant, 2.6g boll weight
and seed cotton yield of 1300kg/ha at plant population of 43000 but at plant population of 10,0000 the
yield was 1900kg/ha. The plant height at this density reduced and remained at 88.7cm. However, boll
number and number of sympodia almost remained the same but boll weight reduced to 2.4g. Ginning out
turn, fibre length, fibre strength and fibre fineness at plant population of 43000 was recorded at 38.0%,
27.7mm, 97.63tppsi and 4.9g/inch respectively and at plant population of 10,0000/ha these values
remained at 37.5%, 27.5mm, 98.0tppsi and 5.1 g/inch respectively. The intermediate growth habit
variety FH-113 produced a yield of 1050kg/ha at 43000 plants per hectare and at 10,0000 plants per
hectare it produced a yield of 1600 kg/ha. It attained a plant height of 100.0 cm at 43000 and 93.0cm at
10,0000 plants/ha. Similarly, for boll weight, almost similar values were found at both plant denstities
but boll number reduced at 10,0000 plants/ha. In terms of fiber quality traits same pattern of variation
existed with little reduction in all the studied traits (Table 7). FH-2015 another compact variety evaluated
at both population densities however it was not successful as it produced a very low yield of 700 and
1100 kg/ha at both populations. Its low yield is evident from less sympodia per plant, less boll number
and reduced boll weight. In terms of fibre quality traits it is most affected by the environment and showed
poor results. Results at both densities showed that FH-114 can be exploited in late sown conditions as it
can tolerate CLCuV to greater extent as compared to other genotypes hence produced more yield. The
current results are in conformity with the results of Krishna et al., (2009) who proposed that by growing
cotton in narrow rows can produce more seed cotton yield than the wider rows.
The same genotypes that were tested for screening against CLCuV also tested for heat stress
tolerance. The genotypes namely, MNH-456, FH-114 and MNH-886 showed boll retention percentage of
82.2, 80.4 and 70.9% respectively (Table 8).These genotypes can tolerate the temperature of 40
0
C and
can be grown most successful at high temperature regions. The remaining genotypes showed variable
range from 29.3% to 75.4% boll retention (Table 8). High temperature results in poor pollen germination
and pollen tube growth affecting yield to greater extent. Temperature beyond 45
0
C severely reduces the
growth and development of cotton plant (Khan et al., 2005). The results are encouraging as the genotypes
like MNH-886, MNH-456 and FH-114 have been developed by keeping in view the improvement in
multi-traits. These varieties not only can cover the threat of CLCuV but also can tolerate heat and have
good fiber quality traits. The further studies are underway to develop better varieties which can withstand
in CLCV conditions and having heat tolerance, good yield and fiber quality.

TABLE 5: MEAN SQUARES FOR SEED COTTON YIELD
Source DF MS (2003-04) MS (2004-05)
Reps 3 135503* 42879*
Doses 2 3479470 4134993
Error 6 12754 11215
Varieties 2 836480* 996353*
Doses x Varieties 4 66963* 18952*
Error 18 16120 16031
TABLE 6: YIELD (KG/HA) PERFORMANCE OF VARIETIES AT DIFFERENT DOSES OF FERTILIZER
Fertilizer Doses Control (0-0-0) Half Dose of Fertilizer(57:27:31) Full Dose of Fertilizer (114:54:62)
Varieties 2003-04 Variety Mean
FH-113 1560 2470 2860 2297
FH-942 1340 2120 2770 2077
FH-1000 1328 1880 2133 1780
Fertilizer doses mean 1409 2157 2588
Varieties 2004-05 Variety Mean
FH-113 1644 2575 2903 2374
FH-942 1390 2237 2685 2104
FH-1000 1160 2010 2250 1807
Fertilizer doses mean 1398 2274 2613
LSD(0.05): For fertilizer doses(2003-04)=112.82
LSD(0.05):For fertilizer doses(2004-05) =106.79
LSD (0.05): For varieties (2003-04) =108.90
LSD (0.05): For varieties (2004-05) =106.60
TABLE 7: PER CENT DISEASE INDEX (PDI), SEVERITY INDEX (SI) AND DISEASE REACTION OF VARIOUS GENOTYPES TESTED AT COTTON RESEARCH INSTITUTE, FAISALABAD
Region Genotypes PDI (%) SI Disease Reaction
Faisalabad
FH-113 63.73 3.82 HS
FH-114 39.27 2.47 S
FH-941 44.84 3.12 S
FH-942 48.05 2.88 S
FH-1067 80.12 4.81 HS
Sahiwal
SLH-284 70.97 4.26 HS
SLH-317 76.12 4.57 HS
Bahawalpur
BH-162 58.85 3.53 HS
BH-167 62.98 3.78 HS
BH-168 49.49 3.06 S
BH-197 74.78 4.49 HS
Multan
MNH-93 58.35 3.5 HS
MNH-149 67.09 4.02 HS
MNH-253 57.55 3.45 HS
MNH-456 19.32 1.74 R
MNH-609 17.82 1.65 R
MNH-700 68.23 4.04 HS
MNH-752 75.25 4.51 HS
MNH-784 63.9 3.83 HS
MNH-787 64.77 3.89 HS
MNH-789 63.14 3.79 HS
MNH-886 18.67 1.69 R
Vehari
VH-144 65.43 3.93 HS
VH-278 76.06 4.56 HS
TABLE 8: PERFORMANCE OF VARIETIES AT DIFFERENT PLANT POPULATION FOR VARIOUS MORPHOLOGICAL AND FIBRE TRAITS
Varieties Plant
Population
ha
-1

Plant
Height
(cm)
Sympodia
Per plant
Bolls
Per
Plant
Boll
Weight
(g)
Seed Cotton
Yield
(kg/ha)
GOT
%
Fibre
Length
(mm)
Fibre
Strength
(tppsi)
Fibre
Fineness
(g/inch)
FH-114 43000 96.4 12.84 12.08 2.61 1200 38.0 27.7 99.6 4.9
10,0000 88.72 11.68 11.48 2.36 1900 37.5 27.5 98.0 5.1
FH-113 43000 100.08 11.04 7 3.32 1050 38.3 28.5 97.3 5.1
100000 93 13 10 3 1600 38 28.0 96.5 5
FH-2015 43000 75.4 7 7 2.10 700 37.0 27.6 94.7 5.2
10,0000 70.4 6 5 1.90 1100 36.3 27.3 94.2 5.4
TABLE 9: BOLL RETENTION PERCENTAGES OF VARIOUS GENOTYPES TESTED FOR HEAT TOLERANCE
Region Genotypes Boll Retention (%)
Faisalabad Region
FH-113 65.0
FH-114 80.4
FH-941 50.0
FH-942 55.2
FH-1067 71.0
Sahiwal
SLH-284 67.2
SLH-317 50.0
Bahawalpur
BH-162 54.4
BH-167 62.1
BH-168 54.1
BH-197 42.2
Multan
MNH-93 45.1
MNH-149 31.7
MNH-253 65.0
MNH-456 82.2
MNH-609 40.0
MNH-700 70.0
MNH-752 42.6
MNH-784 29.3
MNH-787 40.9
MNH-789 65.3
MNH-886 70.91
Vehari
VH-144 75.4
VH-278 50.4
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selected upland cottons. Acta Genetica Sinica 1: 79-85.
Temporal Changes in Metabolically Important

Enzymes and Solutes act as Trigger for Epidermal
Cell Conversion to Fibre Initials in Cotton
Gopalakrishnan N.
1
, A.H. Prakash
2
and Y.L. Balachandran
3
1
Assistant Director General (Commercial Crops), ICAR, New Delhi, India
2
Central Institute for Cotton Research, Regional Station, Coimbatore, India
3
Department of Biotechnology, Bharathiyar University, Coimbatore, India
AbstractAs one of the longest single-cell seed trichomes, cotton fibres act as an excellent model for unraveling
fundamental biological processes such as cell differentiation, cell expansion and cell wall biosynthesis. In order to probe
better cotton fibre production through effective fibre development processes, temporal changes in the oxidative enzymes
and solute contents were studied in near isogenic lines of linted cotton cv. MCU 5 and its lintless mutant MCU5 LL.
The ovules were quantified for Peroxidase (POD), Catalase (CAT), Ascrobate peroxidase (APX) and Superoxide
dismutase (SOD) from ten days pre anthesis to six days post anthesis (DPA). The POD activity in MCU 5 was
maintained around 0.06 units per /g fresh weight ovules pre anthesis and enhanced to 0.08 at anthesis and further to
0.1 units at 2 DPA and later declined, while in MCU 5LL, the POD activity reduced to 0.01units 3 days prior to
anthesis and later increased to 0.04 units at anthesis and further to 0.05 till 6 DPA. The CAT and APX activities were
very low in MCU 5 LL ovules all through, while the MCU 5 ovules had higher activities during pre anthesis (around
1.6 units) and 0.8 units post anthesis. The SOD activity shot up to 4.0 units at anthesis and later maintained at 3.0
units during the fibre initiation process, while MCU 5 LL ovules showed a marginal increment in SOD activity.
Biochemical analysis of the lintless mutant ovules revealed a marked reduction in the synthesis of reducing sugars,
total free amino acids and total soluble protein content, while there was no effect on the proline and phenol content of
ovules from 0 to 5 DPA. The RAPD analysis revealed that two primers were found non-polymorphic with two extra
bands at 204010 bp and 630 10 bp in MCU 5. The failure to trigger the production of anti-oxidants and synthesis of
solutes at anthesis can be assigned as crucial factors for non-conversion of epidermal cells to fibre initials in lintless
mutant and provide enough clues as biochemical determinants for better fibre development process in cotton.
INTRODUCTION
Cotton fibre quality improvement is of vital importance to textile industry. The fiber elongation and
secondary deposition of cellulose are highly sensitive to immediate surroundings. Cotton fibres are a
subset of single epidermal cells that elongate from the seed coat to produce cellulose strands or lint. As
one of the longest single-cell seed trichomes, fibers provide an excellent model for studying fundamental
biological processes such as cell differentiation, cell expansion and cell wall biosynthesis. The molecular
and metabolic mechanisms associated with differentiation of epidermal cells of ovules to trichomes are
still a mystery. In order to enhance the quality attributes of cotton fiber, there is an imperative need to
study the regulation of cotton fiber cell expansion, a vital process controlled by genetic, environmental
and hormonal factors.
Cotton fiber cells are tubular outgrowth of single celled trichomes which arise in near synchrony
from the epidermis of the ovule and may elongate at peak rates in excess of 2 mm per day during the
rapid polar expansion phase of development (Basra and Malik, 1984). Developing cotton seeds are an
excellent system to study diverse patterns of carbon partitioning including cellulose, starch and oil
biosynthesis (Ruan and Chourey, 1998). There has been a substantial progress in our understanding of
cellulose synthesis in developing cotton fibers. However, little is known about the early events
controlling fibre cell initiation. Morphologically, the initiation of each fiber cell is associated with the
spherical expansion and protrusion of one epidermal cell above the ovular surface during anthesis (Basra
and Malik, 1984). Genetic variation between plants for a trait controlled by a single or a few genes
inherited in a single Mendelian fashion is easy to manipulate through breeding. There has been a number
of reliable markers such as protein or, more specifically, allelic variants of several characters such as
7
lipid or sugars must be considered (Prakash et al. 2002). Isozyme numbers are limited and their
expression is often restricted to a specific developmental stage of tissues; and their presence can be
determined by electrophoresis and specific staining.
There are numerous in vitro studies which have thrown light on the basic processes of fiber initiation
and the factors that may have a role thereon. Fiber initiation and development are known to be influenced
by the age of the ovule (Graves & Stewart, 1988), temperature (Xie et al., 1993), plant growth regulators
(DeLonge, 1986) and inorganic nutrients (Eid et al., 1973) in the culture medium. Plant growth
regulators like indole-acetic acid, gibberellic acid, ethylene and abscisic acid play a decisive role in fiber
development (Prakash et al., 2002; Kosmidou-Dimitropoulou, 1986). In the present study, the metabolic
process, biochemical constituents and the RAPD technique were used to study the factors associated in
differentiation of ovular epidermal cells into fiber initials using a normal cotton genotype and its lintless
mutant, as they provide contrasting genetic material for studying the molecular events specific for fibre
development.
The present work aims at investigating the enzymatic changes in the pre and post anthesis period
from -10 to + 6 days anthesis and accumulation of biochemicals in post anthesis for + 60 DAA.
Furthermore RAPD profile technique was used to study the factors associated in differentiation of ovular
epidermal cells into fiber initials using a normal cotton genotype and its lintless mutant, as they provide
contrasting genetic material for studying the molecular events specific for fibre development.
Plant Materials
Cotton (Gossypium hirsutum L.) near isogenic lines-MCU 5, and its lintless mutant MCU 5LL were
grown under field conditions with optimum agronomic practices at Central Institute for Cotton Research,
Regional Station, Coimbatore. The initiated squares were tagged and the ovules were excised at different
growth stages. Harvested ovules at various developing stages were frozen in liquid nitrogen till
extraction. Developing seeds and fibers were analyzed at regular intervals for reducing sugar (Somogyi,
1952), soluble proteins (Lowry et al., 1951) and total phenols (Bray and Thorpe, 1965). The whole
ovules were used for biochemical analysis.
Isolation of Genomic DNA from the Ovules
Fresh ovules (100 mg) were isolated and ground to fine powder with liquid nitrogen and the genomic
DNA was extracted following CTAB method (Doyle and Doyle, 1984). The cotton ovules had higher
polyphenolic compounds which initially interfered in the yield of the DNA which was later removed by
the addition of the PVP- 40 in the extraction buffer which increased the yield and the purity of the DNA
obtained (Porebski et al., 1997). Additional precipitation steps with isopropanol helped in removal of
protein and polysaccharides (Padmalatha & Prasad., 2006.).
RAPD - PCR Analysis
RAPD analyses were carried out according to Sambrook & Russell (2001). One hundred 10 mer
oligonucleotides (Sigma Aldrich) were selected for initial screening of gene specific sequences. 20
primers were selected for the final RAPD PCR analysis (Table 1). Amplification reactions were
performed in a 20 l reaction mixture containing 1.5 l of Taq DNA polymerase (Bangalore Genei,
Bangalore), 0.2 mM dNTPs, 2L of 10X PCR buffer with MgCl
2
, 0.4 M of 10-base primer and 50 ng of
DNA as the template. Amplifications were performed in a Gradient Mastercycler (Eppendorf, Germany.)
by the following program: initial denaturation temperature of 94
o
C for 5 min; 30 cycles each with
denaturing at 94
o
C for 30s, annealing at 55
o
C for 30s and extension at 72
o
C for 1 minute; and final
extension at 72
o
C for 5 min. Reaction products were then loaded on to 1.4% agarose gel for
electrophoresis. Gels were documented under UV following EtBr staining with Alpha Imager systems,
UK. A negative control, without DNA, was included in all reactions.
Temporal Changes in Metabolically Important Enzymes and Solutes act as Trigger for Epidermal Cell Conversion 45
TABLE 1: LIST OF 10-MER PRIMES USED FOR RAPD ANALYSIS OF 2 DAYS POST ANTHESIS OVULES OF MCU 5 AND MCU 5LL.
S. No. Primer Sequence
1 TGAGGGCCGT
2 TCGTTCACCC
3 CATAGAGCGG
4 CCACCACTTC
5 CCTGTCAGTG
6 GGGGAAGACA
7 GTGTCAGTGG
8 CTGGCTCAGA
9 TGCCACGAGG
10 CTGAAGCGCA
11 AAGACCGGGA
12 CCGAGCAATC
13 TGTGGACTGG
14* GAGAGGCTCC
15* TCCGTGCTGA
16 TGCCTGGACC
17 GGCAGGTTCA
18 CTGGTGCTGA
19 GACAGTCCCT
20 TGGTCGGGTG
ESTIMATION OF ENZYMES
Peroxidase activity (POD) was determined by using the method described by Shannon et al. (1966). The
change in absorbance was recorded at 470 nm at an interval of 15 sec for 2 min. The enzyme activity was
expressed as units (mg protein)
-1
. Ascorbate peroxidase (APX) activity was assayed by monitoring the
ascorbic acid-dependent reduction of H
2
O
2
, as described by Anderson et al. (1992). Ascorbate peroxidase
activity was expressed as nmol ascorbate oxidised (mg protein)
1
min
1
. Catalase activity (CAT) was
estimated by the method of Aebi (1984). The absorbance was recorded at 240nm at an interval of 15 sec
for 2 minutes immediately after the addition of enzyme extract. The enzyme activity was expressed as
mmol H
2
O
2
decomposed (mg protein)
1
min
1
. The superoxide dismutase (SOD) enzyme activity was
assayed by the method of Giannopolitis and Ries (1977). The absorbance was read at 560 nm. One unit
of SOD was defined as the level of enzyme activity that inhibited the photoreduction of NBT to blue
formazan by 50 % (expressed as units SOD mg protein
1
).
The normal phenological boll development in MCU 5 and its lintless mutant has been depicted in (Figure
1A& 1B)). Boll bursting stage shows the clear picture of seeds bearing the lint and lintless. Morphology
difference between the bolls at the later stages of anthesis is evident. The difference could be caused due
to the elongation of the fibers in the cultivar MCU 5 in the later stages of the days post anthesis.

Fig. 1: Digital Images of the Various Stages of the Bolls from the Cultivar (a) MCU 5 and its Mutant (b) MCU 5LL
To explore the biochemical changes among the two cultivars MCU 5 and its mutant variety MCU
5LL, total soluble proteins and antioxidant enzymes peroxidase (POD), ascorbate peroxidase (APX),
catalase (CAT) and superoxide dismutase (SOD) were studied in ovules from 10 days prior anthesis to 6
days after thesis. The analysis was done upto 6 days after anthesis based on the earlier observations made
by Hovav et al 2008 and Chaudhary et al 2008, that the oxidant and antioxidant genes are up- or down-
regulated early in fiber development, but not later. Highest number of genes were up-regulated in the
early stage (2 days after anthesis), where at later stages of fiber development, there were only few such
genes (Chaudhary et al 2008).
The total soluble protein concentration was low from -2 days to + 2 days of anthesis in the mutant
variety (MCU 5LL), while MCU 5 showed higher levels at 4, 0 and + 2 days after anthesis and
remained low in -2 and + 4 days anthesis (Fig-2). There was significantly lower level of soluble protein
content in the ovules at anthesis in the lintless mutant (26.36 mg.g
-1
FW) as compared to MCU 5 (46.71
mg.g
-1
FW). With progress in time, the protein content in the mutant seeds increased and by 10 DPA, it
was on par with the normal seeds (Gopalakrishnan et al., 2010). Turley and Ferguson (1996) made a
comparative analysis of the fls mutant SL 171 against an unrelated FLS inbred using two dimensional
PAGE analysis of the total ovular protein. Of the numerous Coomassie blue-stained spots, only five
polypeptides are unique to the mutant. Hence, it is difficult to assign any physiological significance to
these differences because these proteins are of an unknown nature. Similarly, even though there was a
distinct reduction in the protein synthesis in lintless mutant (MCU 5 LL), it is difficult to pin point the
type of protein. Hence, the genomic DNA from the ovules was extracted and subjected further for the
PCR based RAPD.

Fig. 2: Changes in the Total Soluble Proteins in the Cultivars MCU 5 and MCU 5LL
In our study, PCR based RAPD technique was used with randomly generated synthetic
oligonucleotides of 10 bases. The high sensitivity of the method applied to pre-digest DNA permitted the
tagging of specific locus for some characters in nearly isogenic lines of cotton (Arshad & Haidar, 2000).
RAPD PCR analysis among the MCU 5 and its mutant MCU 5LL was initially done with 100 primers.
Majority of the primers failed to amplify in both the genotypes (data not shown) and only 20 primers
gave the best banding pattern. The band size ranged from 2800 bp to 100 bp. The amplified products of
the primer 3` GAGAGGCTCC 5` gave two additional bands of the size of 2040( 10) bp and 630 ( 10)
bp, while a band sized 1160 ( 10) bp of the primer sequence 3`TCCGTGCTGA 5 in the MCU 5 which
are lacking in the MCU 5LL (Figure 4a). All the other eighteen primers gave similar banding pattern
and 100 % polymorphism among the genotypes (Table - 1).
Though many RAPD studies in cotton are done to find their genetic diversity among the cultivar
identification of cotton (Rana & Bhat 2004), there are few studies done to differentiate among the
mutants using RAPD. In the present study, a different banding pattern in two of the primers which gave
extra bands in linted genotype that were absent in the lintless genotype. From the result, we can infer that
these extra bands may have an important role in the fiber initiation and in early fiber elongation process
from 0 days to 5 days of the development of the fibers. Compared with the Arabidopsis trichome, little is
known about the molecular control of the cotton fiber development. So far, a number of genes
differentially expressed during different stages of fiber development have been identified, but their roles
in cotton development are not yet clear (Hulskamp et al., (1994). Ji et al (2003) studied on the genes
which preferably expressed during the early fiber development through cDNA microarray analysis of 0 to
10 DPA ovules, which showed that 172 genes were significantly up-regulated during the course and are
involved in energy metabolism, cell turgor generation and primary and secondary wall biogenesis, which
again showed that turgor pressure is crucial for fibre initiation.
The relationship between the fiber initiation and antioxidant response was studied in ovules from -10
days pre anthesis to + 6 days post anthesis. The highest activity was observed in the -4 and +2 days
anthesis ovules and the lowest with -2 days anthesis ovules Fig-3a). Lintless mutant shows low level of
peroxidase than that of the MCU 5. Maximum decrease in the peroxidase content -2 days anthesis was
seen and recovering in 0 and 2 days to slightly higher levels. Similar pattern of ascorbate peroxidase
activity as such as POD was noticed with MCU 5 (Figure 3b). Increase levels in APX enzymes in days -4
and + 2 days anthesis, with minimal level of APX in days -2 and 0 days anthesis was seen. To the
contrary, lintless mutant showed higher level of APX in days -2 and 0 anthesis, while rest of the ovule
stages displayed somewhat similar levels with not much different changes (Figure 3b). Mutant variety
MCU 5LL showed a low level CAT activity at -10 days anthesis and later increased till 0 days anthesis
(Figure 3c). There was a decline in the activity in +2 and +4 days of anthesis and normalized by +6 days
post anthesis. The CAT activity in MCU 5 showed slight increase during 0 to +2 post anthesis. The
mutant MCU 5LL showed decreased antioxidant enzymes activity during the fiber initial process. Most
of the enzymes maintained low activity after anthesis, while MCU 5 showed higher enzyme activity
during anthesis. The increase of antioxidant activity could be due to the increase in the production H
2
O
2

level. The increase of H
2
O
2
levels has been shown to enhance antioxidant content and antioxidant
enzymes in many plants (Mitler and Tel-Or, 1992 and Lechno et al 1997). Low levels of H
2
O
2
stimulate
expression of necessary genes for the onset of secondary wall cellulose biosynthesis. The elevated levels
of the H
2
O
2
would result in the accumulation of H
2
O
2
- dependent reaction products which could trigger
the programmed cell death response, eventually this could trigger the activation of a cell-death process,
which for the cotton fiber cells constitutes a form of terminal differentiation (Potikha et al 1999). The
role of APX in regulation of intracellular ROS levels by reduction of H
2
O
2
to water using ascorbate as an
electron donor and multiple fold increases in the cotton APX genes was seen till + 5 days anthesis., The
increase in the expression of genes corresponding to POD and APX in + 2 days post anthesis which are
associated with ROS metabolism has also been reported (Chaudhary et al 2009). These reports are in
concurrence to our results, where the highest peroxidase and ascrobate activity was seen in + 2 days
anthesis ovules in MUC 5. Lower levels of these antioxidant enzymes may result in the higher level of
the H
2
O
2
in the ovules resulting in destruction of the cells that trigger fiber initiation and elongation
process.

Fig. 3: Differential Antioxidant Enzyme Activity in Different Ovular Stages form -10 Days Pre Anthesis to + 6 Days Post Anthesis
from of MCU 5 and MCI 5LL Mutant. (a) POD, (b) APX, (c) CAT, and (d) SOD. Values are the Means of Three sets of Observations

Fig. 4: (a) 0.8% Agarose Gel Picture of Genomic DNA of + 2 Days Post Anthesis Ovules of lint (Lane 2 -L) and its lintless (Lane 3-LL) mutant. The lane 1 M
Corresponds to the Size Marker DNA (ECorI / Hind III Double Digested). (b) The RAPD Profile of MCU 5( L) and its Mutant MCU 5LL(LL). The White Arrow
Mark Indicated the Extra Band. 10kbp (Lane 1) and 1kbp(Lane 2) Marker DNA are used as the Ladder
Biochemical accumulation of reducing sugars, total free amino acids, proline and total phenols in
ovules from 0 days anthesis to boll bust was studied in MCU 5 and its lintless mutant 5LL and similar
biochemical compounds were studied in fibers from day anthesis to boll bust in MCU 5. Characteristic
accumulation of reducing sugars (Fig 5a) and total free amino acids (Fig 5b)was noticed in developing
seeds of lintless mutant as compared to linted MCU 5. It has been reported that of larger amount of sugar
molecules is synthesied during secondary wall formation (Fukuda, 1991, 1996). The secondary wall of
developing cotton fiber consists of nearly pure cellulose and is devoid of hemicellulose and lignin (Basra
and Malik, 1984; Ryser, 1985). Rates of cellulose synthesis increase was reported to occur abruptly to
about 100-fold at around 24 days post anthesis of secondary wall formation (Meinert and Delmer, 1977).
Furthermore, development occurs synchronously for nearly all fibers within a boll, with the transition to
secondary wall formation beginning abruptly in varieties of cotton at about 14 to 16 DPA, which is a few
days prior to the cessation of fiber elongation (Meinert and Delmer, 1977). The higher activity in day 25
of ovules and day 15 for fibers of MCU 5 observed in our study is consistent with the above literature
reports. Proline in lintless mutant showed elevated levels in 30 post anthesis. It is reported that the
proline-rich proteins accumulate in later stages during active secondary cell wall formation, indicating
possible regulation at the translational level and function in the secondary cell wall assembly. The proline
content (Fig 5c) was high and ranged around 55 to 65 mg.g
-1
till 15 DAA and there after lesser
accumulation was observed in both fibre (0.5-2.0 mg.g
-1
) and ovules (20-50 mg.g
-1
).

Fig. 5: Measurement of Biochemical Constituents during 0- 60 Days Post Anthesis in Ovules of MCU 5 and MCU 5LL. Changes in Bio-constituents (a)
Reducing Sugars, (b) Total Free Amino Acids, (c) Proline, and (d) Total Phenols in Ovules
CONCLUSION
The process of modification of epidermal cells to fibre initials starts 4-6 days before anthesis that has
been clearly shown by the up and down regulation of biochemical constituents as observed in MCU 5
and its mutant (MUC 5 LL). The final trigger is provided at anthesis where the solutes help in fibre
initiation and elongation processes accounting for the involvement of multiple components. The failure to
trigger the production of anti-oxidants and synthesis of solutes at anthesis can be assigned as crucial
factors for non-conversion of epidermal cells to fibre initials in lintless mutant and provide enough clues
as biochemical determinants for better fibre development process in cotton.
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Study of Interspecific Hybrids (Gossypium hirsutum x

G. barbadense) for Heterosis and Combining Ability
K.P.M. Dhamayanthi
Central Institute for Cotton Research, Regional Station, Coimbatore-641 003, India
AbstractForty nine inter-specific F
1
combinations were made using two tetraploid cotton species Gossypium
hirsutum and G. barbadense. Observations on seed cotton yield, its component traits and ginning per cent were
recorded on parental lines and F
1
hybrids during the year 2007-08 and 2008-09. Among the females BRS-53-53 was
found to be good general combiner for boll per plant and seed cotton yield. Amongst the male parents, ICB-260 and
Pima S4 were found to be the best general combiner on the basis of seed cotton yield and its per se performance,
however B-4 was the best for the majority of the yield components. Five crosses were identified as the best crosses on
the basis of per se performance, combining ability and heterosis. High heterotic crosses viz., which have shown more
than 40% heterosis for seed cotton yield and its component traits could be exploited for increasing yield in inter-
specific cotton hybrids.
INTRODUCTION
Cotton the king of fibre is a premier cash crop of our country grown in about 9 million hectares, which
represent 29 per cent of the world cotton area. India is a pioneer country for the cultivation of cotton
hybrids on commercial scale. Exploiting heterosis is one of the methods to increase cotton yields that
have stagnated in recent years. It has the potential of increasing yield from 10 to 20% and of making
improvements in fibre properties. Increased yield and fibre qualities are vital to keep Indian cotton
competitive with synthetics and foreign production. In India, 40% of cotton production is derived from
intra-specific hybrids of G. hirsutum, and 8% of its production is from G. hirsutum x G. barbadense
hybrids (Singh and Chaudhry, 1997). Yield increase of hybrids over the better parent or best commercial
cultivar (useful heterosis) has been documented earlier showed an average useful heterosis of 21.4% for
F
1
hybrids (Singh et al., 2003). In recent years, breeding progress for increased yield has greatly
decreased. Research on plant breeding needs to address all possibilities to increase yield, including the
use of heterosis. In hybrid development programme, improving the qualitative and quantitative characters
are possible by better commercial exploitation of hybrid vigour (Rajamani et al., 2009). The concept of
combining ability is important in designing the plant breeding programmes. It is highly useful to study
and compare the performance of lines in hybrid combinations. Information concerning to different types
of gene action, relative magnitude of genetic variance and combining ability estimates are significant
markers to shape the genetic make up of a crop like cotton. This important information could prove an
essential strategy to the cotton breeders in screening of better parental combinations for further
enhancement. Cotton breeders have the challenge of finding good combiners by the use of heterosis.
In cotton, high heterosis has been reported at inter-specific and intra-specific levels both in diploid
and tetraploid cotton (Singh and Kalsey, 1983). In recent past, there are hundreds of long and medium
staple hybrids being cultivated in South and Central zone. But in extra long staple category, hybrids are
very limited and the current production of extra long staple cotton is not sufficient to meet the domestic
textile requirement of our country. There is a need to develop suitable high yielding extra long staple
hybrids with desirable fibre qualities to cater the need of the Indian Textile Industry. Hence, in the course
of developing extra long staple inter-specific hybrids, an attempt was made to find out the extent of
heterosis for seed cotton yield and its components in 49 inter-specific F
1
hybrids obtained from seven
diversified G. hirsutum female parent and seven elite male genotypes of G. barbadense.
8
Seven diversified female genotypes of Gossypium hirsutum viz. BRS-53-53, CCH-510-4, CCH-724,
CCH-4, TK-36, ENT- 4, LK-18, and seven male genotypes of G. barbadense viz. B-4, B-5, ICB-20,
ICB-74, ICB-201, ICB-260 and Pima S-4 were identified for better fibre properties to generate 49
hybrids in a line x tester mating design along with 14 parents and a check hybrid DCH-32 in a
randomized block design with three replications. Two rows of each parents and crosses were sown at a
spacing of 90 cm between rows and 60 cm between plants during August 2006. Ten plants were chosen
from two rows of each genotype to record data on seed cotton yield (g), plant height(cm), number of
monopodia/plant, number of sympodia/plant, boll weight (g), number of bolls/plant, ginning outturn, 2.5
percent span length (mm), fibre strength (g/tex) and fibre fineness (/inch). Heterobeltiosis over better
parent and standard heterosis over check DCH-32 were calculated by the following formula;
Heterobeltiosis over BP = (F
1
-BP/BP) x 100; Standard heterosis = (F
1
-SH/SH) x 100
Where F
1
= Mean value of the F
1;
BP = Mean value of the better parent of the particular cross

SH= Mean value over replication of standard hybrids
The analysis of variance showed highly significant differences among the progenies, hybrids and parents
(Table 1). The partioning of hybrid mean square indicated that the variance due to males and females and
an interaction of males x females was significant for all the characters indicating the manifestation of
parental genetic variability in their crosses. Mean, range, coefficient of variation for important ten
characters were studied and presented in Table 2. The maximum variability of 16.38 % was recorded for
seed cotton yield followed by 14.31% of sympodial branches/plant. The coefficient of variability for the
plant height was 12.34 %.
TABLE 1: ANALYSIS OF VARIANCE FOR DIFFERENT CHARACTERS IN INTER-SPECIFIC HYBRIDS (GOSSYPIUM HIRSUTUM L X G. BARBADENSE L )
Source of variation d.f. Pt. Height Boll wt. Bolls/pt. Seed Cotton Yield/pt. GOT %
Replication 2 4.24 0.01 3.17 27.18 0.51
Progenies 62 507.60** 17.08** 78.20** 168.19** 5.33**
Hybrids 48 148.26** 9.27** 82.64** 132.60** 5.27**
Parents vs hybrids 1 7218.03** 173.91** 293.91** 55.18** 3.81**
Lines -females 6 178.63** 147.67** 133.62** 147.27** 1.97**
Tests-male 6 619.34** 28.33 255.13** 543.07** 87.41**
Line x Testers 36 86.29** 162.13** 33.41 97.32** 22.69**
Errors 124 18.29 0.43 11.64 27.56 0.33
*significant at 0.01 level of probabilty
The coefficient of variation among the qualitative characters, ginning outturn, bundle strength and
micronaire value expressed the least value of 2.67, 2.46 and 1.55 respectively. Highest percentage of
heterosis (91.35%) over best parent was recorded in number of bolls/plant followed by seed cotton yield
(73.61%) and number of monopods/plant (73.33%).
TABLE 2: MEAN, RANGE, COEFFICIENT OF VARIATION AND HETEROSIS FOR VARIOUS CHARACTERS OF INTER-SPECIFIC HYBRIDS (G. HIRSUTUM X G. BARBADENSE)
Characters Mean SE CV Range Heterosis Over Best Parent
Seed cotton yield (kg/ha) 984.54 127.42 16.38 719-2435 73.61
Plant height (cm) 119.01 13.56 12.34 110-17-124.60 16.31
Monopods/plant 3.33 0.49 9.60 1.30-6.00 73.33
Sympods /plant 9.41 1.65 14.31 5.13-15.3 31.27
Boll wt. (g) 3.86 0.12 8.32 3.5-5.61 23.81
Bolls/plant 57.66 2.53 9.16 41.24-77.20 91.35
Ginning outturn (%) 30.07 1.03 2.67 28.14-36.01 7.26
2.5% span length (mm) 35.24 0.08 3.5 2 34.23-37.20 4.11
Strength (g/tex) 30.05 0.23 2.46 27.34-33.01 -24.05
Micronaire (/inch) 3.62 0.03 1.55 3.05-4.1 35.37

Study of Interspecific Hybrids (Gossypium hirsutum x G. barbadense) for Heterosis and Combining Ability 53
The per se performance and heterosis over best parent (standard heterosis) for the character under
study is presented in Table 3. The range and mean value among the inter-specific crosses for seed cotton
yield varied from 791-2435 kg/ha. The highest and the lowest seed cotton yield was recorded in the cross
combinations of BRS-53-53 x ICB-260 and LK-8 x B-5 respectively. The hybrid combination CCH-510-
4 x B-4 ranked second in seed cotton yield with 2418 kg/ha, followed by BRS 53-53 x B-4 (2360 kg/ha),
CCH-724 x B-4 (2313 kg/ha), BRS-53-53 x Pima S4 (2286 kg/ha) and CCH-510-4 x ICB-260 (2207
kg/ha). The crosses namely BRS-53-53 x ICB-260 and BRS-53-53 x Pima-S4 exhibited the highest value
for 2.5% span length of 37.3 mm and 37.1 mm respectively. The crosses CCH-510-4 x Pima S-4 (36.8
mm) and CCH-510-4 x B-4 (36.7 mm) also had better 2.5 % span length followed by CCH-724 x B-5
and CCH-4 x ICB-74 each with 36.5 mm length. Maximum number of bolls was recorded in BRS-53-53
x ICB-260 (77.2) followed by CCH-510-4 x B-4 (73.4). The crosses BRS-53-53 x Pima- S4 and CCH-
724 x B-5 recorded the maximum boll weight of 5.6g and 5.1 g respectively. Regarding ginning
percentage, the hybrid CCH-510-4 x B-5 had the maximum ginning outturn of 35.1 % followed by BRS-
53-53 x B-4 (34.5%). The maximum fibre fineness of 4.1/inch was recorded in the crosses BRS-53-53 x
ICB-260 and CCH-510-4 x Pima S-4 followed by CCH-510-4 x ICB-260, CCH-724 x ICB-20 and ENT-
4 x B-5 each with 4.0 /inch.
TABLE 3: PER SE PERFORMANCE OF 49 INTER-SPECIFIC (H X B) HYBRIDS FOR SEED COTTON YIELD AND ITS QUALITATIVE CHARACTERS
S.
No
Crosses SCY
(kg/ha)
Plant
ht (cm)
Sympod/
plant
Bolls/
plant
Boll
wt (g)
GOT
(%)
2.5%
S.L (mm)
B.S/
(g/tex)
Mic
/inch
1 BRS-53-53 x B-4 2360** 121.6 15.3 73.4 4.2 34.5 35.1 30.3 3.7
2 BRS-53-53 x B-5 1982 118.3 13.5 61.8 3.9 30.6 32.7 27.1 3.3
3 BRS-53-53 x ICB-20 1908 116.3 11.9 49.1 3.4 28.3 33 26.9 3.4
4 BRS-53-53 x ICB-74 1893 118.6 13.2 51.3 3.4 27.8 33.2 28.4 3.4
5 BRS-53-53 x ICB-201 1988 119.2 14.3 55.4 3.7 28.1 32.6 26.8 3.6
6 BRS-53-53 x ICB-260 2435** 123.2 13.1 77.2 4.6 33 37.3 31 4.1
7 BRS-53-53 x Pima-S4 2286* 120.4 10.5 69 5.6 33.4 37.1 31 3.6
8 CCH-510-4 x B-4 2418** 124.6 11.4 61.1 4.3 35.1 36.7 30.2 3.6
9 CCH-510-4 x B-5 1965 121.5 13.4 56.7 3.9 30.8 35.1 32.7 3.6
10 CCH-510-4 x ICB-20 2093* 115.9 11.4 63.4 4.1 35.3 35.6 29.6 3.4
11 CCH-510-4 x ICB-74 1901 117.1 13.3 54.2 3.5 29.1 34.2 26.4 3.9
12 CCH-510-4 x ICB-201 1989 114.3 13.7 51.6 3.6 27.5 32.5 28.9 3.5
13 CCH-510-4 x ICB-260 2207** 119.3 13.4 67.2 4.3 34.1 36.4 29.8 4.0
14 CCH-510-4 x Pima -S4 2198* 123.1 10.6 68.1 4.8 34 36.8 33 4.1
15 CCH-724 x B-4 2313** 110.3 11 37.8 3.6 32.5 35.4 28.6 3.8
16 CCH-724 x B-5 1919 118.2 11.2 70.1 5.1 35.2 36.5 30.4 3.5
17 CCH-724 x ICB-20 2010 116.3 12.1 55 4.1 33.4 33.6 27.4 4.0
18 CCH-724 x ICB-74 2096* 119.3 9.8 59.4 4.2 29.1 35.3 30.2 3.6
19 CCH-724 x ICB-201 1976 120.1 11.3 46.5 3.5 28.1 34.6 28.7 3.9
20 CCH-724 x ICB-260 2031* 118.3 9.9 39.7 3.8 29.2 35.4 30.3 3.4
21 CCH-724 x Pima S-4 1977 113.5 10.5 44.6 3.7 30.2 36.5 29.6 3.6
22 CCH-4 x B-4 2002* 112.3 10.5 55.8 4.0 30.3 35.2 31.1 3.6
23 CCH-4 x B-5 1956 116.4 11.2 61.3 3.7 27.8 36.2 28.4 3.3
24 CCH-4 x ICB-20 1999 115.9 9.7 51.7 3.6 29.1 35.7 26.5 3.6
25 CCH- 4 x ICB-74 1866 117.3 11.3 53.7 3.5 28.5 36.5 30.1 3.3
26 CCH- 4 x ICB-201 1885 113.5 12.4 55.7 3.6 28.9 35.2 27.6 3.6
27 CCH- 4 x ICB-260 1961 122.2 14.2 59.6 4.2 31.1 34.9 30.8 4.2
28 CCH- 4 x Pima S-4 2001* 111.3 10.8 51.5 4.0 28.2 35.6 28.4 3.9
29 TK-36 x B-4 2100* 115.2 8.9 48.6 3.6 30.2 34 29.4 3.8
30 TK-36 x B-5 1804 112.6 11.5 59.3 3.5 29.4 36.1 28.5 3.9
31 TK-36 x ICB-20 1965 110.4 10.4 54.8 3.4 27.8 35.3 27.4 3.3
32 TK-36 x ICB-74 2033* 112.6 13.7 58.4 3.4 27.1 34.8 26.7 3.2
33 TK-36 x ICB-201 1968 99.8 12.1 42.8 3.5 26.6 35.1 29.2 3.7
34 TK-36 x ICB-260 2042* 116.3 13.8 64.1 3.7 28.3 36.1 28.6 3.4
35 TK-36 x Pima S-4 1956 120.7 11.3 46.1 4.3 32.0 35.5 30.1 3.6
36 ENT -4 x B-4 2004* 118.2 13.2 56.4 4.0 30.3 36.5 29.5 3.7
Table 3 (Contd.)

Table 3 Contd.
37 ENT -4 x B-5 1993 115.6 12.8 61.3 4.3 28.6 36.7 28.4 4.0
38 ENT -4 x ICB-20 1923 101.3 9.6 55 3.9 27.3 34.2 26.8 3.6
39 ENT -4 x ICB-74 1996 99.8 10.3 56.7 3.8 28.4 35.1 28.3 3.9
40 ENT -4 x ICB-201 1919 110.5 11.6 51.3 3.6 26.8 35.4 26.5 3.5
41 ENT -4 x ICB-260 2090* 113.8 10.5 62.5 4.1 32.4 36.6 28.9 3.7
42 ENT -4 x Pima -S4 1991 110.2 11.1 54.5 3.5 28.1 35.2 27.6 3.6
43 LK-18 x B-4 1955 111.5 10.4 55.3 3.6 27.6 36.4 29.1 3.6
44 LK-18 x B-5 1791 103.5 13.1 58.1 3.9 29.4 35.6 27.6 3.8
45 LK-18 x ICB-20 1907 99.6 10.4 49.3 3.4 28.5 36.7 26.5 3.5
46 LK-18 x ICB-74 1900 110.4 9.9 44.5 3.6 28.6 35.6 28.4 3.7
47 LK-18 x ICB-201 1888 108.5 11 51.7 4.1 29.5 34.5 28.8 3.6
48 LK-18 x ICB-260 1918 110.2 10.6 52.4 3.8 28.7 35.4 27.9 3.5
49 LK-18 x Pima-S4 1882 101.5 9.9 48.2 4.1 29.3 35.7 28.3 3.6
50 DCH-32 (c) 1618 102.4 11.7 41.9 4.1 30.2 36.1 28.6 3.7
CD @ 5% 197.01 32.13 1.68 4.3 0.4 0.78 0.81 0.17 0.34
The partitioning of hybrid mean sum of square indicated that the variance due to male and female
interaction was significant for all the characters. The parents possessed high per se performance coupled
with good gca to be selected for crossing programme. The estimate of parents over the best hybrids
revealed that among the male parents, ICB-260 and Pima S-4 were found to be the best general
combiners on the basis of seed cotton yield. B-4 was the other male parent having good general
combining ability effects for the seed cotton yield (Table 4). The heterosis for number of bolls/plant was
found in positive direction for 6 cross combination out of 49 crosses. The maximum heterosis for number
of bolls/plant was observed in crosses BRS-53-53 x Pima-S-4 (89.26%) followed by BRS-53-53 x B-4
(49.53%). Rajamani et al., (2009) reported high heterosis for bolls/plant in intra hirsutum hybrids. The
heterosis for number of sympodia /plant is in positive direction for cross BRS-53-53 x ICB-260 (19.41
%). The heterosis for ginning out turn is in positive direction in 5 cross combinations out of 49 crosses.
Tuteja et al., (2006) and Verma et al. (2006) reported high heterosis for ginning outturn in G. hirsutum.
The maximum heterosis for ginning percentage was observed in CCH-510-4 x B-4 (26.14%). The
heterosis for boll weight was found in positive direction in 4 combinations out of 49 crosses.
TABLE 4: ESTIMATES OF HETEROSIS OVER BEST PARENT OF 17 PROMISING INTER-SPECIFIC HYBRIDS (H X B) FOR DIFFERENT CHARACTERS
S.
No.
Crosses SCY
kg/ha
Pt. ht
(cm)
Monopod/
Plant
Sympod/
Plant
Bolls/
Plant
Boll
Wt.
(g)
GOT
(%)
2.5%
SL
(mm)
B.S
g/tex
Mic
/inch
1 BRS-53-53x ICB-260 73.64** -9.58 53.24* 19.41* 33.25* 19.08** -3.61 9.79* -16.51 -8.33
2 CCH-510-4xB-4 61.22** -8.56 -52.67 -14.22 7.91 7.31 26.14* 3.19* -13.24 -11.25
3 BRS-53-53xB-4 54.31** -21.62 24.65** 22.55** 49.53* 6.07 -1.83 -4.52 -5.34 33.67
4 CCH-724 x B-4 48.42** -18.13 -29.38 -16.4 -32.61 24.34* 6.4 -4.08 -13.11 -17.66
5 BRS-53-53xPima-S4 37.23** -20.34 48.27** 1.82 89.26* 14.16* -0.97 15.56** -9.17 -15.62
6 CCH-510-4xICB-260 32.56** -2.67 -36.19 -3.34 50.22* -10.25 6.18* 7.63* -18.13 -12.61
7 CCH-510-4xPima-S4 27.41** 9.03 -19.26 -39.41 -28.34 -5.06 -1.39 -5.18 -17.91 -8.27
8 TK-36xB-4 25.60* -21.3 -91.34 -2.11 16.25* 18.12* 0.09 -2.69 -8.24 -10.0
9 CCH-724xICB-74 21.30* -18.24 -47.37 25.37** 3.84 -6.01 16.2* 6.64* -15.81 -3.21
10 CCH-510-4xICB-20 19.04* -16.71 -26.11 2.33 6.59 -11.16 -1.82 -1.08 -15.88 -15.2
11 ENT-4xICB-260 14.56* -22.15 -6.44 -16.74 22.41* 1.02 -1.76 -7.07 -15.09 -33.4
12 TK-36xICB-260 11.34* -7.46 -81.27 -19.54 -5.06 2.16 5.64* 16.03** 3.16* -12.7
13 TK-36xICB-74 8.29* -17.59 -61.48 22.15* -18.54 -7.71 -2.57 -4.1 -10.19 -15.23
14 CCH-724xICB-260 3.66* -4.57 -14.2 -51.6 -6.51 3.05 -21.5 -2.5 -18.31 -16.43
15 ENT-4xB-4 2.58* -16.5 -36.19 -23.18 -36.41 11.01 24.21* 1.08 -23.14 -3.87
16 CCH-4x B-4 2.07* 6.31 -26.28 -26.71 3.44 1.35 -1.04 -3.62 -13.34 -1.07
17 CCH-4xPima S-4 1.94* -2.25 -16.33 -43.62 -42.11 1.08 0.16 -1.07 -11.28 -8.34
SE + 46.37 12.38 0.53 0.94 1.82 0.28 0.51 0.63 0.36 0.09
CD@5% 78.59 23.59 0.79 1.83 4.07 0.57 0.94 0.91 0.78 0.16
CD@1% 112.14 31.17 1.08 2.67 7.08 0.79 1.31 2.15 0.81 0.19

Study of Interspecific Hybrids (Gossypium hirsutum x G. barbadense) for Heterosis and Combining Ability 55
The maximum heterosis for boll weight was observed in cross CCH-724 x B-4 (24.34%). The result
indicated that there is ample scope for developing productive inter-specific hybrids with desirable cross
combinations for seed cotton yield and its components characters having superior fibre quality traits.
Tuteja et al., (2006) and Verma et al., (2006) and Gaurav Khosla et al., (2007) reported similar findings
in intra hirsutum hybrids. The high mean values were associated with high gca effects. However, it was
not in all the cases hence, the choice of parents for crossing should be based on traits that have consistent
and significant gca effects. Similar observation was made by Patel et al., (2009) and Pathak and Parkash
Kumar (2011) in G. hirsutum. The plant height, bundle strength and micronaire showed no desired
heterosis effect in 49 cross combinations. The potential female parents i.e. BRS-53-53 and CCH-510-4
were isolated as the general combiners for seed cotton yield, number of sympodia, no. of bolls/plant and
thus indicated that it could be exploited for further hybridization programme. The estimate of sca effect
revealed that out of 49 crosses, 17 crosses had significant positive sca effect for seed cotton yield. The
crosses BRS-53-53 x ICB-260 and CCH-510-4 x B-4 were identified as the best cross combination for
sca, number of bolls /plant, 2.5 % span length, bundle strength and micronaire. Five promising hybrids
viz. BRS-53-53 x ICB-260, CCH-510-4 x Pima-S-4 were identified on the basis of per se performance,
combining ability and heterosis. High heterotic crosses which have shown more than 40% heterosis for
seed cotton yield and its component traits could be exploited for increasing yield in inter-specific cotton
hybrids. The present study indicated that there is tremendous scope for heterosis breeding for commercial
exploitation and also manifestation of considerable amount of heterosis for improving productive inter-
specific extra long staple cotton hybrids with desirable fibre qualities.
REFERENCES
[1] Gaurav Khosla, Gill, B.S. and Sohu. R. S. (2007). Heterosis and combining ability analysis for plant and seed characters in
upland cotton (Gossypium hirsutum, L). J. Cotton Res. Dev. 21:12-15.
[2] Pathak. R. S. and Prakash Kumar (2011). A study of heterosis in upland cotton (Gossypium hirsutum L.) Theoretical and
Applied Genetics. 47: 45-49.
[3] Patel, K.G, Patel, Rita B., Patel, Madhu I. and Kumar, V.( 2009). Studies on heterosis and combining ability through
introgression in diploid cotton. J. Cotton Res. Dev. 23:23-26.
[4] Rajamani, S., Mallikarjuna Rao, CH and Krishna Naik, R. (2009). Heterosis for yield and fibre properties in upland cotton
Gossypium hirsutum, L. J. Cotton Res. Dev. 23(1): 43-45.
[5] Singh, R.K. and Chaudhry, (1997). Biometrical methods in quantitative genetic analysis (Revised ed.1979). Kalyani
Publisher, New Delhi, pp.191-200.
[6] Singh, P., Loknathan, T. R., and Agarwal, D. K. (2003). Heterosis for fibre properties in intra-hirsutum crosses (Gossypium
hirsutum L.) Indian J. Genet. 63 (4): 325-27.
[7] Singh, P and Kalsey, H.S. (1983). Heterosis in G.hirsutum, L. Indian J. agric. Sci. 53: 624-28.
[8] Tuteja, O.P., Sunil Kumar, Singh. Mahendar and Luthra Puneet (2006). Heterosis for seed cotton yield and fibre quality
characters in cotton (Gossypium hirsutum, L). J. Cotton Res. Dev. 20: 48-50.
[9] Verma, S. K., Tuteja, O.P, Sunil Kumar Ram Prakash, Ram Niwas and Monga, D. (2006). Heterosis for seed cotton yield
and its qualitative characters of Gossypium hirsutum L. in cotton. J. Cotton Res. Dev. 20: 14-17.
Predicting F
1
Performance from Their Parental
Charectaristics in Upland Cotton
(Gossypium hirsutum L.)
R.K. Gumber, Pankaj Rathore and J.S. Gill
Punjab Agricultural University Regional Station, Faridkot-151203, India
AbstractAttempts were made to find out whether various characteristics of parental lines viz., the genetic distance
between parents determined from Amplified Fragment Length Polymorphism (AFLP) data, means of two parents
( P ) i.e. (P
1
+P
2
)/2, and absolute difference between the means of two parents (| P1 - P2 |) can be used to
predict the per se performance and economic heterosis of F
1
hybrids. The line PIL43 with okra leaves was most diverse
from other lines. It was found that genetic distance estimated from AFLP markers was not necessarily associated with
geographical diversity of the parents. The genetic distance was comparatively high between those parental lines that
differ largely for morphological characters. Genetic distance among parents determined from AFLP data proved to be a
good predictor of per se performance and economic heterosis of hybrids for seed cotton yield and boll weight. The means
of the parents were a good predictor for seed cotton yield and boll number. Significant association of absolute difference
between means of the two parents with per se performance and heterosis of F
1
hybrids was observed for seed cotton
yield, ginning out turn and seed index, suggesting that (| P1 - P2 |) is a good predictor for these traits.
INTRODUCTION
In India, for the first time in the world, two hybrids viz., Hybrid 4 (Patel 1971) and Varalaxmi (Katarki
1972) were released in Gujarat and Karnataka states, respectively. Thereafter many hybrids have been
released for commercial cultivation in the country. As a result, the cotton production in India increased
from 2.79 million bales (1 bale = 170 Kg lint) in 1947-48 to 295 million bales in 2009-10 (Anonymous
2010). With the development and cultivation of cotton hybrids, India has become cotton surplus state. In
India, hybrid cotton occupies about 80% of the total cotton area and contributes about 90% to national
annual production. Development of hybrid varieties is considered to be the quickest breeding method for
exploiting the heterosis to improve yield potential of crop plants (Nassimi et al., 2006; Radoev et al.,
2008 and Rameeh 2011). Genetic diversity between parents is important for hybrid breeding and for
maximum usefulness of a cross in pure line breeding. Generally it is assumed that crosses among
genetically diverse parents produce superior hybrids and progenies in the segregating generations
(Rameeh 2011). However, wide crosses suffer from poor adaptation in the target environment and
recombination losses owing to disruption of favourable epistatic gene combinations ( Schill et al., 1998).
Therefore, selection of suitable parents is one of the most important criteria used to allocate resources to
the most promising crosses and to increase the efficiency of breeding program. The identification of
promising F
1
crosses and superior segregants requires the development of a large number of crosses and
their multi-location field evaluation which is very laborious and resource demanding (Melchinger et al
1998). On prediction of test cross variance in maize, Melchinger et al., 1998) reported that the efficiency
of a breeding program may be enhanced if the breeding potential of F
1
crosses could be predicted in
advance. Molecular markers play an important role in crop improvement program and have been used
extensively to predict heterosis and F
1
performance (Gutirrez et al., 2002 and Selvaraj et al., 2010). The
usefulness of molecular genetic distance as a predictor of hybrid performance has been studied in several
crops. Genetic distance estimated using different molecular markers was found to have significant
associations with hybrid performance in maize (Lee et al., 1989; Smith et al., 1990; and Lanza et al.,
1997) and sunflower (Cheres et al., 2000). However, the association between marker based parental
genetic distance and hybrid performance has not been well documented in cotton. Therefore, the present
study was conducted to predict the mean performance and economic heterosis of F
1
hybrids from genetic
distances based on molecular markers, means of the parents ( P ), and the absolute difference between
means of the parents (| P1 - P2 |).
9
Predicting F
1
Performance from Their Parental Charectaristics in Upland Cotton (Gossypium hirsutum L.) 57
Field Experiment
In the present study, 16 F
1
crosses involving 11 diverse parents were evaluated in a randomized complete
block design with three replications at Punjab Agricultural University, Regional Station, Abohar in 2007.
Out of 11 parental lines, nine parents were either approved cultivars or improved lines under cultivation
in different states of India. The parental lines LH 1556, F 1861, F 505, F 846 and LH 900 were the
approved cultivars of American cotton and have been recommended for commercial cultivation in Punjab
state. LH 900 is an early maturing variety having short and compact plant type suitable for late sown
conditions. The parental lines PIL 43 and PIL 8 were the female and male parents, respectively of
approved cotton leaf curl disease resistant hybrid LH 144 (Gill et al., 2008). Likewise, RS2013 is an
approved cultivar from Rajasthan state and has medium size round bolls. GSH 4 is a sympodial cultivar
from Gujarat state, while HS 253 is an advanced line from Haryana. The cultivar Udangsuper is a jassid
susceptible non-descript variety having short stature plant and medium size bolls. The approved hybrid
LHH 144 and open- pollinated cultivar LH 1556 were included as standard checks. Each test hybrid was
accommodated in two rows plot of 8 m length. Rows were kept apart at 67.5 cm while plant-to-plant
spacing was maintained at 75 cm for hybrids and 60 cm for open-pollinated check cultivar.
Recommended agro-managements were carried out. The seeds were sown in the first week of May. The
observations were recorded on five competitive plants for seed cotton yield plant
-1
(g), number of bolls
plant
-1
, boll weight (g), seed index (g), and ginning out turn (%). The economic heterosis in terms of
improvement in per se performance of F1 hybrids over check hybrid LHH 144 and standard cultivar LH
1556 was calculated and expressed as percentage:
Economic Heterosis
F
1
mean

Check mean
x 100

Check mean
Karl Pearsons correlation coefficients (r) were computed as described by Panse and Sukhatame
(1967) to determine the relationship of F
1
performance and economic heterosis with different
characteristics of parental lines.
DNA Analysis
Amplified Fragment Length Polymorphism (AFLP) analysis was performed as per the protocol given by
Vos et al., (1995) to study the molecular diversity among 11 cotton genotypes. For this purpose, a sample
of 250 ng aliquant of genomic DNA was digested with restriction enzymes EcoR1 and Mse1 (1.25 l
-1
)
with incubation at 37
0
C for two hours followed by 70
0
C for 15 minutes to inactivate the enzymes. In the
second step, the following adapter sequences were ligated to the restricted DNA fragments:
5-CTCGTAGACTGCGTACC
CATCTGACGCATGG-3
3-GACGATGAGTCCTGAG
TACTCAGGACTCAT-5
Seven primers were used for the pre-amplification and amplification with the following extensions:
ACT/CAC, ACC/CAC, ACG/CAG, ACT/CTC, AAC/CTG, ACG/CTG, and AGG/CTG,
Where the sequence before the slash refers to the primer extension for EcoR1 and that after the slash
refers to the primer extension for the Mse1. The PCR products were separated by electrophoresis on a
denaturing polyacrylamide gel. After drying, the gels were exposed to phospho-imager plates for 16
hours. The imager plates were scanned with a phospho-imager and polymorphic bands were coded in a
binary form by 1 and 0 for presence or absence in each genotype, respectively.
Estimation of Genetic Distances
The estimates of genetic distances (GD) between all possible combinations of 11 genotypes were
computed using the formula given by Nei and Li (1979):
GD
ij
= (N
i
+ N
j
2 N
ij
) / (N
i
+ N
j
), here GD
ij
is the genetic distance between two genotypes i and j,
Nij is the number of common bands between genotypes i and j, and N
i
and N
j
are the total number of
bands in genotypes i and j, respectively, related to all primer pairs considered in AFLP analysis. Thus GD
reflects the proportion of bands in common between two genotypes and may range from 0 (identical
profiles for two genotypes) to 1 (no common bands).
Estimation of Genetic Distance
The estimation of genetic distances between parental lines using AFLP markers ranged from 0.111
between parental lines F 1861 and Udangsuper to 0.705 between PIL 43 and PIL 8 (Table 1). PIL 43 is a
female parent of an approved hybrid LHH 144 and this line has okra type narrow leaves with deep lobes.
Its bolls are ovate and of very big just like the hybrid LHH 144. On the contrary PIL 8 is the male parent
of hybrid LHH 144 with medium size normal green leaves and elliptic bolls. The genetic distance was
also high (0.679) between PIL43 and Udangsuper. Udangsuper is a local collection from the farmers
field. It has small normal green leaves with medium size round bolls. It has high tolerance to bollworms.
The high genetic distance was reported between parents LH 1556 and GSH 4 (0.52). Low genetic
distance was observed between the parents of LH 1556 and Udangsuper, GSH 4 and Udangsuper,
LH1556 and PIL8, F1861 and PIL8, GSH4 and PIL8, F505 and PIL8, HS253 and GSH4 and Udangsuper
and HS253 (Table 1). There was no association between genetic distance estimated from AFLP markers
and geographical diversity of the parents. The genetic distance was comparatively high between those
parental lines that differ largely for morphological characters.
TABLE 1: ESTIMATES OF GENETIC DISTANCE (GD), MEANS OF THE PARENTS ( P ) AND ABSOLUTE DIFFERENCES BETWEEN MEANS OF PARENTS (| P1 - P2 |) FOR DIFFERENT CHARACTERS
Cross/hybrid Genetic
Distance
(GD)
Seed Cotton
Yield plant
-1

(g)
Number
of Bolls Plant
-1

Boll Weight
(g)
Ginning Out
Turn
%
Seed Index
Mean
| P1 -
P2 |
Mean
| P1 -
P2 |
Mean
| P1 -
P2 |
Mean
| P1 -
P2 |
Mean
| P1 -
P2 |
LH1556x Udangsuper 0.147 105.5 71 34.0 22 2.96 0.11 32.1 -0.5 9.0 1.1
F1861 x Udangsuper 0.111 87.5 35 31.5 17 3.10 0.40 32.5 0.3 8.8 0.2
PIL43 x Udangsuper 0.679 80.0 20 26.0 6 3.50 1.20 32.1 -0.4 9.0 -1.0
GSH4 x Udangsuper 0.131 67.5 -5 23.5 1 3.25 0.70 32.9 1.1 8.6 -0.2
LH900 x Udangsuper 0.281 71.0 2 26.0 6 3.00 0.20 32.6 0.5 8.5 -0.9
F505 x Udangsuper 0.118 79.0 18 28.5 11 2.95 0.10 32.1 -0.4 8.4 0.6
LH1556 x PIL8 0.111 113.0 56 7.5 15 3.07 -1.19 32.6 -1.6 8.5 2.0
F1861x PIL8 0.153 95.0 20 35.0 10 3.75 -0.90 33.0 -0.8 8.6 1.1
PIL43 x PIL8 0.705 87.5 5 29.5 -1 4.15 -0.10 32.7 -1.5 8.8 -0.1
GSH4 x PIL8 0.160 75.0 -20 27.0 -6 3.90 -0.60 33.4 0.0 8.5 0.7
LH900x PIL8 0.309 78.5 -13 29.5 -1 3.65 -1.10 33.1 -0.6 8.3 0.0
F505 x PIL8 0.134 86.5 3 32.0 4 3.60 -1.20 32.7 -1.5 8.2 1.5
HS253 x GSH4 0.152 83.5 37 33.5 19 3.40 -0.40 33.0 -0.8 8.4 0.0
LH1556 x GSH4 0.520 103.0 76 34.5 21 3.31 -0.59 32.6 -1.6 8.9 1.3
RS 2013 x F846 0.272 100.5 11 47.0 10 3.10 -0.40 33.0 -0.8 8.4 0.8
Udangsuper x HS253 0.134 86.0 -32 33.0 -20 3.05 -0.30 32.5 -0.3 8.5 0.2

Predicting F
1
Prediction of Mean Performance and Economic Heterosis
Genetic distance among parental lines estimated by using AFLP markers, means of the parents
(P1+P2/2), and the absolute difference between means of the parents (| P1 - P2 |) were used to predict
the mean performance and economic heterosis of F
1
hybrids. The significant positive correlations of
genetic distance with mean performance (0.472*) and economic heterosis (0.472*) was observed for
seed cotton yield only, indicating that genetic distance between parents is a good predictor of mean
performance and heterosis for this character (Table 2). The associations of genetic distance with mean
and economic heterosis for number of bolls (0.207), boll weight (0.351 & 0.464) and seed index (0.157 &
0.180) were positive but their magnitude was too low to be of any predictive value. In upland cotton,
Solomon et al., (1989) found that AFLP based genetic distance is not a good predictor of heterosis or F
1

performance for yield and most agronomic traits. However, they found positive correlations between
AFLP based genetic distance and F
1
performance for harvest index. Our results are in contrary to that of
Meredith and Brown (1998) who concluded that heterosis for seed cotton yield and boll weight in cotton
can not be predicted from the molecular genetic diversity of the parents. In a study on G. arboreum
(Asiatic cotton), Singla (2008) reported that the mean performance of F
2
populations could not be
predicted from the genetic distance among parental lines. Gutierrez et al .,(2002) found that the
performance of F
2
bulk populations in upland cotton is not always associated with the genetic distance of
the parents but on the genetic background of the parental germplasm. The significant and positive
correlations of means of the parents ( P ) with per se performance (0.529*) and economic heterosis
(0.529*) of F
1
hybrids for seed cotton yield and number of bolls/plant (0.497* each) suggested that
parental means had very high predictive power. For other traits, this association was very weak. The
absolute difference between the means of the parents (| P1 - P2 |) was found to be good predictor of
per se performance and economic heterosis of F
1
hybrids for seed cotton yield, ginning out turn and seed
index. It had significant and positive associations with seed cotton yield for per se performance (0.497*)
and heterosis (0.497*). There was positive and significant correlations with ginning out turn for per se
performance (0.558*) and economic heterosis (0.562*). However, the association for seed index was
negative for per se performance (-0.429*) and heterosis (-0.448*). Although, the correlations for number
of bolls/plant (0.10) was positive but of very low values and has no predictive power.
The result indicated that mean performance and economic heterosis of F
1
hybrids for seed cotton
yield can be predicted accurately from all the three properties of parental lines, viz. genetic distance,
parental means and absolute difference between the means of parents and are supported by theoretical
expectations of Melchinger (1987). Lamkey et al.,(1995) and Melchinger et al., (1998) have reported
that test cross means of F
2
or backcross populations or later selfing generations derived from them are
predictable from the genetic distance and testcross means of the parents in maize. The prediction of F
1

performance from the parental means has advantages- 1) the required information on the performance of
the parents can be obtained within one year across locations 2) the performance of n (n-1) F
1
hybrids can
be predicted from the mean performance of just n parents. However genetic distances estimated by
AFLP markers and the absolute difference between the mean of two parents could not predict the mean
of F
1
hybrids for other characters. The parental means ( P ) for boll weight, ginning out turn and seed
index and | P1 - P2 | for number of bolls per plant and boll weight could not predict F1 performance
and economic heterosis in the present study. The molecular markers based genetic distance provides data
covering the whole genome, whereas genetic variance is composed exclusively of quantitative trait loci
(QTL) effects which are segregating in the populations. Since the distribution of molecular markers and
QTLs responsible for genetic variance are unlinked to any marker and some of the markers are unlinked
to QTLs. Under such circumstances, there is a probability of reduction in the association between genetic
distance and heterosis (Bernardo 1992; Charcosset and Essioux 2004). Melchinger et al., (1998) has
given the theoretical explanations for the absence of correlation between F
1
heterosis and various
predictors. In the present study, we wanted to determine the associations between F
1
performance (F
1
)
and various predictors, (y). However, we actually estimated correlations between the estimates of both
variables (Melchinger et al., 1998). As a result, error in estimation of F
1
means or will reduce the
correlations r(F
1
,) relative to r(F
1
,y). This is supported by high standard derivations for F
1
means and

for most of characters (data not given). This indicated large replication variance for seed cotton
yield/plant, number of bolls/plant, plant height and seed index which could be reduced by increasing the
number of replications. The larger standard error is due to small sample size of F
1
hybrids and their
evaluation in one environment only. Decreasing the standard error of F
1
means and predictors would
require increasing the number of plants in each F
1
hybrid and their evaluation in several environments.
However, this would be extremely resource demanding. Sizeable errors were also associated in
estimation of genetic distance and other predictors.
TABLE 2: ESTIMATES OF CORRELATIONS OF GENETIC DISTANCE AMONG PARENTS (GD), MEANS OF THE PARENTS ( P ) AND ABSOLUTE DIFFERENCE BETWEEN MEANS OF THE PARENTS
(| P1 - P2 |) WITH PER SE PERFORMANCE AND ECONOMIC HETEROSIS FOR DIFFERENT CHARACTERS IN COTTON
Character Correlations with
Genetic Distance
(GD)
Mean of the
Parents( P )
Difference between two
Parents (| P1 - P2 |)
Seed cotton yield/
Plant
Per se performance 0.472* 0.529* 0.497*
Economic heterosis over
LH1556
0.472* 0.529* 0.497*
Number of bolls/
plant
Per se performance 0.207 0.497* 0.100
LH1556
0.207 0.497* 0.101
Boll weight Per se performance 0.351 0.177 -0.048
LH1556
0.464 0.176 -0.049
Ginning outturn Per se performance -0.336 -0.216 0.558*
LH1556
-0.337 -0.210 0.562*
Seed index Per se performance 0.157 0.132 -0.429*
LH1556
0.180 0.146 -0.448*
*Significant at 5% level of significance
CONCLUSION
We conclude from the present study that the per se performance and economic heterosis of F
1
hybrids for
seed cotton yield can be predicted from AFLP based genetic distance between parents, means of the
parents ( P ) and absolute difference between means of the parents (| P1 - P2 |) in upland cotton. The
means of the parents for number of bolls and seed cotton yield; and absolute difference between means of
the parents for seed cotton yield and ginning out turn and seed index are the good predictors of per se
performance and economic heterosis of F
1
hybrids in upland cotton.
REFERENCES
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[2] Bernardo, R., (1992). Relationship between single-cross performance and molecular marker heterozygosity. Theor. Appl.
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[3] Charcosset, A. and Essioux L., (2004). The effect of population structure on the relationship between heterosis and
heterozygosity at marker loci. Theor. Appl. Genet., 89, 336-343
[4] Cheres, M.T., Miller J.F., Crane J. M. and Knapp, S. J. (2000). Genetic distance as a predictor of heterosis and hybrid
performance within and between heterosis groups in sunflower. Theor. Appl. Genet., 100: 889-894.
[5] Gill, M.S., Gumber R.K., Gill J.S., Sohu R.S. and Rathore P., (2008). Varietal impovement program of cotton in Punjab.
Proceedings of the Annual Group Meeting of AICCIP, April 9-10, Ludhiana, pp: 13-24.
[6] Gutierrez, O.A., Basu, S., Saha S., Jenkins, J.N., Shoemaker, D. B., Cheatham, C. L. and McCarty, J.C. Jr., (2002). Genetic
distance among selected cotton genotypes and its relationship with F<sup>2</suup> performance. Crop Sci., 42: 1841-47.
[7] Katarki, B.H., (1972). Varalaxmi: A high yielding hybrid cotton of quality. Indian Farming, 21:35-36
[8] Lamkey, K.R., Schnicker B.J. and and Melchinger A.E., (1995). Epistasis in an elite maize hybrid and choice of generation
for inbred line development. Crop Sci. 35: 1272-1281.
Predicting F
1
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inbred lines and prediction of maize single-cross performance using RAPD markers. Theor. Appl. Genet., 94: 1023-1030.
[10] LeeGodshalk , M., E.B., Lamkey, K. R. and Woodman, W. W., (1989). Association of restriction fragment length
polymorphism among maize inbreds with agronomic performance of their crosses. Crop Sci., 29: 1067-1071.
[11] Melchinger, A.E., (1987). Expectation of means and variances of testcrosses produced from F2 and backcross individuals
and their selfed progenies. Heredity, 59: 105-115.
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variances among F3 progenies of F1 crosses from testcross means and genetic distances of their parents in maize. Theor.
Applied Genet., 96: 503-512.
[13] Meredith, W. R. Jr. and Brown, J. S., (1998). Heterosis and combining ability of cottons originating from different regions
of the United States. J. Cotton Sci., 2:77-84.
[14] Nassimi, A.W., Raziuddin Sardar, A. and Naushad, A., (2006). Study on heterosis in agronomic characters of rapeseed
(Brassica napus L.) using diallel. J. Agron., 5: 505-508.
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Natl. Acad. Sci., USA, 76: 5263-73.
[16] Panse, V.G. and Sukhatme, P. V., (1967). Statistical methods for agricultural workers. ICAR, New Delhi.
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(Brassica napus L.) by quantitative trait locus mapping. Genet., 179: 1547-1558.
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Winter x Spring combinations of rapeseed varieties. Int. J. Plant Breed. Genet., 5: 349-57
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relationship between molecular marker heterozygosity and hybrid performance using RAPD markers in rice (Oryza sativa
L.). African J. Biotech., 9: 7641-7653
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bean (Vicia faba L) II. Genetic effects estimated from generation means in Mediterranean and German environments. Plant
Breeding, 117: 351-359
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molecular level. M.Sc Thesis, Punjab Agricultural University Ludhiana.
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(1995). AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res., 23: 4407-4414.

Thermosensitive Genetic Male Sterility System

in Cotton (G. arboreum L.)
S.M. Palve
1
, V. Santhy
2
, S.R. Bhat
2
, S. Laxman
2
, Rajesh Patil
2
, B.M. Khadi
2
,
Sonali Virkhede
2
and Priyanka Bihariya
2

1
Principal Scientist, Division of Crop Improvement,
Central Institute for Cotton Research, Nagpur440010 India
2
Central Institute for Cotton Research,
Post Bag No. 2, P.O. Shankar Nagar Nagpur440010 India
e-mail: smpalve2k1@yahoo.com
AbstractThermosensitive Genetic Male sterility (TGMS) is a two line method of hybrid seed production where the
sterile flowers are converted into fertile and fertile into sterile at a particular temperature. Once this is stabilized it is
possible to convert any elite variety into TGMS and develop heterotic hybrids. TGMS system in G. arboreum and
photoperiod-sensitive genetic male sterility (PGMS) system in G. hirsutum has been identified for the first time. The
line remains sterile till temperature reaches 24
0
C and show complete pollen fertility at temperature less than 18
0
C.
PGMS lines in G. hirsutum show complete pollen sterility when temperature rises above 40
0
C for continuous period
of time.
TGMS line 1-1 has been characterized and stabilized by continuous selfing. This line was also grown under
phytotron conditions for confirming the phenomenon. Varying minimum/night temperature and a fixed maximum
temperature was provided in the phytotron to study pollen viability in G. arboreum and the phenomenon of reversion
to complete fertility at 16
0
C was confirmed. The period in between the completely fertile and completely sterile phase
which produced partially fertile/partially sterile flowers were observed to be the sensitive stage. The flower behaviour
during normal growing season in the field condition was observed continuously (2007, 2008, 2009 and 2010) in this
lines and correlated with the prevailing weather conditions during the period. During the years 2007, 2008 and 2009,
the flowers produced at the onset (i.e. during last week of August to first week of September depending on the sowing
date) were sterile with very small white anthers. The mean minimum temperature during the period was 24
0
C with
continuous and good sunshine hours. As the temperature reduced towards the end of September and beginning of
October, the flowers started turning fertile with yellow anthers. The flowers turned completely fertile with almost full
yellow anthers when the mean minimum temperature reduced to 18
0
C during the following month. Highest fertility
was recorded during the months of October November which later decreased as the temperature went down.
INTRODUCTION
India is the only country to grow all four species of cultivated cotton Gossypium arboretum, G.
herbaceum, G. barbadense and G. hirsutum. G. hirsutum represents 90% of the hybrid cotton production
in India. India has the largest area under cotton (110 lakh hectares in 2010-11) with an estimated 6
million farms. The rainfed lands produce 65% of Indias cotton while the remaining 35% is attributed to
irrigated lands. The rapid growth in yields between 2002-03 to 2007-08 is attributed to the introduction
and expansion of Bt cotton hybrids and improved cotton varieties, improved crop management practices
and overall favourable weather conditions in most of the states involved.
Indias pioneering role in the successful exploitation of heterosis at the global level on a commercial
scale resulted in the release of first hybrid H4 in 1970 by Late Dr. C.T. Patel in Gujarat. Since then,
followed a release of many hybrids (intra-species and interspecific). The studies on heterosis from early
1950s onwards indicated higher level of heterosis in G. herbaceum G. arboreum than intra herbaceum
crosses. The first ever success story of heterosis breeding in tetraploid cotton encouraged breeders to try
the same principles in diploid cotton which resulted in the development of hybrids namely G. Cot DH 7,
G. Cot DH 9 and DDH 2. These hybrids were based on conventional emasculation and pollinaton. The
10
Thermosensitive Genetic Male Sterility System in Cotton (G. arboreum L.) 63
hand emasculation and pollination (Doak, 1934) was not effective in producing large quantity of
economically viable seeds. Despite these methods, higher level of seed set equal to G. hirsutum was not
possible due to delicate and small flower structure. To overcome this problem of hybrid seed production,
two sources of genetic male sterility (GMS) have been used for seed production.
The two-line systems of hybrid seed production hold promise for breeding specific genotypes bearing
response to temperature and photoperiod fluctuations. However, considering slow increase in yield levels
the male sterility systems have to make a dent in commercialization for larger benefits. There is certain
amount of risk in exploiting heterosis by means of TGMS, if temperature fluctuation occurs at critical
stages of flowering. Therefore, knowledge on critical thermo-sensitive stages for fertility alteration is
useful to determine the most suitable time of sowing for seed multiplication and hybrid seed production.
The present study was therefore undertaken to characterize and stabilize the identified TGMS 1-1 line in
G. arboreum cotton for developing two line-hybrids.
The present investigation was undertaken at Central Institute for Cotton Research, Nagpur, India. The
material consisted of the plants of identified TGMS line 1-1. These plants and progenies of selected
plants were sown for four consecutive years (2007-08 to 2010-11). The line was planted in normal kharif
season (June- July) during the first three years and planted in three staggered sowings during the fourth
year 2010-11 to screen for temperature sensitivity and find out most suitable period for hybrid seed
production.
In field screening, the individual plants were observed daily for pollen sterility/fertility when the
flowering was initiated. At flowering stage, during the first year, the plants which showed fertility
sterility reversion were tagged and others removed. The anthers were collected at anthesis from each
selected plants. For pollen stainability, anthers were smeared in 1 % acetocarmine solution. Round and
dark stained pollen were scored as normal fertile and irregular shaped with empty pollen grain as sterile.
Five randomly selected fields within the portion covered by the coverslip were taken for recording of
data. Plants with no stained pollen were considered completely male sterile, whereas plants having more
than 95 % deeply stained, well filled pollen grains were counted as fertile. The ratio of stained pollen to
total pollen counted was expressed as per cent fertility. At the same time, the selected plants were selfed
for observing boll setting. Shedding of selfed boll was correlated with the pollen fertility/sterility. From
the same plants, seeds were collected boll-wise. Boll to row progenies was raised during successive two
years which were thoroughly monitored for alterations till they were found stable with respect to the
TGMS trait. The parent plants were further continued during summer as rattoons and their reversion to
complete sterility was confirmed.
The flower behaviour during the growing season was correlated with the prevailing weather
conditions especially mean minimum temperature and sunshine hours during the period. The
sterility/fertility of flowers was confirmed by visual observation of flowers as well as by pollen staining
studies under microscope. The flowers showed white anthers during sterility phase and yellow anthers
during fertility phase (Fig 1)


Sterile

Partially Fertile

Completely Fertile
Fig. 1: Sterile, Partially Fertile and Complete Fertile Flowers of TGMS Line
The TGMS line was also raised under controlled conditions in the National Phytotron Facility, Indian
Agricultural Research Institute, New Delhi. TGMS line along with control variety PA 255 under 13 hour
photoperiod with the day/night temperature regime of 30
0
C/25
0
C. The minimum night temperature was
lowered step-wise (2-3
0
C steps) up to 15
0
C. Each of the new temperature regimes was maintained for 8-
10 days and newly opening flowers were checked for pollen viability by fluorescein diacetate (FDA)
staining and observed under fluorescence microscope.
The field grown TGMS 1-1 line was evaluated for sterility and fertility behaviour during the last four
years 2007-08 to 2010-11. The appearance of first flower was recorded in all the plants in the field. The
flowers produced at the onset (i.e during last week of August to first week of September depending on
the sowing date) were sterile with very small white anthers. The mean minimum temperature during the
period was 24
0
C with continuous and good sunshine hours (Table 1 & 2). Mean temperature has been
observed to be the primary factor influencing fertility alteration in TGMS lines of rice (Latha et al.,
2003). The percentage boll set after selfing was zero in the TGMS line during the initial flowering stage
i.e. first week of September. As the temperature reduced towards the end of September and beginning of
October, the flowers started turning fertile with yellow anthers. The flowers turned completely fertile
with almost full yellow anthers when the mean minimum temperature reduced to 18
0
C during the
following months. In rice, the lines with complete pollen sterility under high temperature condition and
more than 30 per cent self seed set under low temperature condition are considered as promising TGMS
lines for commercial exploitation (Lu et al., 1994). These plants when continued as rattoons beyond
February started turning sterile and became completely sterile by the month of April under high
temperature and continuous good sunshine hours. TGMS lines causing male sterility at high atmospheric
temperature and fertility under low temperature have been reported in rice (Ramakrishna et al., 2006).
The pollen sterility was confirmed by visual observation of flowers as well as by pollen staining studies
under microscope. The period in between the completely fertile and completely sterile phase which
produced partially fertile/sterile flowers were observed to be the sensitive stage. Thus three stages critical
fertility phase, critical sterility phase and sensitive phases were identified in the TGMS line 1-1.
TABLE 1: THE DURATION OF SUNSHINE HOURS AND STERILITY/FERTILITY OBSERVED IN TGMS 1-1
Week/ Month Flower Behaviour Weekly Mean Mini Temp (
0
c) Weakly Mean Sunshine Hours
August, 2008
1
st
week Sterile 24.3 0.64
2
nd
week Sterile 24.7 2 ( Sunshine was nil during 4
days)
3
rd
4
th
September 2008
1
st
week Partially fertile 23.8 4.7
2
nd
3
rd
4
th
October, 2008
1
st
week Partially Fertile (40% ) 23.3 7
2
nd
week Fertile (70-80 % ) 21.8 7.6
3
rd
week Completely fertile 18 9
4
th
week Fertile (80%) 15 9
November, 2008
1
st
week Fertile (80%) 14.0 9.0
2
nd
week Fertile (80% ) 15.5 8.5
3
rd
week Completely Fertile 20.3 6.5
4
th
week fertile (60-70%) 12.4 9.8
TABLE2: THE DURATION OF SUNSHINE HOURS AND STERILITY/ FERTILITY OBSERVED IN TGMS 1-1
Week/ Month Flower Behaviour Weekly mean Mini Temp (
0
c) Weakly Mean Sunshine Hours
August, 2009
1
st
week Sterile flowers 25.60 5.2
2
nd
week Partially Fertile 24.80 2.6
3
rd
4
th
September, 2009
1
st
2
nd
3
rd
4
th
week Sterile flowers 25.00 8.7
October, 2009
1
st
2
nd
3
rd
4
th
November, 2009
1
st
2
nd
3
rd
4
th
week Fertile 12.5 8.5
For determining critical temperature for inducing male sterility/fertility in TGMS lines of G.
arboreum plants were raised under controlled condition under Phytotron conditions at IARI, New Delhi
during the year 2008-09. Initially plants were grown at 30
0
C day and 26
0
C night temperatures with 13 hr
photoperiod (Table 4). Under this temperature regime, all the flowers were male sterile in the TGMS line.
The night temperatures were lowered step-wise (2
0
Csteps) and pollen viability was recorded in the
flowers that opened in the next one week. Pollen viability was tested by FDA staining and observed
under microscope (Fig. 2). A night temperature of 20 0C was found to lead partial fertility. The results
revealed that temperature below 16
0
C is necessary for obtaining male fertile plants (Table 4).

Sterile Pollen Grains Partially Fertile Pollen Grains

Normal Fertile Pollen Grains
Fig. 2: Acetocarmine Staining to Determine Sterility and Fertility of Pollen Grains in TGMS Line
TABLE 3: THE WEEKLY MEAN MIN. TEMP (
0
C), WEEKLY MEAN SUNSHINE HOURS AND STERILITY/ FERTILITY OBSERVED IN TGMS 1-1
Week/Month Flower Behaviour Weekly Mean min.
Temp. (
0
c)
Weekly Mean Sunshine Hours
August 2010
1
st
week Partially fertile (6% after pollen staining) 23.0 0
2
nd
week Partially fertile (4% after pollen staining) 24.0 1.7 (Sunshine nil for five days)
3
rd
week Partially fertile (10% after pollen staining) 23.0 1.5 (Sunshine nil for five days)
4
th
week Partially fertile 24.0 2.1 (Sunshine nil for four days)
September 2010
1
st
week Partially fertile (15 % after pollen staining) 24.0 1.7 (Sunshine nil for five days)
2
nd
week (Second
sowing started)
Partially fertile (15 % after pollen staining) 24.0 2.0 (Sunshine nil for five days)
3
rd
week Partially fertile (15 % after pollen staining) 23.0 1.5 (Sunshine nil for five days)
4
th
week Partially fertile (8 % after pollen staining) 23.2 7.0 (Sunshine present on all days)
October, 2010
1
st
week Partially fertile (20% fertility after pollen
staining )
22.9 7.4 ( All Days good sunshine )
2
nd
week Partially fertile (25% fertility after pollen
staining )
22.9 7.2
3
rd
week Partially fertile 23.8 4.0 (Sunshine nil for 3 days)
4
th
week fertile (80% fertility after pollen staining ) 19.7 7.0
November, 2010
1
st
week Fertile (90% fertility after pollen staining) 19.0 7.0
2
nd
3
rd
4
th
week Fertile (80% fertility after pollen staining) 19.0 8.5
TABLE 4: EFFECT OF NIGHT TEMPERATURE ON POLLEN VIABILITY IN TGMS LINE OF G. ARBOREUM
Temperature Regime % Male Fertile Flower Pollen Viability (%) No. Days Tested
30C days/ 26C night 0 0 10
30C days/ 22C night 0 0 7
30C days/ 20C night 9 20-50 7
30C days/ 18C night 37 20-50 7
30C days/ 16C night 41 20-100 7

TABLE 5: STUDY OF TGMS SYSTEM IN DIPLOID COTTON (G. ARBOREUM) UNDER NATURAL FIELD AND PHYTOTRON CONDITIONS
Growth Condition Temperature at Complete Male Sterility Obtained Temperature at Complete
Male Fertility Obtained
Artificial (Phytotron) 22
0
C and above 16
0
C
Natural Field 24
0
Cand above ( As observed for three consecutive years
(2007, 2008 and 2009)
18
0
C
Natural Field (Fourth
Year - 2010)
No complete sterility (for early and late sown) even at 24
0
C
due to cloudy weather with no sun shine consecutively for
several days.
18
0
C

The third sowing date which was in the end of January 2011 started flowering by the end of April
and was at peak flowering by the month of May. The flowers were completely male sterile with
unstained and deformed pollen observed under microscope after aceto-carmine staining (Fig. 2).
Developmental variation in the TGMs line, soon after microspore release and before dehiscence of
anther, it was found that, break down of fertility in the sterile anthers was post-meiotic after the release of
tetrad in the developmental stage (Fig. 3). Similar observation has been recorded Reddy and Reddi
(1974) in pearl-millet that microspores in sterile lines collapsed soon after their release from the tetrads.

Fig. 3: Microspore tetrad Formation in Sterile and Fertile Flowers of TGMS Line
Simultaneously selfing was performed in the TGMS line as well as normal fertile line. The selfed
boll set was absent in the TGMS line till the second week of June where as it was 60-70% in the normal
fertile line. The mean minimum temperature was above 24
0
C during the whole month (Table 3 & 5). In
rice, the TGMS line T 29 was male sterile when exposed to daily mean temperature of 24.1
0
C (Cuong et
al., 2004). Also in maize a novel TGMS line exhibiting full sterility during summer has been discovered
(Tang et al. 2006). In the present study a period of 35 - 40 days with complete male sterility could be
obtained in the third date of sowing unlike other two dates of sowing. The TGMS line developed and
characterized was thus found to be beneficial for hybrid seed production in the month of May and for
maintainance by selfing during the normal kharif season.
Thus, for two-line system using TGMS line, any fertile line can be used as pollen parent, so the
choice of parents in developing heterotic hybrids is greatly broadened. Since there is no need for restorer
genes in the male parents of two-line hybrids, this system is ideal for developing arboreum hybrids.
Identification of proper locations for seed production of hybrid as well as of the TGMS line itself will
play an important role in commercial success of this system.
REFERENCES
[1] Cuong, Van Pham., Seiichi, Murayama., Yukio, Ishimine., Yoshinobu, Kawamitsu., Keiji, Motomura and Eiji, Tsuzuki
(2004) Sterility of thermo-sensitive genic male sterile line, heterosis for grain yield and related characters in F1 hybrid rice
(Oryza sativa L.). Plant Production Science, 7: 22-29.
[2] Doak, C. C. (1934) A new technique in cotton hybridizing suggested changes in existing methods of emasculation and
bagging of cotton flowers. J. Hered. 25: 201-204.
[3] Latha, R. S., Thiagarajan, K. and Thyagarajan, K. (2003) Inheritance of thermo-sensitive genic male sterility trait in rice. J.
Genet. Breed. 57: 89-91.
[4] Lu, X. G., Zhang, Z. G., Maruyama, K., Virmani, S. S. (1994). Current status of two line method of hybrid rice breeding In:
Virmani S. S., editor. Hybrid rice technology: new developments and future prospects. Manila (Philippines) International
Rice Research Institute. p 37-39.
[5] Liu, Y., He, H., Shun, Y., Rao, Z., Pan, X., Huan, Y., Geo, J. and He, X. (1997) Light and temperature ecology of photo-
thermo-sensitive genic male sterile rice and its application in plant breeding. In Proc. Int. Symp. On Two line system of
heterosis breeding in crops pp. 49 -58 (China National Hybrid Rice Research and Development Centre, Changsha, China)
[6] Ramakrishna, S., Mallikarjuna, Swamy., Mishra, B., Virakthamath, B. C. and Ahmed M. Illyas. (2006) Characterization of
thermosensitive geneitic male sterile lines for temperature sensitivity, morphology and floral biology in rice (Oryza sativa)
Asian J. Plant Sciences, 5 : 421 - 428
[7] Tang, J. H. , Fu Z. Y., Hu, Y. M., Li, J. S., Sun, L. L. and Ji, H. Q. (2006) Genetic analyses and mapping of a new thermo-
sensitive genic male sterile gene in maize Theor. Appl. Genet., 113: 11-15.
[8] Reddy, B.B. and M.V. Reddi, (1974) Cytohistological studies on certain male sterile lines of pearl millet (Pennisetum
typhoides. S. and H). Cytologia, 39: 585-589.
[9] Singh, Vrijendra., Deshmukh, S. R., Deshpande, M. B. and Nimbkar, N. ( 2008 ) Potential use of thermosensitive genetic
male sterility for hybrid development in safflower Fourth Int. Safflower Conference, Australia.
[10] Xiaodong, Chen., Dongfa Sun., Rong D. F., Genlou, Sun and Junhua, Peng (2010) Relationship of genetic distance and
hybrid performance in hybrids derived from a new photoperiod-thermo sensitive male sterile wheat line 337 S. Euphytica,
175: 365 371.
[11] Xia, Liu., Xihong Li., Xin, Zhang and Songwen, Wang. (2010) Genetic analysis and mapping of a thermosensitive genic
male sterility gene, tms6(t) in rice (Oryza sativa L.). Genome, 5 3: 119-124.
[12] Yuan, L. P. (1998) Hybrid rice breeding in China. In Advances in Hybrid Rice Technology (Ed. SS Virmani; EA.
Siddique; K. Muralidharan), pp: 27 -33 International Rice Research Institute, Manila, Philippines

Heterosis for Yield and Yield Attributing Traits

in Arboreum Cotton (Gossypium arboreum L.)
S.B. Lalage
1
, N.D. Deshmukh
2
, I.S. Halakude
3
and J.C. Rajput
4

Nirmal Agricultural Research and Development Foundation, Pachora424201 (MS)
1
SR. Scientist (Cotton), Nirmal Agricultural Research and Development Foundation,
Pachora Dist. Jalgaon, India
2
Plant Breeder, Nirmal Agricultural Research and Development Foundation,
3
Research Coordinator, Nirmal Agricultural Research and Development Foundation,
4
Director of Research, Nirmal Agricultural Research and Development Foundation,
AbstractThe studies on heterosis made with 24 specific cross combinations using two diverse genetic male sterile
(GMS) lines viz., NCAGA-4 and NCAGA-26 as female parents and 12 contrasting Arboreum strains as male parents
at R&D, Farm of Nirmal Agricultural Research and Development, Jalgaon. The experiment was laid out in
randomized block design with three replications. The observations were recorded for number of sympodia, number of
bolls/plant, average boll weight (g) and seed cotton yield (g/plant) on five randomly selected plants per replicate from
each genotype. It indicated that the maximum heterosis for seed cotton yield was observed in NACH-433 (121% for
MP and 109.13% for BP) and average boll weight (g) were observed in NACH-433 (55.34% for MP and 53.85% for
BP). Thus, the female parent NCAGA-26 can be used for exploitation of heterosis.
INTRODUCTION
Cotton plays a vital role in the economy of the nation being an important raw material for textile
industry. Gossypium arboreum being resistant to abiotic and biotic stresses, gets well adapted to the
climatic aberrations and also well suited in resource-limited environments. The competitive demand for
fibre warrants to improving the productivity of cotton crop in such situation which is difficult to achieve
through conventional hybridisation and selection. Heterosis breeding seems to be good approach in this
directions. Several studies indicated that in cotton heterosis assisted in gathering the strong genetic
information (Kapoor, et al.,., 2002 and Patil, et al.,., 2009). The present investigation is linked to above
statement.
The experiment was conducted with two diverse genetic male sterile (GMS) lines, namely, (NCAGA-4
and NCAGA-26) used as female parents and 12 contrasting Arboreum strains NSA-12, NSA-13, NSA-
15, NSA-17, NSA-18, NSA-19, NSA-27, NSA-28 NSA-29, NSA-32, NSA-300 and NSA-301 (used as
male parents) in randomized block design with three replications. The 12 male parents were crossed with
each of the GMS line in a line x tester manner to develop 24 hybrids. All these 38 genotypes (24 hybrid +
12 parents + 2 GMS lines) were grown at experimental farm of R&D Division, Nirmal Agricultural
Research and Development Foundation (NARD), Pachora, Dist Jalgaon, India during kharif 2009.
The observations were recorded for number of sympodia per plant, number of bolls/plant, average
boll weight (g) and seed cotton yield (g/plant) on five randomly selected plants per replicate from each
genotype. The data of all the genotypes were pooled and per cent heterosis over mid parent and better
parent was calculated for seed cotton yield and other related traits.
The per cent heterosis over mid parent and better parent for all the traits are given in Table 1. Per cent
heterosis of number of sympodia per plant ranged from 9.26 to 9.89 percent for mid parent and -15.69
11
to 4.17 for better parent. Three hybrids namely, NACH-338 NACH-259 and NACH-7 for mid parent and
NACH-338, for better parent were superior over all the genotypes. The maximum heterosis effect 9.89
and 4.17 per cent was obtained in hybrid NACH-338 over mid parent and better parent respectively.
Per cent heterosis for the number of bolls/plant ranged from -6.67 to 43.00 per cent for mid parent
and -13.27 to 40.40 per cent for better parent. Maximum per cent heterosis was estimated for hybrid
NACH-18 (43.00 % for mid percent) and NACH-12 (40.40 % for better parent), respectively. The
hybrids NACH-18, NACH-12, NACH-433 and NACH-15 exhibited significant positive per cent
heterosis among all the genotypes. Similar heterosis effect for this character was reported by Nanjundan
et al., (1992) and Patil et al., (1991), Chavan et al., (1999), Jain (1996) and Nirania et al., (1992).
Maximum per cent heterosis for boll weight (g) was recorded by hybrid NACH-433 (55.34 % and
53.85 % for mid parent and better parent respectively). The per cent heterosis for this character ranged
from -19.05 to 55.34 per cent for mid parent and -32.89 to 53.85 per cent for better parent. Fourteen
hybrids for mid parent namely NACH-6, NACH-9, NACH-15, NACH-221, NACH-338, NACH-286,
NACH-223, NACH-225, NACH-226, NACH-12, NACH-11, NACH-375, NACH-433, NACH-434 and
Eleven hybrids for better parent namely NACH-15, NACH-221, NACH-338, NACH-286, NACH-223,
NACH-225, NACH-226, NACH-12, NACH-18, NACH-375 and NACH-433 exhibited positive
significunt heterosis over remaining genoypes. Heterosis for this character has also been reported earlier
by Nanjundan et al., (2004) and Chavan et al., (1999).
For seed cotton yield, the per cent heterosis ranged from 0.60 to 121.92 per cent for mid parent and -
12.66 to 109.13 per cent for better parent. Out of 24 hybrids, 18 hybrid for mid parent and 15 hybrid for
better parent showed positive significant heterosis. Maximum heterosis was exhibited by hybrid NACH-
433 (121.92 per cent for mid parent and 109.13 per cent for better parent) followed by NACH-18 (115.34
per cent for mid parent) and NACH-12 (104.74 per cent for better parent). Kapoor et al., (2002), Patil
et al., (2009), Patel (2009) also reported significant positive heterosis for seed cotton yield in cotton
hybrids.
It can be concluded that number of bolls per plant and boll weight were main components for good
yield. Therefore, selection for these characters might results in the improvement of yield and the
promising hybrids like NACH-433, NACH-18 and NACH-12 may be further tested on large plots over
different locations and seasons before recommending them for commercial utilization.
TABLE 1: ESTIMATES OF HETEROSIS OVER MID PARENT AND BETTER PARENT FOR SEED COTTON YIELD AND ITS COMPONENT TRAITS IN DESI COTTON HYBRIDS
Mean MP Mean Mean Mean
1 NACH-7 NCAGA-4 x NSA-12 25.00 4.17 -1.96 64.00 19.63 ** 16.36 ** 2.70 9.09 8.00 172.90 30.76 ** 28.60 **
2 NACH-6 NCAGA-4 x NSA-13 24.00 1.05 -5.88 65.50 20.71 ** 15.93 ** 2.80 15.46 * 12.00 183.55 39.74 ** 38.32 **
3 NACH-9 NCAGA-4 x NSA-15 21.50 -8.51 -15.69 * 64.50 21.70 ** 19.44 ** 2.75 14.58 * 10.00 177.40 39.47 ** 36.46 **
4 NACH-11 NCAGA-4 x NSA-17 23.00 -5.15 -9.80 60.50 11.52 ** 7.08 2.55 3.03 2.00 154.50 15.02 11.43
5 NACH-10 NCAGA-4 x NSA-18 24.00 -4.00 -5.88 64.50 17.28 ** 11.21 * 2.65 4.95 3.92 171.30 23.22 ** 15.70
6 NACH-17 NCAGA-4 x NSA-19 24.00 -5.88 -5.88 56.00 0.45 -5.88 2.60 1.96 0.00 145.80 2.51 -5.60
7 NACH-23 NCAGA-4 x NSA-27 25.00 -3.85 -5.66 67.50 16.38 ** 5.47 2.75 10.00 10.00 185.55 27.97 ** 15.97
8 NACH-24 NCAGA-4 x NSA-28 24.00 -4.00 -5.88 61.00 20.20 ** 17.31 ** 2.55 -0.97 -3.77 156.00 19.63 * 19.27
9 NACH-25 NCAGA-4 x NSA-29 24.50 -2.97 -3.92 61.50 18.84 ** 18.27 ** 2.60 -2.80 -8.77 159.75 15.53 9.01
10 NACH-259 NCAGA-4 x NSA-32 25.50 6.25 0.00 59.50 19.60 ** 14.42 ** 2.45 -5.77 -9.26 145.55 12.65 11.96
11 NACH-415 NCAGA-4 x NSA-300 24.00 -8.57 -11.11 61.00 12.96 ** 8.93 2.55 0.99 0.00 155.00 14.19 9.24
12 NACH-416 NCAGA-4 x NSA-301 24.50 -9.26 -14.04 * 60.50 22.84 ** 16.35 ** 2.55 -19.05 ** -32.89 ** 154.20 0.60 -12.66
13 NACH-15 NCAGA-26 x NSA-12 24.00 3.23 0.00 67.00 29.47 ** 21.82 ** 3.90 54.46 ** 50.00 ** 261.50 100.84 ** 94.50 **
14 NACH-221 NCAGA-26 x NSA-13 23.00 0.00 -4.17 64.50 22.86 ** 14.16 ** 3.80 53.54 ** 46.15 ** 245.25 89.64 ** 84.82 **
15 NACH-338 NCAGA-26 x NSA-15 25.00 9.89 4.17 59.00 15.12 ** 9.26 3.80 55.10 ** 46.15 ** 224.00 78.95 ** 77.85 **
16 NACH-286 NCAGA-26 x NSA-17 23.00 -2.13 -4.17 49.00 -6.67 -13.27 ** 3.70 46.53 ** 42.31 ** 181.50 37.19 ** 30.91 **
17 NACH-223 NCAGA-26 x NSA-18 22.50 -7.22 -8.16 56.00 5.16 -3.45 3.70 43.69 ** 42.31 ** 207.10 51.17 ** 38.89 **
18 NACH-225 NCAGA-26 x NSA-19 24.00 -3.03 -5.88 62.50 15.74 ** 5.04 3.75 44.23 ** 44.23 ** 234.00 66.90 ** 51.51 **
19 NACH-226 NCAGA-26 x NSA-27 23.00 -8.97 -13.21 * 59.00 4.89 -7.81 3.80 49.02 ** 46.15 ** 224.00 56.67 ** 40.00 **
20 NACH-12 NCAGA-26 x NSA-28 24.50 1.03 0.00 69.50 41.84 ** 40.40 ** 3.85 46.67 ** 45.28 ** 268.80 108.61 ** 104.74 **
21 NACH-18 NCAGA-26 x NSA-29 24.50 0.00 -2.00 71.50 43.00 ** 38.83 ** 4.10 50.46 ** 43.86 ** 293.40 115.34 ** 100.20 **
22 NACH-375 NCAGA-26 x NSA-32 21.50 -7.53 -10.42 50.50 5.21 4.12 3.40 28.30 ** 25.93 ** 171.85 35.13 ** 33.84 **
23 NACH-433 NCAGA-26 x NSA-300 24.50 -3.92 -9.26 74.50 42.58 ** 33.04 ** 4.00 55.34 ** 53.85 ** 297.70 121.92 ** 109.13 **
24 NACH-434 NCAGA-26 x NSA-301 26.00 .0.95 -8.77 57.00 20.00 ** 17.53 ** 3.95 23.44 ** 3.95 224.85 48.66 ** 27.36 **
3.34
No. of sympodia/plant Avg. Boll wt (gm)
MP BP
24.96
1.78
3.69
5.01
0.12
0.25
0.34
Seed cotton yield/plant (gm) Sr.
No.
Hybrid
No. of bolls/ Plant
8.89
18.39
1.19
2.46
Cross
S.Em +
CD at 5%
CD at 1%
BP MP BP MP BP

Heterosis for Yield and Yield Attributing Traits in Arboreum Cotton (Gossypium arboreum L.) 71
REFERENCES
[1] Chavan, M. K., Shekar, V. B., Golhar, S. R., Gite, B. D. and Rajput, N. R. (1999). Heterosis studies in the interspecific
crosses of G. arboreum and G. herbaceum, J. Soils and Crops. 9: 195-197
[2] Jain, S.(1996). Heterosis for fibre characters in intra and interspecific hybrid of cotton. J. Cotton Res. Dev. 10: 147-49.
[3] Kapoor, C.J., Singh, M. and Maheshawari, R.V.(2002). Heterosis for yield and yield attributing traits in Desi cotton. J.
Cotton Res. Dev. 16 : 182-183.
[4] Nanjundan, J., Sangwan, B. S., Chhabra, B. S. and Kumar, R., (2004). Heterosis for yield and its component traits in Desi
cotton (Gossypium arbourem L.) J. Cotton Res. Dev. 18 (1): 33-35.
[5] Nirania, K. S., Singh, I. P., Singh, B. P., Jain, P. P. and Tutga, O. P. (1992). Heterosis and combining ability analysis in
Gossypium arboreum L. J. Cotton Res. Dev. 6 (1): 11-17
[6] Patel, K. G., Patel, R. B., Patel, M. I. and Kumar, V. (2009). Studies on heterosis and combining ability through
introgression in diploid cotton. J. Cotton Res. Dev. 23 (1): 23-26.
[7] Patil, F. B., Shinde, Y. M. and Thombre, M. V. (1991). Heterosis in multiple environments for yield components and its
relation with genetic divergence in cotton. Indian J. Genet. 51: 118 - 24.
[8] Patil, S. S., Gavit, A. F., Magar, N. M. and Pawar, V. Y.(2009). Heterosis in hybrid of Gossypium arboreum cotton. J.
Cotton Res. Dev. 23: 209-212.

Multi-Level Determination for Heat Tolerance

of Cotton Cultivars
Nicola S. Cottee
1
, Michael P. Bange
2
, Daniel K.Y. Tan
3
, J. Tom Cothren
4

1
Faculty of Agriculture Food and Natural Resources, The University of Sydney,
Sydney, NSW 2006 Australia
2
CSIRO Division of Plant Industry, Locked Bag 59, Narrabri, NSW 2390 Australia
3
Cotton Catchment Communities Cooperative Research Centre, Locked Bag 1001
Narrabri, NSW 2390 Australia
4
Department of Soil and Crop Sciences, Texas A&M University,
College Station Texas 77843, USA
E-mail: nicola.cottee@csiro.au
AbstractHigh temperatures (>35
o
C) are common throughout the cotton-growing season in many regions and may
adversely affect the growth and development potential of the crop, ultimately limiting yield. Development of stress
screening techniques will enable selection of heat tolerant genotypes for incorporation into future breeding programs.
This study assessed the use of the membrane integrity and enzyme viability assays as biochemical screening techniques
for determination of genotypic difference in heat tolerance under field conditions. These biochemical screens were
evaluated as part of a multi-level approach assessing morphological, physiological, biochemical and molecular
determinants of heat tolerance in response to evaluated temperatures.
High yielding Australian cotton cultivars of known and differing yield performance in hot environments were
evaluated to ascertain whether biochemical screens could be employed to detect differences in their heat tolerance.
Cultivar Sicot 53 was selected as a relatively thermotolerance genotype whilst Sicala 45 was selected as a cultivar with
relatively lower heat tolerance. To simulate elevated temperature in the field, clear plastic tents were constructed above
the cotton crops in Narrabri, Australia and College Station, USA to determine whether a field based high temperature
stress improved the resolution of biochemical screens for heat tolerance and whether differences detected translated to
physiological performance.
This study revealed that field-based elevated air temperatures were not sufficient to resolve cultivar differences in
cell membrane integrity or enzyme viability under ambient field conditions across the three cotton-growing seasons.
However, exposure of leaf tissue to high air temperature using tents resulted in a genotype specific response to heat
stress. Implementation of tents increased the resolution of the membrane integrity assay and changed the response of
enzyme viability for cultivars Sicot 53 and Sicala 45. Cultivar differences for membrane integrity (P=0.007) and
enzyme viability (P<0.001) were consistent in explaining differences for photosynthesis (P=0.046), electron transport
rate (P=0.057) and stomatal conductance (P=0.036) which reflected previously determined differences in yield. This
highlighted the potential for development of rapid biochemical screening methods for heat tolerance to be used in
combination with a multi-level approach also incorporating morphological, physiological and molecular performance
indicators to ensure that genotypes selected would contribute to improvements in economic yield.
INTRODUCTION
Laboratory based assays have been developed to rapidly screen large cotton populations for heat
tolerance (Azhar et al., 2009; Khan et al., 2008; Rahman et al., 2004). Incorporation of rapid biochemical
assays for heat tolerance determination into conventional breeding programs may assist in breeding for
increased yield in hot cotton growing environments (Constable et al., 2001; de Ronde and van der
Mescht 1997; Ismail and Hall 1999; Rahman et al., 2004; Saadalla et al., 1990; Sullivan 1971). Inclusion
of heat tolerance screens into current breeding programme may decrease the intergenerational time and
logistic constraints associated with breeding programs focused on long term selection for yield in warm
and hot environments.
Reductions in performance at temperatures exceeding the thermal kinetic window (23.5 to 32
o
C) for
cotton (Burke et al., 1988) are commonplace in Australian cotton production regions and may be
attributed to decreased plant growth and development, as well as increased fruit shedding (Hodges et al..
12
Multi-Level Determination for Heat Tolerance of Cotton Cultivars 73
1993). At a leaf level, evaluation of processes associated with chronic photosynthetic decline (Reddy et
al., 1991) such as membrane leakage and permeability (Sullivan 1971) and decreased stability of
photosynthetic (Salvucci and Crafts-Brandner 2004a) and respiratory enzymes (de Ronde and van der
Mescht 1997) offer potential for development of rapid and reliable biochemical screens for heat tolerance
determination.
However extension of the principals of these screens to in-situ high temperature stress in the field is
limited, principally due to the difficulties in creating an effective in-situ heat treatment in a field
environment. A recent study by Cottee et al., (2010) indicated that genotypic differences determined
using the laboratory-based cell membrane integrity and enzyme viability assays for heat tolerance
determination reflected higher order physiological performance and yield under elevated temperature
tents in the field. Although biochemical assays have been successfully used to discern heat tolerance in
cotton cultivars using different thermal environments (Ismail and Hall 1999) or by staggering planting
date (Rahman et al., 2004), it is unknown whether laboratory determined biochemical performance
reflects genotypic differences in cell membrane integrity and enzyme viability under in-situ high
temperature stress in the field.
In this study, heat tolerance was ascertained for two high yielding Australian cotton cultivars with
known field performance under high temperatures using the cell membrane integrity and enzyme
viability assays under ambient field conditions and were then validated under elevated temperature tents.
Genotypic differences for biochemical performance were then compared with photosynthesis, stomatal
conductance and electron transport rate under tents to determine whether elevated temperatures similarly
affected leaf level physiological performance.
This paper presents research that indicates that genotypic differences for membrane integrity and
enzyme viability are not able to be detected for material grown under ambient field conditions. However,
exposure of leaf material to in-situ elevated temperatures under tents increases the resolution of the cell
membrane integrity assay and increases the response of dehydrogenase activity in cotton cultivars with a
relatively higher (Sicot 53) and lower (Sicala 45) heat tolerance and these biochemical differences were
consistent with previously determined performance under field conditions and were reflected in higher
order physiological performance under tents.
Genotypes
Normal leaf, non-transgenic, medium maturity and high yielding cultivars of upland cotton (Gossypium
hirsutum L.) were established under field conditions. Sicot 53 was selected as a relatively thermotolerant
cultivar while Sicala 45 was selected as a cultivar with relatively lower heat tolerance. This selection was
based on recent breeding data (not published) in hot and cool environments in experiments conducted by
the Commonwealth Scientific and Industrial Research Organisation (CSIRO) breeding program over a
number of sites and seasons (cf. Reid et al., 1989)
Experimental Design and Plot Management
Cultivars were grown over three consecutive seasons in a randomized complete block design. Season 1
(sown 14
th
October 2005) and 3 (sown 18
th
October 2006) were located at the Australian Cotton Research
Institute, Narrabri, Australia (30
o
S, 150
o
E). Season 2 (sown 25
th
April 2006) was located at the Texas
Agricultural Experiment Station near College Station, Texas, U.S.A. (30
o
N, 96
o
W). Plots were furrow
irrigated every 10 to 14 d, according to crop requirements and high input management and pest control
were maintained throughout the season. Meteorological data were collected less than 2km from the field
sites.

In Situ Heat Stress Treatment
Heat stress treatments were applied in the field to identify whether cultivar differences could be
determined using biochemical heat tolerance assays under in-situ high temperature stress in a field
environment. In-situ high temperature treatment was imposed by the construction of Solarweave tents
above the crop canopy to increase ambient air temperatures (Lopez et al., 2003). Tents were applied
between first square and the cessation of vegetative growth for a period of between 4 and 20 days within
the 3 seasons. Time of application was dependent on environmental and managerial conditions whereby
run 1, 2 and 3 refer to an early, mid and late season tent. Tents were only applied 4 days post irrigation
and hence soil moisture was non-limiting during the time of tent application. Mean temperature under the
tent was higher than external ambient air temperatures for all tent events (Table 1.). The average daily
maximum temperatures were increased by 5.3
o
C under tents, (pooled for all measurements within the 3
seasons).
TABLE 1: MEAN DAILY MAXIMUM, MEAN AND MINIMUM TEMPERATURE UNDER AMBIENT (CONTROL) AND TENT TEMPERATURE REGIMES
IN THE FIELD FOR SEASONS 1 AND 3 IN NARRABRI AND SEASON 2 IN TEXAS. THE MEAN IS TAKEN FROM 3 REPLICATES
Season Run Daily Maximum Temperature Daily Mean Temperature Daily Minimum Temperature
Control Tent Control Tent Control Tent
1 1 37.25 43.92 28.61 31.89 22.35 23.06
1 2 33.04 36.77 24.02 25.79 18.00 19.23
2 1 40.08 46.51 31.05 32.64 23.15 22.66
3 1 34.82 37.31 25.30 26.26 16.09 16.46
3 2 37.00 42.68 27.22 29.06 19.02 18.89
3 3 36.29 42.83 25.09 27.72 16.16 16.41
Mean 36.41 41.67 26.88 28.89 19.13 19.45
l.s.d. 2.44 1.1 0.65
F test P value <.001 <.001 n.s.
Biochemical Assays for Heat Tolerance
In-vitro measurements of membrane integrity and enzyme viability were made on leaf material grown
under ambient field conditions and under tents. Leaves of cultivars Sicot 53 and Sicala 45 were collected
under ambient field conditions or under the tents between 1400 and 1500 h and transported back to the
laboratory in paper bags. For each run, leaves for the ambient field and tent treatments were sampled
once at the end of the tent measurement period and leaf material was taken from the same row within the
same plot for comparison between treatments. Leaf discs with a 10mm diameter were excised from the
interveinal portion of the 3
rd
youngest fully expanded leaf. Discs were triple rinsed to remove exogenous
electrolytes.
For the membrane integrity assay, 5 discs were submerged in 10 mL distilled water in 25mL glass
vials. One set of leaf discs were held for 2 h at 25
o
C (REC
25
) in a thermally controlled water bath. An
additional set of leaf discs from the same parent leaf were held for 2 h at 45
o
C (REC
45
) in an adjacent
water bath. Vials were allowed to cool to room temperature. Initial electrical conductivity (IEC
t
) was
determined at each water bath temperature using a low range (0-1990 S cm
-1
) ECTestr calibrated
conductivity meter (Oakton Instruments, Vernon Hills, IL). Vials were then autoclaved at 121
o
C and 103
kPa for 15 min and cooled to room temperature. Final electrical conductivity (FEC
t
) was determined for
each water bath temperature with the calibrated conductivity meter. Relative electrical conductivity was
determined for each water bath temperature where REC
t
= (IEC
t
/FEC
t
)*100. Relative cellular injury was
then calculated where RCI
45
= (1-((1-(IEC
45
/FEC
45
))/(1-(IEC
25
/FEC
25
))))*100 to account for differences
incurred during the sampling and water bath incubation period, whereby RCI only shows the injury
caused by increased temperature (Sullivan 1971). Increasing REC
t
and RCI
45
indicates decreasing
membrane integrity (Sullivan 1971).
For the enzyme viability assay, 2 leaf discs were taken from the same leaf sampled for the cell
membrane integrity assay and were submerged in 2 mL distilled water in 14 mL glass vials. One set of
leaf discs were held for 2 h at 25
o
C (Abs
25
) in a thermally controlled water bath. An additional set of leaf
discs from the same parent leaf were held for 2 h at 40
o
C (Abs
40
) in an adjacent water bath. Vials were
cooled to 25
o
C and a buffer solution containing 0.01 M phosphate buffered saline (-0.138 M NaCl;
0.0027 M KCl with TWEEN 20 (0.05% v/v)), pH 7.4 at 25
o
C) and 0.8% w/v 2,3,5-triphenyltetrazolium
chloride (TTC) was prepared and 8 mL added to each vial. Leaf discs were held at -33 kPa for 15 mins to
ensure TTC uptake into the leaf and incubated in the dark for 24 h at 25
o
C. Enzyme viability was
determined by spectrophotometric absorbance at 530 nm using 95% ethanol as a reference. Relative
absorbance at 530 nm was calculated where RAbs
40
= (Abs
40
/Abs
25
)*100 to account for differences
incurred during the water bath incubation period. High absorbance indicates high enzyme viability
whereas low absorbance is indicative of impaired enzyme activity and decreased capacity for reduction
of TTC to a red formazan product.
Physiological Measurements
To determine the impact of tents on potential growth, photosynthesis, electron transport rate and stomatal
conductance were simultaneously measured using a Li-6400 portable photosynthesis system (Li-Cor Ltd,
Lincoln, NE) with a pulse-amplitude modulated (PAM) leaf chamber fluorometer sensor head.
Measurements were taken on at least 2 days during the tent measurement period. Measurements were
made on the third youngest fully expanded leaf for 3 different plants per plot and four replicates per
treatment. All measurements were taken between 1000 and 1300 h. Environmental variables were set to
approximately optimal conditions and as such, all differences in physiological data present the
consequences of previous exposure to ambient (control) or elevated (tent) temperatures after returning
leaves to optimal conditions. The leaf chamber block temperature was maintained at 30
o
C, light intensity
was set at 2000 mol PAR m
-2
s
-1
, flow rate was adjusted to maintain vapour pressure deficit between 1.5
and 2.5 kPa and relative carbon dioxide concentration was set at 400 mol CO
2
mol
-1
using a CO
2
mixer.
Data Analysis
To determine cultivar biochemical and physiological performance, under ambient field conditions and
under tents, two way ANOVA (cultivar.tent) was conducted for electrical conductivity, relative cellular
injury, absorbance at 530 nm, relative absorbance at 530 nm, photosynthesis, electron transport rate and
stomatal conductance under ambient field conditions and under tents for seasons 1 and 3 in Narrabri and
season 2 in Texas. Treatments were blocked by replicate nested within run, nested within season
(season/run/replicate).
Results
Genotypic differences were found for both membrane integrity and the enzyme viability assays under the
tents (Table 2). REC
45
(Fig. 1b) and RCI
45
(Fig. 1c) decreased under tents compared with ambient field
conditions (Table 2). REC
45
was lower for Sicot 53 for leaf tissue grown under ambient field conditions
and this difference was greater under the tents (Fig. 1b) indicating relatively higher membrane integrity
compared with Sicala 45. Similarly, RCI
45
was slightly lower for Sicot 53 under tents compared with
Sicala 45 (Fig. 1c).

Fig. 1: Mean Relative Electrical Conductivity at (a) 25
o
C (REC
25
) or (b) 45
o
C (REC
45
) in the Water Bath, (c) Relative Cellular Injury at 45
o
C in the Water
Bath (RCI
45
), Absorbance at 530 nm Measured at 25
o
C (d) (Abs
25
) or 40
o
C (e) Abs
40
in the Water Bath and (f) Relative Absorbance at 530 nm Measured
at 40
o
C (RAbs
40
) in the Water Bath for Cotton Cultivars Sicot 53 and Sicala 45 under Ambient Field Conditions (Control) and under Tents Pooled for all
Measurements in Seasons 1 and 3 in Narrabri and Season 2 in Texas. The Vertical Lines in (b), (c), (e) and (f) Represent the Least Significant
Temperature Treatment by Cultivar Interaction at P=0.050
An interaction between cultivar and tent treatment was determined for the enzyme viability assay at
40
o
C in the water bath (Table 2). For Sicot 53, enzyme viability was higher for leaf material collected
under the tents compared with ambient field conditions at high water bath temperatures (Fig. 1e).
Conversely enzyme viability was lower for leaf material collected under the tents compared with ambient
field conditions for Sicala 45 (Fig. 1e). As such, Abs
40
was lower for Sicot 53 under ambient field
conditions but higher under tents compared with Sicala 45 (Fig. 1e). These genotype and tent treatment
differences were similarly reflected in calculation of RAbs
40
(Fig. 1f).
No genotypic differences were determined for leaf tissue grown under ambient field conditions or
under the tents and subsequently incubated at control (25
o
C) water bath temperatures measured using the
membrane integrity (REC
25
) or enzyme viability (Abs
25
) assays (Table 2). Genotypic differences were
found for all physiological measurements taken on leaves under ambient field conditions and under the
tents (Table ). Photosynthesis (Fig. 2a) and electron transport rate (Fig. 2b) were decreased while
stomatal conductance (Fig. 2c) was increased and under tents compared with ambient field conditions
(Table ). The decrease in photosynthesis under tents compared with ambient field conditions was greater
for Sicala 45 compared with Sicot 53 (Fig. 2a). Similarly, the decrease in electron transport rate was
slightly higher for Sicala 45 compared with Sicot 53 under tents (Fig. 2b). The increase in stomatal
conductance under tents compared with ambient field conditions was greater for Sicot 53 compared with
Sicala 45 (Fig. 2c). No inherent genotype differences were determined for photosynthesis, electron
transport rate or stomatal conductance under ambient (control) field conditions (Fig. 2).
l.s.d. Control Tent
R
C
I
4
5
20
30
40
50
A
b
s
2
5
0.0
0.1
0.2
0.3
0.4
0.5
R
E
C
2
5
0
2
4
6
8
10
12
Sicot 53
Sicala 45
R
E
C
4
5
20
30
40
50
A
b
s
4
0
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
l.s.d. Control Tent
R
A
b
s
4
0
0
20
40
60
80
100
120
140
(a)
(b)
(c)
(d)
(e)
(f)

Fig. 2: Mean (a) Photosynthesis (b) Electron Transport Rate or (c) Stomatal Conductance for Cotton Cultivars Sicot 53 and Sicala 45 under Ambient
Field Conditions (Control) and under Tents Pooled for all Measurements in Seasons 1 and 3 in Narrabri and Season 2 in Texas. The Vertical Lines in (a),
(b) and (c) Represent the Least Significant Temperature Treatment by Cultivar Interaction at P=0.050.
DISCUSSION
In this study, laboratory assays for membrane integrity and enzyme viability were successfully
implemented to resolve differences between genotypes with previously determined heat tolerance under
field conditions. No genotypic differences were determined for leaf material collected under ambient
field conditions (Fig. 1). However, exposure of plants to in-situ high temperatures under tents resolved
genotypic differences for membrane integrity and enzyme viability. Consistent with previously
determined relative heat tolerance under field conditions, Sicot 53 outperformed Sicala 45 for cell
membrane integrity (P=0.007) and enzyme viability (P<0.001) (Table 2) as well as photosynthesis
(P=0.046), electron transport rate (P=0.057) and stomatal conductance (P=0.036) (Table ) when exposed
to high temperatures under tents.
Imposition of tents increased daily maximum air temperatures between 6 and 20
o
C, with a daily
maximum average of 5.3
o
C higher than the control and this increase in temperature was associated with a
genotype specific change in membrane integrity and enzyme viability. Exposure to elevated temperature
under tents improved the resolution of the membrane integrity assay (Table 2) where Sicot 53
outperformed Sicala 45 for REC
45
(Fig. 1b). These cultivar differences were similarly reflected in
calculation of RCI
45
(Table 2). The improvement in genotypic resolution under tents for the membrane
integrity assays indicates that resolution of genotypic differences for biochemical performance are best
detected in the presence of stress, a notion similarly supported by Ashraf et al., (1994), Ismail and Hall
(1999) and Rahman et al., (2004).
l.s.d. Control Tent
S
t
o
m
a
t
a
l

c
o
n
d
u
c
t
a
n
c
e

(
m
m
o
l

H
2
0

m
-
2

s
-
1
)
0.2
0.4
0.6
0.8
Sicot 53
Sicala 45
P
h
o
t
o
s
y
n
t
h
e
s
i
s

(
m
o
l

C
O
2

m
-
2

s
-
1
)
20
25
30
35
E
l
e
c
t
r
o
n

t
r
a
n
s
p
o
r
t

r
a
t
e

(
m
o
l

e
-
1

m
-
2

s
-
1
)
200
220
240
260
280
(a)
(b)
(c)
TABLE 2: F TEST P VALUES FOR RELATIVE ELECTRICAL CONDUCTIVITY (REC
25
, REC
45
), RELATIVE CELLULAR INJURY (RCI
45
), ABSORBANCE AT 530 NM (ABS
25
, ABS
40
)
AND RELATIVE ABSORBANCE AT 530 NM (RABS
40
) FOR COTTON CULTIVARS SICOT 53 AND SICALA 45 GROWN UNDER AMBIENT FIELD CONDITIONS OR UNDER TENTS,
POOLED FOR ALL MEASUREMENTS TAKEN IN SEASONS 1 AND 3 IN NARRABRI AND SEASON 2 IN TEXAS
Biochemical Measurement Cultivar Tent Cultivar. Tent
REC
25
n.s. n.s. n.s.
REC
45
n.s. 0.023 0.007
RCI
45
n.s. 0.005 0.077
Abs
25
n.s. n.s. n.s.
Abs
40
0.004 n.s. <0.001
RAbs
40
0.022 n.s. <0.001
Conversely, the genotypic response for enzyme viability changed under tents compared with ambient
field conditions. Although Abs
40
was lower for Sicot 53 compared with Sicala 45 under ambient field
conditions, Sicot 53 outperformed Sicala 45 under tents (Fig. 1e). This interaction was reflected for
calculation of RAbs
40
(Table 2). These results were similar to those reported by de Ronde et al. (1995)
who attributed higher absorbance for heat stressed cotton compared with the control to a higher affinity
for acclimation to high temperature stress in heat tolerant cotton cultivars. The reversal of genotypic
performance under ambient field conditions and under tents indicates that the enzyme viability test for
heat tolerance determination should be considered under in-situ high temperature stress, or hot growing
regions for which the heat tolerant cultivars are being specifically selected.
To determine whether genotypic differences for biochemical performance translated to higher order
physiology, gas exchange, fluorescence and water flux were evaluated under tents. Decreases in
photosynthesis and electron transport coupled with increases in stomatal conductance under tents indicate
that the temperature stress imposed by tents was sufficient to significantly alter physiological
performance (Fig. 2). Sicot 53 was found to outperform Sicala 45 for both biochemical assays (Table 2)
and these differences are reflected physiological performance under tents (Table ).
TABLE 3: F TEST P VALUES FOR PHOTOSYNTHESIS, ELECTRON TRANSPORT RATE AND STOMATAL CONDUCTANCE MAIN EFFECTS AND INTERACTIONS FOR COTTON CULTIVARS SICOT 53
AND SICALA 45 UNDER AMBIENT FIELD CONDITIONS (CONTROL) AND UNDER TENTS POOLED FOR ALL MEASUREMENTS TAKEN IN SEASONS 1 AND 3 IN NARRABRI AND SEASON 2 IN TEXAS.
Physiological Measurement Cultivar Tent Cultivar. Tent
Photosynthesis n.s. 0.006 0.046
Electron transport rate n.s. <.001 0.057
Stomatal conductance n.s. <.001 0.036
Decreases in net photosynthesis under elevated temperatures may be attributed to limited electron
flow through photosynthetic and respiratory pathways (Wise et al., 2004) as a result of compromised
membrane permeability (Gupta 2007) and a decline in the activity of rate-limiting enzymes, particularly
those associate with photosynthetic and respiratory channels (Bjorkman et al., 1980; Burke et al., 1988;
Salvucci and Crafts-Brandner 2004 b). Alternately, it may be proposed that genotype specific decreases
in membrane permeability and deactivation of respiratory enzymes found in this study are indicative of
an overall decrease in the functionality of a range of proteins and metabolic processes that are decreased
with elevated temperature under tents. However, photosynthetic rates still require delivery of carbon
dioxide to the photosystem and in this study, it is likely that stomata remained open to facilitate gas
exchange and maintain leaf temperature under elevated temperature in the tents, as water was non-
limiting (Lu et al., 1994).
CONCLUSION
In this study, genotypic differences determined for the membrane integrity and enzyme viability assays
were most reflective of field performance when evaluated under in-situ high temperature stress in the
field. Similar to previously determined heat tolerance in the field, Sicot 53 outperformed Sicala 45 under
tents for both membrane integrity and enzyme viability and differences found using biochemical screens
for heat tolerance helped to explain differences in photosynthesis, electron transport rate and stomatal
conductance under tents. Thus, the biochemical assays were successfully implemented to resolve
between a cultivar with relatively high heat tolerance and a cultivar with relatively lower heat tolerance
under elevated temperatures in the field. The findings of this study indicate that biochemical screens for
heat tolerance may be useful for initial screening of large populations but heat tolerance should be
confirmed under contrasting thermal environments considering higher order physiological performance
before selections are made for incorporation into breeding programmes.
ACKNOWLEDGEMENT
Thanks to Darin Hodgson, Jo Price, Jane Caton, Merry Errington, Warren Conaty, Erica Cuell and Aman
Dayal in Narrabri and Texas A&M Aggies Josh Bynum, Ellen Batchelder, Joerdan Kennedy, Justin
Scheiner and Matt Nors for technical assistance, particularly constructing and dismantling the tents. This
work received partial financial support from the Cotton Catchment Communities Cooperative Research
Corporation, the Australian Cotton Research and Development Corporation, The University of Sydney,
Texas A&M University and CSIRO Plant Industry.
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[4] Burke, J. J., Mahan, J. R. and Hatfield, J. L. (1988). Crop-specific thermal kinetic windows in relation to wheat and cotton
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[10] Hodges, H. F., Reddy, K. R., McKinion, J. M., and Reddy, V. R. (1993). Temperature effects on cotton. Mississippi
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morphology in cowpea. Crop Sci. 39, 1762-1968.
[12] Khan, A. I., Khan, I. A., and Sadaqat, H. A. (2008). Heat tolerance is variable in cotton (Gossypium hirsutum L.) and can
be exploited for breeding better yielding cultivars under high temperature regimes. Pak. J. of Botany. 40, 2053-2058.
[13] Lopez, M., Gutierrez, M. V., El-Dahan, M. A. A., Leidi, E. O. and Gutierrez, J. C. (2003). Genotypic variation in response
to heat stress in Upland cotton. In 'Proceedings of the 3rd World Cotton Conference', 2003, Capetown, South Africa, pp.
104-108
[14] Lu, Z., Radin, J. W., Turcotte, E. L., Percy, R. and Zeiger, E. (1994). High yields in advanced lines of Pima cotton are
associated with higher stomatal conductance, reduced leaf area and lower leaf temperature. Physiol. Planta. 92, 266-272.
[15] Rahman, H., Malik, S.A. and Saleem, M. (2004). Heat tolerance of upland cotton during the fruiting stage evaluated using
cellular membrane thermostability. Field Crops Res. 85, 149-158.
[16] Reddy, V.R., Baker, D.N., and Hodges, H.F. (1991). Temperature effects on cotton canopy growth, photosynthesis and
respiration. Agron. J. 83, 699-704.
[17] Reid, P.E., Thomson, N.J., Lawrence, P.K., Luckett, D.J., McIntyre, G.T. and Williams, E.R. (1989). Regional evaluation
of cotton cultivars in eastern Australia, 1974-85. Austr. J. Expe. Agric. 29, 679-689.
[18] Saadalla, M.M., Quick, J.S., and Shanahan, J.F. (1990). Heat tolerance in winter wheat: II. Membrane thermostability and
field performance. Crop Sci. 30 (6), 1248-1251.
[19] Salvucci, M.E., Crafts-Brandner, S.J. (2004a). Mechanism for deactivation of Rubisco under moderate heat stress. Physio.
Planta. 122, 513-519.
[20] Salvucci, M.E. and Crafts-Brandner, S.J. (2004b). Relationship between the heat tolerance of photosynthesis and the
thermal stability of Rubisco activase in plants from contrasting thermal environments. Pl. Physio. 134, 1460-1470.
[21] Sullivan, C.Y. (1971). Mechanisms of heat and drought resistance in grain sorghum and methods of measurement. In
'Sorghum in the Seventies.' (Eds NGP Rao and LR House) pp. 247-284. (Oxford & IBH Publishing Co: New Delhi, India)
[22] Wise, R.R., Olson, A.J., Schrader, S.M., and Sharkey, T.D. (2004). Electron transport is the functional limitation of
photosynthesis in field-grown Pima cotton plants at high temperature. Pl. Cell and Environ. 27, 717-724.
Genetic Variability in Single Plant Selections

for Improving Drought Tolerance
in Upland Cotton
Suman B. Singh, A.H. Prakash and Amol A. Karpe
Central Institute for Cotton Research, Nagpur, India
AbstractDrought tolerant is one of the most important environmental stress that limits the plant growth and
development and in turn productivity worldwide. Work on development of drought tolerant genotypes was initiated
using drought tolerant sources. Screening of 112 single plant selections in two sets of experiment for physiological and
biochemical parameters resulted in identification of few single plant selections. Selections showed significant difference
for seed cotton yield ranging from 408.86 to 1803.86 kg/ha. The lines were separated based on the differential
expression of biochemical factors in control and stress plants. The difference was expressed as per cent
increase/decrease over the control. Considering the holistic approach viz., physiological, biochemical and yield per se
eight lines namely, DTS 39-09, DTS 62-09, DTS 44-09, DTS 67-09, DTS 155-09, DTS 104-09, DTS 100-09 and
DTS 108-09 were found to be tolerant. Selections DTS 67-09 identified as drought tolerant in trial 1 recorded good
yield and fibre quality with 26.9 mm 2.5% span length, 23.5 g/tex fibre strength and 4.2 10
-6
g/in micronaire while
DTS 104-09 in second trial recorded good yield and fibre quality with 27.2 mm 2.5% span length, 18.7 g/tex fibre
strength and 4.2 10
-6
g/in micronaire. The results suggest that selection of genotypes based on drought susceptibility
index was effective in development/identification of drought tolerant genotypes.
INTRODUCTION
Cotton is the most important fibre crop and the basic input to the textile industry. In India, cotton is
grown in about 11.0 million ha of which more than 70 per cent area is rainfed. Maharashtra has an
estimated area of 3.503 million ha under cotton, predominantly under rainfed cultivation (97%). The
rainfed tract of central India receives about 500-1200 mm rainfall annually. Uneven distribution of
rainfall with prolong dry spells during critical crop growth phases considerably affects growth and
development leading to low yield particularly in shrink-swell soils. Thus a rainfed-cotton cultivar needs
to withstand extended period of water stress and able to utilize rain when it occurs. Late maturing
cultivars have shown to meet these requirements (Stiller et. al., 2004) and genotype with okra leaf traits
have also been successful (Stiller et. al., 2004, Thomson, 1994).
Physiological traits associated with WUE or stress tolerance have not been used extensively in plant
breeding. This is due to the difficulties associated with measuring these traits on large number of plants,
low heritability and complex relationships between these traits and yield (Hall et. al., 1994). It has been
well established that plant accumulates a variety of osmo-protectant solutes as an adaptive mechanism to
environmental stress such as salinity (Hayashi et al., 1997), water deficit (Rhodes and Hanson, 1993),
temperature extremes (Hayashi et. al., 1997). Osmo-protectant solutes includes (i) sugar and sugar
alcohols i.e. polyols (Thomas et. al., 1995).(ii) proline (Aspinall and Paley, 1981) and a number of
quaternary ammonium compounds and tertiary sulphonium compounds (Rhodes and Hanson, 1993).
Developing plants with higher levels of natural solute accumulating capacity to increase stress
tolerance by selection between cultivars or isogenic lines and / or by genetic engineering of plants to
accumulate high levels of polyols (Thomas et. al., 1995) and glycine betaine (Hayashi et. al. 1997) is one
way to increase plant tolerance to stress.
In present study genotypes from the 50 germplasm lines screened for water use efficiency (WUE) and
grouped into high, medium and low WUE groups. F
1
crosses were affected using lines with high water
13
Genetic Variability in Single Plant Selections for Improving Drought Tolerance in Upland Cotton 81
use efficiency and genotypes with broad genetic base. Crosses were evaluated for drought tolerance
efficiency (DTE), seed cotton yield and other economic characters. Single plant selections were made in
F
2
of the crosses which recorded high drought tolerance efficiency. The selections were tested in two sets
(set 1 and set 2) of experiment in pots with 49 and 62 single plant selections under simulated drought and
non-stressed conditions in pots in different years. Simultaneously these selections were also tested under
field conditions in randomized block design with three replications. Five random plants were tagged from
each plot for recording observations on number of bolls per plant, boll weight (g) and seed cotton yield
(kg/ha).The data was subjected to statistical analysis by adopting standard procedure of Panse and
Sukhatme (1985).
Observations on physiological (chlorophyll content and membrane stability) and biochemical
parameters (reducing sugars, amino acid and phenols) were recorded during peak flowering stage.
Chlorophyll a and b were estimated as per the method out lined by Arnon (1949) and membrane stability
index was measured by the method described by Sullivan (1971).
Single plant selections (112 no.) tested in two different sets for experiment (in pots) under simulated
drought differed significantly for chlorophyll content, membrane stability, reducing sugars, amino acids
and phenols. Selection DTS 83-09 recorded significantly higher seed cotton yield (2073 kg/ha) followed
by DTS 36-09 (1827 kg/ha), DTS 75-09 (1795 kg/ha), DTS 88-09 (1751 kg/ha), DTS 85-09 (1714 kg/ha)
and DTS 67-09 (1701 kg/ha) under field condition in first set (Table 1). However, under simulated
drought (in pots) DTS 39-09, DTS 67-09, DTS 36-09, DTS 82-09, DTS 85-09, DTS 88-09, DTS 62-09
selections showed their superiority towards higher yields. In second set, DTS 119-09 (1803 kg/ha), DTS
102-09 (1775 kg/ha), DTS 104-09 (1750 kg/ha), DTS 116-09 (1694 kg/ha) and DTS 151-09 (1681 kg/ha)
recorded significantly higher seed cotton yield under field trial (Table 2) while under stressed condition
selections DTS 103-09, DTS 131-09, DTS 155-09, DTS 108-09, DTS 104-09, DTS 107-09 were the
better performers. The results are in agreement with the finding of Singh et.al. (2006) and
Rajarajeswari(1995).
Among biochemical and physiological parameters, the SPS showed considerable variability for these
traits as they were found to increase or decrease with the increase in the stress level. Higher reducing
sugars and amino acid content and less of phenols in a particular genotype is considered as drought
tolerant. Similarly positive chlorophyll value and negative membrane stability index help in rating a
genotype as tolerant or susceptible. The total chlorophyll content varied significantly among the SPS
tested but reduction was recorded after stress. DTS 21-09, DTS 44-09, DTS 39-09, DTS 62-09, DTS 67-
09, DTS 92-09 and DTS 76-09 did not show any variation for total chlorophyll content however, some of
them maintained higher level of chlorophyll under stress (5 to 44 per cent increase) over the control.
Similar results with increase level of chlorophyll content and physiological activities in high yielding
genotypes were reported by Gadallah (1995) and Krasichkova et. al.(1989). High yielding SPS under
field condition namely DTS 88-09, DTS 36-09 and DTS 6-09 had larger effect under stress i.e. -41 to -48
per cent reduction over the non-stressed condition.
CSI (maintenance of higher chlorophyll content under high temperature), a highly desirable
characteristic under moisture stress conditions, was significantly higher in selection namely DTS 44-09,
DTS 01-09, DTS 15-09, DTS 39-09, DTS 62-09, DTS 66-09, DTS 72-09, DTS 70-09, DTS 36-09, DTS
79-09, DTS 80-09,DTS 24-04, DTS 83-09, DTS 84-09, DTS 86-09, DTS 89-09 and DTS 90-09 under
both stressed and non-stressed conditions.
Membrane stability index ranged from 27.1 to 65% and 23.2 to 88.3 per cent under non-stressed and
stressed conditions respectively. The electrolyte leakage has been used successfully to measure
membrane integrity in plants subjected to a variety of environmental stresses (McKersie and Tomes
1980; Blum and Ebercon 1981, McKersie et. al. 1982; Sapra and Anade 1991, Agarie et. al. 1995). Kuo
and collegues (1993) showed that yields of vegetable species with low electrolyte leakage were more
stable in different growing conditions.
In general there was an increase in the membrane stability index, amino acids and reducing sugar
from non-stress to stress condition. Accumulation of solutes such as sugars, amino acids, organic acids
and ions during drought stress have been observed in many crops. One of the most notable changes is the
synthesis and accumulation of low molecular weight, osmotically active compounds such as sugar,
alcohols, amino acids, organic acids and glycine betaine (Turner 1979, Yancy et. al. 1982, Morgan 1984,
Good and Zaplachinski 1994; Bartels and Sunkar 2005. The accumulation of these compounds lead to
osmotic adjustments as indicated by an increase in the intra-cellular osmotic potential of the cell
(Morgan, 1984).
Based on these physiological and biochemical parameters the selections were grouped / rated as
good, moderate and susceptible for individual characters (Table 3). The selections were rated against
LRA 5166 as standard (moderately tolerant to moisture stress). The selections which represented good or
moderate category for the traits were selected. Few promising selections were identified which showed
more than 25% and 90% increase over the check Rajat and LRA 5166. Selection DTS 67-09 (identified
as drought tolerant) also recorded good yield (1701 kg/ha) and good fibre quality with 26.9 mm 2.5%
span length, 23.5 g/tex fibre strength and 4.2 10
-6
g/in micronaire . DTS 25-09, DTS 44-09 and DTS 74-
09 cultures which recorded more than 24.4 g/tex fibre strength and S/L ratio of >0.80 as against LRA
5166 which recorded fibre strength of 21.2 g/tex and S/L ratio of 0.75 (Table 4). In the second set,
selections were identified having more than 50 % and 100% increase over the check Rajat and LRA
5166. Selection DTS 104-09, identified as drought tolerant, also recorded good yield (1750 kg/ha) and
good fibre quality (27.2 mm 2.5% span length, 18.7 g/tex fibre strength and 4.2 10
-6
g/in micronaire).
DTS 96-09, DTS 143-09 and DTS 106-09 had more than 21.4 g/tex fibre strength and S/L ratio of >0.75
as against LRA 5166 which recorded fibre strength of 16.9 g/tex and S/L ratio of 0.69 (Table 5).
Thus, single plant selections namely DTS 39-09, DTS 44-09, DTS 62-09, DTS 67-09, DTS 100-09,
DTS 104-09, DTS 108-09 and DTS 155-09 were identified as tolerant lines through holistic approach of
physiological, biochemical and yield per se was analyzed. These entries can be advanced further for
detailed studies and promotion for multilocation evaluation trials under stress conditions.
TABLE 1: PERFORMANCE OF TOP TEN SINGLE PLANT SELECTION IN FIELD TRIAL OF SET 1
S. No. SPS SCY
(kg/ha)
% Increase over Check B. Wt.
(g)
GOT
(%)
2.5% SL(mm) FS
(g/tex)
Micronaire
(10
-6
g/in) Rajat LRA 5166
1 DTS 83-09 2073.19 55.11 139.11 3.2 39.4 27.9 21.5 4.3
2 DTS 36-09 1827.04 36.69 110.72 3.7 38.9 25.3 22.4 4.0
3 DTS 75-09 1792.93 34.14 106.79 3.3 38.8 24.9 20.6 4.3
4 DTS 88-09 1751.73 31.06 102.04 4.4 40.5 27.5 18.6 4.1
5 DTS 85-09 1714.23 28.25 97.71 3.1 42.1 31.7 19.1 3.8
6 DTS 67-09 1701.16 27.27 96.20 3.6 42.4 26.9 23.5 4.2
7 DTS 69-09 1690.27 26.46 94.95 3.8 38.2 - - -
8 DTS 6-09 1679.20 25.63 93.67 4.3 37.2 27.3 22.5 4.0
9 DTS 82-09 1671.40 25.05 92.77 3.2 39.7 26.3 20.1 3.5
10 DTS 71-09 1669.34 24.89 92.52 3.4 42.0 - - -
Rajat 1336.55 - - 3.3 38.9 - - -
LRA 5166 867.010 - - 2.7 39.0 28.0 21.2 4.6
C.V 20.07
TABLE 2: PERFORMANCE OF TOP TEN SPS IN TRIAL II
S. No. SPS SCY (kg/ha) % Increase over Check B. Wt. (g) GOT (%) MHL (mm)
Rajat LRA 5166
1 DTS 119-09 1803.36 60.19 141.13 2.8 38.9 22.3
2 DTS 102-09 1775.25 57.69 137.37 2.7 39.8 25.0
3 DTS 104-09 1750.64 55.51 134.08 4.0 40.1 27.2
4 DTS 116-09 1694.99 50.56 126.99 3.1 42.6 19.3
5 DTS 151-09 1681.65 49.38 124.86 4.1 40.6 24.5
6 DTS 150-09 1676.04 48.88 124.11 3.5 41.6 21.5
7 DTS 110-09 1672.76 48.59 123.67 3.4 38.7 24.2
8 DTS 103-09 1666.67 48.05 122.85 3.7 42.7 28.8
9 DTS 131-09 1640.35 45.78 119.33 3.3 38.7 24.2
10 DTS 120-09 1616.39 43.58 116.13 4.0 37.7 26.0
Rajat (c ) 1125.73 - - 3.0 41.8 21.1
LRA 5166 ( c ) 747.86 - - 3.0 42.8 26.3
C.V. 19.28
Genetic Variability in Single Plant Selections for Improving Drought Tolerance in Upland Cotton 83
TABLE 3: EVALUATION FOR DROUGHT TOLERANCE PARAMETERS
Characters Good Moderate
Chlorophyll stability
index (%)
DTS 21, 44, 39, 62, 67, 92, 76 LRA5166, 72, 73, 74, 75, 47, 42, 38, 81, 82, 14, 83, 5
Membrane stability index
(%)
DTS40, LRA5166, 44, 41, 1, 39, 65, 7, 6, 77, 78, 81, 24,
86, 88, 91
23, 6, 64, 66, 67, 36, 70, 73, 92, 38, 79, 83, 84,85, 90,
91
Reducing sugar mg/g DTS6,LRA5166,44,41,39,15,66,70,7,79,24,37,84,88,91 25, 40, 64, 72, 73, 47, 42, 78, 43, 80, 89, 90
Amino acid mg/g DTS 25, 23, 65, 66, 72, 42, 76, 78, 80, 82, 85 44,1,62,63,64,67,92,47,38,76,43,79,24,86,87,88,77
Phenols mg/g DTS 44, 1, 39, 15, 19, 62, 63, 65, 74, 92, 75, 77, 14 26, 21, 40, 23, 6, LRA, 41, 64, 66, 67, 36, 70, 73, 92,
42, 76, 7, 78, 43, 80, 84, 86, 87, 89, 90
Yield stability DTS 25, 23, LRA,5166, 41, 1,39, 15, 19, 62, 63, 65, 66,
67, 36, 70, 72, 43, 79, 24, 82, 14, 84, 85, 87, 88, 90, 91, 7
26, 21, 40, 6, 44, 47, 42, 76, 78, 81, 86
TABLE 4: TOP TEN SPS WITH GOOD FIBRE QUALITY IN TRIAL I
S. No. SPS SCY (kg/ha) 2.5%SL (mm) Maturity UR Micronaire (10
-6
g/in) FS (g/tex) Elongation(%) S/L ratio
1 DTS 25-09 756.45 27.3 0.76 48.2 4.2 25.3 6.0 0.93
2 DTS 44-09 1495.75 27.3 0.73 48.6 3.9 24.1 5.4 0.88
3 DTS 74-09 1509.01 25.9 0.84 49.1 4.1 24.4 5.9 0.94
4 DTS 73-09 1399.63 24.1 0.74 52.1 3.7 23.9 6.2 0.99
5 DTS 67-09 1701.16 26.9 0.77 48.9 4.2 23.5 6.0 0.87
6 DTS 87-09 1048.03 27.5 0.82 48.0 4.0 23.5 5.6 0.85
7 DTS 90-09 1475.72 28.7 0.79 49.7 4.4 23.3 5.0 0.81
8 DTS 84-09 1210.46 25.4 0.83 52.4 4.8 23.2 5.4 0.91
9 DTS 91-09 1584.96 28.4 0.72 49.4 3.7 23.2 5.1 0.81
10 DTS 72-09 1325.44 31.9 0.68 45.6 3.6 23.2 5.6 0.72
LRA 5166 867.01 28.0 0.80 47.2 4.6 21.2 5.4 0.75
TABLE 5: PERFORMANCE OF TOP TEN SINGLE PLANT SELECTIONS IN TRIAL I
S. No. SPS SCY (kg/ha) % Increase over Check B. Wt.
(g)
GOT
(%)
2.5% SL
(mm)
UR
(%)
FS
(g/tex)
Micronaire
(10
-6
g/in)
Rajat LRA5166
1 DTS 43-09 1534.31 31.88 113.26 3.6 35.9 30.0 46 19.2 3.6
2 DTS 83-09 1436.11 23.44 99.61 4.1 36.0 27.2 50 17.5 3.8
3 DTS 90-09 1348.10 15.88 87.38 3.7 37.5 28.0 51 20.4 3.9
4 DTS 79-09 1324.07 13.81 84.04 3.1 37.5 25.5 52 19.3 4.1
5 DTS 86-09 1285.88 10.53 78.73 3.4 38.8 24.5 51 17.6 3.7
6 DTS 66-09 1255.55 7.92 74.51 3.7 35.4 24.4 52 18.2 4.2
7 DTS 77-09 1230.86 5.80 71.08 4.1 36.4 26.3 48 22.1 3.8
8 DTS 69-09 1212.97 4.26 68.59 3.9 39.7 26.3 49 17.9 4.3
9 DTS 62-09 1190.06 2.26 65.41 3.0 35.7 25.4 50 18.6 4.3
10 DTS 76-09 1157.05 - 60.82 3.7 36.7 23.3 49 20.0 3.8
LRA 5166 719.44 3.1 35.3 25.4 50 19.2 3.6
Rajat 1163.34 3.7 34.1 25.3 50 19.2 4.3
C.V. 21.40
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15.
[2] Ashraf, M., Saeed, M. M. and Qureshi, M. J. (1994) - Tolerance to high temperature in cotton at initial growth stage. Env.
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L.G. Paleg and D. Aspinall. Academic Press, Sydney. pp. 205-241.
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[5] Devendra Singh, Singh,S. B. and Ahuja, S. L. (2006) - Screening of cotton genotypes for drought tolerance based on stress
susceptibility index. J. Cotton. Res. Dev. 20:(1) 89-90.
[6] Gadalloh, M. A. A. (1995) - Effect of water stress, abscissic acid and proline on cotton plants. J. Arid Envt. 30:(3), 315-325.
[7] Good, A. G and Zaplachinski, S. T.(1994) - The effect of drought stress on fresh amino acid accumulation and protein
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[8] Hall, A. E., Richards, R. A., Condon, A. G., Wright, G. C., and Farquhar. G. D. (1994) - Carbon isotope discrimination and
plant breeding. p. 18-113. In. J. Janick (ed.) Plant Breeding Review, Vol. 12. John Wiley and Sons. New York.
[9] Hayashi, N., Alia, Mustardy, Deshnium, L., Ida P. M. and Murata, N. (1997) - The Plant J. 2. 133-142.

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the early stages of development. Doklady Vsesovuznoi Ordena. Trudovogo Krasnogo Znameni Akademmi Sel skokhozya
istvennykh Nauk. Imen V. I. Lenina. No. 12, 9-11, Institute Fiziologii Bio fiziki. Rastenil, Academy of Sciences, Tadjik
SSR. Dushanbe. Tadjik SSR.
[11] Morgan, J. M. (1984) - Osmoregulation and water stress in higher plants. Annu. Rev. Plant Physiol., 35:299-319.
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Physiology, 9(2): 109-112.
[14] Ranney, T. G., Bassuk, N. L. and Whitlow, T. H. (1991) - Osmotic adjustment and solute constituents in leaves and roots of
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osmotic adjustment in tetreploid wheats. Photosynthetica 35: 129-138.
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[17] Stiller, W. N., Reid P. E., and Constable, G. A. (2004) - Maturity and leaf shape as traits influencing cotton cultivar
adaptation to dryland conditions. Agron. J. 96: 656-664.
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injury and resistance in crops. Crop Sci. Soc. Am. Madison, Wis.
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mariana) progenies: II osmotic adjustment and changes of soluble carbohydrate and amino acids under osmotic stress.
85:645-651.
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Melbourne, V.I.C.
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stems and roots of apple. Aust. J. Plant. Physiol. 22:747-754.
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Genetic Parameters of Physiological Traits for Salinity

Tolerance in Diverse Genotypes of Cotton
(Gossypium hirsutum L. and Gossypium barbadense L.)
Hosseini Gholamhossein
1
and Behdarvand Pejman
2

1
Cotton Research Institute of Iran
2
Department of Botany, University of Pune- 411007, India
AbstractGenetic and physiological parameters of different traits related to salinity tolerance in six cotton
(Gossypium hirsutum L. and Gossypium barbadense L.) genotypes and their progenies were estimated using diallel
crosses which conducted at Agricultural and Resources Research Center of Mazandaran, Iran and Department of
Botany, University of Pune, India during 3 years ending to 2009. Along with nutrition by normal Hoglands solution
in non-saline condition, incremental levels of NaCl to Hoglands solution in a sand culture was added until electrical
conductivity of 24 dSm
-1
was attained inducing salinity stress and caused osmotic shock. Combined analysis based on
two salinity levels revealed significant salinity-level effects for traits. High ratios of
2
A / (
2
A +
2
D) and high
narrow-sense heritability estimates were observed for root length, plant height, Na
+
,

K
+
, Ca
2+,
K
+
/Na
+
, Ca
2+
/Na
+
, root
length/shoot height and Tolerance Index (TOL), indicating the more involvement of genes additive effects in their
genetic control. High differences of [
2
g / (
2
g +
2
e)] - [2
2
gca +
2
sca / (
2
g +
2
e)] were observed for K
+
/Na
+
and
TOL, indicated non-allelic effects in their genetic control. Negative and high index of SI for Na
+
indicated that its
mean in a saline environment was more than two times from the mean in a non-saline environment. Low estimates of
negative SI index for root length, Ca
2+
indicated the effects of a saline environment for these traits were better than the
other traits. The results of factor analysis indicated that selection for morphological traits, specifically; that selection
based on K
+
, Ca
2+
and K
+
/ Na
+
should be more efficient than other traits. Cluster classification of genotypes by means
of principle component analysis method on the basis of value of correlation matrix distinguished genotypes Sindose-80
and Sindose-80 Siokra from other genotypes in salinity tolerance. Low estimates of the tolerance index (TOL) in the
above genotypes are also conforming of their salinity tolerance in early stages of growth.
INTRODUCTION
Cotton is a dual-purpose (fiber and oil) crop. It is moderately salt-tolerant crop with a salinity threshold
level 7.7 dSm
-1
. Its growth and seed yield is severely reduced at high salinity levels. However, inter and
intra-specific variation for salt tolerance in cotton is valuable and thus can be exploited through specific
selection and breeding for enhancing salt tolerance of the crop. Salinity of agricultural lands and
irrigation water is one of the factors of environment, which limits the growth and yield of cotton and
other crops in many arid and semi-arid regions of the world (Postel, 1989). The stresses imposed by
salinity are mainly due to ion compositions and concentrations in rhizosphere and also in plant tissues
(Volkmar et al., 1997). Information on the mechanisms involved in salt tolerance and their genetic
control is essential to facilitate selection for characteristics and to design an efficient breeding
programme for genetic improvement of salinity tolerance (Ashraf, 1994). Plant breeders have focused on
finding new resistant cultivars because of detrimental effects of saline soil and irrigation water to reduce
quantitative and qualitative cotton crop. Identifying proper selection criteria for salinity tolerance is also a
major problem. In this respect, some selection criteria including Geometric Mean Productivity (Fischer
and Murrer, 1978), Stress Tolerance Index (Fernandez 1993) and Tolerance Index (Rosielle and
Hamblin, 1981) have been defined. Significant general combining ability (GCA) and specific combining
ability (SCA) effects of salinity resistance have been reported in Gossypium hirsutum (Saghir and et al.,
2002). The objective of the present study is to determine the important genetic parameters for shoot dry
weight (g/ plot), root length (cm), plant height (cm), shoot fresh weight, Ca
2+
, K
+
, and Na
+
contents (mg
g
-1
),Ca
2+
/Na
+
and K
+
/Na
+,
shoot dry weight / shoot fresh weight(SDW/SFW), root length/shoot height
(RL/SH) and Geometric Mean Productivity (GMP), Stress Tolerance Index (STI) and Tolerance Index
(TOL) for shoot dry weight in order to select a suitable breeding program for cotton breeding line and
cultivars.
14
A complete diallel cross of six cotton genotypes (Gossypium hirsutum L. & Gossypium barbadense L.)
viz Delinter, Sindose-80, Bulgare-539, Termez-14, B-557, Siokra having diverse genetic origin was
conducted over two years to determine the potential for improvement in yield, its components, oil and
fiber quality traits by means of genetic analysis, combining ability, heritability and heterotic effects. The
detailed studies were based on F
1
generations whereas crossed seed in first year were used for F
1

generation in the second year. The successful hybrids recognized and distinguished by morphological
markers such as flower colour, spot position and its colour in petal, leaf colour and it
s shape (Fig.1,2),
fiber colour, seed linter, and also molecular marker (SSR). Successful hybrids also exposed to salinity
and non-salinity condition for selecting the resistant genotypes to saline environments. Six diverse cotton
(Gossypium hirsutum L. and Gossypium barbadense L.) genotypes were crossed in a diallel fashion, and
their 15 F
1
progenies along with their parents were evaluated in sand cultures under normal and saline
environments in a block design with three replications at the greenhouse during 3 years ending to 2009.
Sterilized seeds were germinated in Petri dishes at 22 3C for 4 days. Eight uniform seedlings were
transplanted into plots that separated in two sand boxes filled with washed and sterilized river sand,
covered with polythene beads. After the establishment, five plants were maintained for evaluation.
Temperatures during the experiment were averaged 33.21/ 10.6C (day/night) and relative humidity was
31.43-75.04% and the photoperiod was 14 hrs. Plants were given deionized water up to 10 days after
transplanting and saline and non-saline (control) grown plants were irrigated thereafter every 2 days with
half-strength Hoagland
s nutrient solution (Hoagland and Arnon, 1950) with NaCl (EC = 22 dSm
-1
) and
without it (EC = 0.88 dSm
-1
) and pH = 7. Electrical conductivity of the saline treatment was increased to
the desired level by incremental addition of the salt over 10-day period to avoid osmotic shock to the
seedlings. Plants in both environments were irrigated until saturated, with the excess solution allowed to
drain under sandboxes. The plants in both environments were harvested 45 days after planting at 7-8 leaf
stages. The characteristics such as shoot dry weight (SDW) in gr/plot, root length (cm),plant height (cm),
shoot fresh weight in gr/plot, Na
+
, K
+
, and Ca
2+
contents (mg g
-1
), K
+
/Na
+
, Ca
2+
/Na
+
, shoot dry weight /
shoot fresh weight(SDW/SFW), root length/shoot height (RL/SH), Geometric Mean Productivity(GMP),
Stress Tolerance Index (STI) and Tolerance Index (TOL) for shoot dry weight were determined. The
harvested plants thoroughly washed with distilled water then dried in an oven for 72 h at 80 C to a
constant weight. Plant samples were ground by mill and dried in a furnace at 500 C for 2 hrs for ion
extraction. Plant samples were added to 5mL of 2M HCl for digestion, and the digested solutions were
filtered and diluted by distilled water. The final volume of each sample was 100mL. Sodium and K
+

levels of each sample were measured by flame photometry and Ca
2+
was measured by atomic absorption
spectrophotometry (Isaac and Kerber, 1971). The selection criteria indices, including Geometric Mean
Productivity as GMP = (Yp)(Ys) (Fischer and Murrer, 1978),Stress Tolerance Index as STI = (Yp)
(Ys)/Yp
2
(Rosielle and Hamblin,1981) and Tolerance Index as TOL = Yp Ys ( Fernandez, 1993 ) were
calculated for yield of shoot dry weight in non-saline(Yp) and saline(Ys) environments. Tolerance Index
(TOL) was also calculated for Ca
2+
, K
+
, Na
+
, Ca
2+
/Na
+
, and K
+
/Na. Data subjected to analysis of variance
and means were compared by using the Least Significant Differences (LSD). Variations in general
combining ability (GCA) of the parental lines and specific combining ability (SCA) of crosses for the
measured characteristics were partitioned from the total genetic variance using Griffings Method II,
Model I (Griffing, 1956) . The components of variance ratio as
2
A / (
2
A +
2
D) and high narrow-sense
heritability [
2
A/(
2
A+
2
D+
2
e)] and [
2
A/(
2
g+
2
e)] were computed as a (
2
A,
2
D,
2
e) Estimated from
diallel analysis) and ab(
2
g,
2
e Estimated from Randomized Complete Block Design) method for each
characteristic to determine the relative importance of additive and non-additive gene effects respectively
(Baker, 1978). The differences of variance ratios as [
2
g/(
2
g+
2
e)]
b
-

[2
2
gca+
2
sca /(
2
g+
2
e)]
ab
and
[2
2
gca+
2
sca/(2
2
gca+
2
sca+
2
e)]
a
were computed for each characteristic to determine the non-allelic
effects in their genetic control. High-parent hetrosis was calculated as mean deviation of a cross
performance from the mean of its superior parent (Mather and Jinks, 1982).

Genetic Parameters of Physiological Traits for Salinity Tolerance in Diverse Genotypes of Cotton 87
X1= Delinter X2 = Sindose-80 X3 = Bulgare-539 X4 = Termez-14 X5= B-557
X6 = Siokra Xij = X
X1= Delinter X2 = Sindose-80 X3 = Bulgare-539 X4 = Termez-14 X5= B-557
X6 = Siokra Xij = X
For ion composition, the parents and Siokra with 1.66 g/plot had superior mean for shoot dry weight,
Delinter with 10.90 cm had superior mean for root length, Termez-14 with 25.74 cm had superior mean
for plant height, Termez-14 with 10.81 g/plot had superior mean for shoot fresh weight, Sindose-80 with
15.68 mg g
-1
had superior mean for Na
+
, Bulgare-539 with 24.67 mg g
-1
had superior mean for K
+,

Delinter with 47.74 mg g
-1
had superior mean for Ca
2+.

TABLE 1: ANALYSIS OF VARIANCE FOR SHOOT DRY WEIGHT, ROOT LENGTH, PLANT HEIGHT,
SHOOT FRESH WEIGHT, SHOOT DRY WEIGHT NA
+
, K
+
, CA
2+
, IN A SALINE ENVIRONMENT (MEAN OF THE SQUARES)
S.O.V D.F
S
h
o
o
t

D
r
y

W
e
i
g
h
t

(
S
D
W
)

R
o
o
t

L
e
n
g
t
h

(
R
L
)

P
l
a
n
t

H
e
i
g
h
t

(
P
H
)

S
h
o
o
t

F
r
e
s
h

W
e
i
g
h
t

(
S
F
W
)

Na
+
K
+
Ca
2+

GMP
(SDW)
STI
(SDW)
TOL
(SDW)
Crosses 35 0.187** 11.32** 8.21** 6.11** 21.46** 39.58** 23.63 0.082** 0.33** 0.15**
GCA 5 0.28** 25.19** 15.99** 8.76** 47.77** 98.33** 62.18 0.132** 0.45** 0.46**
SCA 30 0.183** 5.99** 5.96** 5.45** 13.19** 21.15* 10.88 0.056** 0.26 0.24**
Error 70 0.044 1.015 0.966** 0.711 1.99 1.256 0 .98 0.011 0.055 0.014
2
A/(
2
A +
2
D) 0.41 0.66 0.49 0.36 0.65 0.69 0.69 0.45 0.28 0.70
Narrow-sense heritability
a
0.28 0.45 0.43 0.28 0.48 0.54 0.54 0.33 0.25 0.71
Broad-sense heritability
a
0.88 0.92 0.86 0.87 0.83 0.99 0.76 0.79 0.85 0.94
C.V (%) 11.25 10.15 9.55 10.62 8.66 5.69 5.62 8.41 17.10 25.05
Narrow-sense heritability
ab
0.25 0.36 0.36 0.28 0.34 0.36 0.39 0.24 0.19 0.51
ab
0.66 0.64 0.69 0.63 0.65 0.83 0.73 0.69 0.66 0.75
b
0.76 0.79 0.82 0.76 0.74 0.94 0.79 0.72 0.69 0.84
BSH
b
-BSH
ab
0.10 0.15 0.13 0.13 0.09 0.11 0.06 0.03 0.03 0.09
*, ** Significant at P= 0.05and P= 0.01 respectively; A and D as defined in the text refer to additive and dominance genetic effects
respectively. And also a, b and ab as defined in the text refer to estimation of parameters with diallel assumption,without diallel
assumption(RCBD method) and
2
A,
2
D with diallel assumption and
2
P without diallel assumption respectively.
Significant variations in general combining ability and specific combining ability estimates were
observed for shoot dry weight, root length, plant height, shoot fresh weight, Na
+
, K
+
, and Ca2
+
contents
and indices of Geometric Mean Productivity (GMP), Stress Tolerance Index (STI), Tolerance Index
(TOL) for shoot dry weight (Table 1) indicates the importance of both additive and non-additive genetic
effects for these characteristics. High ratios of
2
A/(
2
A+
2
D) and high narrow-sense heritability
estimates of root length, plant height, Na
+
,

K
+
, Ca
2
and Tolerance Index (TOL), indicated the more
involvement of genes additive effects in their genetic control. Therefore, the efficiency of selection based
on these characters is expected to be high but, shoot dry weight, shoot fresh weight and indices of
Geometric Mean Productivity (GMP), Stress Tolerance Index (STI) were controlled pre-dominantly by
non-additive genetic effects (Table 1). When gca effects are not pre-dominant in self-pollinated crops, the
major portion of the variability, is due to additive additive genetic effects or divergence among
progenies in the same parental array and therefore, should be delayed to later generation. The high
differences of variance ratios [
2
g / (
2
g+
2
e)]
b
- [2
2
gca+
2
sca /(
2
g+
2
e)]
ab
were observed for those all
characteristics that had high narrow-sense heritability, indicating correlation between narrow-sense
heritability and non-allelic effects in their genetic control. This is resulted from interaction of many locus
of gene additive action in the quantitative characteristics that produce non-allelic effects. Sairam and
Tyagi (2004) also reported that Salinity stress response is multigenic, as a number of processes involved
in the tolerance mechanism are affected, such as various compatible solutes/osmolytes, polyamines,
reactive oxygen species and antioxidant defense mechanism, ion transport and compartmentalization of
injurious ions. Various genes/cDNAs encoding proteins involved in the above mentioned processes have
been identified and isolated in plants.
REFERENCES
[1] Ashraf, M. (1994). Breeding for salinity tolerance in plants. Critical Revi.3: 1742.
[2] Ashraf, M. and Saghir, A. (2000). Genetic effects for yield components and fibre characteristics in upland cotton
(gossypium hirsutum L.) cultivated under salinized (NaCl) conditions. Agron. J. 20:917-926.
[3] Baker, R. J. (1978). Issues in diallel analysis. Crop Sci.18: 533-536
[4] Fernandez, G. C. J. (1993). Effective selection criteria for assessing plant stress tolerance. In Adaptation of food Crops to
Temperature and Water Stress. Kuo.C.G.,Ed., AVRDC: Shanhua, Taiwan. 257-270.
[5] Fischer, R. A. and Murrer, R. (1978). Drought resistance in spring wheat cultivars .I. Grain yield response
.Aust.J.Agric.Res.29:897-912.
[6] Griffing, B. (1956). Concept of General and Specific Combining Ability in Relation to Diallel System. .Aust.J.Bio.Sci.
9:463-493
[7] Hayman, B.I. (1954). The Theory and Analysis of Diallel Crosses. Genetics 39:789-809
[8] Hoagland, D. R. and Arnon, D. I. (1950). The water culture method for growing plants without soil. Cali. Agric. Exp. Stn.
Cric.307,32 pp.
[9] Isaac, R. A. and Kerber, J.D. (1971). Atomic absorption and flame photometry: techniques and uses in soil, plant and water
analysis. In Instrumental Methods for Analysis of Soil and Plant Tissue, Walsh,L.M.,Ed., Soil Sci. Soc. Am. Madison,
WI:17-37.
[10] Postel, S. (1989). Water for agriculture: Facing the limits. Worldwatch paper 93.Worldwatch Institute. Washington DC.
[11] Rosielle, A. A. and Hamblin, J. (1981).Theoretical aspects of selection for yield in stress and non-stress environments. Crop
Sci.21: 943-946.
[12] Saghir, A., Khan, N. O., Igbal, M. Z., Hussain, A. and Hassan, M . (2002). Salt tolerance of cotton (Gossypium hirsutum L.)
Asian j. Pl. Sci. 1, 6:715-719.
[13] Sairam, R. K., Tyagi, A. (2004). Physiology and molecular biology of salinity stress tolerance in plants .Curr. Sci. 86,
3:407-421.
[14] Volkamar, K. M., Hu, Y. and Steppuhn, H. (1998). Physiological response of plants to salinity: a review. Can. J. Pl. Sci. 78:
19-27.
Marker-Assisted Selection
for Improving Drought Resistance in Cotton
Yehoshua Saranga
1
, Avishag Levi
1
and Andrew H. Paterson
2

1
The Robert H. Smith Faculty of Agriculture, Food and Environment,
The Hebrew University of Jerusalem, P.O. Box 12, Rehovot 76100, Israel

2
Plant Genome Mapping Laboratory, University of Georgia, Athens, GA 30602, USA
AbstractMarker-assisted selection (MAS) is gaining an increasing recognition as an efficient approach for
improving simply inherited traits, however, there are hardly any examples of successful MAS for complex polygenic
traits, such as yield and drought resistance. Quantitative trait loci (QTLs) for yield and drought-adaptive
physiological traits (osmotic potential, carbon isotope ratio - an indicator of water use efficiency, and leaf chlorophyll
content), were exchanged via MAS between elite cultivars of the two cotton species, Gossypium barbadense and G.
hirsutum. The resulting near isogenic lines (NILs) were examined in three field trials to test the effect of the
introgressed QTL alleles on cotton productivity under drought conditions and physiological traits. A considerable
number of NILs exhibited the expected phenotypes in term of greater osmotic adjustment, higher carbon isotope ratio,
and higher chlorophyll content. Moreover, a few NILs exhibited modifications in non-targeted traits such as greater
photosynthetic capacity under severe drought, modified leaf morphology, and considerable changes in metabolic and
mineral profiles. Finally, NILs introgressed with QTL alleles associated with high yield rarely exhibited a yield
advantage over the recurrent parent, consistently with other introgression studies, suggesting that the well balanced
genetic and physiological systems of the recurrent parents may be interrupted by these introgressions. We conclude
that MAS is a useful approach to enhance drought-adaptive traits in cotton, but complimentary recombination and
selection are required to combine these traits with high yield potential.
INTRODUCTION
About one-third of the world's arable land suffers from chronically inadequate water availability for
agriculture, and in virtually all agricultural regions, crop yields are periodically reduced by drought
(Boyer, 1982). Developing drought resistant crop plants is vital to meeting increasing demand for
agricultural products during an environmental shift to greater aridity (Parry et al., 2005). The
development of drought-tolerant crops by traditional breeding has been hampered by the low heritability
of traits such as yield, particularly under drought, and by large genotype x environment interactions.
Modern genomic techniques have aided tremendously in identifying quantitative trait loci (QTLs) and
diagnostic DNA markers in a wide range of crops and paved the way towards more efficient breeding
approaches.
Cotton (Gossypium spp.) with two pre-dominant cultivated species, G. hirsutum L. and G.
barbadense L. (denoted hereafter as GH and GB respectively), is the worlds leading fibre crop and also
an important oilseed. Cotton is an herbaceous warm-season crop and a major consumer of water.
Whether irrigated or not, cotton is often exposed to drought, which adversely affects both yield and lint
quality. In this regard, concerted efforts are required to improve drought resistance of cotton. Both GH
and GB cottons are tetraploid comprised of A and D sub-genomes that diverged from a common
ancestor about 4-11 million years ago and rejoined in a common nucleus about 1-2 million years ago
(Wendel, 1989). Modern cotton cultivars are outcome of intensive selection to produce large amounts of
specific types of fibers. This selection has unintentionally narrowed the genetic variability for drought
resistance (Rosenow et al., 1983). The availability of two domesticated closely-related cotton species that
have evolved independently and retained different genes or alleles for various traits provide an
opportunity to restore some of the desirable alleles "left behind" during domestication, by introgression
between the two domesticated species.
A long term project aimed at improving cotton adaptation to water-limited condition are briefly
reviewed in this paper.
15
Analysis of F
2
and F
3
generations of an inter-specific cotton population (GH var Siv'on x GB var F-177)
indicated a total of 79 QTLs for ten measures of plant productivity and physiological variables (Saranga
et al., 2001, 2004; Paterson et al., 2002). Productivity of cotton in well-watered versus water-limited
conditions was largely accounted for by different QTLs, indicating that adaptation to both conditions can
be combined in the same genotype (Saranga et al., 2001; Paterson et al., 2002). QTL likelihood intervals
for high seed cotton yield and low leaf osmotic potential corresponded in three of four possible genomic
regions (two involving QTLs specific to water-limited conditions), implicating osmotic potential as a
major component of improved cotton productivity under arid conditions (Saranga et al., 2004). These
results provided the first evidence that there appears to exist not only a phenotypic correlation but also a
partly common genetic basis of OA and productivity (Saranga et al., 2001). Two of these three loci
mapped to homeologous (corresponding) locations on the two sub-genomes of tetraploid cotton
(chromosomes 6 and 25), suggesting that a particularly important role of one or more ancestral genes in
that region may have been retained since the A-D genome divergence and polyploid formation. The
finding that the GH allele was favourable at some loci and the GB allele at other loci suggested that
recombination of favourable alleles may form novel inter-specific genotypes that are better adapted to
arid conditions than either of the parental species.
Selected genomic regions containing QTLs for yield and drought related physiological traits (osmotic
potential, carbon isotope ratio - an indicator of water use efficiency, and leaf chlorophyll content), were
exchanged via marker assisted selection (MAS) by backcrossing the source of the favourable allele to the
alternative parent (GH var Siv'on or GB var F-177). The resulting near isogenic lines (NILs) were
examined in three field trials to test the effect of the introgressed QTL alleles on cotton productivity
under drought conditions and the underlying physiological traits (Levi et al., 2009a). Many NILs
exhibited the expected phenotypes including lower osmotic potential or greater OA (5 out of 9), higher
carbon isotope ratio (4 of 6) or higher chlorophyll content (2 of 3). A few NILs exhibited modifications
in non-targeted traits such as leaf size, leaf pubescence, and stomatal density. Two NILs that were
subjected to gas-exchange study exhibited improved photosynthetic variables and one of them showed a
stable net rate of CO
2
assimilation across a wide range of leaf water potentials with a notable advantage
over its recurrent parent under severe drought (Levi et al., 2009b). Increased levels of several solutes
(alanine, aspartic acid, citric acid, malic acid, glycerol, myoinositol, threonic acid, potassium, magnesium
and calcium) were found under drought conditions in one or more NILs as compared with their recurrent
parents, which could contribute to their superior capacity to cope with drought (Levi et al., 2011).
Finally, NILs introgressed with QTL alleles associated with high yield in our mapping study rarely
exhibited a yield advantage over the recurrent parent.
CONCLUSION
Yield is known as a low-heritability complex trait, influenced by multiple gene networks and epistatic
interactions among genetic elements, as well as between genetic and environmental variables (Levi et al
2009a). Breeding for yield under stress conditions is even more complex due to the difficulty to define
and apply a precise set of environmental condition relevant to the range of naturally occurring stress
scenarios. This emphasizes the power of genetic mapping, allowing the dissection of complex traits and
distinguishing common heredity from casual associations (Paterson et al. 1988), which cannot be
achieved by conventional approaches. Targeting specific genomic regions and characterizing their effects
can enable the reconstruction of favourable loci into elite cultivars.
Marker-assisted selection (MAS) is gaining an increasing recognition as an efficient approach for
improving simply inherited traits; however, there are hardly any examples of successful MAS for
complex polygenic traits, such as yield and drought resistance. In the current study, cotton NILs
exhibited an improvement in drought related traits, compared with the recipient parent. From a
physiological point of view, such changes that can be considered as enhanced drought tolerance.
However, NILs containing putatively favourable alleles for yield did not exhibit a clear advantage. Elite
Marker-Assisted Selection for Improving Drought Resistance in Cotton 91
cotton cultivars, such as the recipient parents used in our study, are the outcome of intense selection over
many generations among huge number of individual genotypes. The well balanced genetic and
physiological systems of such elite cultivars may be interrupted by introgressions of large QTL regions
or undetected non-targeted introgression. This possibly is a major reason for the poor success of MAS for
improved yield in previous studies (Cattivelli et al. 2008) as well as in the current study. It is established
that maximal selection efficiency for quantitative traits may be obtained by a combination of molecular
and classical-phenotypic approaches. The successful introgression of QTLs for drought related traits in
this study may serve as a basis for future breeding, however, complimentary dissection of the
introgressed regions and conventional breeding are required to combine the advantageous effect of the
QTL(s) with high yield potential.
REFERENCES
[1] Boyer J.S. (1982). Plant productivity and environment. Science 218, 443-448.
[2] Cattivelli L., Rizza, F., Badeck, F. W., Mazzucotelli, E., Mastrangelo, A. M., Fracia, E., Mare, C., Tondelli, A., Stanca,
A.M. (2008) Drought tolerance improvement in crop plants: An integrated view from breeding to genomics. Field Crop Res
105: 1-14.
[3] Levi, A., Paterson, A. H., Barak, V., Yakir, D., Wang
,
B., Chee, P. W. and Saranga, Y.(2009a). Field Evaluation of Cotton
Near-Isogenic Lines Introgressed with QTLs for Productivity and Drought Related Traits. Molecular Breeding 23:179-195.
[4] Levi, A., Ovnat, L., Paterson, A. H., Saranga, Y. (2009b). Photosynthesis of cotton near-isogenic lines introgressed with
QTLs for productivity and drought related traits. Plant Science 177:88-96.
[5] Levi A., Paterson, A.H., Cakmak, I., and Saranga, Y. (2011). Metabolite and mineral analyses of cotton near-isogenic lines
introgressed with QTLs for productivity and drought related traits. Physiol, Plant. 141:265-275.
[6] Parry M.A.J., Flexas, J., Medrano, H. (2005). Prospects for crop production under drought: research priorities and future
directions. Ann. Appl. Biol., 147, 211-226.
[7] Paterson, A.H., Lander, E.S. Hewitt, J.D., Peterson, S., Lincoln, S.E. and Tanksley, S.D. (1988). Resolution of quantitative
traits into Mendelian factors by using a complete linkage map of restriction fragment length polymorphisms. Nature.335:
721-726.
[8] Paterson, A. H., Saranga, Y., Menz, M., Jiang, C., Wright, R. (2002). QTL Analysis of Genotype x Environment
Interactions Affecting Cotton Fiber Quality. Theor. Appl. Genet., 106:384-396.
[9] Rosenow, D.T., Quisenberry, J.E., Wendt, C.W., Clark, L.E. (1983). Drought tolerant sorghum and cotton germplasm.
Agricultural Water Management, 7:207-222.
[10] Saranga Y., Menz, M., Jiang, C., Wright, R., Yakir, D., Paterson, A.H. (2001). Genomic dissection of genotype x
environment interactions conferring adaptation of cotton to arid conditions. Genome Res., 11:1988-1995.
[11] Saranga Y., Jiang, C., Wright, R., Yakir, D., and Paterson, A.H. (2004). Genetic dissection of cotton physiological
responses to arid conditions and their inter-relationships with productivity. Plant Cell Environ., 27: 263-277.
[12] Wendel, J. (1989). New World tetraploid cottons contain Old World cytoplasm. Proc. Natl. Acad. Sci. USA 86: 4132-4136.

Tak FA84-4: New Jassid Tolerant Cotton Variety

P. Seburuang
1
,W. Sirichumpan
2
,P. Nakapan
3
,S. Thaitad
1
,S. Lapbunjob
1
A. Traisiri
1
,N.T. Khumla
1
,S. Areerak
1
,S.A. Juttupornpong
4
,N. Panlai
5
,
A. Kasivivat
6
,P. Sangsoda
7
,R. Chuekittisak
8
, B. Kumseub
9
,
P. Pulcha
7
andK. Khockakang
7
1
Nakhon Sawan Field Crops Research Center, Tak Fa, Nakhon Sawan60190
2
Khon Kaen Field Crops Research Center, Mueang, Khon Kaen40000
3
Phitsanulok Agriculture Research and Development Center, Wang Thong, Phitsanulok65130
4
Suphan Buri Agriculture Research and Development Center, U Thong, Suphan Buri72160
5
Lop Buri Agriculture Research and Development Center, Khok Toom, Mueang, Lop Buri15210
6
Phetchabun Field Crops Research Center, Mueang, Phetchabun Khon Kaen67000,
7
Loei Plant and Production Technical Service Center, Post Box 1, Mueang, Loei42000
8
Sukhothai Plant and Production Technical Service Center, Si Samrong, Sukhothai64120
9
Nakhon Ratchasima Agriculture Research and Development Center,
Si Khiew, Nakhon Ratchasima30340
E-mail: sebunruangp@yahoo.com
AbstractCotton damage from cotton jassid (Amrasca biguttula Ishida) becomes very serious problem, especially in a
dry period EL Nino, because of 50-100% reduction in cotton yield. Farmers overcome this serious damage by
increasing insecticide dose, resulting in more toxic residual effect to human health and environment. In order to reduce
more systemic insecticide used for jassid control, jassid tolerant hairy leaf cotton variety improvement was conducted
at Nakhon Sawan Field Crops Research Center in 1997. The hairy leaf IRMA1243, exhibiting jassid tolerance, was
crossed with smooth leaf and leaf roll resistant Tak Fa 2, which has high yield and good fiber quality cotton variety.
Thereafter, plants were selected in bulk in F
2
-F
4
generations, followed by plant-to-row or pure line selection in F
5
-F
6

generations for hairy leaf cotton lines with jassid tolerance, leaf roll disease resistance and high yield under non
systemic-insecticide application and artificial leaf roll disease inoculation. Uniform 14 lines with good plant type were
selected for yield potential evaluation in 2003-2008. The promising hairy leaf line, Tak Fa 84-4 was out-standing in
jassid tolerance, high yield, good fiber quality and leaf roll disease resistance.
INTRODUCTION
In the past 30 years, recommended cotton varieties from Department of Agriculture (DOA) had smooth
leaf, more susceptible to cotton jassid (Amrasca biguttula Ishida) than hairy leaf cotton (Boonramphan,
1999). The damage from cotton jassid was more during a dry period when leaves were almost completely
destroyed, resulting in reduction in photosynthesis, yield and fiber quality (Cowland, 1947). Most cotton
farmers overcome this serious damage by increasing systemic insecticide application, resulting in more toxic
residual effect to human health and environment. Some cotton farmers grow local hairy leaf cotton for less
insecticide spraying, especially systemic insecticide. However, they get less income from selling their seed
cotton due to quality (low ginning out turn or fiber percentage, short and coarse/high micronaire fiber or lint).
Besides, their local cotton yields rather low within 5-6 growing period as it is rather late maturing. In order to
produce a good hairy leaf cotton variety with good yield and quality, resistance to important leaf roll disease
and tolerance to the jassid for less insecticide use especially systemic insecticide use for cotton jassid control,
good jassid tolerant hairy leaf cotton variety improvement began at Nakhon Sawan Field Crops Research
Center (NSFCRC) in 1997.
Crossing and Selection
Hairy leaf IRMA 1243, exhibiting jassid tolerance, was crossed with smooth leaf and leaf roll resistant
Tak Fa 2, which has high yield and good fiber quality cotton variety. Thereafter, selection for good hairy
16
Tak FA84-4: New Jassid Tolerant Cotton Variety 93
leaf cotton lines with cotton jassid tolerance, leaf roll disease resistance and high yield under non
systemic insecticide application and artificial leaf roll disease inoculation. Good resistant/tolerant plants
were selected in bulk in F
2
-F
4
generations, followed by plant-to-row or pure line selection in F
5
-F
6

generations. Uniform promising rows/lines with good plant type were selected for yield potential
evaluation (preliminary trial, standard trial, regional trial and farm trial) in 2003 2008. Seed cotton
harvest took place four times with intervals of 7-10 days, beginning at 120 days after emergence.
Evaluation on Reaction to Leaf Roll Disease and Jassid
Leaf roll disease evaluation, in comparison with Tak Fa 2 and Deltapine Smooth Leaf (DPSL) that is
very susceptible, was conducted in the green house of NSFCRC in 2007. Reaction to jassid trial was
conducted at NSFCRC in 2006 under insecticide application once a week. The other trial was conduct
again in 2009 under insecticide application once a week in week1
th
-week6
th
followed by synthetic
pyrethroid application only, which did not have effect on jassid in week7
th
week14
th
.
Cotton Yield and Cotton Fiber Evaluation
Mean seed cotton yield data from preliminary trial, standard trial, regional trial and farm trial evaluation
are shown in Table 1. Tak Fa 84-4 yielded 1,625 kg per hectare of seed cotton, which was in the same
level of Tak Fa 2. Data on fiber quality are shown in Table 2, suggesting superiority of Tak Fa 84-4.
TABLE 1: MEAN DATA ON SEED COTTON YIELD (KG/HA) OF TAK FA 84 4 VARIETY, COMPARED TO TAK FA2 VARIETY IN 20032008 TRIALS.
Variety PT
2/
(2003) ST
3/
(2006) RT
4/
(2007) FT
5/
(2008) Mean
6/
Relative to Tak Fa 2
Tak Fa 84 4 1,938 1,675 a 1,519 a 1,713 a 1,625 98
Tak Fa 2 - 1,644 a 1,606 a 1,731 a 1,663 100
CV (%) 15.3 16.0 17.3 11.4 - -
No. of locations
1/
(2) (3) (6) (6) (15)
Remarks: Means within the same column followed by a common letter are not significantly different at 0.05 probability
level by DMRT.
1/
Number in parenthesis is the number of locations.
2/
Preliminary trial
3/
Standard trial
4/
Regional trial
5/
Farm trial
6/
Averaged from PT, ST, RT and FT in 20032008
TABLE 2: MEAN DATA ON GINNING OUT TURN OR GOT (FIBER OR LINT) PERCENTAGE AND FIBER QUALITY OF TAK FA 844 VARIETY,
COMPARED TO TAK FA 2 VARIETY IN 20062008.
Variety Got (%) Fiber Length
(Inch)
Fiber Bundle Strength
(g./tex)
Micronaire
(Fiber Fineness)
Fiber Uniformity
(%)
ST (3 locations)
Tak Fa 844 37.6 1.24 22.9 3.8 54
Tak Fa 2 36.4 1.20 20.8 3.7 52
RT (6 locations)
Tak Fa 844 38.1 1.21 24.4 3.9 57
Tak Fa 2 36.6 1.17 22.1 3.8 54
FT (3 locations)
Tak Fa 84 4 38.4 1.22 25.5 3.8 56
Tak Fa 2 37.2 1.20 21.6 4.1 54
Mean
1/

Tak Fa 84 4 38.0 1.23 24.6 3.9 56
Tak Fa 2 36.7 1.19 21.7 3.9 53
1/
Averaged from 3 locations of ST (standard trial), 6 locations of RT (regional trial) and 3 locations of FT (farm trial) in
2006 2008
Source: Sebunruang et al. (2009)
Reaction to Jassid
Mean number of jassids/10 cotton plants of Tak Fa 84-4 was lower than that of Tak Fa 2 in 2006 and
2009.(Table 3), this should be due to more hairiness as Tak Fa 84-4 had higher number of hairs on cotton
leaf surface as well as on cotton leaf vein (61.6 and 55.9/ 0.20 cm
2
, respectively).
TABLE 3: MEAN NUMBER OF JASSIDS/ 10 COTTON PLANTS AND NUMBER OF HAIRS ON COTTON LEAF SURFACE AND COTTON LEAF VEIN/0.20 CM
2

(NAKHON SAWAN FIELD CROPS RESEARCH CENTER, 2006 AND 2009)
Variety Number of Jassids Per 10 Plants Number of Hairs on
Leaf Surface Per 0.20 cm
2

Number of Hairs on Leaf
Vein per 0.20 cm
2
2006 2009
Tak Fa 844 97 a 556 a 61.6 a 55.9 a
Tak Fa 2 195 b 760 b 12.9 b 16.2 b
CV (%) 22.1 14.9 63.8 53.3
Means within the same column followed by a common latter are not significantly different at 0.05 probability level by
DMRT.
Source: Traisiri et al. (2006) Traisiri et al. (2009)
Leaf Roll Disease Reaction
Tak Fa 84-4 cotton variety was considerably resistant to leaf roll disease in the same level as Tak Fa 2
(Table 4). It should be very useful for cotton integrated pest control in which less insecticide application
would be needed, comparing to Tak Fa 2 and other smooth leaf cotton variety such as Deltapine Smooth
Leaf cotton variety that was very susceptible to leaf roll disease and failed to be commercially grown.
TABLE 4: REACTION TO LEAF ROLL DISEASE UNDER ARTIFICIAL INOCULATION IN 2007 OF TAK FA 844, TAK FA 2 AND DELTAPINE SMOOTH LEAF COTTON VARIETIES
Variety Leaf roll Disease Infected Plants (%) Disease Reaction
1
Tak Fa 844 0 Resistant
Tak Fa 2 3.3 Resistant
Deltapine Smooth Leaf 93.3 Susceptible
1/
Disease reaction is based on 3 levels as following:
0 10 % Resistant
11 40 % Moderately resistance
41 100 % Susceptible
Source: Lapbunjob et al. (2007)
From overall results and discussion it can be concluded that Tak Fa 84-4 cotton variety had good
adaptation in cotton growing area in the same high level of yield as Tak Fa 2. Due to hairiness of Tak Fa
84-4, growing this variety under less insecticide spraying will result in less reduction of cotton yield and
damage from cotton jassid. Moreover, Tak Fa 84-4, with good fiber quality, was resistant to leaf roll
disease; therefore, it is very useful for cotton farmers to grow under less pesticide use for controlling
jassid and aphid that was the leaf roll disease vector.
REFERENCES
[1] Boonramphan, P. (1999)-Cotton variety improvement for cotton jassid damage resistance. Project of Cotton variety
improvement for hight yield and good fiber quality. Kasetsart University. 10 p. (in Thai)
[2] Cowland, J.W. (1947)-The cotton jassid (Empoasca libyca Berg.) in the Anglo-Egyptian Sudan and experiments on its
control. Bull. Entom. Res. 38 (2): 99-115.
[3] Lapbunjob, S., P., Sebunruang and Thaitad, S. (2007)-Leaf roll disease of cotton lines evaluation,p 15. In: Annual report
2007. Nakhon Sawan Field Crops Research Center. Office of Agriculture Research and Development, Region 5,
Department of Agriculture. (in Thai).
[4] Sebunruang, P., S.Thaitad, P.Sangsoda, P.Pulcha, N.Panlai, R. Chuekittisak, S. Juttupornpong and Kasivivat, A. (2009)-
Farm trial: fiber quality cotton variety, p 13-14. In: Annual report 2009. Nakhon Sawan Field Crops Research Center, Field
Crops Research Institute, Department of Agriculture. (in Thai)
[5] Traisiri A., Sebunruang, P., Khumla, N. and Lapbunjob, S. (2006)-Study on distribution of cotton insect pest, p. 37. In:
Annual report 2006. Nakhon Sawan Field Crops Research Center, Field Crops Research Institute, Department of
Agriculture. (in Thai)
[6] Traisiri A., Sebunruang, P., Lapbunjob, S., Khumla, N. and Thaitad. S. (2009)-Reaction of cotton lines to aphid.
(Bemisia tabaci Gennadius.), p. 20-21. In: Annual report 2009. Nakhon Sawan Field Crops Research Center, Field Crops
Research Institute, Department of Agriculture. (in Thai).
High Boll Weight and High Ginning Outturn

The Major Tools for Breaking Yield Barriers
in Gossypium arboreum
Punit Mohan
1
, S. Manickam
2
, S.K. Verma
3
, D. Pathak
4
,
A.S. Singh
5
and Tarun Kumar Das
6
1
Principal Scientist, Central Institute for Cotton research, Nagpur-440010, India
2
Principal Scientist, Central Institute for Cotton Research, Regional Station, Coimbatore, India
3
Senior Scientist, Central Institute for Cotton Research, Regional Station, Sirsa, India
4
Assistant Plant Breeder, PAU, Ludhiana, India
5
Programme Coordinator, KVK, Tura, India
6
Subject matter specialist, Agri. Extension, KVK, Tura, India
AbstractThe high boll weight, high ginning outturn and optimum boll numbers are the major tools for breaking
yield barriers in cotton. Therefore, a exploratory and expedition survey of the tribal area of West Garo Hills of North
Eastern Hill Region, Meghalaya was conducted in 2000- 2010. The 59 germplasm lines including perennials of
Gossypium race cernuum were collected from where there was no introduction of improved cotton varieties and
commercial hybrids.The collected germplasm accessions were characterized by deep palmate leaf lobes, long petiole (20
cm), elongated acute and acrescent capsule with high boll weight (7.5g) and trilocular ovary, length of bursted locule
loaded with seed cotton (upto 15.5 cm), high locule retentivity and resistance to high wind velocity with minimum
locule shedding. They also possessed high ginning outturn (upto 51%), short staple (21.8 mm) fibre bundle strength
(19.7 g/tex) and coarse fibre (micronaire 8.0).The above germplasm accessions are important source for genetic
improvement of arboreum cottons in terms of locule retention capacity, boll weight and ginning outturn.
INTRODUCTION
Gossypium arboreum L., commonly known as tree cotton or desi cotton is native to India is under
cultivation from time immemorial. Arboreum cottons have wide adaptability and are relatively tolerant to
biotic and abiotic stresses (Singh and Punit Mohan, 2005). In any successful varietal improvement
programme, the availability of adequate variability in basic genetic stocks is most essential and their
utilization in breeding programme for building up improved strains are very necessary. Hence, the
collection, conservation, evaluation, documentation and utilization of diverse germplasm material assume
special significance.
The boll weight and ginning outturn are the basic features of breeding importance in arboreum
cotton. Present investigation was therefore undertaken to study boll weight, ginning outturn and fibre
technological properties of cerunnm race for improvement of yield, boll weight and ginning outturn of
desi cotton.
National germplasm exploration and expedition programme of West Garo Hills, Meghalaya, was jointly
carried out by the Central Institute for Cotton Research, Nagpur and National Bureau of Plant Genetic
Resources, New Delhi in 2000 and 2010. The 59 morphologically variant germplasm lines were
collected and evaluated for morphological characters and fibre technological properties. The important
characters of selected accessions are presented in Table 1.
The germplasm accessions collected had specific morphological characters viz., long petiole, deeply
palmate leaves, cordate base, central lobe once or twice toothed near the sinus, long broad bracts,
17
elongated- ovate acute, acrescent capsule/boll, white and yellow petals, yellow and creamy pollens, tri-
locular ovary, high boll weight (5 - 7.5 g), high locule retentivity, locules with 12-20 seeds free from
each other and firmly bound through interlacing of their wool. They also possessed high ginning outturn
(upto 51%), short staple (17-21.8 mm) and coarse fibre (micronaire upto 8) (Table 1).
High boll weight was recorded in the germplasm accessions GHA-11, GHA-6, GHA-3, GHA-18,
GHA-23, 30826 and 30838 respectively. The ginning out-turn ranged from 38.0 to 51.0 % (Table 1),
however earlier studies (Singh and Nandeshwar 1983; Singh and Raut, 1983; Punit Mohan et al., 1992)
have reported higher ginning outturn (GOT) with high seed density/ boll in selected genotypes of
cernuum race.
Bolls per plant is one of the important components of yield and is positively correlated with it. All
the available reports indicated a significant positive correlation between boll number and yield (Shinde
and Deshmukh, 1985; Aher et al., 1989 and Duhoon, 1989). Boll number was negatively correlated with
boll weight. Therefore, simultaneous improvement of boll number and boll weight may be difficult.
(Butany et al.,1966; Bhatt et al., 1967; Singh et al., 1968 and Singh et al., 1979).
The improvement of genetic make-up of cotton plant for assured yield contributing characters are
boll number, boll weight, and ginning outturn. Extensive genetical investigations of the above three
important yield contributing characters have been carried out by several workers. The findings reported
by them have been summarised in Table 2.
Amount of lint obtained from the seed cotton and staple length are the important criteria that
determine the genetic potential of a cultivar. These two components are correlated with the density i.e.
development of fibre per unit area on seed coat. The ginning outturn is another important component of
lint yield but it is again a polygenic character.
The ginning outturn of arboreum cottons in India ranges from 22 to 52 per cent. Comilla cotton
belonging to race cernuum ginns upto 49 per cent or even more (Anonymus, 1956 a; Barooch and De,
1950). The race bengalense and the race indicum types occupy the second and third position,
respectively. In bengalense cottons those possessing white flowers and narrow central leaf lobes like
Roseum, N. Roseum 6 and 231 Rosea, shows higher ginning (36 per cent to 43 per cent). Therefore,
cernuum and Roseum cottons have been used in hybridization programme, wherever necessary, for
improving the ginning character of arboreum cottons of different tracts . The variability potential in
Gossypium arboreum race cernuum offers immense possibilities in breeding for improvement in boll
weight, fibre strength and ginning outturn of desi cotton.
LOCULE RETENTIVITY
Shedding of seed cotton locules from the fully opened mature bolls is a serious problem in arboreum
cottons. (Singh and Punit Mohan 2004). Most of the released varieties of this species suffer from locule
shedding after boll bursting. As a result the seed cotton loaded locules falls on the ground, get mixed with
leaf bits and soil particles resulting in the deterioration of the cotton quality. This species warrants
immediate picking after boll bursting. The high locule retentivity was observed in race cernuum than race
bengalense and indicum (Singh and Punit Mohan 2004). Locule retentive cultivars help in reducing the
pre-harvest seed cotton loss due to adverse weather in the form of high wind, rain and hailstorm.
Therefore, race cernuum can be utilized in the future breeding programmes for the development of locule
retentive cultivars of Arboreum.
DISEASE RESISTANT
The Grey mildew disease of cotton especially G. arboreum caused by the fungus Ramularia areola Atk
(Ramularia gossypii) has been reported as a disease of great economic importance in India and other
cotton-growing areas in the world. In all 1592 accessions of Gossypium arboreum were screened against
the grey mildew disease (Ramularia areola, synonym Ramularia gossypii), six germplasm lines resistant
High Boll Weight and High Ginning OutturnThe Major Tools for Breaking Yield Barriers in Gossypium arboretum 97
to grey mildew disease have been identified namely G 135-49, 30805, 30814, 30826, 30838 and 30856,
belonging to G. arboreum race cernuum and can be utilized in development if varieties/ donor lines
resistant / tolerant to grey mildew disease.
Morpho- anatomical investigations of resistant lines were carried out which indicates host immunity
and greater variability in anatomical features viz., thick smooth cuticle, interlocked epidermal cells, high
degree of lamina thickness in comparison to commercial susceptible G. arboreum cultivars viz., AKH 4
and AKA 8401 (Punit Mohan et al., 1997). The variability in foliar anatomical features have been
summerised in Table 4. The race cernuum is an ecotype representing the final product of localised
selection tendencies among perennial cottons in North-East India as reported by Simlote (1956).
The results presented in Table 1 shows that significant variability for boll weight, ginning percentage,
fibre properties and their extent of association with several economic traits have been observed.
However, their mode of inheritance need to be studied as it will help in better selection of parental
genotypes and their effective manipulation in breeding programme.
TABLE 1: HIGH GINNING OUTTURN AND HIGH BOLL WEIGHT GERMPLASM ACCESSIONS OF GOSSYPIUM ARBOREUM
Sr.
No.
Name Boll
wt. (g)
Bursted
Locule
Length
(cm)
Ginning
Outturn
(%)
2.5%
Staple
Length
(mm)
Uniformiy
Ratio (%)
Fineness
Micronaire
10
-6
g/in
Bundle
Strength
Tenacity
(g/tex) at
3.2 mm Gauge
Elongation
(%)
1. GHA-1 6.5 14.0 40.1 18.9 52 7.9 15.3 5.7
2. GHA-2 6.3 14.0 40.0 18.2 51 8.0 14.8 5.4
3. GHA-3 7.2 13.5 42.3 18.6 51 8.0 14.8 4.9
4. GHA-4 6.8 12.3 38.5 19.0 51 8.0 15.4 5.1
5. GHA-5 6.8 13.2 40.2 17.9 52 8.0 14.6 4.9
6. GHA-6 7.5 13.5 48.5 19.4 52 8.0 15.4 4.8
7. GHA-7 7.0 14.5 39.5 18.0 51 8.0 14.7 5.3
8. GHA-8 7.2 14.0 39.0 19.6 51 8.0 15.1 5.1
9. GHA-9 6.8 13.3 40.1 19.9 52 7.5 14.8 4.6
10. GHA-10 6.0 13.0 42.0 18.1 54 8.0 14.4 5.3
11. GHA-11 7.5 14.2 41.0 18.9 51 7.9 15.3 5.2
12. GHA-12 7.3 15.0 48.0 18.2 53 8.0 14.4 5.2
13. GHA-13 6.9 14.2 39.5 18.6 53 8.0 14.8 5.2
14. GHA-14 6.0 14.0 40.2 18.2 53 7.9 14.8 5.2
15. GHA-15 5.8 12.4 38.0 18.9 55 7.5 14.8 5.0
16. GHA-16 6.9 13.7 38.3 17.6 51 7.8 14.8 5.5
17. GHA-17 6.3 13.7 44.0 18.0 52 7.5 15.2 5.3
18. GHA-18 7.3 15.3 40.1 17.8 51 7.9 15.0 5.4
19. GHA-19 7.0 14.0 49.3 17.6 52 7.8 15.4 5.5
20. GHA-20 6.3 13.2 40.3 18.5 51 7.3 16.0 5.0
21. GHA-21 5.5 12.8 38.7 18.0 51 7.5 15.4 5.5
22. GHA-22 5.0 13.3 38.0 17.6 52 7.8 14.5 5.1
23. GHA-23 7.3 14.9 40.5 17.9 51 7.8 14.9 5.3
24. G 135-49 5.9 13.0 39.6 21.8 50 7.5 19.7 5.0
25. 30805 6.4 13.0 43.1 20.2 50 7.9 17.9 4.4
26. 30814 6.1 13.0 48.7 17.5 52 7.3 16.4 5.0
27. 30826 7.3 15.3 51.0 18.5 49 7.0 17.0 5.1
28. 30838 7.1 15.5 48.1 20.3 50 7.5 16.4 4.9
29. 30856 6.3 14.3 46.1 17.0 48 7.5 16.0 5.1
Range 5.0-7.5 12.3-15.5 38.0-51.0 17.0-21.8 48-55 7.0-8.0 14.4-19.7 4.4-5.7

TABLE 2: GENE ACTION AND TRAITS
Character Gene Action References
Boll diameter and boll shape Polygenic Control Patel and Patel (1927)
Boll shape and size Multiple gene
Control
Balls (1908), Kearney (1923) Kokuev (1935)
Boll size Multiple gene
control
Ramiah and Bholanath (1947)
Length and breadth of bolls Multiple gene
Control
Afzal (1930)
Variability and correlations for boll
attributes
Multiple gene
control
Singh (1988)
Ginning Outturn Multiple gene
Control
Mc Lendon (1912), Kottur (1923),Patel and Patel
(1927)OKelly and Hull (1930)
TABLE 3: GENETIC SOURCES FOR HIGH GINNING OUTTURN AND HIGH BOLL WEIGHT (GOSSYPIUM ARBOREUM)
1. Khasia Hills Cotton G. Arboreum Race
Cernuum
White Petals, Ginning Outturn 50 %, Fibre Length 18-22mm, Boll
Weight 6 to 7 g.
2. Jaintia Hills Cotton
G. arboreum race
cernuum
Yellow petals, ginning outturn 52 % coarsests cotton of Assam, Lint
white creamy colour.
3.
Haflong Hills
Cotton
G. arboreum race
bengalense
Vernacular name- Khum-Mah, Khum-sab, patsen-grows between
1500 to 2000 feet above sea level. All these are short linted, fairly
soft cotton of creamy, white colour with ginning percentage of 37-
40%.
4. Lushia Hills Cotton
G. arboreum race
bengalense
Consists of three local varieties -Lafang: yellow flower, short lint,
inning outtrurn 37%, halo length -19mm.Lapui- white flower and
coarser, ginning outturn 37% and halo length -22mm.Lauk- Yellow
flower, short linted, rusty brown colour.
5. Mikar Hills Cotton
G. arboreum race
cernuum
Two local known varieties viz. Borkapa and Sorukapa- yellow
flower.
6. Naga Hills Cotton
G. arboreum race
cernuum
White flower, Ginning outturn 40-45 %, coarse fibre and boll weight
5 to 6 g.
7. Abor Hill Cotton
G. arboreum race
cernuum
Yellow flowered, fairly fine, short stapled, maximum halo length 20-
24 mm., monopodial habit, ginning outturn 35-40 % and boll weight
6 g.
TABLE 4: FOLIAR ANATOMICAL FEATURES OF IMMUNE AND SUSCEPTIBLE TREE COTTON (GOSSYPIUM ARBOREUM L.) IN RELATION TO GREY MILDEW DISEASE (RAMULARIA AREOLA ATK)
Germplasm
Accession
Cuticle
Thickness
() *
Lamina
Thickness
()*
Thickness
Covered by
Palisade
Parenchyma() *
Thickness
Covered by
Spongy
Parenchyma() *
No. of
Epidermal
Cell *
No. of
Stomata*
Bangladesh 5.166 115.62 46.90 39.52 19.31 71.19
G 135-49 4.936 113.16 51.82 39.52 19.23 67.50
30805 4.166 113.16 41.98 39.52 19.45 69.31
30814 4.920 113.16 41.98 39.52 19.38 69.28
30826 4.920 113.16 41.98 39.52 19.55 71.53
30838 4.920 110.70 41.98 39.52 21.74 71.46
30856 3.690 113.16 41.98 39.52 19.55 71.81
AKH 4 2.460 108.24 37.06 39.52 14.77 89.44
G 27 2.460 105.75 37.48 39.52 17.53 103.35
Range 3.690 110.70 41.98 39.52 19.23 67.50
(immune) 5.166 115.62 51.82 39.52 21.74 71.81
Range(susceptible) 2.460- 2.460 105.75-108.24 37.06-37.48 39.52-39.52 14.77-17.53 89.44-
103.35
*per microscopic field
REFERENCES
[1] Afzal, M. (1930) - Studies in inheritance of Cotton- Mem. Dept. Agric India., Bot., 17 : 75-115.
[2] Aher, R.P., Sanap, M. M. and Thete, R,Y.(1989) - Genetic parameters and correlation coefficient studies in Deshi Varieties
of Cotton - J. Maharashtra Agri. Univ., 14 (1), 63-64.
[3] Anonymous, (1956) A guide to Indian cottons (Revised) - I.C.C.C., Bombay-74.
High Boll Weight and High Ginning OutturnThe Major Tools for Breaking Yield Barriers in Gossypium arboretum 99
[4] Balls, W. L. (1908) Mendelian Studies of Egyptian Cotton. J. agric. Sci, 2: 346- 379.
[5] Barooch, S. R. and II, B., (1950) Hill Cotton in Assam - I.C.G.R.,4 : 65- 69.
[6] Bhatt, M. G., Singh, R. B. And Mor, B. R. (1967) - Genetic variability in upland cotton (G. hirsutum L.) II Analysis of yield
and its components. Indian J. agric. Sci., 37, 555 - 559.
[7] Butany, W.T., Munshi Singh and Mehra, R. B. (1966) - Interrelationships between some characters in hirsutum Cotton.
Indian J. Genet. and Pl. Breeding, 26,262-268.
[8] Duhoon, S. S. (1989) - variability, correlations and path analysis of nine characters in Gossypium arboreum Cotton. J.
Indian Soc. Cotton. Improv., 14, 39-44.
[9] Kearney, T. H. (1923) - Segregation and Correlation of Characters in an upland Egyptian Cotton hybrid. U.S. Dept. Agric.
Bull., 1164-68.
[10] Kokuev, V. I. (1935) - Inheritance of Certain agronomic and morphological characters in cotton. A Sredoz N. I. Kh. I.
Tashkent, 80: 65-80.
[11] Kottur, G.L. (1923) - Studies on inheritance in cotton .I. History of a Cross between Gossypium herbaceum and
G.neglectum. Mem. Dept.Agric. Indian. Bot., 12: 71-133.
[12] Mc Lendon, C. A. (1912) - Mendelian inheritance in cotton hybrids - Georgia sta. Bull., 99: 141 -228.
[13] OKelly, J.F. and Hull. W. W. (1930) Cotton inheritance studies. Lint percentage -Tech. Bull. Miss, Agric. Expt. Sta.,
18:15.
[14] Patel, M. L. And Patel, S.J. (1927) Studies in Gujarat Cottons. Part IV Hybrids between Broach Desi and Goghari
varieties of Gossypium herbaceum - Mem. Dept Agri. Indian Bot., 14: 131-176.
[15] Punit Mohan, Bhat. M.G., Singh, V. V. and Singh, P (1992) - variability for biomass and harvest index in Asiatic
(G.arboreum L.) and American (G. hirsutum L.) cottons. Adv. Plant Sci. 5: 100 105.
[16] Punit Mohan., Mukewar, P.M., Sheo Raj and Singh, V.V. (1997) - Anatomy of Gossypuim arboreum lines immune to grey
mildew disease, - J. Cotton Res. & Dev. 11: 191-195.
[17] Punit Mohan., Mukewar, P.M., Singh, V.V. and Kairon, M. S. (2000), - Perpetual immunity of arboreum cotton to grey
mildew. - SAIC, News Letter, 10:8.
[18] Ramiah, K and Bholanath (1947) Studies on the cotton boll with special reference to G. arboreum - 3rd - Conf. Cotton.
Gr. Prop. India. I.C.C.C., Bombay, 100-106.
[19] Shinde, V. K. and Deshmukh, M. D. (1985)-Genetic variability for yield and characters association in desi cotton - J.
Maharashtra Agri. Univ, 10, 21-22.
[20] Simlote K.M. (1956) Cotton improvement in India - Simlote Institute of Plant Industry, Indore, 201 P.
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characters in hirsutum cotton - Indian J. Genet. and Pl. Breeding ., 28(2), 216-222.
[22] Singh, B. N., Singh, H. G. and Singh, U.P. (1979) - Path analysis of yield and fibre components in upland Cotton Indian J.
agric. Sci., 49,7 63-765.
[23] Singh, Munshi and Raut, R.N. (1983) Genetic research on cotton and Jute In: Genetical Research in India, 1983, pp.154
171, Pal B.P.(Ed) ICAR, New Delhi.
[24] Singh, P. and Nandeshwar, S.B.(1983) Variability in Gossypium arboreum Linn. Race Cernuum in Garo Hills of India.
Indian J. agric. Sci., 53(7) : 511-13.
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Improv. 30: 75-84.
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cotton J. Indian Soc. Cotton Improv. 24:155-163.
Development of Naturally Coloured Gossypium

hirsutum Cotton Genotypes Suitable for Textile
Industry through Genetic Improvement
Manjula S. Maralappanavar, Vikas V. Kulkarni, Somshekhar, C. Madhura, S.S. Patil,
K. Narayanan, K.J. Sanapapamma and Jyoti V. Vastrad
Agriculture Research Station, UAS, Dharwad580007, Karanataka, India
AbstractBroad use of natural coloured cottons is not effective yet due to their lower fiber qualities in comparison to
conventional white cottons and limited range of natural colours. The low yielding, short fibered coloured cottons were
not suitable for machine spinning; therefore they failed to face the rapid industrial turnover. Off late attention is
diverted to study the possibility for using these eco-cottons commercially but few systematic studies and reports are
available on the breeding programmes and meagre information on the fiber quality of naturally colored cottons. We
report breeding efforts made to improve the naturally coloured G. hirsutum cotton genotypes with respect to yield and
fiber quality through intra-specific, inter-specific and three way crosses between coloured and superior white genotypes
followed by selections independently in three populations. Stable genotypes with uniform colour and high yield
potential and improved fiber qualities were developed. Eight of the 32 advanced coloured genotypes in three
populations tested under three replicated trials were superior to white linted check, Sahana (2138 kg/ha) for seed cotton
yield. Three of these, dark brown (DDB-12 with 2986 kg/ha seed cotton yield), medium brown (DMB-225 with 2934
kg/ha) and green (DGC-78 with 1381 kg/ha) were potential for seed cotton yield and quality in the respective colour.
The genotype DDB-12 had 21.6 mm fiber length and 18.6 g/tex strength; DMB-225(medium brown) had 22.9mm and
20.4 g/tex while green linted DCG-78 had 25.8mm and 22.2g/tex, span length and strength respectively. These
genotypes performed consistently over three years of testing for yield and fiber quality. Yarn and fabrics were
manufactured which were suitable for mill spinning and eco-fabrics of commercially acceptable range could be
produced.
The simultaneous development of medium, dark brown and green shades along with white cotton will help in the
creation of variability in the textile industry. Efforts need to be done to further improve the fibre properties which are
being done using the stable color cotton lines developed in the present study for crossing with superior white cultivars
for introgression and selection. Biotechnological approaches have been initiated to diversify the lint colour.
INTRODUCTION
The vast majority of cotton grown commercially in the world has white lint. However, there are
genotypes / species which produce naturally coloured cotton and most of the wild species of cotton have
coloured lint or fuzz. Though historical evidence like the fossils obtained from the excavations at Huca
Preita in Northern coastal Peru indicated the usage and cultivation of colour cottons with lint colour from
tan to red shades before 2500 B.C., only some of which exist today. It seems others have been lost, as
they have never been described in the botanical literature. The ability of the white cotton to take up any
colour to produce a large range of shades and colours in fabrics has lead to the popularization of white
cotton.
In the recent past the thinking over of the other side of industrial revolution has made man realize
that there is an urgent need to protect the environment. This eco- awareness has led to the revival of
colour linted cottons, which can do away the processes of dyeing and processing. These colour cottons
have once again crawled out with the efforts of an American Scientist, Sally Fox who began to improve
colour cotton since 1982 and has developed naturally Colour Cotton Corporation that markets colour
cotton fabrics (Fox, 1987). Naturally pigmented cottons not only are economical but more ecofriendly
as they can avoid the use of carcinogenic dye chemicals in the fabric and also minimize the affluent of
the dyeing industries, which pollute the land and water resources. Previous studies along with the
advantage of inherent colour have also shown the flame resistance of brown cotton (Kimmel and Day,
2001; Williams, 1994) and colour change (darkening instead of fading) occurring with certain laundering
18
Development of Naturally Coloured Gossypium hirsutum Cotton Genotypes Suitable for Textile Industry 101
methods (Oktem et al, 2003; Vanzandt, 1994). It is also reported that naturally coloured cottons bring
medical remedy for over 50 different somatic and psychosomatic disorders of man (Vreerland, 1993).
The study demonstrates that naturally pigmented cottons have excellent sun - protection properties (high
UV protection factor (UPF values), which are far superior to conventional bleached or unbleached cotton
(Gwendolyn and Patricia, 2005) (green cotton UPF = 30 to 50 +; tan UPF= 20 to 45; brown UPF = 40 to
50+; bleached conventional UPF = 4; unbleached conventional UPF = 8).
On one hand, the knowledge of all these desirable features of the naturally pigmented cottons, there
is no second thought for the adaptation of these cottons; on the other hand history shows that though they
co-existed with white cotton they were left behind in the mid way. The low yielding, short fibered
coloured cottons were not suitable for machine spinning; therefore they failed to face the rapid industrial
turnover. Off late attention is diverted to study the possibility for using these eco-cottons commercially
but few systematic studies and reports are available on the breeding programmes and still meagre
information on the fiber quality of naturally coloured cottons(wich and Frydrych 1998,1999; wich
et al., 1999).
We present a complete study done at UAS, Dharwad encompassing breeding to improve the naturally
coloured G. hirsutum cotton with respect to yield and fiber quality to make these cottons commercially
viable to both the farmer and textile industry. Among the four cultivated species of cotton, colour lint is
reported in G. arboreum among diploid and in G. hirsutum in tetraploid cottons. White G. barbadense
that are inherently superior sources of fibre quality and white G. hirsutum cottons are a source for both
yield and fibre quality have been used for improvement of color cotton.
The dark brown selection DDBS-98, white released varieties, Abadhitha of G. hirsutum and Suvin of G.
barbadense with characters as given Table 1 were used in the crossing programme to develop 3
populations as detailed in figure 1. The green genotype used in the crossing was very low yielding and
had unstable colour expression.
In all the selection cycles from 200001 to 200506 individual plants were selected in field after
peak boll bursting based on morphological observation on lint color, yield per plant and fibre length.
Individual plants were sown in plant to progeny rows and again individual plants were selected. In
advanced generations, during 2006 07 progeny row selections were effected.
Population I consisted of eighteen colour cotton genotypes from the cross DDBS 98 (Dark brown
Hirsutum) x Abadhita (white Hirsutum) and one green from (Green Hirsutum x Abadhitha); population
II, had ten genotypes from DDBS-98 (Dark brown Hirsutum) x Suvin (white Barbadense) and
population III had seven genotypes from three way crosses [three selections from [(colour Hirsutum x
white Hirsutum) x White barbadense] and 4 selections from [(colour Hirsutum x white Barbadense) x
White hirsutum]. These stable genotypes were sown under three independent trials along with colour
parent DDBS 98 and white commercial check, Sahana. The trials were laid in three replications with
two rows per genotype and spacing of 90 x 20 cm. under rainfed condition at ARS, Dharwad Farm.
Recommended agro-management were followed.
The seed cotton yield was analyzed in RBD trial. Observations on boll weight were taken as average
of twenty bolls per genotype. The ginning out turn (GOT) was calculated (using 300g seed cotton yield to
get seed and lint weight) using the standard formula , GOT =[(Lint weight / Seed cotton weight) x 100]
and seed index (SI) was weight of 100 seeds and lint index (LI) was calculated using the formula, LI=
[(SI x GOT) / (100 GOT)].
The fibre quality was analyzed at Central Institute of Research on Cotton Technology (CIRCOT),
Regional laboratory at Dharwad, using HVI machine.

The genotype, DMB225 which had significantly higher yield based on progeny row results of the
year 2006-07 was grown on an area of one acre under isolation in 2007-08 with a spacing of 90 x 20cm
following the same package of practices as in commercial white hirsutum varieties and the lint was tested
for its suitability for commercial yarn and fabric production.
Population I was developed from intra-specific crosses, among the eighteen improved brown linted
genotypes and one green genotype, DG-78 were tested along with white variety Sahana and dark brown
parent DDSG-98. All the test entries were superior for both yield and fiber properties compared to
original dark brown parent DDSG-98. Also some of the genotypes were numerically superior to even the
white commercial cultivar Sahana for seed cotton yield and boll weight, but inferior with respect to lint
index and GOT. Three genotypes DDB-12 (2986 kg/ha) with dark brown, DMB 225 (2934 kg/ha) with
medium brown and DDB-210 (2834 kg/ha) with dark brown color were significantly superior for seed
cotton yield than Sahana (2138 kg/ha) (Table 2 and Fig. 2).
Most of the selected genotypes were better for fibre quality than original parent DDBS-98 but were
not on par with white check Sahana. Among the high yielding genotypes, medium brown coloured
selection DMB-225 had better fiber length of 22.9 span length (2.5 %) and 20.4 g/tex strength and
micronaire of 4.0 (Table 3). As it was very difficult to get stable green colour plants, in each generation
individual plants selected on basis of uniform green shade and fiber length were bulked and advanced to
next generation to repeat the same. The green genotype DG-78 thus developed was highly stable for
green colour although its yield was not comparable to improved brown selections. It had good fiber
quality of 25.8mm span length and 22.2 g/tex strength (Table 3).
In the population II, consisting of stable lines from inter-specific cross between dark brown G.
hirsutum and white G. barbadense, among ten genotypes only one cream coloured selection, DCR-110
was numerically superior to white check, (Sahana) for seed cotton yield, lint index and GOT and even on
par for fiber quality. Most of these selections were of medium brown lint. The genotypes DMB 105 had
very good fiber properties as 26.2mm 2.5% span length and 21.5 g/tex tenacity (Table 4 and 5).
In the third population derived from three way crosses using both white hirsutum and barbadense,
only 7 selections could be advanced to replicated trial as most of them had very light colour. Though
none of them was better than Sahana but were better than coloured parent. The highest yielding
genotypes among colour selections in this population were dark brown, DDB-5 which had 22.8mm span
length and one more medium brown type DMB-3 had the highest strength of 20.6g/tex. The cream
colored genotypes DC-12 and DC-5 had fiber length of 24mm (Table 6 and 7).
Across all the populations using different approaches, intra-specific, inter-specific and three way
cross followed by selections, it was observed that the segregants in inter-specific approach using
G.barbadense had very good fiber quality in the early generation but as selections were made for darker
shades in the advanced generations the fiber quality and the yield were reduced and those selections with
good quality had light shades. In studies with white G. hirsutum cotton improvement by utilizing the G.
barbadense for superior fiber quality similar observations of the reduction in fiber quality in the
selections of advanced generations have been observed. However in the intra-specific population using
G. hirsutum, there was significant increase in the yield and thus through this approach it was possible to
improve seed cotton yield and boll weight along with moderate improvement of fiber quality parameters.
The color in fiber lint is governed by major gene, multiple alleles and modifiers (Khan et al., 2009 and
Waghmare and Koranne., 2000). and the expression of colour depends on the modifiers which may be
present in the same species of G.hirsutum. It is observed that G. hirsutum colour genotypes can be
improved by intra-specific hybridization followed by selection approach that is by using the white
superior G.hirsutum genotypes. Once genotypes with dark brown colour with high yield and medium/
light brown genotypes with good fiber quality parameters are available in the G.hirsutum species, these
can be intermated to combine color, yield and fiber quality into single genotype.
Development of Naturally Coloured Gossypium hirsutum Cotton Genotypes Suitable for Textile Industry 103
The result indicated the progress in the improvement of naturally coloured cottons to make them
commercially viable. Among all the genotypes put together, DDB-12 (dark brown), DMB 225
(medium brown) and DCR 110 (cream) were best in that shade of brown. The green genotype DCG-78
had good fiber traits. The genotypes, DDB-12, DMB-225 and DCG-78 were grown in larger plots over
three years and tested for their productivity and suitability for yarn and fabric manufacture. Dark and
medium brown genotypes were found to be on par with white linted variety, Sahana with respect to yield
and suitable for various fabric productions (Fig.2 & 3). The green genotype DCG 78 though had
comparatively less yield potential but it necessiated to promote this genotype as it had superior fiber
quality traits. The simultaneous development of medium and dark brown and green shades along with
white cotton will help in the creation of variability required for the textile industry. These three genotypes
were presented in the Zonal Research and Extension Committee Meeting of the university during 2009-
10 and have been approved for commercial cultivation of these genotypes under contract farming under
the technical supervision of the university (Proceedings of Rabi/Summer 2010-11).
Efforts need to be done to further improve the fibre properties which are being done using the stable
colour cotton lines developed in the present study for crossing with superior white cultivars for
introgression and selection. Also there is a necessity to diversify the lint colour which is attempted
through biotechnological approaches. The Flavonoid 3-5 hydroxylase gene from petunia and
snapdragon which is the enzyme responsible for turning the pathway of pigmentation towards blue
shades has been cloned. The transformation of these genes in the improved genotypes is initiated. The
expression of these genes in lint may help in creation of a shade not naturally available.
REFERENCES
[1] Apodaca, J.K., (1990). Naturally coloured cotton: A new niche in the Texas Natural Fibers. BBR Working Paper Series
1990 2 Bureau of Business Research, University of Texas at Austin.
[2] Fox, S. (1987). Naturally Coloured Cotton: Spin-off. December, 48-50.
[3] Gwendolyn, H. and Patricia, C. C. (2005). The ultraviolet protection factor of naturally pigmented cotton. The Journal of
Cotton Science, 9, 47-55.
[4] Khan A. A., Azhar F. M., Khan I. A., Riaz A. H. and Athar M. (2009). Genetic basis of variation for lint colour, yield and
quality in cotton (Gossypium hirsutum L.) Plant Biosystems. (143) pp 517-524.
[5] Kimmel, L.B. and Day, M.P., (2001).New life for an old fiber: attributes and advantages of naturally coloured cotton.
American Association of Textiles and Colour Cotton Review 1(10), 32-36.
[6] Oktem, T.A., Gurel and Akdemir, H., (2003). The characteristic attributes and performance of naturally coloured cotton.
American Association of Textiles and Colour Cotton Review, 3(5), 24-27.
[7] Proceedings of Rabi/Summer 2010-11. Combined Zonal Research and Extension Advisory Council and Zonal Research and
Extension Formulation Comitee Meeting (Zone- I & II, III & IV), DATED 02-04 September, 2010, UAS, Dharwad.
[8] Stephens, S.G., (1975). A re-examination of the cotton remains from Huaca Prieta, NorthCoastal Peru. American Antiquity
40(4), 406-418.
[9] wich T. and Frydrych I., (1998). Naturally colored cotton as an element of humanoecology, Architektura Tekstyliw (1):
20-22.
[10] wich T. and Frydrych I., (1999). Naturally colored cottons: Properties of fibres and yarns. Fibres andTextiles in Eastern
Europe, (4), 25-29.
[11] wich T., Frydrych I. and Balcar G., (1999).The assessment of naturally colored cotton properties. Bulletin of Gdynia
Cotton Association, (3), 12-26.
[12] Vanzandt, M.J., (1994). Development of fabric from fox fibre naturally coloured cotton and evaluation of flame-resistant
characteristics. Ph.D. Dissertation, Texas Technical University, Lubbock, TX.
[13] Vreerland, J.M., (1993). Naturally coloured and organically grown cottons.Anthropological and Historical perspective.
Proceedings of Beltwide Cotton Conferences, 1533-1536.
[14] Williams, B., Fox (1994). Fibre naturally coloured cotton, green and brown (coyote) resistance to changes in colour. Ph.D.
Dissertation, Texas Technical University, Lubbock, TX.
[15] Waghmare V. N., Koranne K. D. (2000). Inheritance of lint colour in Desi cotton (Gossypium arboreum L.) Indian Journal
of Genetics and Plant Breeding (60).
Divergent Selection for Yield and Earliness

in Cotton (Gossypium hirsutum L)
P. Michalakopoulos, C. Goulas, A. Katsiotis and S. Rangasamy
Organization of Agricultural Vocational Education Training and Empl,
Agricultural University of Athens, Greece
INTRODUCTION
Knowledge of the nature and the size of genetic influences on the breeding material are necessary so that
a breeding procedure could be decided upon, as the genetic behavior of each trait may vary, depending on
the genetic material and the environment. Therefore, the gene action of quantitative traits must be studied
before the beginning of a breeding program.
The existence of genetic variability and the definition of its nature (additive, non-additive, etc.) are
necessary for the decision upon the most effective breeding methodology, regarding the creation of the
desired variety. The first and, by far, simplest approach is that of divergent selection (high vs low yield or
desired vs undesired genotypes, in general). Its planning permits information retrieval regarding the
existence of genetic variability and the heredity mode of the specific trait. Furthermore, it provides
information on the response to the selection as well as on indirect changes in traits indirectly selected
(Falconer and Mackay, 1996). Divergent selection has been mainly implemented in corn (Chesang-
Chumo, 1993; Masole, 1993; Martin et al., 2004; Pressoir and Berthaud, 2004; Chimenti et al., 2006;
Uribelarrea et al., 2007; Hallauer et al., 2010) and in other crops, such as in wheat (Hucl, 1995; Al
Hakimi et al., 1998), alfalfa (Chloupek, 1999; Lamb et al., 1999; Klos and Brummer, 2000) and cotton
(Verhalen et al., 1975; Radin et al., 1994; Ulloa et al., 2000). The objectives of this study were: (i) to
determine the effect of divergent selection for yield and earliness in diverse segregating populations
derived from six crosses and (ii) to examine the potential use of specific genetic material in planning and
implementing a breeding program for yield and its components and earliness of upland cotton.
The genetic material used consisted of seven commercial varieties; five cultivated in Greece and two in
India. The experiments were conducted during five cultivation periods. In year 2003, crosses were
performed using as a common parent one of the varieties cultivated in Greece. During year 2004, the F
1

hybrids were backcrossed to their parental progenitors.
During the 2005 cultivation period, divergent selection in the F
2
generation and for all six crosses
was performed for yield and earliness (Falconer, 1960). One hundred individual plants, using a grid mass
selection scheme, were analyzed (10 grids and 10 plants per grid) as described (Gardner, 1961). The
distance among the lines was 97cm and among the plants on the line 25cm, meaning four plants per
square-meter. The parents of each cross were alternately sown as testers on the two lines of each grid.
The earliness and the yield in seed cotton were registered for every individual plant (F
2
and the respective
P
1
and P
2
parents). For earliness, the day-span from the sowing until the opening of the first ball (DBO)
was used. The yield of, the seed cotton (weight) was expressed in grams.
In each grid of the ten F
2
plants per cross divergent selection was performed (selection ratio 10%) for
the following traits; a) one high-yielding plant (HY) and one low-yielding plant (LY), b) one earlier plant
(HE) with the later one (LE), c) one earlier plant among those yielding more than 85% of the average of
witnesses (YHE) and the later one among those yielding more than 85% of the average of the witnesses
(YLE). Thus, balanced mixtures of the ten selected plants, HY-LY, HE-LE, YHE-YLE, were created for
each one of the six crosses.
19
Divergent Selection for Yield and Earliness in Cotton (Gossypium hirsutum L) 105
The six (6) F
3
selected populations (HY-LY, HE-LE, YHE-YLE) of each cross (36, in total) were
analyzed during the 2006 cultivation period. The experimental design used was the Randomized
Complete Blocks (RCB) with four replications. An experimental block of one, five-meter long, line was
used. The distance among the lines was 97cm, while the final density after sparsing was 12 to 14 plants
per meter. Earliness (DBO) and seed yield were registered. Random F
4
seed sample was taken from each
selected population and it was used for analysis during the 2007 cultivation period, according to the same
procedure described for the F
3
selected populations.
STATISTIC ANALYSIS
For each one of the six crosses the variance of one hundred plants was estimated for all the traits,
estimating the total phenotypic variance as
p
2
=
g
2

+
e
2
. The variance of the thirty plants per parent (P
1

and P
2
) was also estimated. The average variance of the plants of P
1
and P
2
parents (30+30) was an
estimation of
e
2
. Criterion F was made for the evaluation of variances P
1
and P
2
, as well as for F
2
vs. P
1
,
P
2
and (P
1
+P
2
)/2. Therefore, estimation of
p
2
=
g
2

+
e
2
and of H
2
heredity coefficient was made
feasible. Furthermore, the genetic variability coefficient (GCV) and the selection differential for
divergent selection (high-low), and the expected selection progress R = S x H
2
for yield and earliness
were estimated.
The data concerning the selection effectiveness were analyzed with the conventional RCB Anova
(Steel and Torrie, 1980). The distinction of average rates was made with the least important difference
(Fishers protected LSD) for every trait. The statistic model was fixed and made possible the estimation
of the genetic variance
g
2
among the six genotypes according to the mean rate of each genotype (plot
mean basis).
In year 2005, F
2
yield was practically around parental mean rate, as expected from a qualitative trait like
yield. The genetic variability among F
2
plants ranged from 36.59% in GR
1
x GR
5
to 42.16% in GR
1
x IN
1

in bigger amounts in relation to the genetic variability among parental plants in all crosses. The data of
the divergent selection revealed substantial differentiation between high-yielding and low-yielding plants,
implying indication of existence of genetic variability potential use for effective selection for yield. The
heredity coefficient was satisfactorily high ranging from 0.38 to 0.60 (Table 1).
TABLE 1: DATA OF SELECTION FOR YIELD (G / PLANT) IN DIVERGENT SELECTION IN 2005
Cross Means CV%
G
2
h
2
Selection Differential
Parents F
2
Parents F
2
- +
1. GR
1
x GR
2
111.27 113.29 26.07 25.03 40.04 1242.05 0.60 58 78
118 - 104.53 136
2. GR
1
x GR
3
111.90 118.23 30.32 31.21 40.68 1128.27 0.49 61 88
116.53 - 107.27 149
3. GR
1
x GR
4
105.93 98.81 21.45 36.28 38.56 556.70 0.38 51 61
118.80 - 93.07 112
4. GR
1
x IN
1
106.95 102.29 27.41 29.54 42.16 938.90 0.50 54 83
114.90 - 99 137
5. GR
1
x GR
5
112.03 113.04 27.07 25.32 36.59 846.22 0.49 53 79
113.63 - 110.43 132
6. GR
1
x IN
2
110.23 107.36 27.94 26.27 41.28 1073.40 0.55 57 75
103.03 - 117.43 132
As for earliness, the data indicated the existence of genetic variability and the heredity coefficient
ranging from 0.54 to 0.83, implying indication of effective selection potential. The data of divergent
selection allowed for the differentiation among early and late plants from 14 to 19 days, implying
potential for selection regarding earliness (Table 2).

TABLE 2: DATA OF SELECTION FOR EARLINESS (DBO) IN DIVERGENT SELECTION IN 2005
Cross Means
P
2
(CV%)
G
2
h
2
Selection Differential
Parents F
2
Parents F
2s
- +
1. GR
1
x GR
2
113.5 115.6 2.88 - 2.87 5.1 24.08 0.69 7 9
104.6 - 122.4 16
2. GR
1
x GR
3
114.7 113.6 2.86 - 2.59 4.68 18.52 0.66 7 9
107.4 - 122 16
3. GR
1
x GR
4
106.3 109.9 2.46 - 3.18 5.07 22.00 0.71 5 10
107.1 - 105.4 15
4. GR
1
x IN
1
115.7 114.5 3.97 - 4.10 5.60 24.82 0.60 8 11
108.7 - 122.7 19
5. GR
1
x GR
5
112.7 112.5 3.88 - 2.78 4.90 16.41 0.54 6 8
106.8 - 118.7 14
6. GR
1
x IN
2
106.8 112.1 1.84 - 2.39 5.07 26.90 0.83 6 10
101 - 112.7 16
The combined selection for yield and earliness (Table 3) revealed potential for genotype selection
combining satisfactory yield ranging from 45% to 72% - with respective earliness from five to seven
days compared to the selection for earliness alone, being from eight to eleven days. The combination
between earliness and yield appears to be possible while the combination between yield and maturity
seems to be impossible for the specific genetic material and under the specific circumstances (as shown
in the first cross).
TABLE 3: DATA OF SELECTION FOR YIELD (GR) AND EARLINESS (DBO) IN DIVERGENT SELECTION IN 2005
Cross Means Means Selection Differential
Parents F
2

Yield Earliness Yield Earliness Yield / Early Yield / Late
1. GR
1
x GR
2
114.2 113.5 113.3 115.6 165 109 148/ 121
118 - 110.4 104.6 - 122.4
2. GR
1
x GR
3
118.5 114.7 118.23 113.6 168/ 107 147/ 119
128.7 - 108.3 107.4 - 122
3. GR
1
x GR
4
107.8 106.3 98.81 109.9 150/ 105 133/ 114
116 - 99.7 107.1 - 105.4
4. GR
1
x IN
1
104.5 115.7 102.29 114.5 163/ 107 120/ 120
113.6 - 95.4 108.7 - 122.7
5. GR
1
x GR
5
114.8 112.7 113 112.5 148/ 107 148/ 117
115.6 - 113.9 106.8 - 118.7
6. GR
1
x IN
2
107.8 106.8 107.4 112.1 142/ 107 133/ 117
100.4 - 115.1 101 - 112.7
SELECTION EFFECTIVENESS
Yield
The productive behaviour (yield) of the F
3
genotypes and the check for the year 2006 is shown in Table 4,
whereas variance analysis and estimation of the genetic parameters for yield during the same year are
shown in Tables 5 and 6, respectively. The results indicated that, genotypes differed in five out of the six
crosses on 99% significance level and on 90% in the sixth cross. The CV% variance coefficient ranged
from 2.78 to 8.07% while the heredity coefficient gave values of 0.25 for the GR
1
x GR
2
and higher (0.41
to 0.57) for the rest of the crosses. Finally, the genetic variance coefficient ranged on low levels from
3.17 to 6.59 %.
The genotypes which had been selected as high-yielding (HY) in year 2005, were differentiated from
the low-yielding ones (LY) in the GR
1
x GR
2
, GR
1
x GR
5
and GR
1
x IN
2
crosses (Table 4). The
genotypes combining high-yield and earliness (YHE) were differentiated from the ones combining high-
yield and maturity (YLE) in two crosses; in the GR
1
x GR
4
cross, where the late genotype was the
highest-yielding one, as expected, and in the GR
1
x GR
4
cross, with the opposite result, however; the
earlier genotype was higher-yielding than the late one. As far as earliness alone is concerned, the earlier
genotypes (HE) were differentiated from the later ones (LE) in two crosses (GR
1
x GR
3
and GR
1
x GR
4
),
with the latter out-yielding the first ones, as expected.
TABLE 4: MEANS FOR YIELD (G PLOT
-1
) OF F
3
GENOTYPES IN 2006
Cross
Source GR
1
x GR
2
GR
1
x GR
3
GR
1
x GR
4
GR
1
x IN
1
GR
1
x GR
5
GR
1
x IN
2

HE 2059bc 2110d 1894c 1966abc 2032bc 1963d
HY 2158ab 2456a 2226ab 1868bcd 2179a 2188a
LE 2060bc 2443a 2166b 2137a 1999c 1996cd
LY 1946c 2374ab 2159b 1835cd 2010bc 2066bc
YHE 2329a 2362abc 2176b 1748cd 2178a 2151a
YLE 2200ab 2188cd 2336a 1757cd 1974c 2109ab
Check 2051bc 2198bcd 2223ab 2056ab 2029bc 2109ab
LSD 208 186 156 221 109 84
The behaviour of the F
4
genotypes and the check, variance analysis and the genetic parameters,
respectively, as for yield in the subsequent 2007 year, are presented (Table 7, 8 and 9). The genotypes
differed in all six crosses on 99% significance level. The CV % variance coefficient ranged from 2.51 to
3.83% while the heredity coefficient gave significantly higher values, ranging from 0.73 to 0.90 for all
crosses. The genetic variance coefficient ranged again on low levels from 4.86 to 8.57%.
TABLE 5: ANALYSIS OF VARIANCE FOR YIELD (G PLOT
-1
) IN 2006
Cross
GR
1
x GR
2
GR
1
x GR
3
GR
1
x GR
4
GR
1
x IN
1
GR
1
x GR
5
GR
1
x IN
2

S
Genotypes.
47208 60635.4 57782.2 84253 23182.8 20622.4
S
Error.
20361 16240.6 11426.2 22974 5578 3323.8
CV% 6.76 5.56 4.91 8.07 3.62 2.78
F
test
+ ** ** ** ** **
**p = 0.01, * p = 0.05, + p = 0.10
TABLE 6: ESTIMATIONS OF GENETIC PARAMETERS FOR YIELD (G PLOT
-1
) IN 2006
Cross
GR
1
x GR
2
GR
1
x GR
3
GR
1
x GR
4
GR
1
x IN
1
GR
1
x GR
5
GR
1
x IN
2

G
2
6711.7 11098.7 11589 15319.7 4401.2 4324.6
P
2
27072.7 27339.3 23015.2 38293.7 9979.2 7648.4
h
2
0.25 0.41 0.50 0.40 0.44 0.57
GCV% 3.88 4.59 4.94 6.59 3.21 3.17
TABLE 7: MEANS FOR YIELD (G PLOT
-1
) OF F
4
GENOTYPES IN 2007
Cross
Source GR
1
x GR
2
GR
1
x GR
3
GR
1
x GR
4
GR
1
x IN
1
GR
1
x GR
5
GR
1
x IN
2

HE 1847c 1748c 2057b 1822abc 1777e 1747cd
HY 2094a 1730cd 1855d 1736c 1948c 1857b
LE 1701d 1696cd 1818d 1899a 1836de 1947a
LY 1828c 1581f 1631e 1911a 1987bc 1826bc
YHE 1804c 2006a 1929c 1779bc 2064ab 1946a
YLE 1847c 1681
de
1842d 1529e 1914cd 1740d
Check 1935b 1868b 2088b 1861ab 1830de 1842b
LSD 79 64 72 98 85 81
Progeny means followed by a different letter are significantly different at the P = 0.05.
TABLE 8: ANALYSIS OF VARIANCE FOR YIELD (G PLOT
-1
) IN 2007
Cross
GR
1
x GR
2
GR
1
x GR
3
GR
1
x GR
4
GR
1
x IN
1
GR
1
x GR
5
GR
1
x IN
2

S
Genotypes.
56491.8 69687.9 111498 75805.9 54687.6 35547.5
S
Error.
2914.2 1927.4 2442 4493.7 3419.3 3055.3
CV% 2.86 2.51 2.57 3.83 3.00 2.98
F
test
** ** ** ** ** **
**p = 0.01, * p = 0.05, + p = 0.10.

TABLE 9: ESTIMATIONS OF GENETIC PARAMETERS FOR YIELD (G PLOT
-1
) IN 2007
Cross
GR
1
x GR
2
GR
1
x GR
3
GR
1
x GR
4
GR
1
x IN
1
GR
1
x GR
5
GR
1
x IN
2

G
2
13394.4 16940.1 27264 17828 12817.1 8123
P
2
16308.6 18867.5 29706 22321.7 16236.4 11178.3
h
2
0.82 0.90 0.92 0.80 0.79 0.73
GCV% 6.13 7.43 8.57 7.62 5.82 4.86
The genotypes which were selected as high-yielding (HY) in year 2005 were differentiated from the
low-yielding ones (LY) in year 2007 in four out of the six crosses. However, only in one of them (GR
1
x
GR
2
) was the equivalent differentiation noticed in year 2006. In the other two crosses (GR1 x GR5 and
GR
1
x IN
2
), with differentiation in year 2006, no difference was noticed for yield in year 2007. The
genotypes combining high yield and earliness (YHE) were differentiated from those combining high
yield and maturity (YLE) in five out of the six crosses (except for the GR
1
x GR
2
). Therefore, the
differentiation noticed in year 2006 in the GR
1
x GR
4
and GR
1
x GR
5
crosses was maintained for the next
generation in year 2007. It is noticeable that the early genotype (YHE) out-yielded the late one (YLE) in
all six crosses. As far as earliness alone is concerned, the early genotypes (HE) were differentiated from
the later ones (LE) in three out of the six crosses (GR
1
x GR
2
, GR
1
x GR
4
and GR
1
x IN
2
), with the
second maintaining the differentiation presented in year 2006. Furthermore, in the first two crosses the
early genotype (HE) out-yielded the later one (LE) while in the third cross, the opposite trend was
observed.
Earliness
The earliness behaviour of the F
3
genotypes and the check, variance analysis and the genetic parameters
were reported in Table 10,11 and 12 for year 2006 respectively. The genotypes differed in five out of the
six crosses on 99% significance level, and on the sixth cross on 90% significance level. The CV%
variance coefficient ranged on very good levels in all crosses from 1.17 to 2.06%, while the heredity
coefficient gave high values from 0.71 for the GR
1
x GR
4
to 0.91 for the GR
1
x IN
2
cross. Finally, the
variance genetic coefficient was on low levels, ranging from 3.21 to 4.86%.
TABLE 10: MEANS FOR EARLINESS (DBO) OF F
3
GENOTYPES IN 2006
Cross
Source GR
1
x GR
2
GR
1
x GR
3
GR
1
x GR
4
GR
1
x IN
1
GR
1
x GR
5
GR
1
x IN
2

HE 105c 104cd 98e 108cd 106de 117a
HY 117a 108b 104bc 109c 114b 111b
LE 120a 114a 109a 114b 118a 111b
LY 108bc 102d 102cd 105d 106de 107c
YHE 107c 106bc 99de 109c 104e 107c
YLE 117a 112a 107ab 118a 115b 106cd
Check 108bc 106bc 102cd 113b 111c 105d
LSD 3 3 3 3 3 2
TABLE 11: ANALYSIS OF VARIANCE FOR EARLINESS (DBO) IN 2006
Cross
GR
1
x GR
2
GR
1
x GR
3
GR
1
x GR
4
GR
1
x IN
1
GR
1
x GR
5
GR
1
x IN
2

S
Genotypes.
121.42 53.76 47.82 60.69 99.75 66.40
S
Error.
5 4.66 4.47 5.07 3.60 1.64
CV% 2.01 2.02 2.06 2.04 1.71 1.17
F
test
+ ** ** ** ** **
**p = 0.01,* p = 0.05, + p = 0.10.

TABLE 12: ESTIMATIONS OF GENETIC PARAMETERS FOR EARLINESS (DBO) IN 2006
Cross
GR
1
x GR
2
GR
1
x GR
3
GR
1
x GR
4
GR
1
x IN
1
GR
1
x GR
5
GR
1
x IN
2

G
2
29.11 12.27 10.84 13.91 24.04 16.19
P
2
34.10 16.94 15.31 18.98 27.64 17.83
h
2
0.85 0.72 0.71 0.73 0.87 0.91
GCV% 4.86 3.28 3.21 3.38 4.43 3.66
TABLE 13: MEANS FOR EARLINESS (DBO) OF F
4
GENOTYPES IN 2007
Cross
Source GR
1
x GR
2
GR
1
x GR
3
GR
1
x GR
4
GR
1
x IN
1
GR
1
x GR
5
GR
1
x IN
2

HE 112bc 110def 106cd 112de 112de 110g
HY 122a 113cd 109bc 114cd 119ab 114cd
LE 122a 117a 113a 120ab 121a 114de
LY 113bc 107g 107bcd 110e 111de 112ef
YHE 113bc 112cde 105d 115cd 110e 112efg
YLE 120a 115ab 110b 121a 118bc 119b
Check 115b 114bc 113 116bc 116c 111fg
LSD 3 2 3 4 2 2
TABLE 14: ANALYSIS OF VARIANCE FOR EARLINESS (DBO) IN 2007
Cross
GR
1
x GR
2
GR
1
x GR
3
GR
1
x GR
4
GR
1
x IN
1
GR
1
x GR
5
GR
1
x IN
2

S
Genotypes.
72.65 33.76 23.11 50.13 71.13 57.57
S
Error.
5.63 2.88 4.78 6.29 2.87 1.72
CV% 2.05 1.52 2.02 2.18 1.47 1.15
F
test
** ** ** ** ** **
**p = 0.01, * p = 0.05, + p = 0.10.
TABLE 15: ESTIMATIONS OF GENETIC PARAMETERS FOR EARLINESS (DBO) IN 2007
Cross
GR
1
x GR
2
GR
1
x GR
3
GR
1
x GR
4
GR
1
x IN
1
GR
1
x GR
5
GR
1
x IN
2

G
2
16.75 7.72 4.58 10.96 17.06 13.96
P
2
22.39 10.60 9.36 17.25 19.93 15.68
h
2
0.75 0.73 0.49 0.64 0.86 0.89
GCV% 3.53 2.48 1.98 2.88 3.58 3.27
The genotypes selected as early (HE) in year 2005 were differentiated from the later ones (LE) in all
six crosses in year 2006, with the early genotypes remaining early and the late ones remaining late. This
differentiation ranged from six days for the GR
1
x IN
2
to fifteen days for the GR
1
x GR
2
cross. The
genotypes combining high yield and earliness (YHE) were differentiated from those combining high
yield and maturity (YLE) in five out of the six crosses, with the first (YHE) being earlier than the second
ones (YLE) from six (GR
1
x GR
3
) to eleven (GR
1
x GR
5
) days. The high- (HY) and low- (LY) yielding
genotypes differed in earliness in five out of the six crosses, with the high-yielding ones being later, as
expected.
The same holds true for the behavior of the seven genotypes (F
4
) for earliness in year 2007 (Table
13). The genotypes differed in all six crosses on 99% significance level while the CV% variance
coefficient was on very good levels in all crosses, ranging from 1.15 to 2.18% (Table 14). The heredity
coefficient gave values from 0.49 for the GR
1
x GR
4
to 0.89 for the GR
1
x IN
2
cross and the variance
genetic coefficient ranged from 1.98 to 3.58% (Table 15).
Early genotypes (HE) were differentiated from later ones (LE) in all crosses in year 2007 as well,
with the HE remaining early and the LE late. This differentiation occurred from four for the GR
1
x IN
2
to
ten days for the GR
1
x GR
2
cross. Likewise, the genotypes combining high yield and earliness (YHE)
were differentiated from those combining high yield and maturity (YLE) in all six crosses, with the first
(YHE) being earlier than the latter (YLE) from three (GR
1
x GR
3
) to eight (GR
1
x GR
5
) days. High-(HY)
and low-yielding (LY) genotypes different in earliness in five out of the six crosses, with the high-
yielding ones being later in year 2007 as well.
To sum up, in divergent selection, regarding yield, differentiation between high (HY) and low (LY)
was noticed for three out of the six crosses in both years 2006 an 2007 (F
3
and F
4
genotypes,
respectively). However, only one of the three crosses maintained this differentiation for both years. The
genotypes combining yield and earliness (YHE and YLE) were differentiated in two crosses in year 2006
and in our in year 2007.
As for earliness, the differentiation between early and late genotypes was absolute for all six crosses
during both cultivation periods. In addition, differentiation was noticed between HY and LY genotypes in
the same five out of the six crosses in both years of analysis, with the first being later than the latter, as
expected. The behavior of the YHE and YLE genotypes was differentiated in five in year 2006 and in six
crosses in 2007, with the YHE genotypes being earlier than the YLE ones.
Selection for earliness and yield appears to be possible while selecting for yield and late maturity
seems to be impossible with this specific genetic material and under these specific circumstances.
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[10] Martin, A. S., Darrah L. L., and Hibbard B. E. (2004). Divergent selection for rind penetrometer resistance and its effects
on European corn borer damage and stalk traits in corn. Crop Sci. 44:711717.
[11] Masole, H. (1993). Evaluation of high and low divergent rind penetrometer resistance selection at three plant densities in
maize. M.Sc. thesis. Univ. of Missouri, Columbia.
[12] Pressoir, G., and Berthaud, J. (2004). Population structure and strong divergent selection shape phenotypic diversification in
maize landraces. Heredity. 92: 95101.
[13] Radin, J. W., Lu. Z. M., Percy, R. G., and Zeiger, E. (1994). Genetic variation for stomatal conductance in Pima cotton and
its relation to improvements of heat adaptation. Proc. Natl. Acad. Sci. USA 91: 72177221.
[14] Ulloa M., Cantrell R. G., Richard P. G., Zeiger E. and Lu. Z. (2000). QTL Analysis of stomatal conductance and
relationship to lint yield in an interspecific cotton. J. Cotton Sci. 4:10-18.
[15] Uribelarrea, M., Moose, S. P. and Frederick, E. Below. (2007). Divergent selection for grain protein affects nitrogen use in
maize hybrids. Field Crops Research 100 (1): 82-90.
[16] Verhalen, L. M., Baker, J. L. and McNew, R. W. (1975). Gardners Grid System and Plant Selection Efficiency in
Cotton. Crop Sci.15: 588-591.

Development of Recombinant Inbred Lines

for Fibre Quality Traits in Gossypium hirsutum L
Jagmail Singh
1
, Babita Chaudhary
2
, Preeti Srivastva
3
,
Sapna Tiwari
4
and Mukesh Kumar Sharma
5

1
Principal Scientist,
3
Research Associate,
4
Senior Research Fellow,
5
Technical Officer T5,
Division of Genetics, IARI, N. Delhi110012
2
Senior Scientist, Sunnhemp Research Station, (CRIJAF), Pratapgarh (UP)
INTRODUCTION
Cotton is an important commercial crop and plays a key role in Indian economy. It is a major fibre crop
contributing nearly 75% to the natural fibre production. During 2010 the area under cotton was about 11
million ha with an expected production of 330 to 335 lakh bales. Cotton is grown in tropical and sub-
tropical regions of 80 countries. China, India, USA, Pakistan, Brazil, Uzbekistan, Turkey, Greece,
Turkmenistan and Syria are top 10 cotton producing countries of the world. India ranked first in area
under cotton and second in total production after China.
Since the launching of All India Coordinated Cotton Improvement Project in 1967, considerable
success has been achieved in developing improved cultivars with high yield potential and superior fibre
quality. Cultivars with spinning counts ranging from 6s to 120s counts have been developed. The
research programmes was largely centred on improving fibre length and fineness but fibre strength
attracted relatively less attention. Development of G. hirsutum varieties like G-67 and MCU 5 was a
major mile-stone in improvement of yield and fibre quality. G. barbadense varieties Sujatha and
Suvin could be spun at 100s counts and 120s counts, respectively. Development of inter-specific and
intra-specific tetraploid hybrids like H-4, H-6, H-8, H-10, JKHy-1, JKHy-2, DHH-11, PKVHy-2, NHH-
44, Savita, etc (intra- specific) and Varalaxmi, DCH-32, NHB-12, HB 224, DHB 105, TCHB 213, Sruthi,
etc (inter-specific) also played significant role in increasing production of superior quality cotton.
The adoption of modern high speed ring/open-end spinning system by textile industry has increased
demand for high fibre strength cottons. Most of the presently developed varieties have low fibre strength
of 18 to 20 g/tex. The genetic variation available for the improvement, especially for fibre strength in
Gossypium hirsutum cotton that accounts for 90% of total production is limited. Research was therefore
initiated at Indian Agricultural Research Institute, New Delhi to develop recombinant inbred lines for
fibre quality traits, especially fibre strength and to study the genetic variation in segregating generations
and the feasibility to develop superior lines combining high yield with high fibre strength.
Research work was initiated at Indian Agricultural Research Institute, New Delhi to develop strains
having high seed cotton yield and fibre strength that resulted in the development of the strain P 56-4 for
high fibre strength (Singh and Kaushik 2006; Singh et al., 2007). The mean fibre strength of Pusa 56-4
was found to be 27.8 g/tex against 23.1 g/tex of local check Pusa 8-6 during 5-year period from 2003 to
2007 (Table 1). Likewise, its mean 2.5% span length was 28.4 mm and micronaire 3.9 associated with
fibre uniformity (53.5%), good elongation (6.2%) and low short fibre index (5.0%). During the same
period the mean 2.5% span length of local check Pusa 8-6 was 28.2 mm, mean micronaire value 4.6,
mean uniformity ratio 51.7%, mean fibre elongation 5.8% and mean short fibre index 6.1%. Variety RS
2013 developed by Agricultural Research Station, Sriganganagar was released for commercial
cultivation in Rajasthan state. It belongs to medium staple category with fibre length of about 25 mm and
fibre strength of about 19 g/tex. Besides evaluation at IARI, New Delhi, Pusa 56-4 and RS 2013 were
also evaluated for some fibre quality traits at Central Institute for Cotton Research (CICR), Nagpur and
20
University of Agricultural Sciences (UAS), Dharwad during 2006. Pusa 56-4 showed 27.7 g/tex mean
fibre strength, 28.7 mm 2.5% span length, 3.8 micronaire value and 6.3% fibre elongation. The variety
RS 2013 showed 22.1 g/tex fibre strength, 25.4 mm 2.5% span length, 4.4 micronaire value and 6.1%
elongation (Table 2). Pusa 56-4 and RS 2013 were therefore selected as contrasting parents for
developing recombinant inbred lines for fibre quality traits i.e. fibre strength.
TABLE 1: EVALUATION OF PUSA 56-4 VS LOCAL CHECK PUSA 8-6 FOR FIBRE QUALITY TRAITS FROM 2003 TO 2007
Year Strain/
Variety
2.5% Span
Length (mm)
Fibre
Strength
(g/ tex)
Micronaire
Value
Uniformity
Ratio
(%)
Fibre
Elongation
(%)
Short
Fibre
Index (%)
2003 Pusa 56-4 28.6 29.4 3.7 53.0 6.1 6.8
Pusa 8-6 28.0 24.2 4.7 52.1 5.5 7.8
2004 Pusa 56-4 28.5 26.7 3.8 52.1 6.0 8.0
Pusa 8-6 28.0 23.3 4.7 52.7 4.8 11.7
2005 Pusa 56-4 27.9 26.9 4.0 52.3 6.3 3.2
Pusa 8-6 27.5 22.1 4.7 49.9 6.2 3.5
2006 Pusa 56-4 29.7 28.6 4.0 56.0 6.4 3.5
Pusa 8-6 29.6 22.9 4.6 51.5 6.3 3.5
2007 Pusa 56-4 27.7 27.5 4.0 55.6 6.2 3.5
Pusa 8-6 27.9 23.0 4.5 52.1 6.3 4.2
Mean Pusa 56-4 28.4 27.8 3.9 53.5 6.2 5.0
Pusa 8-6 28.2 23.1 4.6 51.7 5.8 6.1
TABLE 2: PERFORMANCE OF PUSA 56-4 AND RS 2013 FOR IMPORTANT FIBRE QUALITY TRAITS AT DELHI, NAGPUR AND DHARWAD DURING 2006
(a) Fibre Strength (g/tex)
IARI, N. Delhi CICR, Nagpur UAS Dharwad Mean
Pusa 56-4 28.4 28.9 25.9 27.7
RS 2013 19.9 24.8 21.7 22.1
(b) 2.5% Span length (mm)
Pusa 56-4 28.5 29.0 28.6 28.7
RS 2013 25.7 25.7 24.9 25.4
(c) Micronaire value
Pusa 56-4 4.1 3.8 3.6 3.8
RS 2013 4.5 4.5 4.2 4.4
(d) Fibre Elongation (%)
Pusa 56-4 6.3 6.4 6.1 6.3
RS 2013 5.7 6.4 6.3 6.1
Strain Pusa 56-4 was crossed with variety RS 2013 in 2004 to obtain F
1
generation. It was
advanced to F
2
generation during 2005 and was evaluated during 2006. Two hundred and ninety seven
single plants from F
2
generation were phenotyped for fibre strength and other fibre quality traits. Data
were also recorded on seed cotton yield and important yield components. These single plants were
advanced to F
3
, F
4
and F
5
generations through selfing, following single seed descent method and were
evaluated for fibre quality traits in F
3
generation (200708), F
4
generation (200809) and F
5
generation
(2009-10). Data were also recorded on seed cotton yield and important yield components.
Data on mean, range and standard deviation related to fibre quality traits, seed cotton yield and important
yield components in F
2
, F
3,
F
4
, and F
5
generations are given in Table 3.
Fibre Quality Traits
The data indicated wide variation for several fibre quality traits in 4
th
generations, especially in F
2
. The
2.5% span length ranged from 22.7 to 30.6 mm in F
2
; 22.6 to 30.6 mm in F
3
, 23.5 to 31.9 mm in F
4
and
21.0 to 31.8 mm in F
5
.The mean 2.5% span length was 26.8 mm in F
2
, 26.6 mm in F
3
, 27.6 mm in F
4

and 26.4 mm in F
5
. The fibre strength ranged from 18.1 to 33.1 /tex in F
2
; 18.8 to 27.9 g/tex in F
3
, 20.0
to 30.7 g/tex in F
4
and 18.0 to 27.9 g/tex in F
5
generation. The mean fibre strength in F
2
, F
3
, F
4
and F5

generations was 26.6, 23.1, 25.1 and 23.7g/tex, respectively. The micronaire value varied from 3.6 to 4.9
Development of Recombinant Inbred Lines for Fibre Quality Traits in Gossypium hirsutum L 113
in F
2
, 3.2 to 5.7 in F
3
, 2.8 to 5.4 in F
4
and 2.7 to 5.2 in F
5
, with the respective mean of 4.2, 4.4, 3.9 and
4.1. The fibre uniformity ranged from 47.6 to 61.3% in F
2
, 46.6 to 58.5% in F
3
, 45.8 to 60.2% in F
4
and
45.8 to 58.4% in F
5
. The mean fibre uniformity ratio was 55.8%, 52.4%, 53% and 52% in F
2
, F
3
, F
4
and
F5

generations, respectively. There was not much variation for fibre elongation and fibre maturity. The
mean fibre elongation per cent was 6.3, 6.1, 6.1 5.2, respectively in the 4
th
generations, which ranged
from 5.4 to 6.7% in F
2
, 5.5 to 6.6% in F
3
, 5.5 to 6.9% in F
4
and 4.8 to 5.7% in F
5
. All the plants in the 4
generations showed good fibre maturity ranging from 0.8 to 0.9. The short fibre content was found to be
3.5% to 8.8% in F
2
, 3.5 to 11.2% in F
3
, 3.5 to 8.6% in F
4
and 3.5 to 9.5% in F5, with mean of 3.8%,
4.8%, 4.1% and 4.0%, respectively in the 4 generations.
TABLE 3: PERFORMANCE OF THE CROSS PUSA 56-4 AND RS 2013 FOR IMPORTANT FIBRE QUALITY TRAITS AND SEED COTTON YIELD IN F
2
, F
3
, F
4
AND F
5
GENERATIONS AT IARI, N. DELHI
Trait Generation Mean SD Minimum Maximum
2.5% span length (mm) F
2
26.8 1.5 22.7 30.6
F
3
26.6 1.6 22.6 30.6
F
4
27.6 1.7 23.5 31.9
F
5
26.4 1.7 21.0 31.8
Fibre strength (g/tex) F
2
26.6 2.3 18.1 33.1
F
3
23.1 1.6 18.8 27.9
F
4
25.1 2.0 20.0 30.7
F
5
23.7 1.8 18.0 27.9
Micronaire value F
2
4.2 0.3 3.6 4.9
F
3
4.4 0.5 3.2 5.7
F
4
3.9 0.5 2.8 5.4
F
5
4.1 0.5 2.7 5.2
Fibre uniformity (%) F
2
55.8 2.3 47.6 61.3
F
3
52.4 2.5 46.6 58.5
F
4
53.0 2.6 45.8 60.2
F
5
52.0 2.4 45.8 58.4
Fibre elongation (%) F
2
6.3 0.2 5.4 6.7
F
3
6.1 0.2 5.5 6.6
F
4
6.1 0.2 5.5 6.9
F
5
5.2 0.2 4.8 5.7
Fibre maturity F
2
0.9 0.0 0.8 0.9
F
3
0.8 0.0 0.8 0.9
F
4
0.8 0.0 0.8 0.9
F
5
0.8 0.0 0.8 0.9
Short fibre index (%) F
2
3.8 0.7 3.5 8.8
F
3
4.8 1.6 3.5 11.2
F
4
4.1 1.1 3.5 8.6
F
5
4.0 1.0 3.5 9.5
Yield / plant (g) F
2
43.2 31.0 22.0 196.3
F
3
75.9 33.1 25.2 193.1
F
4
163.4 95.7 21.5 581.4
F
5
147.7 69.8 27.4 342.8
Boll weight (g) F
2
3.5 0.7 2.0 5.1
F
3
3.4 0.6 2.1 5.7
F
4
3.9 0.7 2.1 5.8
F
5
3.6 0.7 1.9 5.6
Ginning outturn (%) F
2
30.6 3.3 25.6 41.4
F
3
31.7 2.4 25.6 39.3
F
4
32.6 2.4 26.0 40.4
F
5
33.4 2.7 26.5 44.7
Seed index (g) F
2
9.3 1.5 4.0 13.5
F
3
8.1 1.0 5.6 11.6
F
4
8.9 1.3 3.0 12.2
F
5
8.2 1.2 5.4 12.8
Lint index (g) F
2
3.9 0.7 2.1 5.8
F
3
3.8 0.5 2.5 6.0
F
4
4.3 0.6 2.9 6.4
F
5
4.0 1.0 3.5 9.5
Yield and Yield Components
Wide variation was also observed for seed cotton yield per plant in all the 4 generations, i.e. F
2
through
F
5
. It ranged from 22.0 to 196.3 g in F
2
, 25.2 to 193.1 g in F
3
, 21.5 to 581.4 g in F
4
and 27.4 to 342.8g in
F
5
generation. The mean seed cotton yield was 43.2 g in F
2
, 75.9 g in F
3
, 163.4 g in F
4
and 147.7g in F
5.

The boll weight was found to vary from 2.0 to 5.1 g in F
2
, 2.1 to 5.7 g in F
3
, 2.1 to 5.8 g in F
4
and1.9 to
5.6g in F
5
generation, the mean in the 4 generations being 3.5g, 3.4g, 3.9g and 3.6g, respectively. The
ginning outturn also showed wide variability, ranging from 25.6 to 41.4% in F
2
, 25.6 to 39.3% in F
3
, 26.0
to 40.4% in F
4
and 26.5 to 44.7% in F
5
. The mean ginning outturn in the 4 generations was 30.6% in F
2
,
31.7% in F
3
, 32.6% in F
4
and 33.4% in F
5
. Good variation was also observed for seed index and lint index
in all the 4 generations.
The distribution for fibre strength in F
2
generation was little skewed as 76.5% plants out of 297
showed high fibre strength (above 25 g/ tex). Furthermore, 21 plants (7.0%) in F
2
showed above 28 g/tex
fibre strength which was higher than that of the better parent, suggesting transgressive segregation for the
trait. The range observed for fibre strength in F
3
, F
4
and F
5
generations was relatively less as compared
to F
2
generation. Transgressive segregation for fibre strength was observed as 2.7% plants showed
higher fibre strength than the better parent. Transgressive segregation was also observed for 2.5% span
length with 18.7% plants in F
2
showing higher fibre length than 28.4 mm of better parent Pusa 56-4.
Only 1.3% plants in F
2
showed lower micronaire value (finer fibre) as compared to 3.9 micronaire value
of better parent Pusa 56-4 and 6.3% plants showed higher short fibre index than better parent Pusa 56-
4. The boll weight was found to be higher than 4.5 g of Pusa 56-4 in 21% plants. Transgressive
segregation was also observed for low as well as high ginning percentage. The variation and
transgressive segregation observed for fibre length and strength and other traits has practical implication
for combining high yield and superior fibre quality in G. hirsutum cottons. Percy et al. (2006) also
reported transgressive segregation for high and low lint percentage, with about 5% of lines exceeding
high ginning outturn parent. Similarly, about 25% lines exceeded the height of the taller parent. They
further reported that 20% lines possessed fibre strength equivalent to high fibre strength parent while 4%
exceeded high strength parent. Fibre length and uniformity were reported to be normally distributed,
while 14% lines showed transgressive segregation for lower micronaire value. They also opined shifted
distribution among recombinant inbred lines towards smaller boll size. Reinisch et al., (1994) reported
strongly distorted segregation of molecular markers in early generations while Jiang et al., (2000)
reported skewed transmission in advanced generations of inter-specific hybrids. The transgressive
segregation was also observed for several important traits in our study.
As many as 93 plants out of 297 evaluated in F
4
generation showed above 26.0 g/tex fibre strength
during 200809. The fibre strength of these 93 plants ranged from 26.1 to 30.7 g/tex, 2.5% span length
from 25.1 mm to 31.9 mm and micronaire value from 2.9 to 4.7. Although the overall seed cotton yield
among these 93 plants varied from 32.3g to 459.2 g/plant, nonetheless 30 plants showed per plant yield
of above 200 g/plant. Thus, the F
4
plants derived from the cross Pusa 56-4 RS 2013 showed
valuable genetic variation for simultaneous improvement of yield and fibre quality in G. hirsutum
cottons, suggesting the feasibility of combining high yield and superior fibre quality.
REFERENCES
[1] Jiang, C., Wright, R. J., El-Zik, K. M. and Paterson, A. H. (2000). QTL analysis of leaf morphology in tetraploid
Gossypium. Theoretical and Applied Genetics. 100: 40918.
[2] Percy, R. G., Cantrell, R. G. and Zhang, Jinfa. (2006). Genetic variation for agronomic and fibre properties in an
introgressed recombinant inbred population of cotton. Crop Science. 46: 13117.
[3] Reinisch, M. J., Dong, J., Brubaker, C. L., Stelly, D. M., Wendel, J. F. and Paterson, A. H. (1994). A diallel RFLP map of
cotton, Gossypium hirsutum Gossypium barbadense: chromosome organization and evolution in disomic polyploidy
genome. Genetics. 138: 82947.
[4] Singh, J. and Kaushik. S. K. (2006). High fibre strength strains of American cotton suitable for rotor spinning. ICAR News.
12 (2): 156.
[5] Singh, J., Kaushik, S. K. and Chaudhary, Babita. (2007). Seed cotton yield and fibre quality analysis in American cotton.
Journal of the Indian Society for Cotton Improvement. 32 (3): 18892.
Elite Cotton Varieties in the Zimbawean Private

Sector Research Programme
Mandiveyi Jeremiah Kudzayi
Seed Cotton Breeder, Quton Seed Company (Pvt) Ltd, Box 192, Kadoma, Zimbabwe
AbstractIn the 2009-2010 cropping season, seven elite cotton varieties from Qutons (Quton Seed Company)
Medium Staple Breeding Programme were planted across 10 sites to evaluate field and fiber characteristics. The yield
data indicated that the entry 675-02-1 performed better than others with the average seed cotton yield of 2.33 t/ha
under dryland conditions. Entry 675-02-1 has medium bolls with very good seed cotton retention and early maturity.
Two hundred and sixty four lint samples from the first picking were collected and sent to the Cotton Company of
Zimbawe (Cottco) Laboratory for fiber analysis in terms of fiber length, strength, elongation, maturity and micronaire
(HVI). The latest maturing entry B81-05-1, had the longest fibers associated with strong fiber and a good micronaire.
This variety also had the highest ginning outturn and the heaviest bolls in this season. Another elite line MS98-01-68,
produced the second best fiber strength value of 31.8 g/tex. Strong fibers can be span at higher speeds and meets
demands of modern spinning technologies. Earlier maturing varieties were generally better yielding mainly due to an
end of season drought spell that affected late maturing varieties.
INTRODUCTION
Quton Seed Company (Pvt.) Ltd. (Quton), a subsidiary of Seed Co-Group is the only cotton planting seed
company operating in Zimbabwe. It has a breeding unit within the Department of Research and Specialist
Services based in Kadoma, Zimbabwe. This Department has a strong breeding programme that has
yielded significant results by releasing cotton varieties such as QM 301, a high quality medium staple
variety that meets all stakeholders requirements. The major aim of the cotton variety development
programme of Quton is production of medium-staple (Gossypium hirsutum) varieties suitable for open-
end spinning for Zimbabwe and the region.
Currently, one public and one private Research Institutes are involved in cotton research in
Zimbabwe. The main focus on cotton breeding programs is to develop cotton varieties which produces
high yields of seed cotton and lint of a suitable quality under the wide range of growing conditions in
Zimbabwe (Cotton Outlook 2001 Special Edition). Research works are also carried out for fiber quality,
disease and pest resistance, drought tolerance and early maturity. Development of medium-staple cotton
varieties suitable for modern spinning methods is the main target of cotton research in the private sector
represented by Quton Seed Company. Quton Seed Companys breeding work is mainly divided into two
separate programs that are sub-divided into three maturity groups (Late, Medium and Early Maturity).
The general objectives of each program and current successes are outlined below.
The Medium Staple (MS) Program
This concentrates on breeding of improved cotton varieties in the medium staple range (2.5% Span
Length between 28mm and 32mm) for the main cotton- growing areas of Zimbabwe and other regional
countries. The objectives include wide adaptability, resistance for jassid, bacterial blight, verticillium
wilt, and improvements in plant habit, earliness, ginning percentage, yield potential and fiber quality. The
present Quton variety for this program is Albar Plus QM 301.
Commercially Released Private Sector Varieties
Qutons produced the first non-government bred cotton variety called Albar Plus QM 301 in the history
of cotton production in Zimbabwe in September 2006. This variety was selected as a single plant at
Cotton Training Centre during the 2000-2001 season out of F
3
generation of a population created by
crossing a number of West African and local varieties.
21
Albar Plus QM 301 is a late maturing and indeterminate cotton variety with a potential yield between
1500 and over 4000 kg/ha. It has some improvements in verticillium wilt and bacterial blight tolerance.
However, its leaf pubescence has space and therefore is less tolerant to jassid attack. The fibre quality of
Albar Plus QM 301 is significantly better particularly in terms of fiber length and strength. The lint
colour is white with a reflectance value of 78,5. QM 301 has the following fiber parameters: 2.5% span
length 28.5mm, fiber strength 31.2 g/tex, fiber elongation 7.7%, micronaire 4.4.
Albar Plus QM 301 was released as a new variety with improved fiber quality as an alternative to SZ
9314 in both the middleveld and lowveld.
The Long Staple (LS) Programme
The long staple programme on the other hand is a relatively small program whose main objective is the
development of long staple (up to 34mm) cotton varieties. Current production of the long staple variety is
insignificant since farmers shun the low yield potential of the variety under dryland conditions. The
general aim of the programme is thus to produce high quality cultivars, which also meet the needs of
producers in terms of reliable yield performance. Quton has a couple of elite lines under evaluation but
none have been registered for commercial production yet.
Experimental Procedure
Cotton variety assessment trials were conducted in summer 2009-2010 cropping season at two on-station
sites and 10 off-station sites in major cotton-growing areas of Zimbabwe. Seven most advanced varieties
from the Quton breeding programme were evaluated against the current commercial variety, SZ 9314
(bred by Cotton Research Institute, a government owned breeding unit based in Kadoma). These cotton
varieties were planted and evaluated for field characteristics (yield, ginning %, boll weight, earliness) and
fiber characteristics (2.5% span length, strength, elongation, maturity and micronaire). Acid delinted
seeds were planted to ensure good germination. Five seeds were planted per hill and thinned to one plant
per station within the first three weeks of crop emergence.
All trials were in dryland areas and were sown between mid- November and mid-December. Trials
were laid out in Randomised Complete Blocks with eight treatments replicated thrice at eight locations
(Anfield, Bindura, Bluegrass, Dande, Kadoma, Muzarabani, Raffingora, and Shamva). Gross plot size
ranged between 17.1 square metres and 22.8 square metres depending on site. Prior to sowing, compound
L (5:18:10) at 250 kg/ha had been banded adjacent to sowing furrows. At all sites trials were sown at a
standard spacing of 1.0 m x 0.30 m giving a plant population of 33 333 plants per hectare. Top dressings
of ammonium nitrate (34.5%N) were done 8 weeks after crop emergence at the rate of 100 kg/ha. At all
sites hand-hoeing and/ or mechanized cultivation were used to control weeds. At Kadoma and Bindura,
herbicides Codal Gold 412.5 DC was applied pre-emergence, at the rate of 4.0 litres per hectare. Scouting
and spraying, wherever possible, followed methods and recommendations laid down in the relevant
sections of the Cotton Handbook. Where trials were conducted on farmers fields, scouting and spraying
were left to the grower to use his own discretion, but chemicals were provided by Quton.
Harvesting was spread over a period from early March to late July across all sites. The number of
picks taken varied from a minimum of two at some sites to a maximum of three. For all trials, boll
samples were collected at the first pick stage and were used as the ginning sample. The boll samples were
standardized by taking 100 fully open undamaged bolls per plot. The first two plants on either side of the
plot were excluded. Ninety gram lint samples were taken from the ginning of each plots seed cotton. The
lint samples were sent to The Cotton Company of Zimbabwe (Cottco) for fiber testing using HVI
Spectrum from Ulster Technologies, USA. Most of the areas that hosted our trials received normal to
above normal rainfall with a poor distribution thereby affecting mainly the late-planted trials. There was
a prolonged mid-season drought spanning between January and March. The crops that were established
early yielded quite well compared to late-planted trials.

Elite Cotton Varieties in the Zimbawean Private Sector Research Programme 117
Field Characteristics
Seed cotton yields from individual sites are presented in Table 1. The variety 675-02-1 was outstanding
in overall performance across both high and low input cotton-growing areas with a 14% yield advantage
over the main control variety SZ 9314 (Table 1). This variety will benefit farmers due to its superior
earliness if introduced into commercial production. At one of the sites, 675-02-1 produced some tight
locs. Although MS99-03-176 produced a very satisfactory average seed cotton yield of 2300 kg/ha, its
productivity was poor under low input conditions (the predominant conditions in which Zimbabwean
cotton is grown). The latest maturing entry, B81-05-1 with an earliness index of 54% (Table 2) produced
the heaviest boll and the highest ginning percentage compared to all test varieties. Although the variety
MS99-01-68 has smaller bolls than SZ 9314 and QM 301, it is superior to both in yield and earliness.
MS98-01-176 produced better seed cotton and lint yield than QM 301 because of and late maturity than
controls (Table 2).
TABLE 1: SEED COTTON YIELD (KG PER HA) OF MS VARIETY
Treatment Variety Raffingora Seedco Bluegrass Bluegrass Bluegrass Bindura 1 Bindura 2 Shamva Anfield Mean
(High) (High) (Low) (Low) (Low) (High) (Low) (High) (High)
1 675-02-1 2659 3058 1378 1410 1728 3037 1799 3648 2290 2334
2 679-02-1 2813 2539 1420 1237 1310 2976 1685 3788 2316 2232
3 772-03-6 3097 2954 1359 1185 1517 3051 1297 3453 2569 2276
4 B81-05-1 2715 2399 1682 1542 1504 2440 1229 3267 2901 2187
5 MS98-01-68 2946 2817 1662 1308 1506 2819 1562 3343 2350 2257
6 MS99-03-176 2930 2996 1338 1304 1457 2911 1646 3665 2456 2300
MEANS 2860 2794 1473 1331 1504 2872 1536 3527 2480 2264
CHECKS(1) :
8 SZ 9314 2379 2335 1472 1250 1481 2685 1164 3489 2214 2052
CHECKS(2) :
7 QM 301 2482 2796 1568 1304 1378 2684 1460 3458 2136 2141
OVERALL:
MEANS 2753 2737 1485 1318 1485 2825 1480 3514 2404 2222
STD ERR 241.1 158.9 11906 154.1 143.8 133.5 172.9 100.3 201.1 1468.0
5% LSD 731.2 482 362.8 467.5 436.2 405 524.3 304.4 609.9 480.4
C.V. 15.2 10.1 14 20.3 16.8 8.2 20.2 4.9 14.5 13.8

*Trials at 11 site having net plots size between 17.1 and 22.8 sq. m under RBD with three replications
TABLE 2: FIELD CHARACTERISTICS OF MS VARIETY ASSESSMENT TRIALS AT NINE SITES, 2009-2010 SEASON.
Variety Total Seed Ginning Lint Yield Boll Weight Seed Earliness Plant Population
Cotton (kg/ha) % (kg/ha) (g) Weight % Height (cm) (000s)
675-02-1 2334 40.79 953 5.93 11.32 72 132 33.65
679-02-1 2231 39.37 900 5.92 10.93 68 130 33.17
772-03-6 2276 40.72 921 6.23 11.53 66 131 33.92
B81-05-1 2187 42.19 926 6.82 12.94 53 129 33.32
MS98-01-68 2257 41.94 942 5.96 11.33 71 135 33.28
MS98-03-176 2300 41.20 947 5.97 10.91 64 133 33.26
SITES 9 9 9 9 9 9 9 9
MEANS 2264 41.03 931 6.14 11.49 66 132 33.43
CHECKS(1):
SZ 9314 2052 41.48 844 6.20 11.66 65 129 33.39
MEANS 2052 41.48 844 6.20 11.66 65 129 33.39
CHECKS(2):
QM 301 2141 42.80 916 5.84 10.55 70 128 33.44
MEANS 2141 42.80 916 5.84 10.55 70 128 33.44
OVERALL:
MEANS 2222 41.31 919 6.11 11.40 66 131 33.43
Fiber Characteristics
Fiber strength is now the most important fiber parameter in both rotor and friction spinning. The fiber
strength data from 11 trials (Table 3) show that most entries produced stronger fibers than the controls.
The entry with the best fiber strength value (32.12 g/tex), B81-05-1 also had the highest fiber length
value of 31.61 mm (2.5% span length). The fiber strength values of three best lines (675-02-1, B81-05-1
and MS98-01-68) are inherently superior (Table 3) across both high and low potential areas, while the
variety 772-03-6 consistently gave poor strength values across all sites. Another entry, 675-02-1 with
very good field characteristics had a fiber length value equal to QM 301 but its fiber strength was better
than QM 301 by 2.5%. Although 679-02-1 inherently has the shortest fiber, it produced a good fiber
strength value of 31.5 g/tex. B81-05-1 is high grade cotton with long and strong fiber (Table 4).
TABLE 3: FIBER STENGTH VALUES OF MS VARIETY
Variety Raffingora Seedco Seedco Bluegrass
1
Bluegrass
2
Bluegrass
3
Bindura
1
Bindura
2
Shamva Anfield Anfield Mean
High High Low Low Low low High low High High Low
675-02-1 31.80 33.43 29.77 30.47 30.57 32.10 32.87 34.10 31.13 29.30 31.23 31.52
679-02-1 31.40 32.23 32.03 29.07 32.43 31.20 32.57 33.47 30.73 31.30 30.07 31.50
772-03-6 31.07 31.77 30.03 29.10 29.07 31.67 30.67 33.43 29.43 29.47 30.50 30.56
B81-05-1 31.03 35.93 30.80 29.23 32.40 33.57 32.23 34.10 32.53 30.47 31.03 32.12
MS98-01-68 29.50 33.77 30.67 31.43 30.20 32.00 33.73 35.50 31.50 30.83 30.37 31.77
MS99-03-176 32.00 33.23 31.27 28.60 31.27 31.87 31.03 32.80 29.73 29.80 31.80 31.22
MEANS 31.13 33.39 30.76 29.65 30.99 32.07 32.18 33.90 30.84 30.20 30.83 31.45
CHECKS(1) :
SZ 9314 30.80 33.60 28.70 30.23 29.30 30.77 31.77 33.50 28.13 31.20 30.20 30.75
CHECKS(2) :
QM 301 31.23 31.67 30.53 29.50 30.53 30.50 31.63 34.40 28.53 28.90 30.67 30.74
OVERALL:
MEANS 31.1 33.5 30.5 29.7 30.7 31.7 32.1 33.9 30.2 30.1 30.7 31.3
STD DEV 1.7 2.6 1.6 1.7 1.6 1.6 1.6 1.5 1.5 1.6 1 1.6
C.V. 5.6 7.8 5.3 5.6 5.3 5 5 4.3 5.1 5.2 3.4 5.2
*Assessment Trials across eleven site during 2009-10 season having 17.1 and 22.8 sq. m plot size under RBD with three
replications.
TABLE 4: FIBER CHARACTERISTICS OF MS VARIETY
Variety Length (mm) Strength Elongation Uniformity Micronaire Maturity
675-02-1 29.96 31.52 5.86 84.80 4.13 0.88
679-02-1 29.64 31.50 6.04 84.34 4.18 0.88
772-03-6 30.21 30.56 5.96 84.56 4.09 0.87
B81-05-1 31.61 32.12 5.78 85.27 4.03 0.88
MS98-01-68 30.61 31.77 5.91 85.01 4.08 0.88
MS98-03-176 31.01 31.22 5.89 84.91 4.03 0.87
MEANS 30.51 31.45 5.91 85 4.09 0.88
CHECKS(1):
SZ 9314 29.66 30.75 6.18 84.41 4.23 0.88
CHECKS(2):
QM 301 30.00 30.74 6.26 84.86 4.04 0.87
OVERALL:
MEANS 30.34 31.27 5.99 85 4.10 0.88
*Assessment Trials across eleven sites during 2009-2010 season
Fiber elongation values were average for the most promising entries and were less than 7%. The
micronaire, uniformity and maturity values were within acceptable ranges. The three varieties (B81-05-1,
MS98-01-68 and MS99-03-176) combined major gains in both fiber length and fiber strength parameters.
Pests
Major pests observed across the ten sites were termites, jassids (leaf hoppers), aphids, bollworms (red,
Heliothis), cotton strainers, and leaf eaters (mainly elegant grasshoppers and semi loppers). Pest menace
was high and an average of 10,6 sprays were applied during the season with an average of 7,1 against
bollworms, 4.0 against aphids, 1.8 against red spider mite and 1.0 against jassids. At some trial sites,
insect damage caused serious plant population losses.

Elite Cotton Varieties in the Zimbawean Private Sector Research Programme 119
All the varieties were assessed for jassid resistance using a method to build up high jassid numbers in
an unsprayed block. Reasonable levels of jassid resistance are essential for varieties grown in the
smallholder sector. Jassid build up was poor and was also compounded by the mid-season drought. No
meaningful data was obtained this season and thus no scores are presented in the field data.
CONCLUSION
Zimbabwe mainly grows cotton for lint exports to overseas markets. The competitiveness of its lint that
is traded based on quality parameters. The MS breeding program has made huge gains in the
improvement of fiber quality parameters especially fiber length and strength particularly against the main
commercial and government owned variety, (SZ 9314). The field data of Qutons elite lines also shows
that the MS breeding program has diverse varieties in different maturity groups surpassing the yield
potential of SZ 9314.
REFERENCES
[1] Cotton Outlook, 2001 Special Edition, International Cotton Advisory Committee, 60
th
Plenary Meeting, Victoria Falls,
Zimbabwe.
[2] Cotton Handbook of Zimbabwe.
[3] Cotton Research Institute Annual Report, 1983-1984, Department of Research and Specialist services, Government Printer,
Harare, 1985.
[4] The ICAC Recorder, International Cotton Advisory Committee, Technical Information Section, Volume. XX No.
1 March 2002
Development of Biotic Stress Resistance Transgenic

Diploid Cotton Utilizing Agrobacterium
and Shoot Apical Meristem Cells
Sukhadeo Nandeshwar, Pranjib Chakraborty, Kanchan Singh,
Mithila Meshram, Bipinchandra Kalbande
Division of Crop Improvement, Central Institute for Cotton Research, Nagpur-MS. India
INTRODUCTION
Cotton is a global crop with world hectarage of 32.4 m and production of 87.4m bales. It is one of the
major source of fibre, cattle feed and edible oils. The productivity of cotton is adversely affected by
biotic and abiotic stresses. Major concern world over has been to protect it from bollworm (Helicoverpa
armigera) a single pest capable of causing damage to the extent of 40% besides infestation of fungal
pathogens.
In past two decades, extensive research efforts have been made for transformation, regeneration and
genetic enhancement of cotton, especially G. hirsutum. Gene conferring agronomic advantages has been
introduced through Agrobacterium and particle gun and the plants were regenerated through callus based
somatic embryogenesis (Firoozabady et. al., 1987; Umbeck et al., 1987; Perlak et al., 1990; Finer and
Mc Mullen 1990; Bayley et. al., 1992; Thomas et al., 1995; Rajshekharan et al., 1996; Zhao et al. 2006).
All these workers however mostly employed Coker (G. hirsutum) sister lines and their derivatives for
transformation. Shoot tip meristem and apex culture offers unique advantage to regenerate plants directly
from inoculated shoots on simple MS medium. The procedure has been used to regenerate and transform
many cultivars of cotton (Gould et al., 1991a; Gould and Megallanes-Cedeno 1998; Zapata et al., 2002,
Kategari et al., 2007 and Nandeshwar et al., 2009) has reported successful transformation in diploid
cotton G. arboreum by transferring cry IAc gene. This protocol has also been used in crop plant like
maize (Gould et al., 1991) and pine (Gould et al., 2002). It has also been observed that the incidence of
genetic mutation and somaclonal variation were low in plant regeneration from shoot tip explants (Gould
and Megallanes-Cedeno 1998). Recently new transformation methods have been reported which
minimizes the steps of in vitro regeneration and transformation which also increases transformation
frequency.
Diploid cotton is widely cultivated in at least six states in India covering approximately 7% of the
cotton-growing area. The importance of diploid cotton especially G. arboreum is increasing owing to the
development of long staple and high yielding varieties combined with its inherent tolerance to number of
diseases such as leaf curl, new wilt etc. and drought. However plants are often badly damaged by
bollworm and grey mildew. In this study we describe two efficient and simple protocols of
transformation and development of transgenic diploid cotton by using cry IAc and chitinase genes
respectively.
Plant Material
Seeds of G. arboreum cv RG 8 and PA255- popular commercial varieties were selected for
transformation. Seeds were delinted with concentrated sulphuric acid followed by treatment with lime
water and several rinses with running tap water. They were sun-dried and stored for further use. Seed
sterilization was carried out with doubled distilled water added with few drops of teepol. Seeds were
rinsed several times in this solution and washed 3-4 times with doubled distilled water. Seeds were then
22
Development of Biotic Stress Resistance Transgenic Diploid Cotton Utilizing Agrobacterium 121
transferred in the solution of bavistin (1 % w/v) and agitated on orbital shaker at 90 rpm for 20 min,
followed by 3-4 washes with autoclaved distilled water. Seeds were treated with 0.1% solution of
mercuric chloride for 7 min followed by continuous washes with autoclaved doubled distilled water till
the traces of mercuric chloride are completely removed and were collected in petri plates and inoculated
in conical flask containing half strength MS medium from germination.
Bacterial Culture
An Agrobacterium tumefaciens strain EHA 105 carrying the binary vector pBinBt3 was used. The vector
was obtained from the National Research centre on Plant Biotechnology, New Delhi, India. The plasmid
harboured the cry IAc gene and the nptII gene each driven by CaMv 35s promoter. With regard to
chitinase gene, binary plasmid pBin-AR was used as a vector for transformation. The plasmid harboured
the chitinase gene isolated from cotton G.hirsutum and nptII (Chakraborty personal communication).The
bacterial culture was maintained on YEM medium containing 50mg/L Kanamycin. For inoculation of
explants, a single colony of bacterial cells was grown overnight in liquid YEM broth at 28
0
c.
Transformation and Co-cultivation
Seven days old geminated seedling (fig.1.a) were used for isolation of shoot tip explants. Both the
cotyledonary leaves were gently removed from the seedling with the help of sterile forceps and an
oblique excision was made at the base of the shoot tips just below the cotyledonary leaf as described by
Nandeshwar et al., (2002). All extraneous tissue surrounding the meristem dome was removed for
efficient Agrobacterium infection.
Shoot tip explants were inoculated with an overnight grown culture of Agrobacterium for 60 minutes
(fig.1.b). The infected explants were collected in petri-plate overlaid with sterile filter paper and co-
cultivated for 4 days separately on simple MS medium. Following co- cultivation, explants were
transferred to the same medium for 2 weeks containing 500mg/L carbenicillin. Subsequently, explants
were subjected to screening on kanamycin medium (50mg/L). To promote shoot elongation, shoots (or
explants with developing shoots) were transferred to MS medium supplemented with 50mg/l kanamycin,
0.1mg/L kinetin, and 0.1 mg/l gibberellic acid (fig.1.c) or 2mg/L BA and 1 mg/L kinetin and 50 mg/L
kanamycin for regeneration of shoot directly or multiple shoot formation (fig.1.d). Shoot (desired from
direct shoot regeneration or multiple shoot induction) were separated and transferred in MS medium
containing 15g/L glucose and NAA (0.1 mg/L). Rooted plants were established (fig.1.e) in glass house
for further analysis. The T
o
plants were first tested for gene amplification by PCR and positive plants
were selfed to collect T
1
seeds for raising T
1
plant progenies.
For in-planta transformation method, surface sterilized seeds were sown directly in earthen pot
containing sand:soil mixture. Seven days healthy seedlings were selected and they were vertically
bisected along the line of cotyledonary leaf attachment. Twenty four hours grown 200 L of
Agrobacterium culture pre-mixed with modified 100 L vir induction medium diluted with MS broth was
applied at the bisected exposed portion of the apical meristem. The plants were covered with polythene
bags for 10 days. The matured T
o
plants (fig.3) were tested for the presence of gene by PCR. The PCR
positive plants were selfed and T
1
progenies on boll basis was harvested.
Molecular Analysis of Gene Integration
Extraction of genomic DNA was extracted following protocol developed in our lab (Chakra borty et al.,
2007). Young leaves of putative transgenic cotton plants were frozen in liquid nitrogen and ground in a
pre-cooled mortar. Ground tissue (0.5g) was extracted in 15 ml extraction buffer (100mM Tris HCl pH 8,
20 mM EDTA, 1.4 M NaCl, 2% CTAB and 0.5M glucose) in a 50 ml centrifuge tube storing remaining
powder at -70 C for later use. The mixture was homogenised and incubated at 60 C for 30 min with
occasional shaking. The DNA was extracted once each in equal volume of phenol: chloroform:
isoamylalcohol, (25:24:1) and chloroform:isoamyl alcohol (24:1) each time producing an emulsion by
gently inverting the tube few times and transferring the upper aqueous layer to a fresh tube following
centrifugation at 6000g for 8 min. at 4 C. The DNA was precipitated with 0.1 volume of 3M sodium
acetate (pH 5.2) and 1 volume of isopropanol at room temperature. Following precipitation, the DNA
was spooled gently with the help of bent Pasteur pipette after allowing the tube to stand at 20 C for 30
min. Following spooling of DNA, the remaining solution was centrifuged at 14000 rpm for 5 min. The
spooled and the pelleted DNA was rinsed with 70% alcohol, dried briefly and dissolved in 200l of
sterile distilled water. Purity of DNA was checked by obtaining absorbance ratios at A260/280 and
A260.230. Concentration of DNA in the sample was calculated by the value of absorbance at A260.
Detection of Cry IAc and Chitinase Gene by PCR
Presence of the transgene was ascertained by polymerase chain reaction (PCR) amplification of 1 kb
fragment of cry IAc gene using a set of primers cry IAc F (5 CCCAGAAGTTGAACTTGGTGG 3) and
cryIAc R (5 CCGATATTGAAGGGTCTTCTGTAC). The PCR was performed in 50 l reaction
volumes. Each reaction consisted of 2l of 100ng/l template, 5l of 10xPCR buffer, 0.5l of 25mM
dNTPs, 1.5 l of 15mM MgCl
2,
1l each of 10M reverse and forward primer, 0.25l of 5U/l Taq DNA
polymerase (MBI Fermentas, USA), and 38.7 l sterile distilled water. The PCR protocol was
standardized with the initial denaturation at 94 C for 4 C followed by 39 cycles consisting of
denaturation at 94 C for 45 sec. primer, annealing at 60 C for 30 sec, and primer extension at 72 C for 1
min and a final extension at 72 C for 5 min. Negative controls were maintained for non-specific
amplification, if any.
Presence of Chitinase gene was confirmed using the same method as followed for CryIAc gene by
using gene specific primers Chitinase F (5 GGGGTACCATGAGCTTTCTTCAGGCCTT3) and
Chitinase R (5 GGGGTACCGCACGTTTACTGATTTCATTT 3).The PCR protocol was modified for
Chitinase with initial denaturation at 95 C for 5 minutes followed by 10 cycles of denaturation at 95 C
for 45 sec. primer annealing at 55 C for 30 sec and primer extension at 72 C for 60 sec. Next twenty
cycles consisting of denaturation at 95 C for 60 sec annealing at 58 C for 30 sec and primer extension at
72 C for 60 sec. Final extension was done at 72 C for 5 min.
Detection OF Transgene BY Southern Hybridization
Genomic DNA was extracted from transgenic cotton plant by the above-described method. The total
DNA (10 g) was subjected to digestion with EcoRI restriction enzyme. Following electrophoresis, the
DNA was stained with ethidium bromide and examined under UV light. The digested DNA fragments
were transferred on positively charged nylon membrane (Biodyne B, Pall Gelman, USA) by alkaline
transfer using 0.4N NaOH, as transfer solution (Sambrook et al., 1989). The cry1Ac gene fragment (1 kb)
present in plasmid pGemT-cryAc-1 (PCR amplified cry1Acgene fragment cloned in 3.0 kb plasmid
pGEM-T marketed by Promega, USA) was used as DNA probe for Southern hybridization. The fragment
was excised from pGemTcryAc-1 and pGEM-T chitinase (4.0 kb) with PstI and SacII and purified using
gel extraction kit (Qiagen, Germany) as per manufacturers protocol. Probe was labelled with
digoxigenin DNA labelling and detection kit (Roche, Germany) following manufacturers protocol.
Southern blot containing the genomic DNA was hybridized to DIG labelled cry1Ac probe and Chitinase
probe at 68C for 24 h in hybridization oven. The hybridized probes were immuno-detected on the nylon
membrane in a visual reaction catalyzed by anti-DIG-AP conjugate in the presence of chromogenic
substrate NBT/BCIP, following manufacturers protocol.
Expression of Cry I Ac Gene
Two discs were excised from terminal leaf of the putative transformants and crushed in 0.5 mL extraction
buffer in a 1.5-mL eppendorf tube with the help of micro-pestle. Qualitative expression of CryIAc protein
in Bt-transgenic plants was ascertained by appearance of a purple band on Bt-express immune detection
strips, while quantification of insecticidal protein in the sample was done using Bt Quant ELISA kit
(Kranthi et al., 2005). In general, the plants found positive for expression of Bt protein in qualitative
assay were further analyzed for quantitative expression by enzyme-linked imunosorbent assay (ELISA).
Both assay kits were developed in this institute, while it is commercialized and marketed in India by
Innovative Biosciences Pvt., Nagpur, India.
RESULTS
Out of 3772 explants co-cultivated with Agrobacterium containing Cry 1Ac gene12 plants were
established. The transformation frequency was 3.6%. It was observed that during first and second cycles
of passage, large number of explants survived on kanamycin selection medium but during subsequent
passages, drastic reduction in the survival of explants was observed (data not shown). The shoots
emerging from the surviving explants though robust, nevertheless, slow growing during initial stages.
These were transferred immediately to fresh MS medium supplemented with 0.1 mg/L kinetin and 500
mg/L carbenicillin. The shoot apices after third cycle of selection were transferred to regeneration
medium. The regeneration was carried out by direct shoot organogenesis and induction of multiple
shoots. In a later case, the meristematic cells (in the axils of cotyledonary node) proliferated within 20
days. The proliferation was accompanied by lateral expansion of the explants accompanied by
degeneration of primary shoot. Within 46 weeks, a huge mass containing large number of shoot buds
differentiated (Fig. 1). The average number of shoots produced per explants was 1012 shoots. The shoot
growth in both the processes of regeneration was slow. The kanamycin-resistant shoots were excised and
transferred on rooting medium after 3 months. In all, 64 independent, putatively transformed shoots were
selected out of 3772 explants used in co-cultivation. Finally only six putatively transformed shoots were
survived at T
0
generation. The rooted plants were liquid-hardened and established in the clay pots
containing sand and soil mixture (1:1) in glass house.
With regard to in-planta transformation, 78 seedling were raised in earthen pots and their shoot tips
were bisected vertically and applied with Agrobacterium suspension premixed with vir induction
medium. Of the 78 seedlings, 50 seedlings survived and reached to maturity and 17 plants were PCR
positive. At flowering, all the PCR positive plants were selfed and seeds were collected to rise T1
generation.
Molecular Characterization of Transformed Plants
Successful transformation was ascertained through PCR amplification of 1 kb DNA fragment of cry1Ac
gene in transgenic plant (fig.2.a). The fragment was not amplified with the gene-specific primer in
negative control. Southern hybridization of genomic DNA of transgenic plant was done by using a 1 kb
DNA fragment of cry1Ac gene as probe. The probe hybridized with genomic DNA fragments of 2.4 and
2.0 kb in Bt-RG8-16, while it hybridized with a single genomic DNA fragment in case of Bt-RG8-16
(fig.2.b), showing them to be the product of two independent transformation events.
In case of Chitinase, out of 50 plants which were survived after transformation 17 were found to be
PCR positive. In all the PCR positive plants there was amplification of 1.3 kb gene fragment (fig.4.a).
Sothern hybridization of genomic DNA of transgenic plants was confirmed by using 1.3 kb DNA
fragment of Chitinase as probe (fig.4.b).
Gene Expression Analysis
The progenies of PCR positive self pollinated T
0
plants were evaluated for expression of insecticidal Cry
IAc protein. The concentration of Bt protein evaluated by Bt ELISA in putatively transgenic plant ranged
from 0.24 to 4.36 g/g fresh weight. The quantitative expression of Bt protein was evaluated for 117 out
of a population of 238 plants at 60 and 120 days of the plant growth. Only 35 plants expressed Bt protein
of 3g/g fresh weight and above, while 47 plants exhibited expression between 1 and 3 g/g fresh
weight.
Transgenic plants containing Chitinase gene, gene expression analysis was carried out in T
1
progenies by fungal spore inoculation method followed by Chitinase activity assay

DISCUSSION
Efficient transformation system is essential for genetic manipulation and improvement of cotton through
genetic engineering. The cotton is highly recalcitrant to regeneration by somatic embryogenesis due to
number of impediment including genotype specificity (Trolinder and Goodin1987). However, cotton
belonging to Coker series of G. hirsutum in general has been found amenable to regeneration by somatic
embryogenesis (Firoozabady et al., 1987; Umbeck et al., 1987; Perlak et al., 1990). Besides Coker,
genotype Acala of G. hirsutum has also been used successfully for genetic transformation (Rajasekaran et
al., 2001). Successful transfer of regenerative trait from such transformed genotypes to recalcitrant elite
cultivar by back-crossing often leads to introgression of undesirable characters (Kumar et al., 1998).
Therefore in the absence of regeneration by somatic embryogenesis, transformation using shoot apices
has been the best choice for gene transfer. Gould and Magallanes-Cedeno(1998) and Zapata et al., (1999)
have developed a method to directly transform elite genotypes of cotton by shoot-tip culture. The
meristem-based method has been successfully used in Agrobacterium-mediated transformation of petunia
(Ullian et al., 1988), pea (Hussey et al., 1989), corn (Gould et al., 1991b), banana (May et al. ,1995), and
rice (Park et al., 1996). Trolinder et al., (2006) used chilled apical shoot tips to develop transgenic plants.
A new method was also developed using a tissue culture independent in-planta protocol which was
standardized to develop transform plants. Such protocols have also been used earlier in crops like
buckwheat (Kojima et al., 2000), mulberry (Ping et al., 2005), kenaf (Kojima et al., 2004), soybean
(Chee et al., 1989) and rice (Supertana et al., 2005). In all these methods, Agro bacterium is targeted to
the wounded apical meristem of the differentiated seed embryo.
In shoot tip-based method seven-day-old in-vitro germinated seedlings were used as source of
explants. The explants are processed so as that maximum number of meristematic cells get exposed to
Agrobacterium, harboring cry1Ac gene during co-cultivation. Healthy seedlings, removal of excess
tissue, and incorporation of acetosyringone in the medium during agro-infection, besides temperature,
were the key factors for successful transformation. Acetosyringone pre-induction of Agrobacterium
and/or inclusion of acetosyringone in the co-cultivation medium can enhance transformation (Veluthambi
et al., 1989; Sunilkumar et al., 1999). Satyavathi et al., (2002) reported high transformation efficiency
above 60% in three Indian varieties, using embryo axis as explants, while Kategari et al., (2007) obtained
transformation frequency of only 0.2% in Bikaneri Narma, using embryo axis.
Selection of explants on kanamycin medium is an important initial step in the development of
transgenic plants in meristem-based method of gene transfer. Maintenance and restoration of
meristematic activity of shoot apical meristem is also essential for successful transformation. This can be
accomplished by adding low level of kinetin in the medium. Incorporation of tolerable levels of
kanamycin during initial stages of selection allowed proliferation of greater number of shoots. After
considerable elongation of shoots, level of kanamycin was enhanced to 50 mg/L during subsequent
passages for proper selection of putative transform ants. Rooting was induced in kanamycin-free MS
medium as the antibiotic impeded the development of normal roots. The transgenic plants regenerated
through the procedure described by us were phenotypically normal. The plants were obtained within 6
months, compared to those regenerated by embryogenesis based transformation system, which takes 1
year or more to obtain fertile plants.
In-planta transformation method offered a simple and unique procedure for gene transfer. In that
meristem cells were exposed through vertical bisection. These cells were targeted to Agrobacterium
containing Chitinase gene. The shoot formed from such cells may either be chimeric or completely
transgenic depending on the type of the cells which have been transformed. The transgenic nature of the
plants was confirmed in T
0
and T
1
plant progenies. It has been observed that compared to somatic
embryogenesis and shoot tip-based organogenic method which usually takes longer longer time to obtain
promising transformants. The present in-planta transformation and shoot tip-based organogenic protocol
are simple, less time consuming and potentially useful approach for crops which are recalcitrant to
regeneration. Transformation of the vertical bisected seedling obviates essential tissue culture

requireme
Moreover
Trans
Fig. 1: Transfo
Direct Sh
Regeneration
Fig. 2: a PCR A
49, Bt-RG8
Trans
Develo
ent that nee
r, this method
formation of
ormation of Shoot
hoot Organogenes
of Putative Trans
Amplification of cr
8Lines 12, 16, 18,
cry1Ac Gene
formation of
opment of Biotic
ed good ski
d eliminates
f CryIAc gen
ttip Explants of G.
is. A In vitro Germ
formed Shoots on
ry1Ac Gene Fragme
37, 59, and 82. b
e as DNA Probe. M
f Chitinase g
c Stress Resista
ill, expense
chances of s
ne in G. arbo
Arboreum cv. RG8
minated Seedling o
n MS Medium Conta
Plant In vitro. g. E
ent in Transgenic
Southern Hybridiz
M, -HindIII Ladder
gene in G. ar
ance Transgenic
and labour
somaclonal v
oreum cv. RG
8 by Agrobacteriu
f cv RG 8. b Shoot
aining Kanamycin.
Establishment of T
G. arboreum cv. R
zation of Genomic
r; 2, Transformed P
rboreum cv. P
c Diploid Cotton
r to produce
variation indu
G8
m Containing cry1
t Tip Explants c. Sc
e. Multiple Shoot
Transformed Plant
RG8. M,1 kb ladder;
DNA of Transgenic
Plant, 4, CryIAc Po
PA255
n Utilizing Agrob
e handful o
uced through
Ac Gene and Rege
creening of Explan
Induction f. Regen
t
1, Positive Contro
c G. arboreum cv.
ositive, 6, Non-Tra
bacterium
of transform
h in vitro cult

eneration of Trans
nts on Kanamycin M
neration of Putativ
ol; 2, Negative Con
RG8 Cotton using
ansgenic ve.
125
med plants.
ture.
genic Plant by
Medium. d.
e Transformed

trol; 3, Non-Bt;
g Fragment of
126
F
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Cloning and Characterization of Cellulose Synthase

Genes from Arabidopsis thaliana
Balasubramani G., Amudha J., Sahare S. and Kranthi K.R.
Central Institute for Cotton Research, P.B. No. 2. Shankar Nagar P.O. Nagpur, India
E-mail: bala77bio@gmail.com
SUMMARY
Cellulose allied genes viz., Rsw1 and AthA from Arabidopsis, which brings about rapid conversion of
carbon to UDP glucose to facilitate the synthesis of cellulose and / or callose as energy-efficient process.
The full length sequence of AthA and Rsw-1 genes were amplified from A. thaliana. The amplified
product was characterized as 5.3kb and 6.kb for AthA and Rsw-1 respectively. The amplicons were
cloned into pJET 1.2/blunt vector and transformed into E. coli (DH5 Alpha). The clones were confirmed
by restriction analysis and PCR. The positive clones were subjected to nucleotide sequence analysis.
Since the sequence was colossal we used primer walking method to obtain full length sequence
information and then aligned with help of bioinformatics tool BLAST-Align. The results yielded the full
length sequence of AthA and rsw-1 and BLAST analysis showed that homology the AthA gene has 99%
with CesA2 (AthA) gene and the sequence was submitted to GenBank (Acc. No. FJ687279). In addition
to these any successful fiber specific expression and localization demands delineation of promoters that
would facilitate expression of gene in secondary wall synthesis (16-40DPA) phase. Thus, fiber specific
promoter was isolated from cotton. Inverse primers were designed to amplify upstream regulatory region
in G. hirsutum cultivar, which yielded ~1.8kb amplicons. The amplicon was cloned and sequenced by
primer walking and homology results showed the similarity with CesA4 promoter region. The nucleotide
sequence was annotated and submitted to GenBank (Accession No. HM142347). This loom for
enhancing high fibre strength coupled with other desirable traits genotypes through transgenic approach
to meet the requirement of modern textile industry.
INTRODUCTION
Cotton fibers provide humankinds most important renewable textile fiber. Approximately 100 million
families rely on cotton production as a major source of their income and 150 countries either import or
export cotton lint (Chen et al. 2007). The rapid increase of the worlds population and the severe
decrease in arable land raise serious concerns about our continued ability to meet global demands for
cotton. It is even more challenging to improve both yield and fiber quality simultaneously. The yield of
cotton fibers known as cotton lint, is usually negatively associated with fiber quality (Meredith, 1984).
With the modernization of textile industry and high speed spinning technology strong and long fiber is in
high demand. Mills using open-end rotor and friction spinning have given improved fiber strength as
highest priority. Plant cell shape is a key determinant in plant morphogenesis and is in turn strongly
influenced by the organization of the cell wall. Within the cell wall, cellulose microfibrils constitute the
major load bearing structures, and their oriented deposition confers differential extensibility to the wall
(Carpita and Gibeaut, 1993; McCann and Roberts, 1994). Presumably under the influence of the
microtubular cytoskeleton, microfibrils in the primary wall of growing cells are deposited
perpendicularly to the prospective elongation axis (Wyatt and Carpita, 1993; Wymer and Lloyd, 1996).
Biologically, cotton fiber is an excellent model system for the study of plant cell elongation and cell wall
and cellulose biosynthesis (Kim and Triplett, 2001). The >90% cellulose content in the secondary wall
and the >2.25 cm fiber length of domesticated cotton fiber are key characters that have made this unusual
seed epidermal hair especially useful to humans for over 7 000 years (Ryser 1985; Dillehay et al. 2007).
23
The fiber is composed of nearly pure cellulose, which is also the largest component of plant biomass.
They differentiate over a period of 4560 days, beginning with expansion of individual cells above the
plane of the ovule outer integument near the day of flower opening. Cellulose, a paracrystalline form of
H-bonded - (1, 4)-Glc chains (McCann and Roberts, 1991; Carpita and Gibeaut, 1993), is thought to be
synthesized by membrane-bound complexes. Cellulose synthase genes have been eventually discovered
genetically in bacteria, and divergent homologs in plants have been identified (Saxena and Brown, 2000).
Cellulose biosynthesis in plants has gained ground during recent past. The isolation of plant cDNA
clones encoding cotton homologs of the bacterial cellulose synthase catalytic subunit is a significant
achievement, which promises the elucidation of cellulose biosynthesis. Two cDNA clones of the bacterial
celA genes that encode the catalytic subunit of cellulose synthase are GhCesA-1 and GhCesA2 that are
highly expressed at the onset of secondary wall when the rate of cellulose synthesis in vivo raises over
1000 fold. GhCesA-1 and GhCesA-2 genes have been believed to possess four conserved sub domains
critical for catalysis and / or binding of substrate UDP glucose new insights into biochemistry of
cellulose biosynthesis and identifying of different subunits of GhCesA-1 and GhCesA-2 complex are
likely to stem from Arabidopsis cellulose biosynthetic mutants such as radial swelling1 (rsw1) and their
homologues of cotton.
Recent investigation by Betancur et al (2010) reported that 10 member cellulose synthase (CesA)
gene family in Arabidopsis provides a useful reference point for comparing cell wall biosynthetic
processes between cotton fibers and Arabidopsis shoot trichomes (Somerville 2006). Although their
precise role in the biochemical pathway of cellulose synthesis is still undefined, the cellulose synthases
are UDP-glucose: 1, 4--D-glucosyltransferase enzymes in the glycosyltransferase family-2. In
Arabidopsis, members of a triplet of CesA isoforms (AtCesA1, 3, 6, or a 6-like protein) have non-
redundant roles in primary wall cellulose synthesis (Persson et al. 2007; Desprez et al. 2007), whereas
members of another triplet (AtCesA4, 7, 8) have non-redundant roles in secondary wall cellulose
synthesis in xylem cells (Taylor et al. 2003) For convenience, the six clades that Arabidopsis CesAs form
with their orthologs from other seed plants have been designated P1 (Primary1), P2, and P3 (defined by
AtCesA1, 3, and 6, respectively) and S1 (Secondary1), S2, and S3 (defined by AtCesA 4, 7, and 8,
respectively) (Haigler and Roberts 2009). The remainder of the ten Arabidopsis CesA genes appear to be
relatively recently derived within P1 (AtCesA10) and P3 (AtCesA2, 5, and 9) in the crucifer lineage. The
Arabidopsis members of P3 are functionally interchangeable (Persson et al. 2007; Desprez et al. 2007)
and AtCesA10 has limited expression (Doblin et al. 2002). Based on available evidence, the division of
function between these traditionally defined primary- and secondary wall CesAs is broadly conserved in
other angiosperms even if the number of CesA genes has increased (Tanaka et al. 2003; Djerbi et al.
2005; Kumar et al. 2009). Other genes/proteins may be required for both primary and secondary wall
cellulose biosynthesis, e.g. an endo-glucanaseKorrigan, or alternatively may parallel the CesA family in
having isoforms specialized for primary or secondary wall biosynthesis (Somerville 2006). In the present
investigation full length sequence of AthA and Rsw-1 genes were amplified, characterized and cloned
from A. thaliana. We also successfully cloned and characterized fiber specific promoter belonging to
CesA4 gene family of cotton.
Plant Material
Arabidopsis thaliana (Thale cress), Columbia plants were established initially in pots containing an
autoclaved 1:1:2 mix of peat: soil: sand, (v/v) and later raised at in vitro condition using Murashige and
Skoog (1962) medium containing 0.75% (w/v) agar. Plants were cultured in a photoperiod of 16 hr light
and 8 hr dark cycle. After 12 days of growth seedlings were taken aseptically for DNA isolation.

Cloning and Characterization of Cellulose Synthase Genes from Arabidopsis thaliana 131
Genomic DNA Isolation
Total genomic DNA was isolated by Dellapotta et al. (1983) method using fresh healthy young leaves of
A. thaliana. The DNA was purified with Plant DNA purification kit (Sigma). The genomic DNA was
checked quantitatively and qualitatively by Qubit Fluorometer and agarose gel electrophoresis
respectively.
Primer and PCR Amplification
Amplification of the target genes was carried out using forward and reverses primer designed with
software FastPCR. The forward sequence 5- ATTGTCGATTCGGTTTATTTCGT-3 and reverse
sequence 5- ACAAAGAAGGTGTAAAACAACAC -3 was synthesized by MWG, Bangalore. The
purified genomic DNA was used for target gene amplification. PCR was performed in 20 l reaction
mixture containing 1.0 l of 100 ng DNA, 4.0 l 5 fusion buffers (HF), 1.0 l of 10 mM dNTPs, 1.0 l
of 100 nM of each forward and reverse primer, and 0.2 l of 1U Pfu DNA polymerase for larger DNA
fragment (>10kb). PCR program was standardized using TProfessional Gradient Thermal cycler
(BIOMETRA
). The following PCR conditions of 98C for 30 seconds, followed by 35 cycles of 98C
for 10 seconds, 57.8C for 30 seconds, 72C for 2 min and 10 min of final extension at 72C were used.
The amplified products were electrophoresed on 1.5% w/v agarose gel and documented.
Cloning of Amplicons
The amplicons were cloned into pJET 1.2/blunt vector and transformed to host bacteria (DH5). The
transformed bacteria were screened in the selection media containing IPTG (isopropyl beta-D-
thiogalactopyranoside) with x-gal (5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside) as per the
supplier protocol. The positive bacterial colony was selected for sequence analysis. The white colony was
demonstrated as recombinant bacteria.
Amplification of Promoter
A detailed understanding of the key steps in the fiber development process provides opportunities to
improve fiber quality either by molecular breeding or by genetic engineering technology. The second
technology uses extremely important tools such as cloning the targeted nucleotide sequences, making
desirable gene cassette and transfer to the targeted cells etc. For expressing a new character (gene) in
cotton fibers there is great need of a module (promoter) in expression cassette (transgene construct)
which will specifically express the character in cotton fiber cells. Therefore simultaneously, the upstream
region of fiber gene was isolated and the sequence information confirmed that the fiber specific region
belonging to CesA4 gene family.
Genomic DNA Isolation
DNA isolation was carried out using 20DPA boll of G. hirsutum cultivars LRA -5166, following
Paterson et al (1993) method and purified as mentioned above.
Primer and PCR Amplification
The cellulose synthase gene was used to design primer sequence to amplify upstream region i.e.
regulatory sequence by inverse primers. The primers were designed using FastPCR s/w. The forward
sequence 5-GCTATGTGGTAGGGACAATCTGGTC-3 and reverse sequence 5-
CTCAATACCTTTGTGTCTTGTTGTG-3 was synthesized by MWG, Bangalore. The genomic DNA
was restricted with stringent reaction of Hind-III restriction digestion and allowed for circularization of
DNA using ligase reaction. The ligated reaction was used for amplification of upstream region. Gradient
PCR (Biometra) was performed with reaction mixture components (dNTPs, MgCl, Taq polymerase,
forward and reverse 2 primers, ddH
2
O) and genomic DNA. The amplified product was characterized and
the expected size amplicon was eluted and reamplified with primers. Then the amplicon was eluted from
the gel using Roche agarose gel extraction kit and further subjected to cloning and transformation using
kit. The transformed single colony was sub-cultured and pure recombinant bacteria were subjected to
sequence analysis. After sequencing the nucleotide were BLAST searched for sequence homology and
annotated sequence was submitted to GenBank.
Cotton fibers are a type of trichome and researchers have frequently looked for analogies with vegetative
trichomes, especially those in the model plant Arabidopsis. An alternative model cell type for
investigating the function of genes expressed in cotton fiber would be particularly useful given the
substantial time and resources required for stable cotton transformation. The term trichome denotes a
filamentous outgrowth such as a plant epidermal hair. Trichome now refers to outgrowths of the plant
epidermis exclusive of cells of sub-epidermal origin; mainly leaf and stem (shoot) trichomes, cotton seed
hairs (fiber), and root hairs. These morphologically diverse cells and multicellular structures have been
shaped by natural selection for a wide range of functions, including water absorption, thermoregulation,
UV protection, both repellent and attractant components of defence, and seed dispersal (Evert 2006). The
seed epidermal trichomes that presently dominate world cotton commerce were derived through
domestication of wild cotton species with ancestral fibers that probably evolved to aid seed dispersal
and/or germination (Wendel et al. 2009).
In the present investigation, the cellulose synthase genes and fiber specific promoter were cloned and
sequence information was established. Expressive gene specific primers were designed and amplified
directly from the total genomic DNA of Arabidopsis thaliana. The full length sequence of AthA and Rsw-
1 genes were isolated from Arabidopsis thaliana by using high fidelity Pfu polymerase. The amplified
product was characterized as 5.3kb and 6.kb for AthA and Rsw-1 respectively (Fig-1). The amplicons
were cloned into pJET 1.2/blunt vector and transformed to host bacteria (DH5) and positive colony was
selected after 24hrs of culture (Fig-2). The clones were confirmed by restriction analysis and PCR. The
positive clones were subjected to sequence analysis. Since the sequence was colossal nature, primer
walking method was used to obtain full length sequence information and then Aligned with help of
bioinformatics tool (BLAST-Align). The sequence information showed that the AthA has homology of
99% with CesA2 (AthA) gene and the sequence was submitted to GenBank (Acc. No. FJ687279). The
other cellulose biosynthetic gene such as radial swelling1 Rsw-1 gene specific primers amplified the full
length sequence and the homology analysis showed similarity with rsw1 gene and the sequence was
annotated and submitted to Gen Bank.

Fig. 1: PCR Amplified Amplicons Were Resolved in Agarose Gel Showing 6.0kb and 5.3kb Fragments. Lane 1:
1kb Ladder, 2: RSW-1 Amplicon and 3: AthA Amplicon

Fig. 2: The Amplicons Were Cloned Into pJET 1.2/blunt Vector and Transformed to Host Bacteria (DH5). White Colonies are Transformed E.coli

Fig. 3: PCR Amplified Amplicons from LRA 5166 Cotton Cultivar was Resolved
in Agarose Gel Showing 1.8kb Fragments. Lane 1: Cesa Amplicon, 2: 1kb ladder
Concurrently, to express these cellulose biosynthetic genes in tissue specific, cotton fiber specific
promoter was isolated. Inverse primers were designed to amplify upstream region of G. hirsutum
cultivars. Well characterized single amplicon of PCR product was selected. The size was found ~1.8kb
amplicon (Fig-3). The amplicon was cloned and sequenced by primer walking and homology results
showed the similarity with CesA4 promoter region. The nucleotide sequence was annotated and
submitted to GenBank (Accession No. HM 142347).
Assessing the function of individual CESA genes will require the identification of the null-mutant
phenotypes and of the gene expression profiles for each gene (Delmer, 1999). Beeckman et al., 2002
reported four of 10 CESA genes, CESA1, CESA2, CESA3, and CESA9 are significantly expressed in the
Arabidopsis embryo. They further identified two new mutations in the RADIALLY SWOLLEN1
(RSW1/CESA1) gene of Arabidopsis that obstruct organized growth in both shoot and root and interfere
with cell division and cell expansion already in embryogenesis. In our study we identified similar full
length sequence of RSW1/CESA1 gene. In the embryo, CESA1, CESA2, CESA3, and CESA9 are
expressed in largely overlapping domains and may act cooperatively in higher order complexes. The
embryonic phenotype of the presumed rsw1 null mutant indicates that the RSW1 (CESA1) product has a
critical, nonredundant function, but is nevertheless not strictly required for primary cell wall formation as
reported by Beeckman et al., (2002).
In all species, cellulose synthase catalytic subunits, encoded by CesA genes, share a common
structure that includes several putative transmembrane helices, and a cytoplasmic loop containing four
conserved regions (U1 through U4) predicted to be involved in substrate binding and catalysis (Delmer
1999). CesACesA interactions (Taylor et al. 2000, 2003; Kurek et al. 2002; Gardiner et al. 2003) have
revealed that CesA proteins also play a role in maintaining the association of the particles that compose
terminal complexes (Doblin et al. 2002).Divergence in both terminal complex organization and CesA
gene sequence among groups of organisms in which both have been characterized also indicates that
terminal complex organization, and thus microfibril dimensions, are influenced by CesA structure.
However, the evolutionary relationships among these organisms, which include seed plants (Delmer
1999). Although seed plants are thought to have only rosette terminal complexes, those species that have
been examined have numerous CesA genes.
In A. thaliana, analysis of mutant phenotypes and gene expression have revealed that some of the 10
members of the CesA gene family serve distinct functions in primary and secondary cell wall
synthesis(Taylor et al. 1999; Fagard et al. 2000; Holland et al. 2000; Scheible et al. 2001). However,
some groups of AtCesAs appear to have identical expression patterns and genetic complementation and
co-precipitation experiments have indicated that up to three distinct CesA subunits are required for
assembly of a functional rosette (Taylor et al. 2000, 2003; Scheible et al. 2001; Burn et al.2002; Desprez
et al. 2002; Doblin et al. 2002;Gardiner et al. 2003). Expression analysis revealed similar patterns in
other seed plant species (Tanaka et al. 2003; Burton et al. 2004; Liang and Joshi 2004). Furthermore,
CesA are members of a larger gene superfamily that includes the CesA-like (Csl) genes, some of which
may function in the synthesis of non-cellulosic cell wall polymers (Richmond and Somerville
2001;Dhugga et al.,2004). Thus, the differences between CesAs specialized for primary and secondary
cell wall deposition could relate to the ways in which they interact with each other or the cellular
structures that control terminal complex localization or movement.
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Cotton Transgenic with DRE-binding Transcription

Factor Gene (DREB1A) and Zinc Finger Gene (ZF1)
Confers Enhanced Tolerance to Drought
Amudha J., G. Balasubramani, A.H. Prakash, Shweta C.
K.C. Bansal and K.R. Kranthi
Central Institute for Cotton Research, Post Bag No. 2,
Shankar Nagar440 010, Maharashtra, India
E-mail: jamudhacicr@gmail.com
AbstractDrought tolerant transgenics were developed in elite genotypes LRA 5166 and LRK 516 with Prd29:
AtDREB 1A and PLEA1:Bc ZF1 gene constructs through Agrobacterium mediated transformation. Embryonic axes
were excised from 48 hr grown cottonseeds used as explants and the shoots were screened in the MS medium with 50
g/ml kanamycin. The regenerated shoots were transformed in the MS medium containing auxin 1mg/l and cytokinin
1mg/l. The transgenics were confirmed for the presence of the gene by PCR using specific primers for DREB 1A, ZF1,
npt II and the copy number by Southern analysis. RT-PCR study was carried by isolating mRNA from transgenic
plants and cDNA was synthesized and amplified with npt II primer and DREB 1A gene specific primer produced the
700 bp and 540 bp fragments. T
o
plants were hardened in the polyhouse. The transgenics subjected for drought
tolerance, physiological and biochemical studies viz., PEG (polyethylene glycol), proline, reducing sugar, amino acid,
protein and phenols at regular interval.
INTRODUCTION
Environmental abiotic stresses such as drought are major limiting factors that influence plant growth and
crop production. Abiotic stress mediated gene expression has been shown to be regulated by different
transcription factors of which, Dehydration responsive element binding (DREB 1A) protein plays a key
role. The transcription factors DREB1A are important in the ABA-independent drought tolerant
pathways that induce the expression of stress response genes. DREB transcription factors bind to
dehydration responsive element (DRE/CRT) of genes at promoter region. DRE contains one sequence
A/GCCGAC which was identified as cis acting promoter element which regulates the expression of
downstream genes. It is involved in the transcriptional regulation of a dynamic network of genes
controlling various biological processes, including abiotic and biotic stress responses and finally
enhances plant stress tolerance.
Responses to abiotic stress require the production of important metabolic proteins such as those
involved in synthesis of osmoprotectants and of regulatory proteins operating in the signal transduction
pathways, such as kinases or TFs. Given that most of these responses imply control of gene expression,
TFs play a critical role in the abiotic stress response (Chaves and Oliveira, 2004). TFs are proteins with a
DNA domain that binds to the cis-acting elements present in the promoter of a target gene. They induce
(activators) or repress (repressors) the activity of the RNA polymerase, thus regulating gene expression.
Transcription factors play critical roles in the regulation of cellular and physical changes in response to
environmental stresses in plants. The C-repeat binding factor/dehydration responsive element binding
factor (CBF/DR0EB), NAM, ATAF, and CUC (NAC) transcription factor, zinc finger protein and other
transcription factors have been described as important regulators in plant responses to environmental
stresses. TFs can be grouped into families according to their DNA binding domain (Riechmann et al.,
2000). A group of genes controlled by a certain type of TF is known as a regulon. In the plant response to
abiotic stresses, at least four different regulons can be identified (1) the CBF/DREB regulon; (2) the
NAC (NAM, ATAF and CUC) and ZF-HD (zinc-finger homeodomain) regulon; (3) the AREB/ABF
(ABA-responsive element-binding protein/ ABA-binding factor) regulon; and (4) the MYC
(myelocytomatosis oncogene)/MYB (myeloblastosis oncogene) regulon. The first two regulons are ABA
24
Cotton Transgenic with DRE-binding Transcription Factor Gene (DREB1A) and Zinc Finger Gene (ZF1) 137
independent, and the last two are ABA dependent. It is explained below how these regulons are
controlled and how TFs may be involved in the regulation of photosynthesis as a response to abiotic
stress. The C2H2-type zinc finger proteins have a variety of roles in plant reproductive development and
abiotic stress responses (Ciftci-Yilmaz et al., 2008; Huang et al., 2007). Although their functions in stress
responses are still largely unknown, a number of stress-responsive C2H2 zinc finger proteins have been
identified in various plant species (Huang et al., 2007). In our study we have gene constructs with two
regulons, the CBF/DREB and ZF-HD (zinc-finger homeodomain regulon).
The CBF/DREB regulon is mainly involved in cold stress response. It is conserved throughout the
plant kingdom, including in plants that do not cold-acclimate (e.g. cotton, tomato and rice) (Dubouzet et
al., 2003). In 1994, Yamaguchi-Shinozaki and Shinozaki identified a novel cis-acting element that, in
addition to the ABA-responsive element (ABRE), is also present in the promoter of the RESPONSIVE
TO DEHYDRATION 29A (RD29A), a gene induced by drought, high salinity and cold. This new
element was named C-repeat/dehydration-responsive element (CRT/DRE) and characterized as ABA
independent. The core motif of this cis-acting element is CCGAC and the TFs that bind to it were named
CRT-binding factors or DRE-binding proteins 1 (CBF/DREB 1) (Gilmour et al., 1998; Liu et al., 1998).
CBF/DREB 1 gene expression is quickly and transiently induced by cold stress, and in turn CBF/DREB1
TFs activate the expression of several other genes (e.g. encoding proteins involved in production of
osmoprotectants and antioxidants). The over-expression of CBF/DREB1 genes in Arabidopsis resulted in
plants with improved survival rates when exposed to salt, drought and low temperatures (Jaglo-Ottosen et
al., 1998; Kasuga et al., 1999). This improved tolerance was correlated with both altered relative
abundance of transcripts encoding proteins associated with stress adaptation and increased sugar contents
(Gilmour et al., 2004). When CBF/DREB1 genes from Arabidopsis were over-expressed in other plants,
the result was similar to that in Arabidopsis (Hsieh et al., 2002a; Pellegrineschi et al., 2004), revealing a
conserved signaling and response mechanism even between dicots and monocots.
Various studies have demonstrated that improved stress tolerance by over-expression of
CBF/DREB1 genes is associated with sustained photochemical efficiency and photosynthetic capacity as
compared with wild-type plants (Hsieh et al., 2002b; Savitch et al., 2005; Oh et al., 2007). To overcome
growth retardation, CBF/DREB1 genes have been expressed in transgenic plants under the control of a
stress-inducible promoter, RD29A (Kasuga et al., 2004). These plants have also shown enhanced abiotic
stress tolerance without totally compromising the yield (Pino et al., 2007). However, it seems that the use
of the Arabidopsis RD29A promoter is more efficient in driving the expression of CBF/DREB1 genes in
dicots rather than in monocots, or at least in rice (Ito et al., 2006).
DREB1A and Drought Tolerance
Water deficit stress that known as drought stress has many physiological effects on the plants; include
reduction in vegetative growth, leaf expansion and transpiration. As a secondary effect, ABA
concentration has increased and stomata close to prevent the transpirational water loss that results in
limited photosynthesis due to decline in Rubisco activity (Bota et al., 2004). On the other hand, decline
in intracellular CO
2
levels results in the generation of ROS components that they may lead to photo-
oxidation and finally they can cause extensive peroxidation and de-esterification of membrane lipids, as
well as lead to protein denaturation and mutation of nucleic acids (Bowler et al., 1992).
On the cellular level, removal of water from the membrane disrupts the normal bilayer structure and
results in membrane damage, displacement of membrane proteins and reduced activity ion transporters or
may even complete denaturation of enzymes and cytosolic and organelle proteins, that those can cause
disruption of cellular metabolism. Plant tend to protection of the membranes as well as macromolecules
by synthesis of large number of osmolytes compounds such as proline, glutamate, glycine-betaine,
carnitine, mannitol, sorbitol, fructans, polyols, trehalose, sucrose, oligosaccharides and inorganic ions
like K
+
(Ramanjulu and Bartels, 2002). The primary function of compatible solutes is to maintain cell
turgor and thus the driving gradient for water uptake. Some studies indicate that compatible solutes can
also act as free-radical scavengers or chemical chaperones by directly stabilizing membranes and/or
proteins (Diamant et al., 2001, Garg et al., 2002).
Analyses of the expression of dehydration-inducible genes have shown that at least four independent
signaling pathways function in the induction of stress-inducible genes in response to dehydration: two are
ABA dependent (Uno et al., 2001; Velasco et al., 1998) and two are ABA independent (Savoure et al.,
1997). Dehydration responsive element (DRE) or C-repeat (CRT), a cis-acting element, plays an
important role in regulating gene expression in response to stress in an ABA independent manner
(Yamaguchi-Shinozaki et al., 2002). DRE/CRT exists in many dehydration responsive genes and
DREB1A, as a DRE/CRT-binding transcription factor, can up-regulate them. Consequently, over-
expression of DREB1A gene will significantly introduce expression of dehydration responsive genes and
it can results in high tolerance to drought stress in transgenic plants.
Seeds of cotton variety LRA 5166 and LRK 516 were obtained from the gene bank, Central Institute for
Cotton Research, Nagpur.
Bacterial Strain and Vector
Agrobacterium mediated transformation with pCambia 2300 - Prd29: DREB1a: nos and pBinAR
PLEA 1: BcZF1 : nos gene constructs were used for transformation (Fig 1 & 2). Both the constructs has
npt II gene as selection marker in the T-DNA driven by Cauliflower mosaic virus (35S CaMV) promoter
and NOS-terminator in the vector system. Bacterial culture was maintained on YEMA medium (1.0%
w/v Yeast extract, 1.0%w/v Mannitol, 0.1%w/v Sodium chloride, 0.2% w/v Magnesium sulphate, pH-
7.0) containing 50mg/l kanamycin and 25mg/l rifampicin. For inoculation, one single colony was grown
overnight on liquid YEMA at 28C with appropriate antibiotics.
Transformation of Cotton Plants
Cotton variety LRA 5166 and LRK 516 seedlings were raised aseptically on half- Murashige and Skoog
(MS) medium. The embryonic axes were excised and trimmed from both the sides and used for co-
cultivation with A.tumefaciens. The explants were co-cultivated in the half-MS liquid medium with
actively growing culture of A.tumefaciens at 1.0 OD and 100mM acetosyringone. After overnight co-
cultivation, shoots were decontaminated in the half MS medium containing cefotaxime 250 mg/l. The
explants were then transferred to selection medium containing kinetin 0.1mg/l, BAP 0.1mg/1 and
kanamycin 50mg/l. The kanamycin resistant shoots were sub-cultured in a media containing 0.1 mg/l
BAP for root induction. Rooted plants were rinsed well and transferred to pots containing peat, soil and
sand in 1:1:1 ratio. Plants were covered with plastic bags and then to a pot with soil for hardening for 15
days before transferring to the greenhouse under natural condition. T
1
plants were raised in the green
house.
Screening for Transformed Plants using PCR
Genomic DNA was isolated by the method of Paterson et al.,(1993) from the young leaves of T
0
plants
grown in the Polyhouse. The template DNA was used for PCR amplification with the DREB 1A gene
specific primer (5-3) F-AAG AAG TTT CGT GAG ACT CG and R- CTTCTGCCATATTAGCCAAC,
ZF1 gene specific primer (5-3) F- ATG GTT GCT ATT TCA GAG ATC and R- TAA CCT TCT GGA
CAA ACA ACT and npt II specific primer F-GAGGCTAATTCGGCTATGACTG and R-
ATCGGGAGAGGCGATACCGTA was carried out to check the presence of the transgene. PCR was
performed in 20l (total volume) reaction mixture containing 1.0 l of 100ng DNA, 2.0 l 10X reaction
buffer, 2.0 l of 10mM dNTPs, 1.0 l of 100nM of each forward and reverse primer, 3.0 l of 25mM
MgCl
2
and 0.5 l of 1U of Taq DNA polymerase. The following PCR conditions of 94C for 5min, then
35 cycles of 94C for 30sec, 56C for 1min, 72C for 1 min and 5 min of final extension at 72C was
maintained in a thermo cycler (BIOMETRA). The amplified products were resolved on 1.5 % w/v
agarose gel and documented.
RT-PCR
The mRNA was isolated using mRNA capture kit (Roche, Germany) from young leaves of T
0
plants. The
mRNA was used for cDNA synthesis by Transcriptor high fidelity c DNA kit (Roche, Germany).
SOUTHERN HYBRIDIZATION OF TRANSFORMED PLANTS
To confirm the gene integration Southern blotting method was used in the transgenic plants (T
0
plants).
Genomic DNA was isolated from the leaves of the T
0
plants. For Southern hybridization 10g of total
genomic DNA from the putative transgenics was digested with Bam HI and resolved on 0.8% agarose.
The probe was labeled with non- radioactive DIG labeling kit (Roche, Germany).
RNA DOT BLOT ANALYSIS
The independent events of T1 plants were grown in the poly house for best event selection and gene
expression analysis under drought stress condition. RNA was isolated from the young leaves of the
independent transgenic events and blotted on the nitrocellulose membrane and with the specific DREB
1A and ZF 1 probe.
PHYSIOLOGICAL STUDIES
Transgenic events of LRA 5166 and LRK 516 with DREB 1A and BcZF1 resp. were subjected for
drought tolerance physiological studies. Moisture was withheld after 75 days after transplanting and
plants started growing under receding moisture. The reducing sugars, amino acids and proline (Bates et
al., 1973) were quantified at regular interval using standard procedures. The leaf discs from non-stressed
plants of both transformed and non-transformed were placed on PEG medium with varying degrees of
stress (0, 0.4, 0.6, and 0.8 MPa). The biochemical changes induced due to stress was quantified at 0, 7 &
15 days after inoculation.
Plant Transformation and Regeneration
Cotyledons of 2-3 day old were trimmed and embryonic axes measuring about 5-10 mm was used for co-
cultivation. After co-cultivation for overnight the explants were decontaminated with Carbenicillin
50ug/50 ml. The explants were inoculated and selected in MS medium with kanamycin 50 mg/l as
selection marker which allows only the transformants to grow. The shoot induction was observed after 10
-15 days in the MS medium. The putatively transformed shoots were sub-cultured twice in the shooting
medium and then transferred to rooting medium after shoots attained a height of 5-6 cm. Rooted plants
were rinsed well and transferred to pots containing peat, soil and sand in 1:1:1 ratio . The number of
embryonic axes used for the experiment and the transformation frequency was given in the table 1. T
1

plants are grown in the green house under controlled condition.
TABLE 1: TRANSFORMATION FREQUENCY OF THE PUTATIVE TRANSFORMANTS
Genotypes Gene Construct Number of
Explants Used
Putative
Transformants
PCR Positive
for npt II
Transformation
Frequency (%)
LRA 5166 At DREB 1A 450 45 4 0.88
LRA 5166 PLEA1:BcZF1 650 50 6 0.92
LRK 516 At DREB 1A 545 46 4 0.73
LRK 516 PLEA1:BcZF1 550 46 5 0.90

PCR Analysis of the Transgenic Plants
The PCR analysis with the npt II gene and DREB 1A, ZF 1A gene specific primers was carried out with
the genomic DNA as template of individual plants (Fig 3&4). The independent transgenic events positive
for npt II gene and DREB 1A gene is given in the Table 2.
TABLE 2: TOTAL NUMBER OF TRANSGENICS EVENTS POSITIVE FOR AT DREB 1A AND BCZF1 GENE.
Sr. No Variety Gene Construct No. of To Plants
Survived in Kanamycin
Medium
No. of npt II Positive
Putative
Transformants
No. of
Specific
Positive
1 LRK 516 At DREB 1A 42 38 15
2 LRA 5166 At DREB 1A 17 13 11
3 LRK 516 PLEA1:BcZF1 10 4 3
4 LRA 5166 PLEA1:BcZF1 8 6 5
Southern Hybridization by Non-radioactive Labeling Method
Southern hybridization was carried out in the PCR positive T
0
transgenics by using Roche DIG labeling
kit. Genomic DNA was isolated from the transgenic plants and they were restricted with the BamHI
enzyme. The restricted DNA was resolved on 0.8% agarose gel electrophoresis and then blotted on to
nitrocellulose membrane. The DREB 1A gene was amplified from the gene construct with the specific
primer and eluted after purification and the probe DNA was labeled with the dig labeling kit. DNA
samples of the independent transgenic plants of cotton was digested with BamHI and hybridized with
DREB 1A gene and ZF 1 gene probe has shown the integration of single copy gene ( Fig 5 & 6 ).
RNA Dot Blot Analysis
The independent events of T
1
plants were grown in the poly house for best event selection and gene
expression analysis under drought stress condition. RNA was isolated from the young leaves of the
independent transgenic events and blotted on the nitrocellulose membrane and hybridized with the
specific DREB 1A and ZF 1 probe using Roche DIG labeling kit. The dot blot assay result has shown
positive in the T
1
transgenic plants (Fig 7).
RT-PCR Study
The mRNA from transgenic plants were isolated and cDNA was synthesized using the transcriptor high
fidelity cDNA synthesis Roche kit .The cDNA of the transgenic plants were used for amplification with
npt II primer and DREB 1A gene specific primer. The amplified product was checked by agarose gel
electrophoresis.
PHYSIOLOGICAL STUDIES
Transgenic events of LRA 5166 and LRK 516 with DREB 1A and BcZF1 resp. were subjected for
drought tolerance physiological studies. Sixteen transgenic plants of LRA 5166 and LRK 516 with
DREB 1A and BcZF1 raised in pots along with one control plant of LRA 5166. The plants were grown
under optimum growth conditions till flowering. Moisture was withheld after 75 days after planting and
plants started growing under receding moisture. All physiological and biochemical constituents were
quantified at regular interval. The leaves of both non-transformed and transformed plant started to droop
within seven days after stress induction. The transformed plants would show recovery during night and
leaves were turgid in early part of the day. This helped the plant to recover and grow normally. The leaf
water potential also was maintained at higher level due to accumulation of solutes, which was not
observed in case of control plants.

Over Expression of DREB 1A Improved Drought Tolerance of Transgenic Plants
T
1
transgenic plants at the vegetative stage were deprived of water for 15 days exhibited increased
tolerance to drought stress, compared to control plants. T
o
evaluate physiological changes in two-month-
old transgenic plants deprived of water for 15 days, soluble sugar contents in the leaves of wild type and
different transgenic lines were compared. Under control conditions, the soluble sugar contents in leaves
were similar for wild type and different transgenic plants, that is, around 2.56 mg/g FW. However,
following drought treatment, the soluble sugar levels in transgenic plants increased after 5 days, and
reached 3.64 to 7.01 mg/g FW at 20 days, whereas those in control plants at the same developmental
stage reached only 3.803 g/g FW at 20 days. The amino acid content in transgenic plants after drought
treatment increased after 5 days, and ranged between 0.344 to 0.985 mg/g FW at 20 days, whereas those
in control plants at the same developmental stage reached only 0.750 mg/g FW at 20 days. After drought
treatment, the phenol levels in transgenic plants increased after 5 days, and ranged between 1.879 to
3.498 mg/g FW at 20 days, whereas those in control plants at the same developmental stage reached
only 2.355 mg/g FW at 20 days. The statistical analysis showed that soluble sugar, phenol and amino
acids contents of transgenic plants were significantly higher than those of the control plants at 5, 10, 15
and 20 days (P\0.05)(Table 3).
Analysis of Stress Tolerance and Physiological Changes in Transgenic Cotton by PEG Treatments
In this method of screening, the leaf discs from non-stressed plants of both transformed and non-
transformed were placed on PEG medium with varying degrees of stress (0, 0.4, 0.6, and 0.8 MPa). The
biochemical changes induced due to stress was quantified at 0, 7 & 15 days after inoculation. The data
showed that there was an inherent tolerance developed in transgenic plants due to DERB 1A and BcZF1
genes (Fig 8). The control plants showed a immediate burst in synthesis of reducing sugars, amino acids
and proline but declined by seven days, while the transformed plants showed a gradual increase in solute
accumulation and maintained high even after 7 days. The non-transformed discs produced very high
phenolics which led to death of the tissues with stress. Over expression of DREB 1A and BcZF1
improved drought tolerance in transgenic cotton and after stress alleviation the solute content reached
normal and was similar to control plants.
All three common abiotic stresses, drought, low temperature and high salinity, cause an accumulation
of compatible solutes and antioxidants, such as sugars, proline (Hasegawa et al. 2000; Fukutaku and
Yamada 1984). There are some overlaps in the regulation pathways of gene expression between different
environmental stresses (Rabbani et al. 2003). Because of the various adverse effects of abiotic stresses on
plant growth and productivity, it is very important to improve stress tolerance of the crop plants to
maintain growth and productivity and increase crop yield under stress conditions. Although the tolerance
of plants to abiotic stresses is well known to be a multigenic trait; plant improvement using genes that
play a role in the abiotic stress response is frequently insufficient to improve stress tolerance
significantly. To overcome this, transcription factors that regulate several stress-responsive genes (e.g.
the DREB1s family), often have been used to manipulate plants in order to have a broader response and
maintain the function and structure of cellular components (Saibo et al. 2009).
Under drought temperature conditions, many plants produce osmolytes such as free proline and
various soluble sugars, which may function as osmoprotectants in stress-tolerant plants (Ishitani et al.
1996; Igarashi et al. 1997; Taji et al. 2002). Overexpression of CBF3/DREB1A in transgenic
Arabidopsis plants produced a higher level of free proline than in control plants under stress conditions.
(Gilmour et al. 1998), and over expression of OsDREB1A in transgenic rice plants was associated with
increased soluble sugars levels compared with wild-type plants under drought, high salt and low
temperature conditions (Ito et al. 2006). In our study, rd29A:: DREB 1A transgenic plants accumulated
higher levels of soluble sugar than wild-type plants under drought stress conditions, suggesting that
overexpression of DREB activated the expression of downstream genes involved in sugar biosynthesis,
which in turn, enhanced tolerance to drought stress in transgenic plants. Overexpression of the DREB1A
gene in transgenic Arabidopsis and rice plants activated the expression of many stress-inducible genes
and improved drought, high salt and freezing tolerance (Liu et al. 1998; Kasuga et al.1999, Dubouzet et
al. 2003). Likewise, 35S::DREB1A transgenic cotton plants in our study showed increased tolerance to
drought stress.
In plants, the EAR (ERF-associated amphiphilic repression)-motif was first identified in the C-
terminal region of class II AP2/ERFs and Cys2/His2-type zinc-finger proteins (Ohta et al. 2001). Zinc
finger proteins as members of transcriptional factors were further grouped into the subfamilies of TFIIIA,
WRKY, Dof, LIM, and RING finger. In plants zinc finger proteins are involved in growth, development
and responses to environmental stresses. bZIP proteins contain a region of basic amino acids followed by
a region containing at least three to four repeats of Leu or another hydrophobic amino acid. The
hydrophobic region mediates homo dimer formation, whereas the basic region is involved in DNA
binding. Several zinc finger proteins containing DLN-box/EAR-motifs were revealed to have
transcriptional repression activities by transient analyses in plants, such as petunia ZPT2-3 (Sakamoto et
al., 2004), Arabidopsis STZ/ZAT10 and AZF2 (Sugano et al., 2003). The DLN-box/EAR-motif is not the
only determinant of transcriptional repression activity of C2H2-type zinc fingers. Other unknown
domains may also have an impact on the transcriptional activation/repression activity. Alternatively, the
transcriptional repression activity of C2H2 zinc finger proteins may require post-translational
modifications that are only performed in plants. Overexpression of a petunia zinc finger protein gene,
ZPT2-3, in transgenic petunias increased the plants tolerance to drought stress (Sugano et al., 2003).
When plants suffer from drought stress, the ABA signal transduction pathway is activated and ZFP
accumulates quickly. ZFP promotes the accumulation of free proline by activating the expression levels
of pyrroline-5-carboxylatesynthetase and proline transporter genes, and enhances the ROS-scavenging
ability in plant cells by activating the ROS-scavenging enzymes. Our data suggest that ZFP contribute to
the drought tolerance of transgenic plants by regulating their proline levels and ROS-scavenging abilities
with enhanced tolerance to abiotic stress. In the present study, all the transgenic lines grew normally,
suggesting that DREB genes from a monocotyledon plant might not affect the growth and development
of a dicotyledonous (Chen et al. 2003). Nevertheless, the basis of underlying abnormal growth and
development of transgenic plants with DREB genes might be very complicated and possibly related to a
range of factors, such as gene structure, uncertain insertion sites of the exogenous genes, and instability
of gene expression caused by copy numbers (Finnegan and McElroy, 1994).
DREB1A is a transcription factor that induced by abiotic stress, strongly up-regulates many
downstream genes; result in adaptation of plants to stress conditions and exercise specific tolerance
mechanisms. Many studies indicated that over-expression of DREB1A in crop plants can result in high
tolerance to abiotic stress and thereby increased efficiency of plants production. Taken together suggest
we used gene transfer by DREB 1A and ZF1 strategy to prevent of large and widespread yield reductions
under stress conditions and obtain more products in agronomic development. However DREB1A has a
key role in tolerance to abiotic stress such as drought, salinity, heat and low temperature stresses. Given
that the molecular control mechanisms of abiotic stress tolerance are similar and DREB1A effected on
majority of them, therefore it is possible that over-expression of DREB1A gene leads to tolerance in
transgenic plants. It is hoped in the future that these efforts will help to prevent global-scale
environmental damage that is resultant from these stress.
L- 100
516 (ZF 1)
Fig. 4
Cotton Tran
Fig. 3: PCR A
bp ladder, C-C
4: PCR Analysis of
sgenic with DRE
Analysis of T
0
Tran
Control, 1-4: LR
Putative Transfor
E-binding Trans
Fig. 1: Gene Const
Fig. 2: Gene Co
nsgenics with npt
RA 5166 (DREB
mants with DREB
scription Factor
truct and Restricti
nstruct and Restr
t II Gene Specific P
B 1A); 5-9: LRA
1A and ZF1 Gene Sp
Gene (DREB1A
ion Map of DREB 1A
riction Map of Zf1
rimer Produced th
A 5166 (ZF 1),
pecific Primer Am
A) and Zinc Fing
A
he Expected 700 b
10-12: LRK 51
plified the Expecte
ger Gene (ZF1)

bp Fragment.
16 (DREB 1A);
ed 540 bp, 480 bp
) 143

; 13-15: LRK
p Resp.

Lane 1: 1 KB Ladder, Lane 2&3: Control, Lane 3-5: DNA samples of the independent transgenic plants of cotton.
Fig. 5: Southern Analysis of DREB 1A Transgenic Plants. Genomic DNA (10ug) Was Digested with Bam HI
and Hybridized with DREB 1A Gene Probe Labeled with Dig - Labeling Kit.

Lane 1: 100bp Ladder, Lane 2-5: DNA samples of the independent transgenic plants of cotton.
Fig. 6: Southern Analysis of ZFP 1 Transgenics. Genomic DNA (10ug) was Digested with Bam HI and Hybridized
with Gene Probe Labeled with Dig-labeling Kit
1-3: R
independent

Cotton Tran
RNA sample fr
t transgenic pla
Fig
o
Fig. 8: Compara
sgenic with DRE
from independe
ants (ZF 1), 7 :+
g. 7: RNA Dot Blot A
of the Independent
ative Expression o
E-binding Trans
ent transgenic
+ve control(ZF
Assay of the T1 tra
t Transgenic Even
of LRA 5166 Transf
scription Factor
plants (DREB
1)
ansgenic Plants. RN
ts and Blotted with

formed and Non-tr
Gene (DREB1A
B 1A), 4: cont
NA Was Isolated fr
h the Specific DRE
ransformed Leaf D
A) and Zinc Fing
trol (DREB 1A
rom the Young Lea
EB 1A and ZF 1 Prob
Disc to PRG Stress
ger Gene (ZF1)

A)5-6: RNA s
aves
be
s and Alleviation
) 145
sample from

146
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Study of Heterosis in Inter Varietal Crosses

of Asiatic Cotton (Gossypium herbaceum L)
N.N. Patel, D.U. Patel, D.H. Patel, K.G. Patel, S.K. Chandran and V. Kumar
Main Cotton Research Station, Navsari Agricultural University,
Athwa Farm, Surat-395007, India
AbstractThe present investigation was carried out using line x tester analysis consisting of four line and ten
pollinator parents to have information on extent of heterosis for seed cotton yield and yield components in Asiatic
cotton (G. herbaceum). High parent heterosis was observed for seed cotton yield ranging from 23.31 to 29.25 whereas
standard heterosis against check G.Cot.DH-9 ranged from 45.07 to 18.21. Three hybrids G.Cot-23 x GBhv-191,
G.Cot-23 x GBhv-178, G.Cot-23 x GBhv-215 exhibited significant and positive heterosis for seed cotton yield over
standard check. Seven cross combinations namely G.Cot-23 x GBhv-191, G.Cot-23 x GBhv-178, G.Cot-23 x GBhv-
215, Jaydhar x GShv-820/91, G.Cot-17 x GShv-384/92, G.Cot-17 x GShv-820/91 and G.Cot-17 x GBhv-201 showed
heterobeltiosis in desired direction for seed cotton yield. The magnitude of heterobeltiosis and standard heterosis was
high for seed cotton yield, boll weight and ginning percentage, medium for plant height and number of bolls per plant
and low for 2.5 % span length. The result indicated that, in general high heterotic hybrids was involved at least one
good donor parent for seed cotton yield and its components. The hybrids, G.Cot-23 x GBhv-191, G.Cot-23 x GBhv-
178, G.Cot-23 x GBhv-215 expected to provide better segregants in subsequent generation as these crosses are
associated with parents having significant and positive gca effects for majority of the characters.
INTRODUCTION
Considerable progress has been made for commercial exploitation of hybrid vigour in Hirsutum, but it is
not so for Herbaceum. The desi or diploid cotton varieties are still preferred in the low rainfall areas
because of their resistance to disease and pest, drought tolerance and suitability under rainfed conditions.
These aroused the interest for developing superior hybrids in Asiatic cotton. The choice of appropriate
breeding procedure for the development of high yielding varieties in intra-specific hybrids of desi-cotton
depends on the nature and magnitude of genetic variation present in the material with respect to yield and
its component traits through line x tester mating design.
The seed of 40 F
1
hybrids developed from line x testers mating design by crossing 4 lines/females and 10
testers/males was sown in randomized block design with three replications at Regional Cotton Breeding
Research Station, Navsari Agricultural university, Bharuch during kharif 2001-02. Each plot consisted of
single row of 5.4 m. length spaced at 120cm apart with plant-to-plant spacing 45 cm. The hybrids were
grown along with parent and standard check (G.Cot-DH-9). The recommended dose of fertilizer was
applied. The insects and pests were controlled with proper insecticides. The data for seed cotton yield and
its components viz; boll weight, number of bolls per plant, plant height, number of monopodia, ginning
percentage and 2.5 % span length were recorded from five competitive plants in each replication. The
heterosis over better parent and hybrid check G.Cot-DH-9 was estimated using standard procedure.
(Panse and Sukhatme (1978) and Singh and Choudhari (1979).
The analysis of variance (Table1) indicated that differences among genotypes were statistically
significant for all the characters, indicating considerable amount of variability. Heterosis over better
parent and standard hybrid variety are presented in Table 2. The heterobetiosis and standard heterosis
over seed cotton yield ranged from 23.31 to 29.25 and 45.07 to 18.21 per cent respectively. Only three
crosses (hybrids) viz.,G.Cot.23 x GBhv-191 (18.21), G.Cot.23 x GBhv-178 (14.37) and G.Cot.23 x
GBhv-215 (12.67 per cent)exhibited significant heterosis over standard check for seed cotton yield.
Similar results were also reported by Tuteja et al., (2004), Preetha and Raveendaran (2008) and Rajamani
25
et al., (2009). For boll weight 15 and 22 hybrids expressed significant positive heterosis over better
parent and standard check respectively. The highest positive heterosis was observed by the cross G.Cot-
23 x GShv-531/92 (32.37 %) over standard check for boll weight. Similarly, for number of bolls per plant
the hybrids Digvijay x GBhv-178 and Jaydhar x GShv-820/91 recorded significant positive heterosis of
17.50 and 13.49 % over better parent and standard check respectively. For ginning percentage, the hybrid
G.Cot-17 x GShv-613/97 (27.31%) recorded maximum standard heterosis. The significant positive
heterosis (23.14%) was exhibited by the hybrid G.Cot-17 x GBhv-201 over better parent and 14.61 %
heterosis was recorded by the hybrid Digvijay x GShv-695/93 over standard check for plant height
(Tuteja and Singh 2001, Kapoor et al., 2002). None of the hybrid had positive heterosis for 2.5% span
length over long staple standard hybrid check (Patel et al., (2000).
TABLE 1: ANALYSIS OF VARIANCE (MEAN SQUARE) FOR EXPERIMENTAL DESIGN FOR DIFFERENT CHARACTERS IN COTTON
Source of
Variation
d.f. Seed Cotton
Yield
Per Plant
Boll
Weight
(g)
Number
of Bolls Per
Plant
Ginning
Percentage
Plant Height
(cm)
2.5 % span
Length (mm)
Replications 2 177.85 0.01 19.71 1.36 25.78 0.24
Treatments 54 1868.80 0.32 172.59 18.38 1423.26 4.17
Parents 13 959.97** 0.32** 109.88** 14.53** 1182.65** 3.02**
Females 3 2928.92** 0.91** 173.35** 42.16** 1769.68** 2.94**
Males 9 407.78** 0.11** 50.78** 5.38** 1062.55** 2.95**
Females vs Males 1 22.88 0.43** 451.35** 14.01** 502.45** 3.87*
Hybrids 39 1928.65** 0.31** 176.27** 19.02* 1511.66** 1.99**
Parents vs Hybrids 1 11451.0** 0.77** 703.91** 2.45 982.25** 0.05
Control vs Rest 1 1767.22 0.26 313.11 59.13 1544.60 108.44
Error 108 57.87 0.002 13.70 1.28 30.09 0.66
S.Em. + 4.39 0.03 2.14 0.65 3.17 0.47
*Significant at 5% ** Significant at 1%
TABLE 2: ESTIMATES OF HETEROSIS IN PERCENTAGE OVER BETTER PARENT (BP) AND STANDARD CHECK (SC) G.COT.DH-9 FOR DIFFERENT CHARACTERS
Hybrids
Seed Cotton Yield Per Plant Boll Weight No. of Bolls Per Plant
BP BP BP SC BP SC
Digvijay x GShv-384/92 8.23 -15.97** 6.13** -4.15* 5.71 -7.98*
Digvijay x GShv-695/93 2.76 -26.39** 1.64 2.90 -6.30 -26.08**
Digvijay x GShv-820/91 4.26 -20.96** -5.59** -6.64** 7.01 -14.64**
Digvijay x GShv-613/97 7.71 -24.53** 7.10** -4.15* 0.00 -19.60**
Digvijay x GShv-531/92 1.38 -25.86** 1.62 4.15* 4.27 -17.74**
Digvijay x GBhv-178 13.31** -6.39 4.27* 1.24 17.50** 0.09
Digvijay x GBhv-191 -9.81* -20.99** -2.07 4.98** -4.34 -21.82**
Digvijay x GBhv-198 6.60 -10.88** -5.43** 5.81** -0.56 -21.56**
Digvijay x GBhv-201 16.95** -15.48** -1.28 6.64** -7.09 -26.70**
Digvijay x GBhv-215 -14.75** -25.48** -8.68** 1.66 -8.11 -26.62**
G.Cot-17 x GShv-384/92 22.96** 2.00 9.35** 6.64** 6.61 -2.74
G.Cot-17x GShv-695/93 -13.34** -28.03** 4.10* 5.39** -15.66** -23.07**
G.Cot-17 x GShv-820/91 21.31** 0.75 3.91* 2.90 3.60 -5.49
G.Cot-17 x GShv-613/97 2.52 -14.91** 9.63** 7.05** -8.27 -16.31**
G.Cot-17 x GShv-531/92 11.32** -7.61* 11.35** 14.11** -2.82 -11.35**
G.Cot-17 x GBhv-178 14.80** -4.72 13.31** 10.79** 4.57 -4.60
G.Cot-17 x GBhv-191 -1.74 -13.92** 2.33 9.54** -9.73* -17.65**
G.Cot-17 x GBhv-198 16.11** -2.93 -2.96 8.71** -0.10 -8.86*
G.Cot-17 x GBhv-201 21.26** 0.64 -0.77 7.47** 8.27 -1.23
G.Cot-17 x GBhv-215 13.62** -0.68 -9.68** 0.83 7.20 -2.21
G.Cot-23 x GShv-384/92 -1.22 -1.33 -13.18** 5.81** 7.24 -6.66
G.Cot-23x GShv-695/93 -17.67** -17.77** -3.18* 17.84** -12.93** -28.29**
G.Cot-23 x GShv-820/91 0.30 0.19 5.68** 28.63** 3.34 -14.91**
G.Cot-23 x GShv-613/97 0.15 0.03 -2.27 19.09** 4.20 -14.18**
G.Cot-23 x GShv-531/92 6.05 5.93 8.86** 32.37** -0.32 -17.91**
G.Cot-23 x GBhv-178 14.51** 14.37** 4.32** 26.97** 8.33 -7.71
Table2(Contd.)
Study of Heterosis in Inter Varietal Crosses of Asiatic Cotton (Gossypium herbaceum L) 151
Table2Contd.
G.Cot-23 x GBhv-191 18.35** 18.21** 6.36** 29.46** 16.92** -3.72
G.Cot-23 x GBhv-198 -3.20 -3.31 1.36 23.24** 2.69 -15.44**
G.Cot-23 x GBhv-201 4.95 4.82 3.64* 26.14** 5.60 -13.04**
G.Cot-23 x GBhv-215 12.80** 12.67** 6.36** 29.46** 7.65 -11.35**
Jaydhar x GShv-384/92 -10.29** -30.35** -8.28** -17.43** -9.88* -8.52*
Jaydhar x GShv-695/93 -23.31** -45.07** -13.39** -12.45** -30.94** -29.89**
Jaydhar x GShv-820/91 29.25** -2.01 -8.38** -9.13** 11.80** 13.49**
Jaydhar x GShv-613/97 -7.33 -35.07** 3.40 -7.47** -26.31** -25.19**
Jaydhar x GShv-531/92 5.12 -23.13** -6.49** -3.73* -17.57** -16.32**
Jaydhar x GBhv-178 -13.95** -28.91** -10.26** -12.86** -13.81** -12.51**
Jaydhar x GBhv-191 -16.59** -26.93** -21.71** -16.18** -7.17 -5.76
Jaydhar x GBhv-198 14.51** -4.26 -15.31** -4.98** 0.96 2.49
Jaydhar x GBhv-201 15.79** -16.31** -17.90** -11.20** 0.87 2.39
Jaydhar x GBhv-215 8.53* -5.13 -10.92** -0.83 4.63 6.21
S.Em + 4.39 4.39 0.03 0.03 2.14 2.14
CD at 5 % 12.17 12.17 0.08 0.08 5.92 5.92
CD at 1 % 15.99 15.99 0.11 0.11 7.80 7.80

Hybrids Ginning Percentage Plant Height 2.5 % Span Length (mm)
BP SC BP SC BP SC
Digvijay x GShv-384/92 -0.87 16.62** 7.49** -7.22** 1.28 -19.74**
Digvijay x GShv-695/93 -0.09 17.53** 6.90** 14.61** -3.46 -24.38**
Digvijay x GShv-820/91 -12.09** 4.39 18.94** -18.70** -1.47 -16.69**
Digvijay x GShv-613/97 -1.56 15.79** -9.26** -6.59* -1.58 -22.89**
Digvijay x GShv-531/92 -4.42 12.43** -16.03** -15.78** -0.70 -19.74**
Digvijay x GBhv-178 -8.48** 7.63** 8.19** 8.51** 0.55 -18.05**
Digvijay x GBhv-191 3.64 21.91** 4.79 5.04 2.68 -17.94**
Digvijay x GBhv-198 -0.78 16.71** 8.96** 9.28** 0.00 -16.69**
Digvijay x GBhv-201 -9.61** 8.24** -1.36 -1.06 3.58 -18.28**
Digvijay x GBhv-215 6.58** 25.35** 4.81 5.11* 1.35 -15.44**
G.Cot-17 x GShv-384/92 -0.41 22.21** -2.46 -5.27* 1.28 -19.74**
G.Cot-17x GShv-695/93 -3.40 18.54** -28.49** -23.33** 0.00 -21.43**
G.Cot-17 x GShv-820/91 3.07 26.48** -19.54** -31.58** -3.60 -18.49**
G.Cot-17 x GShv-613/97 3.73 27.31** -31.76** -29.75** -6.61* -26.62**
G.Cot-17 x GShv-531/92 -0.08 22.61** -21.47** -22.02** -0.56 -19.64**
G.Cot-17 x GBhv-178 -0.75 21.81** -13.14** -21.51** -6.23* -23.57**
G.Cot-17 x GBhv-191 -4.65* 17.01** -2.81 -24.51** -1.27 -21.09**
G.Cot-17 x GBhv-198 -6.80** 14.35** -1.31 -23.73** -6.91* -22.45**
G.Cot-17 x GBhv-201 -2.16 20.07** 23.14** -3.50 3.15 -18.63**
G.Cot-17 x GBhv-215 -5.56* 15.88** -18.31** -29.34** -8.80** -23.91**
G.Cot-23 x GShv-384/92 -5.34* 17.32** -12.20** -14.74** -1.39 -19.84**
G.Cot-23x GShv-695/93 2.47 27.00** -9.95** -3.48 -2.78 -20.99**
G.Cot-23 x GShv-820/91 -12.33** 8.67** -4.47 -9.29** -4.41 -19.17**
G.Cot-23 x GShv-613/97 -6.08** 16.40** -23.71** --25.48** -0.56 -19.17**
G.Cot-23 x GShv-531/92 -9.78** 11.82** -24.65** -25.17** -1.81 -20.18**
G.Cot-23 x GBhv-178 -6.82** 15.49** 1.35 -3.77 -5.54* -23.02**
G.Cot-23 x GBhv-191 -7.07** 15.18** 15.08** 9.26** 3.75 -15.68**
G.Cot-23 x GBhv-198 -12.65** 8.24** -16.80** -21.01** -4.20 -20.18**
G.Cot-23 x GBhv-201 -3.20 19.98** -6.07* -10.82** -2.64 -20.86**
G.Cot-23 x GBhv-215 -5.01* 17.72** -25.98** -29.73** -1.76 -18.05**
Jaydhar x GShv-384/92 -1.96 7.14* -28.11** -30.19** 3.13 -18.28**
Jaydhar x GShv-695/93 -8.25** -0.30 -19.42** -13.62** 3.95 -22.79**
Jaydhar x GShv-820/91 -9.26** 7.76** -15.01** -27.74** -3.20 -18.15**
Jaydhar x GShv-613/97 -1.02 8.24** -28.02** -25.91** 1.20 -23.70**
Jaydhar x GShv-531/92 -2.55 9.16** -30.68** -31.17** -2.37 -21.09**
Jaydhar x GBhv-178 -9.30** 1.34 -2.89 -12.26** -7.06* -24.24**
Jaydhar x GBhv-191 -9.09** 1.83 -2.30 -24.12** -5.23 -24.24**
Jaydhar x GBhv-198 -5.56* 3.78 7.55* -16.89** -11.25** -26.07**
Jaydhar x GBhv-201 -17.09** -0.70 -6.11 -26.42** -2.00 -22.69**
Jaydhar x GBhv-215 0.19 8.37** -2.07 -15.29** -6.50* -22.01**
S.Em + 0.65 0.65 3.17 3.17 0.47 0.47
CD at 5 % 1.80 1.80 8.77 8.77 1.30 1.30
CD at 1 % 2.37 2.37 11.51 11.51 1.71 1.71
Identification of superior heterotic hybrids are mostly done on the per se performance. The
performance of hybrids indicated that high seed cotton yielding hybrids also performed well in at least
one or more yield attributing characters. Among females (G.Cot-23) and among males (GBhv-191 and
GBhv-215) appeared promising for seed cotton yield as well as for most of the yield components. The
hybrid G.Cot-23 x GBhv-191 expressing highest positive heterobeltiosis and standard heterosis for seed
cotton yield also had significant heterosis for yield component namely boll weight, ginning percentage
and number of bolls per plant. The results are in agreement with the report of Bhatade et al., (1979);
Waldia et al., (1979); Khadi et al., (1992).It will be a boon to the breeder to screen such parental
combination to exploit them for a successful heterotic breeding.
From the above discussion it is interesting to note that generally high heterotic hybrids involved at
least one good donor parent for seed cotton yield or its components. Therefore, the hybrid G.Cot-23 x
GBhv-191, G.Cot-23 x GBhv-178 and G.Cot-23 x GBhv-215 expected to provide better segregants in
subsequent generation.
REFERENCES
[1] Bhatade, S.S., Shobhane, M.R. and Unchegaonkar, P.K. (1979). Studies on heterosis in inter-varietal crosses of Desi
(Gossypium arboreum L.) cotton. J. Indian. Soc. Cotton Improve., 4 (1) : 28-36.
[2] Kapoor, C.J.; Singh, M. and Maheshwari, R.V. (2002). Heterosis for yield and yield attributing traits in desi cotton. J.
Cotton Res. Dev.,16(2) : 182-183.
[3] Khadi, B.M.; Janagoudar, B.S.; Katageri, I.S. and Eshanna, M.R. (1992). Desi hybrids cotton for rainfed conditions. J.
Cotton Res. Dev., 6(2): 105-110.
[4] Panse, V.G. and Sukhatme, P.V. (1978). Statistical methods for Agricultural workers. I.C.A.R., New Delhi.
[5] Patel, J.C.; Patel, U.G.; Patel D.H.; Patel, R.H. and Patel, M.V. (2000). Performance of intra hirsutum and asiatic hybrids
based on male sterility system. Indian J. Genet., 60: 111-115.
[6] Preetha, S. and Raveendran T. S. (2008). Combining ability and heterosis for yield and fibre quality traits in line x tester
crosses of upland cotton. International J. Plant Breed. Genet., 2(2): 64-74.
[7] Rajamani, S., Rao C. M. and Naik R. K. (2009). Heterosis for yield and fibre properties in upland cotton (G. hirsutum L.).
J. Cotton Res. Dev., 23(1): 43-45.
[8] Tuteja, O.P., and Singh, D.P. (2001). Heterosis for yield and its components in desi cotton hybrids based on GMS system
under varied environments. Indian J. Genet., 61 (3) : 291-292.
[9] Tuteja, O.P.,Verma S.K. and Ahuja, S.L.(2004). Estimation of heterosis for seed cotton yield and its component characters
in Gossypium hirsutum L. J. Cotton Res. Devlop.18 : 38-41.
[10] Singh, R.K. and Choudhary, B. D. (1979). Biometrical methods in quantitative genetic analysis (Revised ed.1979). Kalyani
Publisher,New Delhi, pp.191-200.
[11] Waldia, R.S., Mor, B.R. and Lather, B.S.P. (1979). Heterosis and combining ability in desi (G. arboreum L) cotton. J.
Indian Soc. Cotton Improve., 8 (1) : 36-037.
Assessment of Genetic Diversity for Improved Fibre

Quality Traits in G. barbadense Accessions
to Widen Cotton Gene Pool
Amala Balu P., D. Kavithamani and S. Rajarathinam
Department of Cotton, Centre for Plant Breeding and Genetics
Tamil Nadu Agricultural University, Coimbatore641003
AbstractThe knowledge on the presence of genetic diversity and breeding potential of Gossypium spp., accessions
are vital source for the genetic improvement of cotton fibre quality and productivity. Hence G. barbadense germplasm
is known existing source for availability of substantial variation of fibre quality traits. The objective of this study was
to assess the genetic diversity and relationship among the G. barbadense accessions using multivariate Mahalanobis
D
2
statistics. Fifty five G. barbadense accessions and one G. hirsutum (for forming different clusters) accession were
utilized in this study. Grouping of accessions into different clusters was independent of the geographical origin of
accessions. G. hirsutum accession formed a separate cluster from the G. barbadense accessions. The analysis of
variance showed highly significant differences among genotypes for all the characters studied. Cluster I was largest
consisting of 36 accessions followed by cluster V with five accessions. The composition of clusters indicated the
existence of relationship between genetic diversity and geographical distribution. The inter cluster distances were
maximum when compared to intra cluster distances, which indicated wide genetic diversity among the genotypes of
different groups than those of same cluster. The results inferred that there is need to broaden the genetic base of
individual clusters by introducing different alleles from the germplasm accessions of other clusters. Seed cotton yield,
GOT, boll weight, bundle strength, S/L ratio were the major characters contributing towards divergence.
Keywords: Genetic divergence, Gossypium barbadense, inter cluster, intra cluster multivariate analysis.
INTRODUCTION
A better knowledge about the genetic diversity of cotton is warranted for exploitation of existing
variability. It also plays an important role in the manifestation of heterosis. Hybrids between genotypes
of diverse origin display a greater heterosis than those hybrids involving closely related parents.
Multivariate analysis employing Mahalanobiss D
2
analysis has been extensively used as a quantitative
measure to identify diverse genotypes. Present investigation was carried out to estimate the nature and
magnitude of genetic diversity.
The study was conducted in the department of cotton, Tamil Nadu Agricultural University, Coimbatore
during winter 2010. Fifty five Gossypium barbadense cotton genotypes obtained from Central Institute
for cotton Research (Regional station) Coimbatore along with on G.hirsutum genotype were planted in
randomized block design with three replications. Uniform spacing of 90 cm x 45 cm and all the
recommended field operations were carried out. In each replication five competitive plants were
randomly selected and observations were recorded for 14 characters viz., days to 50 per cent flowering,
no. of bolls /plant, boll weight, lint and seed index, ginning out turn, seed cotton yield / plant, lint yield /
plant. 2.5 per cent span length, bundle strength, micronaire value and fibre strength / length ratio. The
genetic divergence was worked out by using Mahalanobiss D
2
statistic as described by Rao (1952). Based
on the D
2
value, evaluated genotypes were grouped into different clusters by employing Tochers method
as outlined by Rao (1952).

26
RESULT AND DISCUSSION
The analysis of variance showed highly significant differences among genotypes for all the characters
studied and infers existence of considerable genetic diversity among genotypes. Hence, further analysis
was carried out for relative magnitude of D
2
values for all the characters and all the genotypes were
grouped into ten clusters (Table 1).
TABLE 1: GENOTYPES INCLUDED IN DIFFERENT CLUSTERS
Cluster Number Genotypes No. of Genotypes

I
NDGB7, NDGB12, NDGB22, NDGB29,NDGB32, NDGB33, NDGB35,
NDGB41, NDGB43, NDGB44, NDGB45, NDGB46, NDGB48, NDGB49,
NDGB89, NDGB90, NDGB92, NDGB93,ICB1,ICB3,ICB61and ICB73
36
II ICB124 and ICB238 2
III ICB104 and ICB165 2
IV ICB172 and ICB180 2
V ICB85,ICB87,ICB128,ICB 137and ICB164 5
VI ICB169,ICB 171 and ICB207 3
VII ICB 235 and CCB1 2
VIII ICB173 and ICB 242 2
IX SUVIN 1
X MCU 13 1
Among the clusters obtained the maximum number of germplasm accessions (36) were included in
cluster I followed by cluster V (5) and cluster VI (3), while clusters, II,III,IV,VII and VIII included two
genotypes each and the lowest in cluster IX (1) and X (1). Genotypes from diverse eco-geographical
regions of the country were grouped in the cluster I. Thus, the grouping of genotypes into different
clusters was at random and no relationship between the genetic divergence and geographical origin of the
genotypes was observed. Generally geographical diversity has been considered as a measure of genetic
diversity. However, this is an inferential criterion and it may not be so effective in quantifying or
differentiating various populations. The present pattern of grouping of genotypes indicated that the
genetic diversity was not fully related to the geographical diversity. These results are in agreement with
the earlier studies viz., Single and Bains (1968), Singh and Gill (1984), Sumathi and Nadarajan (1994),
Amudha et al. (1997), Pushpam et al., (2004), and Gopinath et al., (2009). Hence, it indicated that the
geographic diversity though important, might not to be the only factor in determining genetic divergence.
It may be the outcome if several other factors such as genetic drift, natural selection forces and diverse
environmental conditions within the country. Therefore, choice of the parents for hybridization should be
decided on the basis of genetic diversity rather than geographical diversity.
TABLE 2: AVERAGE INTRA (DIAGONAL) AND INTER (OFF- DIAGONAL) CLUSTER DISTANCES BASED ON D
2
VALUES
Clusters I II III IV V VI VII VIII IX X
I 42.00 59.55 42.30 79.88 57.20 119.76 122.16 41.29 133.04 150.34
II 5.01 21.46 21.52 67.78 42.02 67.81 54.32 93.22 147.81
III 20.64 34.32 56.74 66.67 95.87 44.60 124.53 127.14
IV 20.71 83.14 38.23 121.53 74.99 143.91 189.39
V 48.85 135.46 136.77 49.12 165.42 228.36
VI 63.34 130.48 112.76 140.69 191.71
VII 36.91 106.28 53.95 146.98
VIII 41.99 120.08 173.15
IX 0.00 122.39
X 0.00
In the present study, inter cluster distances were found to be greater than intra cluster distances,
revealing considerable amount of genetic diversity among genotypes (Table 2). The highest intra cluster
distance was observed in cluster VI (63.34) followed by cluster V (48.85), cluster I (42.00) cluster VIII
(41.99), cluster VII (36.91), cluster IV (20.71), cluster III (20.64) and cluster II (5.01) where as the
highest inter cluster distance was observed in between the clusters X and V(228.36) followed by cluster
Assessment of Genetic Diversity for Improved Fibre Quality Traits in G. barbadense Accessions 155
X and VI(191.71), cluster X and IV(189.39), cluster X and VIII (173.15) cluster IX and V (165.42),
cluster X and I (150.34), cluster X and II (147.81), cluster X and VII (146.98), cluster IX and IV
(143.91), cluster IX and cluster VI (140.69) and the lowest in between cluster III and II (21.46).
Maximum genetic divergence between the clusters indicates that the hybridization between the
genotypes of distant clusters would produce potential and meaningful hybrids and desirable segregants
also. Use of genetically distance genotypes as parents to get the most promising breeding material had
also been suggested by Singh and Singh (1984), Sambamurthy et al., (1995), Pushpam et al., (2004) and
Gopinath et al., (2009). However, Arunachalam and Bandopadhyay (1984), Alther and Singh (2003) and
Kulkarni et al.,(2011) experimentally proved that more number if heterotic combinations with higher
level of heterosis were from the parents grouped into moderate divergent groups like clusters IX and III.
The results obtained from clustering pattern are in agreement with hypothesis of moderate divergence for
the best heterotic combinations.
Divergence reflecting in the material was also evidenced by an appreciable amount of desirable
variation among cluster means for different characters (Table 3). The component of cluster means for
seed cotton yield / plant was the highest for cluster X (1587 kg/ha) followed by cluster V (1164kg/ha)
and the lowest for cluster IX (656 kg/ha). A minimum day to 50 per cent flowering was observed for
cluster X (60 days) followed by cluster I (71 days) a maximum mean value for cluster II and IV (75
days). For plant height, the highest cluster mean was recorded in cluster VI (209 cm) followed by cluster
IV (203 cm) and the lowest in cluster IX (118cm).
TABLE 3: COMPONENTS OF INTRA CLUSTER D
2
OF COTTON
Cluster Different plant characters
1 2 3 4 5 6 7 8 9 10 11 12 13 14
I 71 128.42 14 19 3.86 5.48 10.56 34.17 987.23 338 32.17 27.52 4.36 0.81
II 75 175.66 19 24 4.09 6.07 11.88 33.80 1034.00 350 34.10 30.33 4.10 0.86
III 72 162.41 16 18 4.23 6.79 12.02 36.07 958.75 349 33.17 28.88 4.40 0.82
IV 75 203.16 17 28 4.10 6.05 11.53 34.35 813.00 280 30.47 27.98 4.63 0.89
V 75 128.56 13 17 3.92 6.01 11.11 35.01 1163.80 404 32.72 30.83 4.19 0.89
VI 74 209.00 19 30 3.52 5.10 9.83 34.07 811.50 279 33.20 29.87 4.15 0.88
VII 74 130.16 23 28 4.32 6.18 12.29 33.43 738.50 246 36.90 28.98 3.95 0.76
VIII 74 123.08 15 21 3.76 5.05 10.35 32.61 735.25 245 33.25 28.83 4.55 0.82
IX 73 118.17 24 31 3.20 4.93 11.30 30.40 656.00 200 31.80 35.40 3.45 0.94
X 60 119.50 21 31 4.35 5.55 9.50 36.95 1587.00 586 30.00 22.55 4.25 0.75
1- Days to 50 per cent flowering; 2- Plant height (cm); 3- Symbodia/plant; 4- Bolls/plant; 5- Boll
weight (g); 6. Lint index; 7- Seed index; 8- Ginning Outturn; 9- Seed Cotton Yield (kg/ha); 10- Lint
yield(kg/ha); 11- 2.5 per cent span length (mm); 12- Bundle strength (g/tex); 13- Micronaire value; 14-
Strength/Length ratio.
For sympodia /plant, cluster IX (24) exhibited the highest cluster mean, while cluster V (13) had
minimum cluster mean. The highest cluster mean for number of bolls / plant was exhibited by cluster IX
and X (31) and the lowest was recorded by cluster V (17). The cluster X had the highest cluster mean for
boll weight (4.35g) followed by cluster VII (4.32g) and the lowest cluster IX (3.20g). For lint index,
cluster III (6.79) registered the highest mean value and cluster IX recorded the lowest (4.93) cluster
mean. The highest cluster mean for seed index was exhibited by cluster VII (12.29) and the lowest in
cluster (9.50). For ginning outturn cluster X (36.95) exhibited the highest cluster mean, while cluster IX
(30.40) has minimum cluster mean. The highest cluster mean for lint yield was exhibited by cluster X
(586 kg/ha) followed by cluster V (404 kg/ha) and the lowest in cluster IX (200 kg/ha).
For 2.5 per cent span length, cluster VII possessed the highest cluster mean of 36.90mm followed by
cluster II (34.10mm) and the lowest was registered by cluster X (30.00 mm). For fibre strength, the
highest cluster mean was exhibited by cluster IX (35.40 g / tex), followed by cluster II (30.33 g / tex) and
lowest was in cluster X (22.55g/tex). The highest cluster mean for Micronarie value was exhibited by
cluster VIII (4.55) and lowest was recorded by cluster IX (3.45). For strength / length ratio, the maximum
cluster mean was registered by cluster IX (0.94) and minimum ratio was recorded by cluster X (0.75).
Strength / length ratio (32.40%), seed cotton yield (11.80%), sympodia / plant (9.09), ginning outturn
(8.57%) and boll weight (7.53%) collectively contributed 69.39% towards the total divergence (Table 4)
and the other characters that influenced the divergence were having minimum value (30.09 percent).
TABLE 4: CONTRIBUTION OF DIFFERENT CHARACTERS TO GENETIC DIVERSITY
S. No. Characters Percent Contribution
1 S/L ratio 32.4
2 Seed cotton yield 11.80
3 Sympodia / plant 9.09
4 Ginning outturn 8.57
5 Boll weight 7.53
6 2.5 per cent span length 4.74
7 Bundle strength 4.74
8 Bolls / plant 4.03
9 Micronaire value 2.92
10 Days to 50 per cent flowering 2.47
11 Lint index 2.21
12 Lint yield 2.21
13 Plant height 0.97
14 Seed index 0.48
The better genotypes selected for all the characters under consideration were presented in table 5.
Among these, NDGB 41 included in cluster I possessed the highest seed cotton yield (1806 kg/ha).
Similarly, genotype MCU 13 of cluster X showed minimum days to 50 per cent flowering. In case of
G.barbadense genotypes NDGB 32, NDGB 67 and NDGB 68 of cluster I had a minimum period of 68
days required for 50 per cent flowering. For plant height the highest mean value of 216.33 cm was
recorded by ICB 171 of cluster VI. The genotype NDGB 64 of cluster I exhibited the highest mean value
for number of symbodia / plant (14). The highest mean performance for number of bolls / plant was
recorded by ICB 169 (35 bolls) of cluster VI. For considering boll weight, the highest mean boll weight
of 4.69 g was registered by ICB 3 of cluster I. Likewise, the highest mean value for lint index and seed
index were showed by ICB 104 (7.18) of cluster III and NDGB 64 (14.35) of cluster I. The highest
mean value for ginning out turn of 39.40 per cent was recorded by ICB 87 of cluster V. For considering
lint yield NDGB 41 (cluster I) had showed highest mean value of 619 kg/ha. However, the genotype
CCB 1 of cluster VII was found superior for 2.5 per cent span length (38.55 mm) and genotype suvin of
cluster IX for fibre strength (35.40 g/tex). The genotype NDGB 44 of cluster I for highest micronaire
value (4.9) and ICB 85 of cluster V for strength/ length ratio (0.98) were also observed.
TABLE 5: DESIRABLE GENOTYPES FOR THE IMPORTANT TRAITS OF COTTON
S. No Characters I II III IV V VI VII VIII IX X
1. 50 % flowering NDGB 67 ICB 238 ICB 165 ICB 180 ICB 85 ICB 171 ICB 235 ICB 173 Suvin -
2. Plant height NDGB 78 ICB 124 ICB 104 ICB 180 ICB 128 ICB 171 ICB 235 ICB 173 Suvin MCU 13
3. Symbodia NDGB 64 ICB 238 ICB 104 ICB 180 ICB 164 ICB 207 ICB 235 ICB 242 Suvin MCU 13
4. Bolls NDGB 41 ICB 238 ICB 165 ICB 172 ICB 137 ICB 169 ICB 235 ICB 242 Suvin MCU 13
5. Boll weight ICB 3 ICB 238 ICB 104 ICB 180 ICB 87 ICB 207 ICB 235 ICB 242 - MCU 13
6. Lint index NDGB 64 ICB 238 ICB 104 ICB 180 ICB 87 ICB 207 CCB 1 ICB 242 - MCU 13
7. Seed index NDGB 64 ICB 238 ICB 104 ICB 180 ICB 85 ICB 207 ICB 235 ICB 242 Suvin MCU 13
8. GOT % NDGB 48 ICB 238 ICB 165 ICB 180 ICB 87 ICB 207 CCB 1 ICB 242 - MCU 13
9. Seed cotton
yield
NDGB 41 ICB 124 ICB 104 ICB 242 ICB 85 ICB 207 ICB 235 ICB 242 - MCU 13
10 Lint yield NDGB 41 ICB 124 ICB 104 ICB 172 ICB 85 ICB 207 ICB 235 ICB 242 - MCU 13
11. 2.5% length NDGB 80 ICB 124 ICB 165 ICB 180 ICB 128 ICB 207 CCB 1 ICB 173 Suvin -
12. Strength ICB 61 ICB 124 ICB 165 ICB 180 ICB 85 ICB 171 ICB 235 ICB 173 Suvin -
13. Micro value NDGB 44 ICB 124 ICB 104 ICB 172 ICB 85 ICB 207 CCB 1 ICB 242 - MCU 13
14. S/L ratio NDGB 62 ICB 238 ICB 165 ICB 180 ICB 85 ICB 171 ICB 235 ICB 242 Suvin MCU 13
Based on the present study of genetic divergence and its component analysis, it can be concluded that
inter crossing among the genotypes of genetically diverse clusters showing superior mean performance
may be helpful for obtaining desirable segregants with higher yield and better fibre quality. The
genotypes, NDGB 41, NDGB 64 and ICB 3 of cluster I; ICB 104 of cluster III; ICB 180 of cluster IV and
Assessment of Genetic Diversity for Improved Fibre Quality Traits in G. barbadense Accessions 157
ICB 85, ICB 137 of cluster V expressed superiority for more than two characters. They may be utilized
as parents in hybridization programme for obtaining desirable combinations. It is also evident that cross
combinations between the genotypes falling in cluster X and V; X and VI; and IV may be most
compatible considering the high inter- cluster distances among them for getting inter specific
hybrids(G.hirsutum X G. barbadence). Incase of G. barbadence accessions, cluster IX and V; IX and VI;
VII and V; VI and V for getting desirable combinations based on the high inter cluster distances among
them.
Intra cluster distances, inter cluster distances, cluster mean for all the characters studied and cluster
wise performance of all the genotypes suggest that the genotypes selected for improvement of yield and
quality components were NDGB 41, ICB 85, NDGB 67, MCU 13, ICB 137, ICB 235, CCB1, NDGB 80
and ICB 128. The hybridization programme with the selected genotypes by considering inter cluster
distances may produce high magnitude of heterosis or desirable segregants, which would be meaningful
for improvement in yield and fibre quality traits.
REFERENCES
[1] Altaher, A.F. and Singh, R.P.2003. Genetic diversity studies in upland cotton (Gossypium hirsutum L.) using two
methods of clustering. J.Indian Soc.Cotton Improv.28:158-63.
[2] Amudha, K., Raveendran, T.S. and Krishnadoss, D. 1997. Genetic diversity in coloured linted cotton varieties. Madras
agric. J. 84: 334-37.
[3] Arunachalam,V. and Bandopadhyay, A. 1984.Limits to genetic divergence for occurrence of heterosis: Experimental
evidence from crop plants. Indian J.Genet. Plant Breeding 44: 548 -54.
[4] Kulkarni, A.A., H.C.Nanda and Patil, S.G. 2011. Studies on genetic divergence in upland cotton (Gossypium hirsutum L.).
J. Indian Res. Dev.25 (1): 9-13.
[5] Pushpam, R., Raveendran, T.S., Devasena, N. and Ravikesavan, R. 2004. Studies on genetic diversity in upland cotton
(Gossypium hirsutum L.). J.Indian Soc. Cotton Impro.29:80 -85.
[6] Rao, C.R. 1952. Advanced Statistical Methods in Biometrical Research. John Wiley and Sond, New York.
[7] Sambamurthy, J.S.V., Reddy, M.D. and Reddy, K.H.G.1995. Studies on the nature of genetic divergence in upland cotton
(Gossyoium hirsutum L.). Annals Agric.Rec.16: 307-210.
[8] Singh, R.B. and Gill, S.S. 1984. Genetic diversity in upland cotton under different environment. Indian J. Genet. 44: 506-
73.
[9] Singh, R.B and Bains, S.S. 1968.Genetic divergence for ginning outturn and the components in upland cotton. Indian J.
Genet. 28: 26268.
[10] Singh, V.V. and Singh, A.K.1984. Studies on genetic diversity in upland cotton. J. Indian Soc. Cotton Improv. 9: 41-45.
[11] Sumathi, P. and Nadarajan, N.1994. Genetic divergence in upland cotton (Gossypium hirsutum L.) J. Indian Soc. Cototn
Improc. 3: 36- 42.

Cotton Protection
Survival of Helicoverpa armigera on Bt Cotton

Hybrids in IndiaCan We Buy the Interpretations
A. Prabhuraj
1
, Y.B. Srinivasa
2
and K. Muralimohan
3

1
Department of Agricultural Entomology, College of Agriculture,
University of Agricultural Sciences, Raichur 584 101, India
2
Institute of Wood Science and Technology, PO Malleswaram, Bangalore560003, India
3
Krishi Vigyan Kendra (University of Agricultural Sciences, Bangalore), Magadi,
Ramanagara District562120 India
E-mail: prabhusha@hotmail.com
AbstractA couple of years since commercialization of transgenic Bt cotton in India, reports on survival of
bollworms have appeared from different parts of the country. The first came in 2004 from Central India claiming a
mere 50% reduction in bollworm population. Upto 9% damage was reported on some of the hybrids in Guntur region
of Andhra Pradesh in 2009. Sizeable populations of pink bollworm were recorded on Bollgard-II
from Gujarat in
2010. In the same year, field survival of Helicoverpa armigera and ~9% boll damage was recorded from Raichur belt of
Karnataka on both Bollgard
and Bollgard-II
. The latter study further demonstrated survival and reproduction of the

pest insect for two complete generations on Bt cotton hybrids. This study created considerable national debate over the
credibility of the technology as such. Here, we put forth two prominent public views expressed on the results of the
study and the many interpretations. Interpreting field survival and completion of two generations as development of
resistance in the population has been mooted prominently. The arguments arise because the study per se does not
quantify resistance among the surviving individuals. One side of the argument is that the results of the study, in
addition to development of resistance, be attributed to a situation where plants produce less-than-lethal levels of toxins.
The other side dismisses the low-toxin argument as the hybrids have been demonstrated, during their approval for
commercialization, that the concentration of the toxins is lethal despite variations across plant parts. Opinions such as
disparity in environment leading to low toxin production are unsubstantiated and may be rejected. The second point of
contest is related to survival of the second generation of the insect on Bt plants. As the F1 was the result of mating
between individuals bred on Bt plants, it has been argued that it does not reflect the assumed reality where individuals
from Bt plants mate with those from non-Bt plants. Therefore the experiment may not gel with the concept of refugia.
On the contrary, this argument fails to reason out considerable survival recorded among the first generation
population, whose parents had ample opportunities to mate with partners from non-Bt plants. Additionally, as the
results of the study show that survival of the individuals bred on Bt plants in their F1 generation was equal to that of
the control, it has been expressed that the surviving individuals were considerably resistant.
INTRODUCTION
The cotton plant has been genetically modified for protection against insect pests, particularly from boll
feeding caterpillars. It happened through the introgression of Cry genes derived from the soil bacterium,
Bacillus thuringiensis, which code for insecticidal proteins in plant tissues (commonly referred as Bt
cotton). Today, several hundred hybrids of Bt cotton are present in India that contain either one or two
bacterial genes in their chromosomes (APCoAB, 2009).
This new war between man and insects is being observed with a lot of interest. One of the reasons
being the anticipated development of resistance in bollworms to Bt toxins. Developing resistance is not
uncommon to species like Helicoverpa armigera, the American bollworm; assays have proved its
resistance against most of the conventional insecticides and protein toxins of Bt (Armes et al., 1992; Saad
et al., 2005). Resistance has been a foregone conclusion, and strategies were applied for delaying the
development of resistance, not preventing it, from the time of introducing Bt cotton for commercial
cultivation; for example, implementation of the concept of refugia. Nevertheless, reports claiming
survival of the pest on Bt cotton started trickling from different parts of India within the first couple of
years of commercialization. The first was published in 2004 from Central India; it declared a mere 50%
reduction in bollworm population (Bambawale et al., 2004). Up to 9% damage by bollworms was
reported on some of the hybrids in Guntur region of Andhra Pradesh in 2009 (Prasad, 2009). During
27
2010 field survival of H. armigera with ~9% boll damage was reported from Raichur on Bt hybrids with
single and double genes. This brings us to the question that what should society gather from these
reports? Are the evidences sufficient to show that the pest is able to overcome Bt toxins, or outsmart
current technology? The interpretations are numerous. In this paper we have summarized the
interpretations and communicated our take on them.
A list of publications reporting the occurrence/survival of H. armigera on Bt cotton is provided under
Table 1. It lists reports on the occurrence of H. armigera on various Bt cotton hybrids in different parts of
India. Most incidences are reported from experimental fields laid exclusively for evaluation of Bt cotton
hybrids against bollworms, while some are reported from demonstration plots in farmers fields. With the
exception of one, all others report incidence on hybrids carrying single gene; Ranjith et al., (2010) report
natural populations of H. armigera on cotton hybrids containing two genes (Cry 1Ac and Cry 2Ab). In
all, boll damage ranged from 0.00 to 17.3%. It is important to note that the listed studies merely record
occurrences of caterpillars of H. armigera; none investigate either the reasons for their occurrences or
confirm the fate of the population. Recently, Ranjith et al. (2010) sampled naturally surviving population
of H. armigera on Bt cotton hybrids and established that these populations were capable of completing
the generation and producing viable progeny able to survive on Bt cotton. At the outset, there remains
little doubt over the current ability of H. armigera to survive and breed on commercial Bt cotton
containing one and two Cry genes.
Interpretations provided for the recorded survival of H. armigera can be categorized into those
related to 1) expression of Bt toxin and 2) pest biology. Expression of Bt toxins is known to vary with
age, pedigree and part of the plant (Kranti et al., 2005). The expression is inversely proportional to age,
suggesting that the probability for survival of H. armigera increases with plant age. Given that H.
armigera attacks mid to late-age cotton plants, it appears that the technology can be greatly
compromised. However, this can also be taken care by over expressing the gene such that the
concentration of the toxin in the plant tissue is never below the levels required to kill bollworms. In
addition, second generation Bt hybrids with two Cry genes have been commercially introduced in 2006.
The second gene is expected to support the existing gene in terms of delaying the development of
resistance. However, proof of the ability to survive on both types of hybrids (Ranjith et al., 2010) clearly
suggests that variation in toxin expression with respect to plant age may not have been sufficiently taken
care to quell the pest.
Toxin expression is also influenced by genetic stock of the hybrid (John et al., 2001; Kranthi et al.,
2005). Not all the hybrids exhibit the same level of Bt toxin in a given situation leading to claims that
survival of the pest may be due to poor pedigree of the hybrid. If this is one of the reasons for under-
performance of any Bt cotton hybrid, then utmost care should be taken during the release of hybrids for
commercialization, as this could jeopardise the interest of the farmer as well as the technology. However,
data on expression levels are provided for each hybrid before commercial approval, which should take
care of the potential problem. This is important because several hundreds of new hybrids have entered the
market during the last four years.
Toxin expression in different plant parts is known to be different, which is another reason given for
survival of pest population (Kranthi et al., 2005; Gore et al., 2001; Gore et al.,2003). Among plant parts,
the expression is highest in leaves followed by flowers, squares and bolls. Hence, it is suggested that
chances for survival of H. armigera increases as it mainly feeds on squares and bolls. However this may
not be accurate as 1) young caterpillars feed on tender leaves and flowers before shifting to bolls, thus
exposing themselves to high concentration of toxins, and 2) Ranjith et al (2010) have reared caterpillars
of all developmental stages on leaves of Bt hybrids before obtaining survival. Although this reason may
suit survival of species like the pink bollworm that feeds exclusively within bolls, it fails to explain
survival of H. armigera.
Expression of Bt toxin in the plant is claimed to vary with planting dates, and agronomic practices
like irrigation and soil nutrition (Yang et al., 2005; John et al., 2001; ). Bt expression decreases with
Survival of Helicoverpa armigera on Bt cotton Hybrids in IndiaCan We Buy the Interpretations 163
advancement in date of planting. This is directly related to the performance of the plant itself, which
declines when planting dates are delayed. This automatically translates to insufficient expression of the
toxin and promoting pest survival. Further, moisture and soil nutrition play a major role in plant health
(John et al., 2001). All hybrids respond positively to application of inorganic fertilizers and regular
irrigation, which is further expected to encourage toxin expression (Hallikeri et al., 2009). Therefore
incidence of H. armigera should decrease when crops are managed well. As mentioned earlier, the listed
reports (Table 1) on survival of H. armigera are from either experimental plots of established research
stations or demonstration fields which are obviously managed well. Therefore, we believe that scope for
faulty agronomic practices that encourage pest survival is minimal.
H. armigera has been one of the prominent pests of cotton since the last few decades. The capability
of this pest to attain the notoriety is mainly attributed to its wide host range, high fecundity, several
overlapping generations in a year and its ability to develop resistance to majority of insecticides including
Bt. In fact populations from different regions of India have exhibited varied levels of tolerance to Bt
toxins. Therefore, it may be obvious that some of the populations might have been able to quickly
develop tolerance to Bt toxin and survive on Bt hybrids.
Many of the interpretations provided for survival of H. armigera tend to focus on conditions that
might promote low toxin concentration in plant tissues. However, one has to carefully consider them as
data on toxin concentration provided during approval of each hybrid shows more-than-enough levels
throughout the crop period. There are no data on the extent of variation in toxin concentration for each
hybrid with respect to the factors discussed earlier. Therefore, at present, there are only two options that
remain for interpreting survival of H. armigera on Bt cotton 1) pest population developing tolerance to
the toxins, or 2) vulnerability of the current technology to variations in agronomic and other natural
factors.
TABLE 1: REPORTS OF THE OCCURRENCE OF HELICOVERPA ARMIGERA ON VARIOUS BT COTTON HYBRIDS IN INDIA SINCE ITS COMMERCIALIZATION
Sl.
No
Year of
Incidence
Hybrids States Square/boll
Damage (%)
Reference
1 2002 MECH-162 Andhra Pradesh 50 Bambawale et al, 2003
2 2002-03 RCH2, RCH 20 Maharashtra Unimodel Vennila et al, 2004
3 2002-03 MECH-162 Karnataka 0-4.3 Kengegowda et al, 2005
4 2002-04 MECH-12, MECH-162 and MECH-184 Andhra Pradesh 5.4-17.3 Sharma & Pampapathy et al,2006
5 MECH-162 Andhra Pradesh 9 Prasad et al, 2009
6 2002-03 RCH2, RCH20, RCH144 Andhra Pradesh 0-8.88 Prasad, 2009
7 2003-05 MECH-184 Maharashtra & Karnataka 0.8-2.8 Bambawale et al, 2010
8 2009 MRC 6918, NCS-145 BGII, MRC-7918BGII Karnataka 4.3-8.6 Ranjith et al, 2010
REFERENCES
[1] Armes, N. T., Jadhar, D.R. Bond. G.S. and King, A.B.S. (1992) - Insecticide resistance in Helicoverpa armigera in South
India - Pesticide Science. 34: 335-364.
[2] Asia-Pacific Consortium on Agricultural Biotechnology (APCoAB) and Asia-Pacific Association of Agricultural Research
Institutions (APAARI), 2009, Bt cotton in India: A status Report, 2nd Edn. p 1-49.
[3] Bambawale, O. M., Tanwar, R. K., Sharma, O. P., Bhosle, B. B., Lavekar, R. C., Patil, S. B., Dhandapani, A., Trivedi, T.
P., Jeyakumar, P., Garg, D. K., Jafri, A. A. and Meena, B. L. (2010) - Impact of refugia and integrated pest management on
the performance of transgenic (Bacillus thuringiensis) cotton (Gossypium hirsutum). Indian Journal of Agricultural
Sciences. 80: 8, 730-736.
[4] Bambawale, O. M. et al., (2003) - Performance of Bt-cotton (MECH-162) under irrigated pest management in farmers
participatory field trial in Nanded district, Central India. Current Science. 86: 1628-1633.
[5] Gore J., Leonard, B. R., Adamczyk, J. J. (2001) - Bollworm (Lepidoptera:Noctuidae) survival on Bollgard and Bollgard II
cotton flower bud and flower components. J. Econ. Entomol. 94 (6):1445-1451.
[6] Gore J., Leonard BR., Gable RH. (2003) - Distribution of bollworm, Helicoverpa zea (Boddie), injured reproductive
structures on genetically engineered Bacillus thuringiensis var. kurstaki Berliner cotton. J. Econ. Entomol. 96: 699-705.
[7] Hallikeri S. S., Halemani H. L., Katageri I. S., Patil B. C., Patil V. C. And Palled Y. B. (2009) - Influence of sowing time
and moisture regimes on Cry protein concentration and related parameters of Bt cotton. Karnataka Journal of Agricultural
Sciences. 22(5): 995-1000.
[8] John J., Adamczyk, Jr. and Douglas V. Sumerford. (2011) - Potential factors impacting season-long expression of Cry1Ac
in 13 commercial varieties of Bollgard cotton. Journal of Insect Science. 1: 1-6. 164 World Cotton Research Conference
on Technologies for Prosperity
[9] Kengegowda, N., Patil, B. V. and Bheemanna, M. (2005) - Population dynamics of insect pests on Bt, non-Bt and popular
hybrid cotton in Tungabhadra project area of Karnataka State. Karnataka Journal of Agricultural Sciences., 18 (2): 383-388.
[10] Kranthi, K.R., Naidu, S., Dhawad, C.S., Tatwawadi, A., Mate, K., Patil, E., Bharose, A. A., Behere, G.T., Wadaskar, R. M.
and Kranthi, S. (2005) - Temporal and intra-plant variability of Cry1Ac expression in Bt-cotton and its influence on the
survival of the cotton bollworm, Helicoverpa armigera (Hbner) (Noctuidae: Lepidoptera). Current Science. 89: 291-298.
[11] Prasad N.V.V.S.D., Mallikarajuna Rao. and Hariprasad Rao N., (2009) - Performance of Bt cotton and non-Bt cotton
hybrids against pest complex under unprotected condition. Journal of Biopesticides, 2:107-110
[12] Ranjith. M. T., Prabhuraj. A and Srinivasa Y. B. (2010) - Survival and reproduction of Helicoverpa armigera on Bt cotton
hybrids in Raichur, India. Current Science. 99 (11):1602-1606.
[13] Ranjithkumar L, Patil, B. V. and N G Vijaykumar. (2011) - Impact of irrigation and fertilizer Levels on Cry1Ac Protein
Content in Bt. Cotton. Research Journal of Agricultural Science. 2(1): 33-35.
[14] Saad Mousa, Trilochan Mohapatra and Govind, T. Gujar (2005) - Monitoring of Bacillus thuringiensis Cry1Ac Resistance
in Helicoverpa armigera (Hbner) (Noctuidae: Lepidoptera). 6th Asia-Pacific Rim Conference on the Biotechnology of
Bacillus thuringiensis and its Environmental Impact, Victoria BC. Ct, J.-C., Otvos, I.S., Schwartz, J.-L. and Vincent, C.
(eds): 74-77.
[15] Sharma, H. C. and Pampapathy, G. (2006) - Influence of transgenic cotton on the relative abundance and damage by target
and non-target insect pests under different protection regimes in India. Crop Protection, 25: 800-813.
[16] Vennila, Biradar, S., Gadpayle, V. K., Panchbhai, J. G., Ramteke, P. R., Deole, M. S., and Karanjkar, P. P (2004) Field
evaluation of Bt transgenic cotton hybrids against sucking pests and bollworms. Indian Journal of Plant Protection. 32 (1):
1-10
[17] Yang.C. Xu. L and Yang. D. (2005) - Effects of nitrogen fertilizer on the Bt protein content in transgenic cotton and
nitrogen metabolism mechanism. Cotton Science. 17 (4): 227-231
Field Performance of F
1,
-F
2
and

non-Bt of BG-II
(MRC-7017 Bt) and JKCH-1947 Bt Against Bollworms
of Cotton
G.T. Gujar, G.K. Bunker, B.P. Singh and V. Kalia
Division of Entomology, Indian Agricultural Research Institute, New Delhi 110012
E-mail:gtgujar@yahoo.com
AbstractStudies were undertaken to evaluate comparative performance of F1, F
2
and non-Bt versions of dual
stacked Cry1Ac and Cry2Ab transgenic cotton hybrid, MRC7017Bt and the single stacked Cry1Ac transgenic
JKCH1947Bt during kharif season. The results proved that lowest damage of terminal bud, fruiting bodies green boll,
open boll; incidence of Earias and Pectinophora gossypiella was recorded on MRC 7017Bt F1, JKCH 1947Bt F1 and
MRC7017Bt F
2
as against MRC7017 non-Bt, JKCH1947 non-Bt and JKCH1947Bt F
2
. The highest seed cotton yield
(27.00 q/ha) was recorded in MRC7017Bt F
1
followed by JKCH1947Bt (23.15 q/ha), MRC7017Bt F
2
(22.76 q/ha),
JKCH1947Bt F
2
(21.08 q/ha), JKCH non-Bt (16.08 q/ha) and MRC non-Bt 7017 (15.91 q/ha). The MRC7017Bt F
1

was found to be significantly superior among the entire hybrid tested, whereas, JKCH 1947Bt F
1
was statically at par
with MRC7017Bt F
2
regarding yield, both had significantly more yield than the non-Bt counterparts of JKCH1947Bt
and MRC7017Bt. These results are discussed vis--vis sustainability of Bt cotton in relation to the threat of resistance
evolution to its most important pest, H. armigera.
Keywords: Bt cotton, JKCH1947Bt, MRC7017Bt, bollworms,
INTRODUCTION
Bt cotton expressing Cry1Ac toxin derived from Bacillus thuringiensis was first commercialized in USA
in 1996 and later in many other countries. It proved quite successful in controlling lepidopteran pests
especially bollworms which are the main constraints in cotton productivity (Perlak et al., 2001). In India,
Bt cotton with cry1Ac gene was introduced in 2002 by Mahyco Monsanto Biotech Ltd., Jalna with 3-
cotton hybrids viz., MECH-12, MECH-162, MECH-184 grown over an area of about 38, 000 hectares.
Bt cotton area later increased rapidly to the present level of about 9.4 million ha in 2010. Concurrent with
area increase was the development of Bt cotton hybrids from 3 in 2002 to more than 780 in 2010. The
dual stacked Bt cotton expressing Cry1Ac and Cry2Ab were introduced in 2006 to control sporadic key
lepidopteran defoliator, Spodoptera litura which is tolerant to Cry1Ac and to contain the threat of
resistance evolution in the target bollworms. Presently more than 18 Bt cotton hybrids belonging to
Monsantos 15985 event of Bollgard II series are being grown; with area under dual stacked Bt cotton
being more than the single stacked Bt cotton with Cry1Ac. Cotton production has increased from about
14 million bales in 2002 to about 34 million bales in 2010. Despite increase in the cotton production,
cotton productivity of about 560 kg/ha is well below that of world average of about 690 kg/ha. Further,
there are quite a wide range of regional differences in cotton productivity from the lowest of 336 kg/ha in
Maharashtra to the highest of 774 kg/ha in Gujarat. The productivity differences are attributed to various
biotic and abiotic stresses. Further, one of the reasons of relatively low productivity is the use of
substandard seeds of Bt cotton hybrids and their F
2
and F
3
seeds by the farmers in view of seed cost
(Jayaraman, 2004; Herring, 2008). Besides, biotic stress due to sucking pests and the emergence of
tolerance/resistance in bollworms pose the threat to the sustainability of Bt cotton.
The bollworm complex is considered as the major constraint for the low level of productivity for
non-Bt cotton (Dhawan et al., 1988, 2004; Patil, 1998) and poses a threat to Bt cotton in view of wide
range of insect susceptibility as reported in the cotton bollworm, Helicoverpa armigera (Kranthi et al.,
2001; Gujar et al., 2000; 2008). In recent years, other bollworms like spotted and pink bollworms are
becoming important pests of non-Bt cotton and the latter is damaging the Bt cotton especially after 120
days wherein farmers are aiming for third picking of cotton by irrigating Bt cotton crop after the main
28
cotton season. The sucking insect pests are also important pests and are estimated to cause loss to the
extent of 46.5% (Panchabhavi et al., 1990). In Bt cotton, sucking pests like mealy bug, Phenacoccus
solenopsis is becoming a major pest. It has reportedly caused loss of about Rs 1590 millions in 2007-08,
the highest ever in the recent times (http://ncipm.org.in/Mealybugs/mealybug.htm). Thus, Bt cotton
despite its ability to control bollworm complex needs timely and limited application of insecticides as
some time pest complex is above the threshold levels (Bambawale et al., 2004). Thus, the changing pest
scenario and threat of Bt resistance looming large in view of wide range of Bt cotton hybrids, legal and
illegal, and also F
2
seeds are the main concern of sustainable Bt cotton cultivation (Monga 2008). Hence,
studies were undertaken to evaluate the comparative performance of Bt cotton, single stacked
JKCH1947Bt and dual stacked MRC7017 Bt, with their non-Bt and F
2
counterparts and under the
selection pressure of H. armigera under cage conditions.
Field experiment was laid out in randomized block design (RBD) with 6 treatments, and replications
during kharif, 2007 at transgenic field, IARI, New Delhi. The seeds of JKCH-1947 Bt-F
1
, Bt-F
2
and Non-
Bt were obtained from JK Agri-Genetics Ltd. Hyderabad. Whereas, MRC-7017 Bt-F
1
(BG-II), Bt-F
2

(BG-II) and Non-Bt were obtained from Mahyco Seeds Ltd. Jalna (Maharashtra).
Seeds were treated with imidacloprid 70 WS (Gaucho
) @ 7 g/kg seed before sowing crop The crop

was sown in a plot size of 4.5 x 4.2 sq m. at 75 x 60 cm
2
spacing on dated 26.06.2007 and 42 plant were
maintained in each plot All the agronomical package and practices was uniform throughout season.
Similarly, seed cotton pickings were done thrice in second season on 29.10.2007 (127 DAS), 5.12.2007
(163 DAS) and 10.12.2007 (168 DAS). The observation on the terminal bud damage fruiting damage,
green boll damage and loculi damage caused by Earias spp and H. armigera was recorded. The terminal
bud damage was recorded by counting damaged number of plants in each plot. Similarly, damaged
fruiting bodies were recorded by counting the total number of reproductives fallen down and the number
of damaged fruiting bodies in each plot. The damaged green boll in each variety was recorded by
counting the total number of green boll and damaged green boll on five plant selected randomly. The
population of Earias spp and H. armigera was also recorded by dissecting twenty bolls and counted the
locule damage and larval population. The pink boll worms damage in each variety was recorded by
counting the number of damaged open bolls and total number of open bolls before each picking. The
loculi damage was recorded in damaged open boll by counting the total number of damaged open boll on
five plants selected randomly. The data were calculated on per cent basis and computed The weight of
seed cotton of all pickings was pooled together for each variety separately and yield per hectare was
computed.
From each replicate, 5 plants were tagged and caged with nylon net (1.8 m x 1.2 m x 1.65 m size)
with support of 4 bamboo sticks and another 5 plants were tagged outside of the cage in the same plot at
50 DAS in 2007.
During 2007 at 91 DAS, late second to early third instar larvae of H. armigera weighing from 10 to
15 mg each, were released @ 5/plant on the same tagged plants under caged and un-caged conditions. H.
armigera larval survival (%), fruiting bodies (20 shedding/plot) damage (%) and green boll damage (%)
on plant were recorded 12 days after release from caged and un-caged conditions. Two uniform sprays,
one of acetamiprid 20 % SP (Pride
) @ 20 g a.i./ha and another of imidacloprid 17.8 % SL (Confidor
)
@ 125 ml/ha were made at 25 DAS and 57 DAS, respectively.
The F
1,
F
2
and non-Bt cotton of Bollgard-II MRC-7017 and JKCH1947Bt were included in the field
experiment conducted during kharif, 2007, for studying the comparative performance of these hybrids
against the major boll worms of cotton. The results of the field investigation during 2007 on different
parameters have been presented given below (Table 1 4).
1,
-F
2
and

non-Bt of BG-II (MRC-7017 Bt) and JKCH-1947 Bt Against Bollworms of Cotton 167
TERMINAL BUD DAMAGE (%) AT DIFFERENT DAYS AFTER SOWING (DAS)
The data presented in Table 1, indicated the lowest terminal bud damage was recorded on MRC Bt-F
1

cotton (00.00%), followed by JKCH Bt-F
1
(0.60%), MRC Bt-F
2
(1.20%), and JKCH Bt-F
2
cotton
(1.88%) and these were found statistically at par to each other at 30 DAS. However, it was significantly
highest recorded on MRC non-Bt cotton (20.50%) and JKCH non-Bt cotton (13.95%). Further at 60
DAS, lowest terminal buds damage was recorded on MRC Bt-F
1
cotton (0.60%), followed by JK Bt-F
1

(1.20%), MRC Bt-F
2
(1.80%) and JK Bt-F
2
cotton (3.90%) and all these were significantly superior to JK
non-Bt (29.39%) and MRC non-Bt cotton (29.31%). The mean terminal bud damage was lowest on MRC
Bt-F
1
(0.3%), followed by JK Bt-F
1
(0.9%), MRC Bt-F
2
(1.5%) and JKCHBt-F
2
cotton (2.89%), and the
highest on MRC non-Bt (24.91%) and JK non-Bt cotton (21.67%).
FRUITING BODIES (SHEDDING) DAMAGE (%) AND EARIAS SP. INCIDENCE AT DIFFERENT DAS
At 65 DAS, MRC Bt-F
1
cotton was significantly superior in term of lowest shedding damage (2.50%) as
compared to all other hybrids tested. However, MRC Bt-F
2
(10%) and JKCH Bt-F
1
cotton (10%) were at
par to each other, but significantly superior over MRC non-Bt (42.50%) and JK non-Bt cotton (30%)
(Table 1). At 95 DAS, minimum shedding damage was recorded on MRC Bt-F
1
cotton (7.50%) followed
by JKCH Bt-F
1
(13.75%), MRC Bt-F
2
(20%) and JKCH Bt-F
2
(30%) and all these were found superior
over MRC non-Bt (47.50%) and JKCH non-Bt cotton (38.75%) except JKCH Bt-F
2
cotton.
The mean fruiting body damage was lowest on MRC Bt-F
1
cotton (5%) and found significantly
superior over all the hybrids used. However, JKCH Bt-F
1
(11.87%) and MRC Bt-F
2
cotton (15%) were
found at par to each other and significantly superior over MRC non-Bt (45%) and JKCH non-Bt cotton
(34.38%) (Table1).
TABLE 1: COMPARATIVE PERFORMANCE OF MRC 7017BT AND JKCH 1947BT AGAINST INFESTATION OF EARIAS SPP. AT DIFFERENT DAS DURING KHARIF, 2007
Hybrids

Terminal Bud Damage (%) at DAS* Fruiting Bodies Damage (%) at DAS* Earias sp. Larvae/20 Shedding**
30 60 Mean 65 95 Mean 65 95 Mean
JKCH 1947Bt
Non-Bt 13.95
(21.62)
29.39
(32.74)
21.67 30.00
(33.17)
38.75
(38.35)
34.38
(35.85)
1.25
(1.31)
1.50
(1.4)
1.38
(1.37)
Bt-F
2
1.88
(7.83)
3.90
(11.18)
2.89 23.75
(28.91)
30.00
(33.02)
26.87
(31.19)
0.50
(0.97)
0.50
(0.97)
0.50
(0.98)
Bt-F
1
0.60
(5.27)
1.20
(6.48)
0.9 10.0
(18.14)
13.75
(21.65)
11.87
(19.94)
0.00
(0.71)
0.00
(0.71)
0.00
(0.71)
MRC 7017Bt
Non-Bt 20.50
(26.84)
29.31
(32.73)
24.91 42.50
(40.66)
47.50
(43.57)
45.00
(42.12)
1.25
(1.31)
1.50
(1.40)
1.38
(1.36)
Bt-F
2
1.20
(6.48)
1.80
(7.70)
1.5 10.00
(18.14)
20.00
(26.48)
15.00
(22.64)
0.25
(0.84)
0.25
(0.84)
0.25
(0.85)
Bt-F
1
0.00
(4.06)
0.60
(5.27)
0.3 2.50
(8.49)
7.50
(15.68)
5.00
(12.71)
0.00
(0.71)
0.00
(0.71)
0.00
(0.71)
SEm 1.95 1.41 2.98 3.69 1.86 0.14 0.13 0.097
CD at 5% 4.16 3.01 6.36 7.87 3.96 0.29 0.287 0.21
*Figures in parentheses are transformed by arc-sin transformation,
**Figures in parentheses are transformed by x + 0.5 transformation
At 65 DAS, Earias sp. larval population was none on MRC Bt-F
1
and JKCH Bt-F
1
cotton; MRC Bt-
F
2
(0.25 l/plant) and JKCH Bt-F
2
cotton (0.50 l/plant), whereas on MRC non-Bt (1.25 l/plant) and JKCH
non-Bt cotton had significantly higher (1.25 l/plant) from all Bt cotton hybrids. Similar trend was also
recorded at 95 DAS. Mean incidence of Earias sp. larvae was recorded zero on MRC Bt-F
1
and JKCH
Bt-F
1
cotton, while 1.38 l/plant recorded on both MRC non-Bt and JKCH non-Bt cotton.

Green Boll Locule Damage (%) and Bollworm Incidence at Different DAS
Green boll locule damage, Earias sp. and P. gossypiella larval incidence were recorded at 70, 90 and 105
DAS along with mean observations (Table 2).
TABLE 2: COMPARATIVE PERFORMANCE OF MRC7017BT AND JKCH 1947BT AGAINST LOCULE DAMAGE AND BOLLWORMS INCIDENCE AT DIFFERENT DAS DURING 2007
Hybrids

Green Boll Locules Damage (%) at DAS * Earias sp. Larvae/Bolls** P. gossypiella Larvae/ Bolls**
70 90 105 Mean 70 90 105 Mean 70 90 105 Mean
JKCH 1947Bt
Non-Bt 13.09
(21.11)
34.47
(35.46)
22.43
(28.15)
17.37
(24.57)
1.50
(1.40)
2.00
(1.56)
2.75
(1.79)
2.09
(1.60)
0.50
(0.97)
2.80
(1.79)
2.75
(1.79)
1.75
(1.49)
Bt-F
2
9.33
(17.58)
10.53
(18.88)
17.58
(24.69)
12.48
(20.63)
1.25
(1.31)
1.25
(1.31)
1.25
(1.31)
1.25
(1.32)
0.25
(0.84)
1.25
(1.31)
1.25
(1.31)
1.00
(1.19)
Bt-F
1
6.76
(15.014)
2.99
(14.03)
13.03
(20.95)
8.59
(1697)
0.50
(0.97)
0.50
(0.97)
0.25
(0.84)
0.42
(0.94)
0.00
(0.71)
0.25
(0.84)
0.25
(0.84)
0.42
(0.95)
MRC 7017Bt
Non-Bt 26.02
(28.95)
34.78
(35.68)
24.88
(29.83)
13.37
(24.58)
1.00
(1.18)
1.75
(1.48)
1.75
(1.48)
1.50
(1.40)
0.25
(0.84)
1.75
(1.48)
1.75
(1.48)
1.09
(1.25)
Bt-F
2
6.25
(14.10)
7.30
(15.11)
14.20
(22.12)
8.12
(16.53)
0.75
(1.10)
0.50
(0.97)
0.00
(0.71)
0.42
(0.95)
0.25
(0.84)
0.00
(0.71)
0.00
(0.71)
0.49
(0.99)
Bt-F
1
2.75
(9.02)
4.21
(11.60)
11.41
(19.57)
5.30
(13.27)
0.00
(0.71)
0.25
(0.84)
0.00
(0.71)
0.08
(0.76)
0.00
(0.71)
0.00
(0.71)
0.00
(0.71)
0.08
(0.76
SEm 5.32 8.35 2.39 1.43 0.18 0.18 0.15 0.14 0.17 0.15 0.15 0.14
C.D. at 5% 11.35 17.79 5.10 3.04 0.39 0.38 0.33 0.29 0.36 0.33 0.33 0.29
*Figures in parentheses are transformed by arc-sin transformation.
Green Boll Locule Damage (%)
At 70 DAS, lowest green boll locule damage was recorded on MRC Bt-F
1
cotton (2.75%) followed by
MRC Bt-F
2
(6.25%), JKCH Bt-F
1
(6.76%) and JKCH Bt-F
2
cotton (9.33%) and significantly high on
MRC non-Bt (26.02%) and JKCH non-Bt cotton (13.09%). Almost similar trend was also recorded in
term of locule damage at 90 DAS. Similarly, significantly lowest damage was noted on MRC Bt F
1

cotton (11.41%) as compared to other hybrid tested at 105 DAS. The locule damage (13.03% and
14.20%) was recorded in JKCH Bt F
1
and MRC Bt F
2
cotton, respectively, but both were statistically at
par regarding locule damage. But, significantly higher locule damage was recorded on MRC non-Bt
(24.88%) and JKCH non-Bt cotton (22.43%) compared with all Bt hybrids tested (Table 2).
The lowest locule damage was recorded on MRC Bt-F
1
cotton (5.30%) as compared to other
treatments. The locule damage (8.12 and 8.59%) was observed in MRC Bt-F
2
and JKCH Bt F
1
cotton
respectively, both were found at par but significantly higher locule damage was recorded on JKCH Bt-F
2

(12.48%), MRC non-Bt (13.37%) and JKCH non-Bt (17.37%).
Earias sp. Larval Incidence
The larval population was almost negligible on MRC Bt-F
1
cotton (Table 2), while least larval population
was recorded on JKCH Bt-F
1
cotton (0.50 l/boll) followed by MRC Bt-F
2
cotton (0.75 l/20 bolls), at 70
DAS. The highest larvae were recorded on JKCH non-Bt cotton (1.50 l/20 bolls) followed by that of
MRC non-Bt cotton (1.00 l/20 bolls)
At 90 DAS, lowest Earias sp. larvae were recorded on MRC Bt-F
1
cotton (0.25 l/20 bolls) and found
at par with MRC Bt-F
2
cotton (0.50 l/boll), JKCH Bt-F
1
(0.50 l/boll) and JKCH Bt-F
2
cotton (1.25
l/bolls). All Bt cotton hybrids were found significantly superior to JKCH non-Bt and MRC non-Bt cotton
with a mean of 2.75 and 1.75 l/20 bolls, respectively.
On the basis of seasonal dynamics of Earias sp., larval population was lowest on MRC Bt-F
1
cotton
(0.08 l/boll) and at par with MRC Bt-F
2
(0.42 l/boll) and JKCH Bt-F
1
cotton (0.42 l/boll), whereas, it was
found significantly superior to JKCH Bt-F
2
cotton (1.25 l/boll), MRC non-Bt (1.50 l/boll) and JKCH
non-Bt cotton (2.09 l/boll) (Table 2).
1,
-F
2
and

P. gossypiella Larval Incidence
P. gossypiella larval incidence was none on MRC Bt-F
1
and JKCH Bt-F
1
cotton, while least larval
population was recorded in MRC Bt-F
2
(0.25 l/20 bolls), JKCH Bt-F
2
(0.25 l/20 bolls), MRC non-Bt
(0.25 l/20 bolls) and JK non-Bt cotton (0.50 l/20 bolls) and were at par with each other at 70 DAS (Table
2).
At 90 DAS, no P. gossypiella larvae were recorded on MRC Bt-F
1
and MRC Bt-F
2
cotton but least
recorded on JKCH Bt-F
1
cotton (0.25 l/boll). Significantly higher larval population was recorded on
MRC non-Bt (1.75 l/20 bolls) and JKCH non-Bt cotton (2.80 l/boll) from Bt cotton hybrids tested.
Similar trend was also recorded regarding P. gossypiella larval incidence on different Bt and non-Bt
cotton hybrids at 105 DAS.
The larval population of P. gossypiella was lowest on MRC Bt-F
1
cotton (0.08 l/boll) and was at par
with JKCH Bt-F
1
cotton (0.42 l/boll) and Bt-F
2
(0.49 l/boll). The significantly higher population of P.
gossypiella was recorded in MRC Bt F
2
and JKCH Bt F
2
followed by that of JKCH non-Bt (1.75 l/boll)
and MRC non-Bt cotton (1.09 l/boll) (Table-2).
Fruiting Bodies (Shedding) Damage (%), Green Boll Damage and H. Armigera Survival
under Caged and Un-caged Conditions During 2007
H. armigera larvae were released under caged and un-caged condition coincided with natural infestation
in un-caged plants of different Bt cotton and non-Bt cotton hybrids. To evaluate the performance of F
1

and F
2
of MRC7017Bt and JKCH1947Bt against H. armigera under high pressure of artificial infestation
was recorded in term of fruiting bodies damage (%), green boll damage (%) and H. armigera larval
survival (%) at 12 days after larval released (Table 3).
TABLE 3: COMPARATIVE PERFORMANCE OF BGII AND JKCH-1947 UNDER ARTIFICIAL RELEASE OF H. ARMIGERA DURING, KHARIF, 2007
Hybrids

Damage (%) at 12 DAR* H. Armigera Survival (%) at 12
DAR* Shedding Green Bolls Damaged (%)
C UC Mean C UC Mean C UC Mean
JKCH 1947Bt
Non-Bt 37.50 (37.71) 36.25 (37.01) 36.87 14.00 (21.71) 10.56 (18.96) 12.26 14.00 (21.71) 9.0 (17.13) 11.50
Bt-F
2
16.25 (23.59) 16.25 (23.59) 16.25 7.00 (14.29) 5.21 (13.04) 6.10 7.00 (14.29) 4.0(10.89) 5.50
Bt-F
1
6.25(12.33) 6.25 (12.33) 6.25 0.00 (4.06) 1.85 (7.81) 0.92 0.00 (4.06) 0.00(4.06) 0.00
MRC 7017Bt
Non-Bt 41.25 (39.94) 40.00 (39.20) 40.62 13.00 (20.88) 12.84 (20.81) 12.92 13.00 (20.88) 6.00(13.72) 9.50
Bt-F
2
11.25 (19.52) 6.25 (12.33) 8.75 3.00 (9.67) 1.38 (6.60) 2.19 3.00 (9.67) 3.00(9.67) 3.00
Bt-F
1
2.50 (7.65) 2.50 (7.65) 2.50 0.00 (4.05) 0.29 (4.20) 0.15 0.00 (4.06) 0.00(4.06) 0.00
SEm 4.07 4.44 3.12 1.06 3.12 2.67
CD at 5% 8.67 9.47 6.65 2.26 6.65 5.69
Shedding Damage (%)
Under caged conditions, minimum shedding damage due to the artificial release of H. armigera was
recorded with a mean of 2.5% in MRC Bt-F
1
cotton and JKCH Bt-F
1
cotton (6.25 %), respectively, both
at par to each other. Similarly medium shedding damage was recorded in MRC Bt-F
2
(11.25 %) and
JKCH Bt-F
2
cotton (16.25%). While significantly higher shedding damage (41.25 and 37.50%) was
recorded on MRC non-Bt and JKCH non-Bt cotton, respectively than all the Bt cotton hybrids tested.
Under un-caged condition, significantly least shedding damage was recorded on MRC Bt-F
1
cotton
(2.5%), as compared to other treatments. Medium shedding damage (6.25%) was recorded in MRC Bt-F
2

and JKCH Bt-F
1
cotton, while, significantly higher shedding damage was recorded in JKCH Bt-F
2
cotton
(16.25%) followed by that on JKCH non-Bt cotton (36.25%) and MRC non-Bt (40.62%).

Green Boll Damage (%)
Almost negligible green boll damage (%) was recorded on MRC Bt-F
1
and JKCH Bt-F
1
cotton under
caged conditions but both were found at par with MRC F
2
cotton (3%), whereas, these were significantly
higher than JKCH Bt-F
2
(7.00%),

JKCH non-Bt (14%) and MRC non-Bt cotton (13%).
Similarly, lowest green boll damage observed on MRC Bt-F
1
cotton (0.29%) followed by that of
MRC Bt- F
2
1
cotton (1.85%) under un-caged condition. However, these were
found significantly superior in term of green boll damage with the mean of 5.21 10.56 and 12.84%
observed in JKCH Bt F
2
, JKCH non-Bt cotton (10.56%) and MRC non-Bt, respectively.
On the basis of mean of caged and un caged condition minimum green boll damage was recorded on
MRC Bt-F
1
cotton (0.15%), followed by JKCH Bt-F
1
(0.92%), MRC Bt-F
2
2

cotton (6.10%). However, higher green boll damage was recorded on MRC non-Bt (12.92%) and JKCH
non-Bt cotton (12.26%).
H. armigera Larval Survival (%) at 12 Days after Release
The larval survival (%) of H. armigera was none on MRC Bt-F
1
and JKCH Bt-F
1
cotton, under caged
conditions, while lowest recorded on MRC Bt-F
2
cotton (3%) followed by JKCH Bt-F
2
cotton (7%) both
were found at par regarding larval survival (%). However, larval survival was observed highest on MRC
non-Bt (13%) and JKCH non-Bt cotton (14%).
Under un-caged conditions, H. armigera larval survival (%) was none on MRC Bt-F
1
and JKCH Bt-
F
1
cotton, whereas, lowest recorded on MRC Bt-F
2
(3%) followed by JKCH Bt-F
2
cotton (4%) However,
significantly higher larval survival (%) was recorded on MRC non-Bt (6%) and JKCH non-Bt cotton
(9%). Larval survival on JKCH non-Bt cotton was significantly higher from all other hybrids even MRC
non-Bt cotton. In contrast the larval survival was higher under caged conditions compared with un-caged
conditions.
Mean H. armigera survival was recorded zero on MRC Bt-F
1
and JKCH Bt-F
1
cotton, whereas, least
recorded on MRC Bt-F
2
cotton (3%), followed by JKCH Bt-F
2
(5.5%), MRC non-Bt (9.5%) and JKCH
non-Bt cotton (11.50%). Data revealed that the Bt-F
1
cotton hybrids were superior over Bt-F
2
and non-Bt
cotton hybrids with zero % H. armigera survival. However, Bt-F
2
hybrids were found significantly
superior over non-Bt hybrids in term of H. armigera survival.
Open Boll Locule Damage (%) and P. gossypiella Larval Incidence at Different DAS
MRC Bt-F
1
cotton had minimum open boll damage (4.71%) which was at par with JKCH Bt-F
1
cotton
(6.12%), and both were found significantly to JKCH Bt-F
2
2
cotton (10.83%),
JKCH non-Bt cotton (25.36%) and MRC non-Bt (28.37%) at 127 DAS (Table 4).
TABLE 4: COMPARATIVE PERFORMANCE OF MRC 7017BT AND JKCH 1947BT AGAINST INFESTATION AND LARVAL POPULATION OF P. GOSSYPIELLA AND SEED COTTON
Hybrids

Open Boll Locules Damage (%) at DAS* P. gossypiella Larvae/ Open Bolls** Seed Cotton
Damage (%)*
Seed Cotton
Yield (q/ha) 127 160 Mean 127 160 Mean
JKCH 1947Bt
Non-Bt 25.36 (30.06) 27.13 (31.27) 26.25 (30.78) 0.75 (1.09) 1.00 (1.18) 0.87 (1.14) 11.72 (19.88) 16.08
Bt-F
2
10.81(18.94) 13.80 (21.61) 12.31 (20.384) 0.50 (0.97) 0.75 (1.09) 0.63 (1.06) 6.38 (14.55) 21.08
Bt-F
1
6.12 (14.10) 8.45 (16.79) 7.29 (15.56) 0.00 (0.71) 0.25 (0.84) 0.12 (0.78) 3.72 (11.08) 23.15
MRC 7017Bt
Non-Bt 28.37 (32.06) 34.05 (35.63) 31.21 (33.89) 0.75 (1.10) 1.25 (1.31) 1.00 (1.21) 14.52 (22.23) 15.91
Bt-F
2
10.83 (19.04) 10.78 (18.96) 10.81(19.04) 0.00 (0.71) 0.50 (0.97) 0.25 (0.85) 3.47 (10.33) 22.76
Bt-F
1
4.71(12.37) 9.01(17.40) 6.80 (15.04) 0.00 (0.71) 0.00 (0.71) 0.00(0.71) 2.23 (8.48) 27.00
SEm 2.20 2.58 1.80 0.14 0.17 0.12 1.91 1.01
CD at 5 % 4.69 5.49 3.85 0.30 0.37 0.26 4.07 2.14
1,
-F
2
and

The lowest locule damage was recorded on JKCH Bt-F
1
cotton (8.45%), followed by MRC Bt-F
1

2
(10.78%) and these were found to JKCH Bt-F
2
cotton (13.80%), JKCH non-Bt
(27.13%) and MRC non-Bt (34.05%) at 160 DAS.
The mean locule damage (6.80 and 7.29%) was lowest on MRC Bt-F
1
cotton and JKCH Bt-F
1
,
respectively. Similarly, medium infestation of locule damage (10.81 and 12.31%) was observed on MRC
Bt-F
2
and JKCH Bt-F
2,
respectively. However, significantly higher locule damage was recoded on MRC
non-Bt (31.21%) and JKCH non-Bt cotton (26.25%) from all the Bt cotton hybrids tested.
There was no single larvae of P. gossypiella was observed on MRC Bt-F
1
, MRC Bt-F
2
, JKCH Bt-F
1
cotton but, least larval population was recorded on JKCH Bt-F
2
cotton (0.50 l/boll) followed by that of
MRC non-Bt (0.75 l/boll) and JKCH non-Bt cotton (0.75 l/boll) at 127 DAS and these were found at par
to each other in relation to larval population. Almost similar results were obtained at 160 DAS regarding
larval population of P. gossypiella but, least population were recorded on MRC Bt- F
2
(0.50 l/boll)
followed by MRC non-Bt (1.00 l/boll) and JKCH non-Bt (1.25 l/boll).
On the basis of the mean of larval population/bolls, there were no single larvae observed on MRC Bt-
F
1
, and JKCH1947Bt F
1
, but lowest recorded on MRC Bt-F
2
(0.25 l/boll). However, significantly higher
larvae were recorded on JKCH Bt-F
2
cotton (0.63 l/boll), JKCH non-Bt (0.87 l/boll), MRC non-Bt (1.0
l/boll) (Table 4).
Seed Cotton Damage (%) and Seed Cotton Yield (q/ha)
The data were recorded on seed cotton damage and seed cotton yield (q/ha) for comparative yield
performance of different Bt and non-Bt cotton hybrids (Table-4).
The seed cotton damage (2.23, 3.47, 3.77, 16.38, 14.32 and 19.80%) was recorded on MRC Bt-F
1

MRC Bt-F
2
JKCH Bt-F
1
and JKCH Bt-F
2,
MRC non-Bt and JKCH non-Bt cotton, respectively. The F
1,
F
2

of MRC Bt and JKCH Bt were found significantly superior to non-Bt cotton regarding damaged seed
cotton due to bollworms infestation.
The highest seed cotton yield (27.00 q/ha) was recorded in MRC Bt F
1
followed by JKCH Bt (23.15
q/ha), MRC Bt F
2
(22.76 q /ha, JKCH Bt F
2
(21.08 q/ha), JKCH non-Bt (16.08 q/ha) and MRC non-Bt
7017 (15.91 q/ha). The MRC Bt F
1
was found to be significantly superior among the entire hybrid tested,
whereas, JKCH F
1
was statistically at par with MRC Bt F
2
regarding yield, both had significantly more
yield than the non-Bt of JKCH and MRC (Table 4).
During these studies, less infestation due to bollworms complex like Earias sp., H. armigera and P.
gossypiella was observed on MRC Bt- F
1
cotton and JKCH Bt F
1
and both the hybrid found significantly
superior to other treatments. Similarly infestation of bollworms was recoded in terms of terminal bud
damage, fruiting bodies damage, green boll locule damage, open boll locule damage; seed cotton %
damage was observed less in MRC Bt-F
1
cotton as compared to JKCH Bt-F
1
cotton. The MRC Bt-F
2

cotton showed less bollworms incidence and damage to the cotton plants or different parts than JKCH Bt-
F
2
and non-Bt of both cotton. The seed cotton yield (27 q/ha) was significantly higher in MRC Bt-F
1
than
JKCH Bt-F
1
, whereas seed cotton yield of MRC Bt-F
2
cotton was at par with JKCH Bt-F
1
and Bt-F
2
, but
higher than those of both non-Bt cotton. These results are in agreement with the findings of Jackson et al.
(2003) who reported the effectiveness of Bollgard-II cotton against H. zea incidence regarding less
squares and bolls damage. Udikere et al. (2003) showed that the three Bt cotton hybrids, MECH-12Bt,
MECH-162Bt and MECH-184 were able to reduce larval populations of H. armigera up to 40%, E.
vitella up to 30-40% and P. gossypiella up to 60-80% in south India. The dual gene cotton like MRC
7017Bt with cry1Ac + cry2Ab genes found superior, as compared to Bollgard cotton against bollworms
incidence (Adamczyk et al., 2001) and He et al. (2004) reported that Bt cotton SGK-321 with cry1Ac +
cpTi genes was most effective against O. furnacalis (in artificial release) than GK2Bt (with cry1Ac gene)
and non-Bt counterparts. Bt cotton with dual stacked genes showed better efficacy against pink bollworm
and tobacco caterpillar in India (Badiger et al., 2011).
The H. armigera larval survival after artificial release under caged and un-caged conditions was very
less on MRC Bt-F
1
cotton than JKCH Bt-F
1
, and Bt-F
2
and non-Bt of both the cotton hybrids tested.
Halcomb et al. (2000) reported that Bt cotton showed lower fruiting bodies damage and less larval
survival of H. virescens and H. zea after 48 hr after infestation, compared with non-Bt-cotton. Parker and
Luttrell (1999) reported interplant movement of H. virescens larvae from Bt cotton Event 531 (with
cry1Ac) to non-Bt (Coker 312) cotton planted in pure and mixed form and less fruiting bodies damage on
Bt cotton than non-Bt cotton..
In the present studies, MRC Bt-F
1
cotton was found superior over all the Bt-F
1
, Bt-F
2
and non-Bt
cotton hybrids tested with lower bollworms incidence, higher seed cotton yield and least P. gossypiella
larvae carry over for next season in the field.
Surulivelu et al. (2004) reported significantly low P. gossypiella incidence (0-0.5 larva/20 green
bolls) in RCH2Bt, RCH20Bt, RCH144Bt and MECH162Bt cotton than non-Bt (1.3 to 2.2 l/boll) cotton
hybrids and conventional cotton. The Bt cotton lines viz., Coker-312 Bt, -62 Bt, -65 Bt and -82 Bt with
cry1Ac gene was found superior with 95% less rosetted blooms and 97-99% less seed damaged than non-
Bt (MD51 ne) cotton (Wilson et al., 1992). Liu et al. (2001) reported that Bt cotton with cry1Ac gene
was found more effective against susceptible and resistant strains of P. gossypiella. Ameta and
Sarangdevot (2007) reported that highest seed cotton yield 30.14 q/ha was recoded in RCH- 134 followed
by MRC- (27.96 q /ha), Ankur 09 (26.43 q/ha) and Ankur 651 (24.65 q/ha) and also observed that
among the bollworms, population of Earias spp and H. armigera was very low and they did not inflict
damage to square flower and green bolls, but open boll and locule damage was ranged from 4.25 in RCH
134 to 11.15 % in Ankur 651 Bt. Success of Bt crops in delaying substantial resistance evolution in the
target pests could be attributed to many ecological factors and resistance management tactics (Tabashnik
et al., 2003; 2009).
Acknowledgements: Authors are grateful to the Director, IARI for providing infrastructure and
encouragement for the research work reported herein.
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Intrinsic Rate of Increase and Life Parameters

of Cotton Leaf Eating Caterpillar Spodoptera litura
on Bollgard II Hybrids
Golla M.V. Prasada Rao
1
, T. Sujatha
2
, G.A.D. Grace
3
,
N.V.V.S.D. Prasad
4
and V. Chenga Reddy
5

1,4
(Entomology),
5
Principal Scientist (Cotton)All India Coordinated Cotton Improvement Project,
Regional Agricultural Research Station,
Lam, Guntur, Andhra Pradesh522 034
2,3
Department of Entomology, Regional Agricultural Research Station, Lam,
Guntur, Andhra Pradesh522 034
AbstractThe Bt cotton area in India increased from 0.5 lakh ha in 2002-2003 to 11.00 million lakh ha in 2010-11.
The Bt technology has been highly effective against major bollworms of cotton. However, Bt cotton hybrids, containing
single (Cry 1 Ac) and double (Cry 1 Ac and Cry 2 Ab) genes, are not very effective against Spodoptera litura which is
causing damage to Bt cotton. With this background study was conducted during 2010 season to know the intrinsic
rate of increase of S. Litura on four popularly grown Bollgard II cotton hybrids and life parameters of the pest were
computed in comparison with their counterpart Bollgard and NBt hybrids. The methods suggested by Morris and
Miller and Chaudhary and Bhattacharya were used for constructing the life tables. For computing various life
parameters a computer programme was developed using MS-Excel. All the life parameters of the pest were
significantly and negatively affected on Bollgard II hybrids. Net reproductive rate (Ro) on Bollgard II hybrids ranged
from 16.36 to 22.53, whereas it ranged from 43.89 to 61.17 on Bollgard hybrids and it ranged from 86.79 to 108.22 on
NBt hybrids. Potential fecundity (Pf) of the pest was also negatively affected and pf ranged from 635 to 880 on
Bollgard II hybrids, while it ranged from 906 to 1120 on Bollgard hybrids. Highest Pf was recorded on NBt hybrids
(1182 to 1333). Mean generation time was higher on Bollgard II hybrids followed by Bollgard and NBt hybrids.
Lowest mean generation time (Tc) of 49.92 days was recorded on Bunny NBt and highest Tc of 57.67 day was
recorded in Mallika BG II. The intrinsic rate of increase (r
c
) ranged from 0.05 to 0.09 females / female / day. Lowest r
c

was recorded in Bollgard II hybrids and highest r
c
of 0.09 was recorded in NBt hybrids. In Bollgard hybrids r
c
ranged
from 0.07-0.08. Insect could double its numbers around week days in NBt, nine days in Bollgard and around two
weeks in Bollgard II hybrids. Lowest weekly multiplication rate of 1.40 times was recorded in Mallika BGII and
highest rate of 1.91 times was recorded in Tulasi NBt. Among the four hybrids tested, the effect of toxins on the
growth and development of the pest was highest in Mallika hybrids. The Bollgard II hybrids have significantly
negative effect on the growth and development of S. Litura.
Cotton is an important fibre of India and cultivated in an area of 11.00 million ha with a production
of 325.50 lakh bales and productivity of 503 kg/ha (AICCIP 2010-11). Introduction of Bt cotton has
revolutionized the cotton production in India as the cotton production prior to 2002 suffered huge losses
due to its susceptibility to insect pests (CICR, 2009). Transgenics of cotton with Bacillus thuringiensis
var. kurstaki genes encoding Cry1Ac proteins have been first introduced with the expectations of
reduction in number of pesticide applications, increase in natural enemies, and reduction in farmers
exposure to pesticides. (Gianessi and Carpenter, 1999). The Cry 1 Ac provides control of the American
bollworm Helicoverpa armigera, pink bollworm, Pectinophora gossypiella and spotted bollworm, Earias
vitella. However, it is less effective for the control of beet armyworm, Spodoptera exigua and fall
armyworm, Spodoptera frugiperda (Macintosch et al., 1990; Adamczyk et al., 1998). Likewise, the
tobacco caterpillar Spodoptera litura is also less susceptible to Bollgard cotton containing Cry 1 Ac
(Bheemanna et al., 2008, Lakshmi Sowjanya, 2008 and Sivasupramaniam et al., 2008). Bollgard II
cotton (Cry 1 Ac and Cry 2 Ab) was commercially released and was found to have superior insecticidal
activity compared to bollgard and in particular to augment late season insect control (Akin et al., 2001
and Jackson et al., 2002). The dual gene technology is being considered as an improvised pest
management method not just for its enhanced efficacy but also as an efficient resistance management
29
Intrinsic Rate of Increase and Life Parameters of Cotton Leaf Eating Caterpillar Spodoptera litura on Bollgard II Hybrids 175
strategy. The addition of other Cry proteins stacked with Cry 1Ac has improved the efficacy against army
worms (Adamczyk et al., 2001; Stewart et al., 2001; Adamczyk & Gore 2004). The present study aims to
understand the effect of Bollgard cotton hybrids on the growth and development of S. litura through
laboratory life table studies.
MATERIAL AND METHODS
A laboratory culture of S. Litura was maintained on fresh leaves of NBt cotton at 25 2
0
C and 75 3 %
relative humidity. The adults were fed a liquid diet containing honey and sugar and paired in separate
containers where they were allowed to lay eggs on folded papers. The hatchability of eggs of a cohort
laid on a single day was initially determined by counting the number of eggs hatched in three replicates.
Hundred larvae that hatched from one cohort of eggs were transferred to freshly plucked leaves of
Bollgard, Bollgard II and NBt cotton hybrids viz., Mallika, Bunny, Tulasi and Rasi and maintained in
ventilated plastic containers with wet cotton and filter paper to retain the turgidity of leaves. The larvae
were maintained in groups of 20 for the initial five days to cater to their gregarious nature. Later, the
larvae were maintained individually till death or pupation. Data on mortality were recorded everyday till
all the adults died. The methods suggested by Morris and Miller (1954) and Chaudhary and Bhattacharya
(1986) were used for constructing the life tables and for computing various life parameters a computer
programme developed using MS-Excel was used for processing the data.
Age specific survivorship and mortality of S .litura on different Bollgard II, Bollgard and NBt cotton
hybrids are presented in figures 1-12. Survival of early instars was very low when reared on Bollgard II
hybrids which dropped sharply by 10
th
day and ranged from 46 to 62 %. Lowest survival per cent of 46
was recorded on Mallika, followed by in Tulasi (53 %) and highest survival per cent 62% was recorded
in Rasi and Bunny hybrids. While, Bollgard and NBt hybrids survival per cent ranged from 70 to 88 and
78 to 94 respectively. Thus, toxin expression in Bollgard II hybrids proved very toxic to the early instar
larvae of the test insect. Among the different Bollgard II hybrids tested Mallika BGII is more toxic than
Tulasi, Rasi and Bunny. Among different hybrids age specific female survivorship was the highest in
NBt hybrids (0.25-0.34) followed by Bollgard (0.17 to 0.26) and Bollgard II (0.07-0.11) indicating very
low pupation and adult emergence in Bollgard II hybrids (Fig. 13-24). Lowest survivorship was recorded
in Mallika BG II (0.07), followed by in Rasi BG II (0.10) and Tulasi BG II (0.11) and Bunny BG II
(0.11).
A comparison of life parameters of S. Litura on Bollgard II, Bollgard and NBt cotton hybrids of
Mallika, Tulasi, Rasi and Bunny has been presented in Table 1. The lowest net reproductive rate (R
0
) of
S. Litura was recorded on Bollgard II followed by Bollgard and NBt hybrids. Similar trend was recorded
with regard to other life parameters like potential fecundity, intrinsic rate of increase, finite rate of
increase, annual rate of increase and weekly multiplication rate. Mean generation time (T) was the
longest on Mallika BGII (57.67 days). Insect could double its numbers around week days in NBt, nine
days in Bollgard and around two weeks in Bollgard II hybrids. Lowest weekly multiplication rate of 1.40
times was recorded in Mallika BGII and highest rate of 1.91 times was recorded in Tulasi NBt. Though
the reaction of the pest to Bollgard II hybrids in terms of life parameters was found to be similar, the
effect of toxin expression was stronger in Mallika hybrids compared to other test hybrids viz., Tulasi,
Rasi and Bunny.

Fig. 1: Age Specific Survivorship (Ix) and Mortality (dx) of Spopdoptera Litura on Mallika BGII

Fig. 2: Age Specific Survivorship (Ix) and Mortality (dx) of Spodoptera Litura on Mallika BG

Fig. 3: Age Specific Survivorship (Ix) and Mortality (dx) of Spodoptera Litura on Mallika NBt

Fig. 4: Age Specific Survivorship (Ix) and Mortality (dx) of Spodoptera Litura on Tulasi BGII
0
20
40
60
80
100
120
1 3 5 7 9 1113 1517 1921 2325 2729 31 3335 3739 4143 4547 4951 5355 5759 61
Age in days
I
x
0
2
4
6
8
10
12
14
16
18
20
d
x
lx
dx
0
20
40
60
80
100
120
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49 51 53 55 57 59
Age in days
I
x
0
2
4
6
8
10
12
14
16
18
20
d
x
lx
dx
0
20
40
60
80
100
120
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49 51 53 55 57
Age in days
I
x
0
2
4
6
8
10
12
14
16
18
20
d
x
lx
dx
p p
0
20
40
60
80
100
120
1 3 5 7 9 11 1315 17 1921 23 25 2729 31 3335 37 39 4143 45 4749 51 53 5557 59 61
Age in days
I
x
0
2
4
6
8
10
12
14
16
d
x
lx
dx

Fig. 5: Age Specific Survivorship (Ix) and Mortality (dx) of Spodoptera Litura on Tulasi BG

Fig. 6: Age Specific Survivorship (Ix) and Mortality (dx) of Spodoptera Litura on Mallika NBt

Fig. 7: Age Specific Survivorship (Ix) and Mortality (dx) of Spodoptera Litura on Rasi BGII

Fig. 8: Age Specific Survivorship (Ix) and Mortality (dx) of Spodoptera Litura on Rasi BG
Tulasi BG
0
20
40
60
80
100
120
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49 51 53 55 57 59 61
Age in days
I
x
0
2
4
6
8
10
12
14
16
18
20
d
x
lx
dx
0
20
40
60
80
100
120
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49 51 53 55
Age in days
I
x
0
2
4
6
8
10
12
14
16
18
20
d
x
lx
dx
0
20
40
60
80
100
120
1 3 5 7 9 1113 15 1719 21 2325 27 29 3133 35 3739 41 4345 47 4951 53 5557 59 61
Age in days
I
x
0
2
4
6
8
10
12
14
16
18
d
x
lx
dx
0
20
40
60
80
100
120
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49 51 53 55
Age in days
I
x
0
5
10
15
20
d
x
lx
dx

Fig. 9: Age Specific Survivorship (Ix) and Mortality (dx) of Spodoptera Litura on Rasi NBt

Fig. 10: Age Specific Survivorship (Ix) and Mortality (dx) of Spodoptera Litura on Bunny BGII

Fig. 11: Age Specific Survivorship (Ix) and Mortality (dx) of Spodoptera Litura on Bunny BG

Fig. 12: Age Specific Survivorship (Ix) and Mortality (dx) of Spodoptera Litura on Bunny NBt
on Rasi NBt
0
20
40
60
80
100
120
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49 51 53 55
Age in dyas
I
x
0
2
4
6
8
10
12
14
16
18
20
d
x
lx
dx
y
0
20
40
60
80
100
120
1 3 5 7 9 11 1315 1719 2123 2527 293133 3537 3941 4345 4749 5153 555759 61
Age in days
I
x
0
5
10
15
20
d
x
lx
dx
0
20
40
60
80
100
120
1 3 5 7 9 11 13 1517 19 21 23 25 27 29 31 33 3537 39 41 43 45 47 49 51 53 5557 59
Age in days
I
x
0
2
4
6
8
10
12
14
16
18
20
d
x
lx
dx
0
20
40
60
80
100
120
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45 47 49 51 53 55
Age in days
I
x
0
2
4
6
8
10
12
14
16
18
20
d
x
lx
dx

Fig. 13: Age Specific Female Survivorship and Natality Rate of S. Litura on Mallika BGII

Fig. 14: Age Specific Female Survivorship and Natality Rate of S. Litura on Mallika BG

Fig. 15: Age Specific Female Survivorship and Natality Rate of S. Litura on Mallika NBt

Fig. 16: Age Specific Female Survivorship and Natality Rate of S. Litura on Tulasi BGII
0
0.005
0.01
0.015
0.02
0.025
56.5 57.5 58.5 59.5
Pivotal age in days
I
x
0
50
100
150
200
250
300
350
400
m
x
Ix
mx
0
0.01
0.02
0.03
0.04
0.05
0.06
0.07
0.08
0.09
54.5 55.5 56.5 57.5 58.5
Pivotal age in days
I
x
0
50
100
150
200
250
300
350
400
450
m
x
Ix
mx
0
0.01
0.02
0.03
0.04
0.05
0.06
0.07
0.08
0.09
0.1
51.5 52.5 53.5 54.5 55.5 56.5
Pivotal age in days
I
x
0
50
100
150
200
250
300
350
400
m
x
Ix
mx
0
0.01
0.02
0.03
0.04
0.05
0.06
1 2 3 4 5
Pivotal age in days
I
x
0
50
100
150
200
250
300
m
x
Ix
mx

Fig. 17: Age Specific Female Survivorship and Natality rate of S. Litura on Tulasi BG

Fig. 18: Age Specific Female Survivorship and Natality Rate of S. Litura on Tulasi NBt

Fig. 19: Age Specific Female Survivorship and Natality rate of S. Litura on Rasi BGII

Fig. 20: Age Specific Female Survivorship and Natality rate of S. Litura on Rasi BG
0
0.02
0.04
0.06
0.08
0.1
0.12
49.5 50.5 51.5 52.5 53.5 54.5 54.5 54.5
Pivotal age in days
I
x
0
50
100
150
200
250
300
350
m
x
Ix
mx
u as N t
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
49.5 50.5 51.5 52.5 53.5 54.5
Pivotal age in days
I
x
0
50
100
150
200
250
300
350
400
450
m
x
Ix
mx
0
0.01
0.02
0.03
0.04
0.05
0.06
55.5 56.5 57.5 58.5
Pivotal age in days
I
x
0
50
100
150
200
250
300
m
x
Ix
mx
0
0.01
0.02
0.03
0.04
0.05
0.06
0.07
0.08
0.09
49.5 50.5 51.5 52.5 53.5 54.5
Pivotal age in days
I
x
0
50
100
150
200
250
300
350
400
m
x
Ix
mx

Fig. 21: Age Specific Female Survivorship and Natality Rate of S. Litura on Rasi NBt

Fig. 22: Age Specific Female Survivorship and Natality Rate of S. Litura on Bunny BGII

Fig. 23: Age Specific Female Survivorship and Natality Rate of S. Litura on Bunny BG

Fig. 24: Age Specific Female Survivorship and Natality Rate of S. Litura on Bunny NBt
0
0.02
0.04
0.06
0.08
0.1
0.12
49.5 50.5 51.5 52.5 53.5
Pivotal age in days
I
x
0
50
100
150
200
250
300
350
400
450
500
m
x
Ix
mx
y
0
0.005
0.01
0.015
0.02
0.025
0.03
0.035
0.04
0.045
54.5 55.5 56.5 57.5
Pivotal age in days
I
x
0
50
100
150
200
250
m
x
Ix
mx
y
0
0.02
0.04
0.06
0.08
0.1
0.12
49.5 50.5 51.5 52.5 53.5 54.5
Pivotal age in days
I
x
0
50
100
150
200
250
300
350
m
x
Ix
mx
Bunny NBt
0
0.02
0.04
0.06
0.08
0.1
0.12
48.5 49.5 50.5 51.5 52.5 53.5
Pivotal age in days
I
x
0
50
100
150
200
250
300
350
400
450
m
x
Ix
mx
TABLE 1: LIFE TABLE PARAMETERS OF SPODOPTERA LITURA ON DIFFERENT COTTON GENOTYPES
S.
No.
Life Parameter Bunny Mallika Rasi Tulasi
NBt BG BGII NBt BG BGII NBt BG BGII NBt BG BGII
1 Net Reproductive Rate
(R
0
)
89.43 61.17 18.54 90.55 60.67 16.36 86.79 43.89 22.53 108.22 52.1
2
20.36
2 Potential Fecundity (Pf) 1263 924 635 1312 1065 880 1333 1120 763 1182 906 737
3 Intrinsic rate of increase
(approximate) - r
c

0.09 0.08 0.05 0.09 0.07 0.05 0.09 0.07 0.06 0.09 0.08 0.05
4 Mean Generation time (T
c
)
in days
49.92 50.45 55.84 52.80 55.57 57.67 50.61 50.76 56.40 50.60 50.5
5
56.78
5 Finite rate of increase () 1.09 1.08 1.06 1.09 1.08 1.05 1.09 1.08 1.06 1.10 1.08 1.05
6 Doubling time (DT) 7.71 9.51 13.26 8.12 9.41 14.30 7.86 9.30 12.55 7.49 8.86 13.06
7 Annual rate of increase
(ARI)
1.86X10
1
4

8.44X10
1
2

1.9X
10
8

3.36X
10
13

4.81X10
8

4.81
X 10
7
9.5X
10
13

6.4X
10
11

5.6X
10
8

4.74X
10
14

2.5X
10
12

2.5X
10
8

8 Weekly multiplication 1.88 1.77 1.44 1.82 1.68 1.40 1.85 1.68 1.47 1.91 1.73 1.45
Adamczyk et al., (1998) and Stewart et al., (2001) reported little effect of Bt on various biological
parameters of S. exigua, S. frugiperda and S. littoralis. Yu Yueshu et al., (2004) reported non significant
differences between transgenic Bt cotton Kemian 1 and non-transgenic Bt cotton Yumian 1 in the rate of
damage to leaves, flowers and buds due to S. Litura. Basavaraj et al ., (2008) also reported non-
significant differences between Bt and NBt genotypes at 80, 120 and 140 days of crop age for various
biological parameters viz., larval period, larval weight, larval survival, pupal period, pupal weight and
Adult emergence. Whereas, Chitkowski et al., (2003) reported that the mortality of both S. frugiperda
and S. exigua were significantly greater on Bollgard II plant material than on either Bollgard or
conventional cotton. Similarly, Bheemanna et al., (2008), Lakshmi Sowjanya (2008) and
Sivasupramaniam et al., (2008) also reported that Bollgard II traits were more efficacious against S.
Litura with nil larval population compared to BG and non-Bt cotton traits. Adamczyk et al., (2008)
evaluated Bollgard (Cry 1 Ac), Bollgard II (Cry 1 Ac + Cry 2 Ab) and Wide strike (Cry 1 Ac + Cry 1 F)
technologies against beet armyworm, S. exigua and fall armyworm S. frugiperda. They reported that both
dual gene traits are more efficacious against these armyworm species than Bollgard.
Thus, from the present study it is concluded that Bollgard II hybrids have significant negative effect
on the growth and development of S. Litura which could delay the development of resistance to Cry
toxins in the near future.
ACKNOWLEDGEMENT
The authors are highly thankful to the Associate Director of Research, Regional Agricultural Research
Station, Lam, Guntur, for providing necessary facilities to carry out the research work.
REFERENCES
[1] AICCIP, Annual Report (2010-2011) - All India Coordinated Cotton Improvement Project, Coimbatore, Tamil Nadu-641
003.
[2] Adamczyk, Jr. J.J., Adams, L. C. and Hardee, D. D. (2001) - Field efficacy and seasonal expression profiles for terminal
leaves of single and double Bacillus thuringiensis toxin cotton Genotypes - J. Econ. Entomol, 94 (6):1589-1593.
[3] Adamczyk, Jr. J.J., Mascarenhas, V.J., Church, G.E., Leonard, B.R. and Graves, J. B. (1998) - Susceptibility of
conventional and transgenic cotton bolls expressing the Bacillus thuringiensis Cry 1 Ac endotoxin to fall armyworm
(Lepidoptera: Noctuidae) and beet armyworm (Lepidoptera: Noctuidae) injury- J. Agric. Entomol, 15:163-171.
[4] Adamczyk, Jr., Greenberg, S., Armstrong, J.S and Mullins, W.J. (2008) - Evaluation of Bollgard II and widestrike
technologies against beet and fall armyworm larvae (Lepidoptera: Noctuidae) - Florida Entomologist, 91 (4):532-536.
[5] Adamczyk, Jr., J.J and Gore, J. (2004)- Laboratory and field performance of cotton containing Cry 1Ac, Cry 1F, and both
Cry 1Ac and Cry 1F (WideStrike) against beet army worm and fall armyworm larvae (Lepidoptera: Noctuidae) -Florida
Entomol., 87: 427-432
[6] Akin, D. S., Stewart, S. D and Knighten, K. S. (2001) - Field efficacy of cotton expressing two insecticidal proteins of
Bacillus thuringiensis. In: Dugger, P.A., Richter, D. (Eds.) - Proceedings of the Beltwide cotton conference- Vol.2. National
Cotton Council, Memphis, USA. 1041-1043.
[7] Basavaraja, H., Chillar, B. S and Ram Singh.(2008)- Impact of transgenic Bt cotton on biological parameters of Spodoptera
litura (Fabricius) (Lepidoptera: Noctuidae) - Journal of Entomological Research, 32 (3):183-186.
[8] Bhemanna, M., Patil, B. V., Hosamani, A. C. and Bansi, A. B. (2008) - Comparative performance and economics of BG II
Bt cotton under irrigated conditions- J. cotton res. and development, 22 (1): 118-121.
[9] Chaudhary, R. R. P and Bhattacharya, A. K. (1986) - Bioecology of lepidopterous insects on winged bean, Psophocarpus
tetragonolobus (Linnaeus) De condole- Memoirs of the Entomological society of India, 11: 130p.
[10] Chitkowski, R. L., Turnipseed, S. G., Sullivan, M. J. and Bridges, W. C. (2003) - Field and Laboratory cultivation of
transgenic Cottons expressing one or two Bacillus thuringiensis Var Kurtasaki, Berliner proteins for management of
Noctuid pests - J. Economic Entomology, 96 (3):755-762.
[11] CICR (2009) - Cotton Database. Central Institute of Cotton Research, Nagpur. http://www. cicr.org.in.
[12] Gianessi, L. P and Carpenter, J. E. (1999) - Agricultural biotechnology: Insect control benefits - Report from National
Centre for Food and Agricultural Policy, Washington: 63.
[13] Jackson, R. E., Bradley, J. R., Burd, A. D., Van Duyn, J.W., (2002) - Field and greenhouse performance of bollworm on
Bollgard-II genotypes. In: Dugger, P.A., Richter, D. (Eds.), Proceedings of the Beltwide cotton conference, Vol.2. National
Cotton Council, Memphis, USA. 1048-1051.
[14] Lakshmi Sowjanya, P. (2008) - Effect of Bt toxins (Cry1Ac and Cry 1Ac + Cry 2Ab) on the development and management
of Bollworm complex with special reference to Pectinophora gossypiella (Saunders) and Spodoptera litura (Fabricius) on
cotton - Ph.D (Ag.) Thesis, ANGRAU, Hyderabad.
[15] Macintosch, S.C., Stone, T. B., Sims, S. R., Hunst, P. L., Greenplate, J.T., Marrone, P.G., Perlak, F. J., Fischhoff, D. F. and
Fuchs, R. L. (1990) - Specificity and efficacy of purified Bacillus thuringiensis proteins against agronomically important
insects - J. Invertebr., Pathol., 56 : 258-266.
[16] Morris, R. F. and Miller, C. A. (1954) The development of life tables for the spruce bud worm - Canadian Journal of
Zoology., 32: 283-301.
[17] Sivasupramaniam, S., Moar, W. J., Ruschke, L. G., Osborn, J. A., Jiang, C., Sebaugh, J. L., Brown, G. R., Shappley ,Z. W.,
Openhuizen, M. E., Mullins, J. W and Greenplate, J, T. (2008) - Toxicity and Characterization of Cotton expressing
Bacillus thuringiensis Cry 1Ac and Cry2Ab2 proteins for control of Lepidopteran pests- J. Economic Entomology.,
101(2):545-554.
[18] Stewart, S.D., Adamczyk, Jr. J.J., Knighten, K.S. and Davis, F.M. (2001) - Impact of Bt cotton expressing one or two
insecticidal proteins of Bacillus thuringiensis Berliner on growth and survival of noctuid (Lepidoptera) larvae - J. Econ.
Entomol., 94: 752-760.
[19] Yu Yueshu Kang Xiaoxia Lu Yanhui Liang Jiangya Wang Hong Wu Jieyun Yang Yizhong. (2004)-effects of the transgenic
bt cotton on the increase in population of spodoptera litura fabricius - Jiangsu J. Agrl. Sci., 20(3): 169-172.
Field Efficacy of Widestrike Bt Cotton, Expressing

Cry1Ac and Cry1F Proteins, Against Lepidopteran
Pests in India
Moudgal R.K.
1
, Chetan Chawda
1
, Gajendra Baktavachalam
1

Sundara Rajan
1
and Gary D. Thompson
2
1
DowAgroSciences India Pvt Ltd, Mumbai, India
2
Dow AgroSciences LLC, Indianapolis, USA
E-mail: rmoudgal@dow.com
AbstractDow Agro Sciences India Pvt Ltd (DAS) has been developing Wide Strike
TM
Bt cotton, a breeding stack of
Cry 1F (event 281-24-236) and Cry1Ac (event 3006-210-23) in India since 2005. In this process, DAS has conducted
Bio-safety Research Level (BRL) trials since 2008 in South and Central Zones of India with two cotton hybrids
expressing the WideStrike traits namely WS103 and WS106. This article reports efficacy of WideStrike Bt cotton
against Heliothis armigera and Spodoptera litura observed in the BRL trials. The trials demonstrate that WideStrike
cotton provides effective, season-long, whole plant protection from lepidopteran insects leading to increased crop yields
and productivity. Upon regulatory approval, introduction of WideStrike cotton in India will diversify the choice of
insect protection traits available to the Indian farmers and enable effective management of H. armigera and S. litura in
cotton. The availability of additional cotton insect traits will encourage more breeding efforts and contribute to
resistance management and the longevity of all cotton insect traits. This product has already been approved for
commercial cultivation in USA and Brazil and for feed/food import in Australia, Canada, Japan, Korea and Mexico.
Keywords: WideStrike, Transgenic Insect resistant Cotton, H. armigera, S. litura
INTRODUCTION
Insect resistant transgenic cotton hybrids introduced in India have revolutionized cotton production
taking it up from 136 lakh bales in 2002-03 to 325 lakh bales in 2010-11 with lint productivity going up
from 302 to 503 kg/Ha during the same period, as per Cotton Corporation of India
(http://www.cotcorp.gov.in/statistics.asp#area). Productivity increase is mainly attributed to effective
control of bollworms especially H. armigera through wide spread adoption of Bt cotton. Out of the
estimated 11 MM Ha of cotton in India in 2010, 86% or 9.5 MM Ha was planted with Bt cotton hybrids
which is a remarkably high proportion of Bt cotton in a fairly short period of nine years equivalent to an
unprecedented 188-fold increase from 2002 to 2010 (James, 2010). Currently, commercial transgenic
cotton in India expresses either one of these Bt proteins: Cry1Ac (MON531), Cry2Ab (MON15985),
Truncated Cry1Ac (Event1), Cry1Ab/Cry1Ac (GFM Cry1Ac), Truncated Cry1Ac (Dharwad Event) and
Cry1C (MLS9124).
Many surveys and estimates have shown unequivocal tangible economic benefits to farmers who
grew Bt cotton and intangible benefits to farmers and society in terms of reduced use of pesticides in the
environment and reduced exposure of farmers to pesticides. Subsequent to introduction of Bt cotton in
India productivity nearly doubled from 308 kg per hectare in 2001 to 568 kg/ha in 2009. These gains
resulted from average yields in Bt cotton hybrids that were approximately 50% higher than conventional
cotton. To cite specific examples, Padaria et al (2009) reported that cultivation of Bt cotton in two key
cotton states, Punjab and Andhra Pradesh, increased yield by approximately 67%; increased income by
142%, and reduced the frequency of insecticide spray by 62%. Savings from Bt cotton also includes
reduced labor cost to produce the crop. At the aggregate level, Bt cotton is estimated to have enhanced
Indian farm income by US$ 5.1 billion in the period 2002 to 2008 and by US$ 1.8 billion in 2008 alone
(Brookes and Barfoot, 2010). At the individual farmer level, gains from Bt cotton in India have been
estimated at US$ 156 per hectare. Additional aggregate benefits are 50% reductions in insecticide sprays
and increased regional employment opportunities (Subramanian and Qaim, 2009).
30
Field Efficacy of Widestrike Bt Cotton, Expressing Cry1Ac and Cry1F Proteins, Against Lepidopteran Pests in India 185
In India, Dow AgroSciences India Pvt Ltd (DAS) has been developing WideStrike Bt cotton, a
breeding stack of Cry1F (event 281-24-236) and Cry1Ac (event 3006-210-23) for controlling
lepidopteran insect pests, since 2005. In this process, DAS had conducted Biosafety Research Level
(BRL) trials since 2008 in South and Central Zones with two cotton hybrids expressing the WideStrike
traits namely WS103 and WS106. This article reports efficacy of WideStrike Bt cotton against the H.
armigera and S. litura observed in these BRL trials conducted in India.
Planting Material
The experimental material consisted of two WideStrike cotton hybrids (WS103 and WS106 which
express the insecticidal proteins, Cry1Ac and Cry1F), their non-transgenic counterparts namely non-
WS103 and non-WS106 along with commercial Bt cotton check, non-Bt hybrid check and non-Bt
varietal check recommended for respective regions. Further details on Biosafety Research Level trials
(BRLT) are furnished in Table 1.
TABLE: 1: DETAILS OF BIOSAFETY RESEARCH LEVEL TRIALS (BRLT) CONDUCTED BY DOW AGRO SCIENCES INDIA PVT. LTD. IN INDIA SINCE 2008
Details Biosafety Research Level 1 Trial Biosafety Research Level2 Trial
Aurangabad
(Maharashtra)
Vadodara
(Gujarat)
Dharwad
(Karnataka)
Hyderabad
(Andhra
Pradesh)
Attur
(Tamil Nadu)
Dharwad
(Karnataka)
Guntur
(Andhra
Pradesh)
Year and
season
Kharif 2009 and
Kharif 2010
Kharif 2009
and Kharif
2010
Kharif 2008 and
Kharif 2009
Kharif 2008
and Kharif
2009
Kharif 2010 Kharif 2010 Kharif 2010
Date of
Sowing
11.07.2009 and
13.07.2010
07.07.2009
and
16.07.2010
04.07.2008 and
29.07.2009
08.07.2008 and
17.07.2009
05.10.2010 04.09.2010 02.09.2010
Transgenic
test hybrids
WS103 and
WS106
WS103 and
WS106
WS103 and
WS106
WS103 and
WS106
WS103 and
WS106
WS103 and
WS106
WS103 and
WS106
Non-
transgenic
test hybrids
Non-WS103 and
Non-WS106
Non-WS103
and Non-
WS106
Non-WS103 and
Non-WS106
Non-WS103
and
Non-WS106
Non-WS103
and
Non-WS106
Non-WS103
and Non-
WS106
Non-WS103 and
Non-WS106
Bt check Bunny BGI and
Bunny BGII
RCH-2 BGI
and RCH-2
BGII
Bunny BGI RCH-2 BGI RCH-2 BGI and
RCH-2 BGII
Bunny BGI and
Bunny BGII
Bunny BGI and
Bunny BGII
Non-Bt
hybrid check
H-8 and NHH-
44
H-8 and H-12 Bunny Non-Bt
and DHH-11
Bunny Non-Bt
and LAHH-5
RCH-2 Non-Bt Bunny Non-Bt Bunny Non-Bt
Non-Bt
varietal check
PKV Rajat G.Cot 16 Sahana Narasimha Surabhi Sahana Narasimha
Spacing 90 X 60 cm 120 X 45 cm 90 X 60 cm 90 X 90 cm 90 x 90 cm 90 x 90 cm 90 x 90 cm
Plot size 48.6 sq. mt 64.8 sq. mt 48.6 sq. mt 48.6 sq. mt 335.3 sq. mt. 495.7 sq. mt. 491.3 sq mt.
Experimental
design
RCBD RCBD RCBD RCBD Non replicated
large plots
Non replicated
large plots
Non replicated
large plots
Replications 3 3 3 3 Not applicable Not applicable Not applicable
Row length 9 m 9 m 9 m 9 m 23 m 36 m 34 m
Trial
condition
Rainfed Irrigated Rainfed Irrigated Irrigated Rainfed Irrigated
Evaluation of Insect Damage
The lepidopteran pest population and damage was recorded from 10 plants per replication in the
Biosafety Research Level 1 (BRL-1) trials and 30 plants per plot in Biosafety Research Level 2
(BRL-2) trials across different locations during 2008-2010.
American Bollworm
(H. armigera: Larval counts were recorded at weekly intervals from the 8th to 21st week (~ 60 to 150
Days After Sowing).
Fruiting bodies damage: Fruiting bodies damage was recorded starting from the 8th week to the 21st
week after sowing.
Open-boll and locule damage: The total and damaged number of open bolls and locules were
observed during all pickings.
Tobacco caterpillar (S. litura): Number of larvae per plant was recorded weekly from 60 to 150
Days after Sowing.
DATA ANALYSIS
The Biosafety Research Level1 (BRL-1) trial data was subjected to Analysis of Variance (ANOVA)
using ARM 7.4.0 statistical package. The significance of difference between the treatments was tested by
F-test, while the treatment means were compared by Least significance difference (LSD) at p=0.05.
However, as the Biosafety Research Level2 (BRL-2) trials being non-replicated, statistical analysis
(ANOVA) could not be done, but, the data was used to calculate the mean and its standard error.
RESULTS
American Bollworm and Tobacco Caterpillar Population and Damage
in Biosafety Research Level-1 (BRL-1) Trials Conducted during 2009 and 2010 in Central Zone
The population of American bollworm, H. armigera was widely observed both in 2009 and 2010 BRL-1
trials in the Central Zone. However, when compared to 2010 the population of H. armigera was
generally low in 2009 across the season at both the locations (Figure 1 & 2). It also correlated with higher
pest damages in 2010 compared to the previous year. Among the locations, bollworm damage was more
in Vadodara especially during 2010. During both the years of BRL-1 trials in Central Zone, negligible
population of S. litura was observed across the entire observation period, however, the WideStrike
hybrids were completely free from S. litura infestation.

Fig. 1: Population of H. Armigera on Non-transgenic Cotton in Aurangabad

Fig. 2: Population of H.armigera on Non-transgenic Cotton in Vadodara
-2
0
2
4
6
8
7
-
S
e
p
1
4
-
S
e
p
2
1
-
S
e
p
2
8
-
S
e
p
5
-
O
c
t
1
2
-
O
c
t
1
9
-
O
c
t
2
6
-
O
c
t
2
-
N
o
v
9
-
N
o
v
1
6
-
N
o
v
2
3
-
N
o
v
3
0
-
N
o
v
7
-
D
e
c
N
u
m
b
e
r
/

5

p
l
a
n
t
s
2009 2010
-2
0
2
4
6
8
10
12
14
7
-
S
e
p
1
4
-
S
e
p
2
1
-
S
e
p
2
8
-
S
e
p
5
-
O
c
t
1
2
-
O
c
t
1
9
-
O
c
t
2
6
-
O
c
t
2
-
N
o
v
9
-
N
o
v
1
6
-
N
o
v
2
3
-
N
o
v
3
0
-
N
o
v
7
-
D
e
c
N
u
m
b
e
r
/

5

p
l
a
n
t
s
2009 2010
Infestations of American bollworm and the resulting damage were significantly less in WS hybrids
compared to their non-transgenic counterpart hybrids across both the locations. In 2009 and 2010 the
square damage ranged between 9.06 and 14.63% in non-WS103 and non-WS106 hybrids while the
damage in WS hybrids was negligible (0.13 to 0.82%) (Table 2).
TABLE 2: PERCENT SQUARE AND GREEN DAMAGE ACROSS TWO LOCATIONS IN CENTRAL ZONE DURING KHARIF 2009 AND 2010
No. Hybrid/Variety Square Damage (%) Green Boll Damage (%)
2009 2010 2009 2010
1 WS103 0.13* (2.07)
e 0.15 (2.18) c 0.08* (0.94)

d 0.22 (2.69) c
2 Non-WS103 8.87 (17.33) b 14.63 (22.47) ab 5.93 (14.05) b 7.73 (16.10) a
3 WS106 0.82 (5.16) d 0.24 (2.81) c 0.44 (3.65) c 0.25 (2.88) c
4 Non-WS106 9.06 (17.51) b 14.06 (22.02) b 8.91 (17.36) a 6.33 (14.54) ab
5 Bunny Bt/ RCH2 Bt
0.24 (2.78) e 0.17 (2.39) c 0.06 (1.12) d 0.18 (2.42) c

6 H 8 Non Bt 10.64 (19.03) a 15.31 (23.02) ab 5.39 (13.41) b 5.20 (13.13) b
7 NHH-44/ H12 5.90 (14.01) c 16.23 (23.75) a 5.04 (12.92) b 5.49 (13.54) b
8 PKV Rajat/ G.Cot16 9.90 (18.33) ab 14.58 (22.44) ab 6.15 (14.33) b 8.06 (16.41) a
LSD (p=0.05) 1.02 1.31 2.12 2.08
CV 4.82 4.96 12.47 11.62
Bollgard (Cry1Ac) and Bollgard II (Cry1Ac+Cry2Ab) were used in 2009 and 2010, respectively
*Mean of Aurangabad and Vadodara across the season and Means followed by same letter do not significantly differ
Figures in parentheses are arc-sin transformed values

Green boll damage was less than 10% during both 2009 and 2010 across different entries. However,
significant differences in damage between WS and its non-transgenic counterpart hybrids were clearly
observed. The average green boll damage across the season ranged from 5.93 to 8.91% in non-WS
hybrids while it was only 0.08 to 0.44% in WS hybrids (Table 2).
Open boll and locule damage was observed during harvest in both 2009 and 2010 seasons. In
WideStrike hybrids open boll damage was significantly lower (0.14-0.95%) than in non-WS hybrids (6.22
11.41%). Likewise, locule damage in WideStrike hybrids was also significantly less (00.40%) compared to
their non-transgenic comparators (2.826.48%) (Table 3)
TABLE 3: PERCENT OPEN BOLLAND LOCULE DAMAGE ACROSS TWO LOCATIONS IN CENTRAL ZONE DURING KHARIF 2009 AND 2010
No. Hybrid/Variety Open Boll Damage Locule Damage
2009 2010 2009 2010
1 WS103 0.14* (1.72)
c 0.69 (4.73) d 0.00 (0.00) d 0.34 (3.34) c

2 Non-WS103 7.28 (15.65) a 11.41 (19.74) a 2.82 (9.67) a 6.48 (14.72) a
3 WS106 0.14 (1.76) c 0.95 (5.53) d 0.05 (0.99) d 0.40 (3.51) c
4 Non-WS106 6.22 (14.42) a 9.44 (17.75) abc 3.18 (10.25) a 5.26 (13.13) ab
5 Bunny Bt/ RCH2 Bt
0.05 (0.71) c 0.66 (4.58) d 0.01 (0.33) d 0.43 (3.64) c

6 H 8 Non Bt 4.09 (11.64) b 7.98 (16.37) bc 1.41 (6.80) c 4.66 (12.44) ab
7 NHH-44/ H12 4.16 (11.76) b 6.98 (15.30) c 1.69 (7.48) bc 4.15 (11.75) b
8 PKV Rajat/ G.Cot16 5.55 (13.59) ab 10.03 (18.43) ab 2.41 (8.86) ab 5.85 (13.97) ab
LSD (p=0.05) 2.14 2.67 1.39 2.17
CV 13.74 11.88 14.34 12.97
Bollgard (Cry1Ac) and Bollgard II (Cry1Ac+Cry2Ab) were used in 2009 and 2010, respectively
*Mean of Aurangabad and Vadodara across the season and Means followed by same letter do not significantly differ
Figures in parentheses are arc-sin transformed values

in Biosafety Research Level-1 (BRL-1) Trials Conducted during 2008 and 2009 in South Zone
The population of H. armigera was widely observed both in 2008 and 2009 BRL-1 trials conducted in
South Zone. However, when compared to 2008 the population of H. armigera was generally low across
the season in 2009, particularly in Dharwad trial site (Figure 3 & 4). Infestations of H. armigera and the
resulting damage were significantly less in WS hybrids compared to their non-transgenic counterpart
hybrids. During both the years of BRL-1 trials in South Zone, there was negligible population of S. litura
observed across the entire observation period; however, the WideStrike hybrids were completely free
from S. litura infestation.

Fig. 3: Population of H. Armigera on Non-transgenic Cotton in Dharwad

Fig. 4: Population of H. Armigera on Non-transgenic Cotton in Hyderabad

Fig. 5: Population of H. Armigera on Non-transgenic Cotton in South Zone during Kharif 2010

Fig. 6: Population of S. Litura on Non-transgenic Cotton in South Zone during Kharif 2010

0.0
0.5
1.0
1.5
2.0
2.5
N
o
.

o
f

l
a
r
v
a
e
/

p
l
a
n
t
Atur Dharwad Guntur
-1.0
0.0
1.0
2.0
3.0
4.0
5.0
N
o
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o
f

l
a
r
v
a
e
/

p
l
a
n
t
Attur Dharwad Guntur

Fig. 7: Population of American bollworm (H.armigera) (No./ plant) in South Zone during Kharif 2010

Fig. 8: Population of S. Litura (No./Plant) Across three Locations in South Zone during Kharif 2010
Across both locations, there was significantly higher damage of fruiting bodies on non-WS hybrids
when compared with their transgenic counterparts. In Hyderabad, the square damage on non-WS103
hybrid was 8.27 and 12.92% and on non-WS106 it was 5.22 and 12.85% during 2008 and 2009,
respectively, while the square damage in WS hybrids was negligible. In Dharwad, WS103 and WS106
recorded very limited square damage while their non-transgenic counterparts recorded a significantly
higher damage of 9.17 and 8.00% in 2008 and 2.51 and 2.03% in 2009, respectively (Table 4).
This low level of damage in non-transgenics may be due to the low pest incidence at this
location coupled with unfavourable climatic conditions.
TABLE 4: PERCENT SQUARE DAMAGE IN SOUTH ZONE DURING KHARIF 2008 AND 2009
No. Hybrid/Variety Hyderabad Dharwad
2008 2009 2008 2009
1 WS103 0.08 (1.33)* c 0.00 (0.00) b 1.33 (6.46) b 0.94 (5.50) b
2 Non-WS103 8.27 (16.69) a 12.92 (20.27) a 9.17 (17.53) a 2.51 (9.04) a
3 WS106 0.05 (1.01) c 0.06 (0.00) b 1.09 (5.89) b 0.52 (4.12) b
4 Non-WS106 5.22 (13.18) b 12.85 (21.41) a 8.00 (16.23) a 2.03 (8.17) a
5 Bunny Bt/ RCH2 Bt 0.13 (2.05) c 0.13 (0.65) b 1.94 (7.82) b 0.88 (5.34) b
6 Bunny Non Bt 7.67 (16.07) a 14.09 (21.73) a 7.30 (15.62) a 1.97 (7.99) a
7 DHH-11/LAHH-5 5.53 (13.59) b 12.60 (21.15) a 8.56 (16.75) a 2.03 (8.12) a
8 Sahana/ Narasimha 7.09 (15.41) a 12.87 (22.76) a 6.39 (14.62) a 2.71 (9.41) a
LSD (P=0.05) 1.32 3.01 3.80 2.03
Means followed by same letter do not significantly differ (P=0.05)
*Figures in parentheses are arc-sine transformed values.
Green boll damage was significantly lower on WS103 and WS106 during both 2008 and 2009
seasons. In Hyderabad, the green boll damage was high in 2009 compared to 2008. The damage during
2009 in non-transgenic hybrids ranged from 15.55 to 21.65% whereas it was less than 0.5% in WS
hybrids exhibiting very good efficacy against bollworms. In Dharwad, the green boll damage was
generally low both in 2008 and 2009 (Table 5).
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
WS103 Non-WS103 WS106 Non-WS106 Bt Check BGII Check Non-Bt Variety
check
Non-Bt Hybrid
check
N
o
.

o
f

l
a
r
v
a
e
/

p
l
a
n
t
Attur Dharwad Guntur
0.0
0.2
0.4
0.6
0.8
1.0
1.2
WS103 Non-WS103 WS106 Non-WS106 Bt Check BGII Check Non-Bt Variety
check
Non-Bt Hybrid
check
N
o
.

o
f

l
a
r
v
a
e
/

p
l
a
n
t
Attur
Dharwad
Guntur
TABLE 5: PERCENT GREEN BOLL DAMAGE IN SOUTH ZONE DURING KHARIF 2008 & 2009
2008 2009 2008 2009
1 WS103 0.07 (1.26)*c 0.00 (0.00) c 0.22 (2.51) c 0.14 (2.09) d
2 Non-WS103 2.52 (8.96) a 15.55 (23.21) b 2.75 (9.53) ab 2.39 (8.88) ab
3 WS106 0.05 (1.01) c 0.48 (2.30) c 0.19 (2.26) c 0.20 (2.48) d
4 Non-WS106 1.30 (6.54) b 16.22 (23.68) b 1.87 (7.74) ab 2.90 (9.76) a
5 Bunny Bt/ RCH2 Bt 0.12 (1.99) c 0.40 (2.10) c 0.47 (3.62) c 0.32 (3.18) d
6 Bunny Non Bt 1.29 (6.41) b 18.05 (25.11) ab 3.24 (10.03) a 1.92 (7.95) bc
7 DHH-11/LAHH-5 0.95 (5.59) b 21.65 (27.72) a 2.5 (9.05) ab 1.51 (6.97) c
8 Sahana/ Narasimha 1.55 (6.93) b 19.13 (25.88) ab 1.69 (7.46) b 2.08 (8.29) bc
LSD (P=0.05) 1.88 3.68 2.48 1.34
Open boll and locule damage was observed only in 2009. In WideStrike hybrids open boll damage
was significantly lower (0.07 - 0.58%) than their non-WS hybrids (2.68 5.85%). Likewise, locule damage
in WideStrike hybrids was significantly less (0.01 0.24%) compared to their non-transgenic comparators (1.01
1.83%) (Table 6)
TABLE 6: PERCENT OPEN BOLL AND LOCULE DAMAGE IN SOUTH ZONE DURING KHARIF 2009
Open boll Locule Open boll Locule
1 WS103 0.07 (0.88)* b 0.01 (0.38) c 0.58 (4.35)* c 0.24 (2.77) cd
2 Non-WS103 5.06 (12.96) a 1.68 (7.44) a 2.71 (9.41) a 1.01 (5.76) a
3 WS106 0.18 (2.00) b 0.03 (0.85) bc 0.52 (4.13) c 0.16 (2.31) d
4 Non-WS106 5.85 (13.90) a 1.83 (7.69) a 2.68 (9.37) a 1.08 (5.94) a
5 Bunny Bt/ RCH2 Bt 0.20 (2.04) b 0.08 (1.58) b 1.01 (5.75) c 0.76 (4.68) ab
6 Bunny Non Bt 6.11 (14.29) a 1.66 (7.40) a 2.72 (9.48) a 0.96 (5.63) ab
7 DHH-11/LAHH-5 6.35 (14.59) a 2.18 (8.49) a 2.28 (8.67) ab 0.82 (5.18) ab
8 Sahana/ Narasimha 5.96 (14.09) a 1.90 (7.91) a 1.69 (7.44) b 0.52 (4.14) bc
LSD (P=0.05) 2.49 1.14 1.59 1.45
in Biosafety Research Level-2 (BRL-2) Trials Conducted During Kharif 2010 in South Zone
TABLE 7: PERCENT SQUARE DAMAGE IN SOUTH ZONE DURING KHARIF 2010
No. Hybrid/Variety Attur Dharwad Guntur
1 WS103 0.37 0.12
0.10 0.06 0.12 0.04

2 Non-WS103 6.32 0.99 10.13 1.29 5.85 1.50
3 WS106 0.32 0.12 0.12 0.06 0.08 0.03
4 Non-WS106 6.32 0.86 9.91 1.65 4.51 1.06
5 Bunny BGI/RCH-2 BGI 0.19 0.05 0.17 0.08 0.22 0.09
6 Bunny BGII/RCH BGII 0.20 0.06 0.11 0.07 0.15 0.07
7 Surabhi/Sahana/Narasimha 7.39 1.32 9.02 1.12 5.13 1.38
8 Bunny non-Bt/RCH-2 non-Bt 6.68 0.97 9.09 1.02 5.72 1.34

Mean standard error of mean
American bollworm, H. armigera, was observed across the season at all the three locations in the BRL-2
trials conducted in South Zone. The H. armigera larval population was high in Attur and Dharwad while
it was low in Guntur. The population was high during 90 to 120 days of the crop growth in Attur and
Dharwad. The population of S. litura was higher at Guntur compared to Attur and Dharwad (Figures 5 &
6). The highest population of S. litura was recorded during 70 to 90 days of the crop growth. The
lepidopteran population also correlated well with higher fruiting body damage in Attur and Dharwad
compared to Guntur. Population of American bollworm and its damage was much less in WS hybrids
compared to their non-WS hybrids, across all the three locations. The population of H. armigera larvae
was less than 0.03 larvae per plant on WS hybrids, while it ranged from 0.09 to 0.55 per plant on non-WS
hybrids. Similarly, S. litura population was negligible (< 0.02 larva/ plant) on WS hybrids, while it
ranged between 0.15 and 0.72 larva per plant on non-WS hybrids (Figures 7 & 8).
The square damage was recorded to be the highest in Dharwad followed by Attur and Guntur. It
ranged between 4.51 and 10.13% in non-WS hybrids while the damage in WS hybrids was negligible
(0.08 to 0.37%) (Table 7)
Green boll damage was higher in Dharwad and Attur when compared to Guntur. The damage ranged
from 2.11 to 9.35% in non-transgenic hybrids whereas it was less than 0.23% in WideStrike hybrids
across locations and season (Table 8)
TABLE 8: PERCENT GREEN BOLL DAMAGE IN SOUTH ZONE DURING KHARIF 2010
No. Hybrid/Variety Attur Dharwad Guntur
1 WS103 0.23 0.10
0.12 0.06 0.00

2 Non-WS103 7.63 1.46 9.35 2.07 3.47 0.96
3 WS106 0.16 0.06 0.08 0.05 0.00
4 Non-WS106 8.66 1.89 6.69 1.76 2.11 0.74
5 Bunny BGI/RCH-2 BGI 0.23 0.09 0.10 0.06 0.05 0.02
6 Bunny BGII/RCH BGII 0.27 0.07 0.09 0.06 0.03 0.01
7 Surabhi/Sahana/Narasimha 7.16 1.36 7.37 2.13 2.80 0.80
8 Bunny non-Bt/RCH-2 non-Bt 7.95 1.54 7.05 2.26 2.22 0.77

Moderate to high level of open boll and locule damage was observed during harvest at all locations.
In WideStrike hybrids open boll damage was lower (0.23 2.75%) than their non-transgenic counterpart
hybrids (12.20 18.85%). Likewise, locule damage in WideStrike hybrids was also much less (0.10
1.27%) compared to their non-transgenic counterparts (4.63 16.37%) (Table 9)
TABLE 9: PERCENT OPEN BOLL AND LOCULE DAMAGE IN SOUTH ZONE DURING KHARIF 2010
No. Hybrid/Variety Open Boll Damage Locule Damage
Attur Dharwad Guntur Attur Dharwad Guntur
1 WS103 2.75 0.37
0.36 0.07 0.34 0.19 1.27 0.15 0.11 0.03 0.11 0.06
2 Non-WS103 17.95 3.26 16.99 1.04 14.27 1.00 16.37 2.05 9.24 1.11 4.63 0.39
3 WS106 1.73 0.41 0.35 0.08 0.23 0.23 1.15 0.16 0.11 0.02 0.10 0.10
4 Non-WS106 18.25 1.94 12.20 0.19 15.33 0.24 13.99 1.77 7.42 0.25 5.21 0.19
5 Bunny BGI/RCH-2 BGI 1.93 0.56 0.49 0.06 0.44 0.09 1.29 0.04 0.09 0.02 0.11 0.03
6 Bunny BGII/RCH BGII 1.27 0.44 0.41 0.00 0.31 0.01 0.91 0.18 0.07 0.03 0.08 0.02
7 Surabhi/Sahana/Narasimha 14.07 1.02 13.83 1.63 15.20 0.14 12.03 0.99 8.23 1.35 5.89 0.15
8 Bunny non-Bt/RCH-2 non-Bt 15.57 2.17 11.10 0.53 13.05 0.93 13.28 1.10 6.46 0.47 4.85 0.17

DISCUSSION
Dow AgroSciences India Pvt Ltd conducted Biosafety Research Level 1 (BRL-1) trials at two locations
during Kharif 2008 and 2009 in South Zone; at two locations during Kharif 2009 and 2010 in Central
Zone; and Biosafety Research Level 2 (BRL-2) trials at three locations during Kharif 2010 in South
Zone. Efficacy data recorded during these BRL-1 and BRL-2 trials on American bollworm (H. armigera)
population, tobacco caterpillar (S. litura) population and fruiting body damage clearly demonstrated that
the WideStrike trait offered high levels of protection to leaves, squares and bolls from American
bollworm and tobacco caterpillar in WS103 and WS106 hybrids expressing the insecticidal proteins
Cry1Ac and Cry1F. The population of H. armigera was prominent during all the years of BRL trials
while S. litura was observed in the BRL-2 trials conducted during 2010 in south zone. It was further
observed that the fruiting body damage on WideStrike hybrids was greatly reduced as compared to the
non-transgenic hybrids indicating superior efficacy of WideStrike in controlling the target pests. This is
the first published report on the field efficacy of WideStrike cotton hybrids from India.
The reduction in population of H. armigera and its damage in WideStrike hybrids may be contributed
by Cry1Ac protein because Cry1Ac has been reported to be the most effective toxin among different Cry
toxins tested against H. armigera (Chakrabarti et al., 1998; Akhurst et al., 2003; Avilla et al., 2005).
Other researchers have also reported the efficacy of Bt cotton hybrids expressing Cry1Ac protein in India
(Kranthi et al., 2005; Gujar et al., 2010). Literature also suggests that Spodoptera spp. is relatively
tolerant to most of the known Bt endotoxins including Cry1Ac (Regev et al., 1996; Singh et al., 2004).
However, WideStrike cotton which expresses a novel Cry1F protein in addition to Cry1Ac which
controls S. litura effectively and also has activity on Heliothine species. Similarly, Tindall et al.,
2009 reported that Cry1F protein in transgenic cotton and maize provided very effective control
of Spodoptera spp. Apart from the above field efficacy data, laboratory assays also confirmed
that WideStrike hybrids and the individual proteins in diet assays controlled H. armigera and S.
litura effectively (Moudgal, unpublished information).
Upon regulatory approval, introduction of WideStrike dual Bt cotton in India will diversify the
choice of insect protection traits available to Indian farmers and enable effective management of
bollworms and improved control of S. litura in cotton. The additional diversity in insect proteins can
contributed to a delay in resistance development and therefore help preserve the technology of Bt cotton.
Globally, this event has been approved in USA (2004) and recently in Brazil (2009) for cultivation and in
Australia, Canada, Japan, Korea and Mexico for feed/food import, after completing necessary safety
assessments.
REFERENCES
[1] Akhurst, R. T., James, W.J., Bird, L.J. and Beard, C. (2003) - Resistance to the Cry1Ac -endotoxin of Bacillus
thuringiensis in the cotton bollworm H. armigera (Lepidoptera: Noctuidae), J. Econ. Entomol. 96:12901299
[2] Avilla, C., Vargas-Osuna E., Gonzlez-Cabrera J., Ferr, J., and Gonzlez-Zamora, J. E. (2005) - Toxicity of several -
endotoxins of Bacillus thuringiensis against H. armigera (Lepidoptera: Noctuidae) from Spain. J. Invertebr. Pathol., 90: 51
54.
[3] Brookes, G. and Barfoot, P. (2010) - GM Crops: Global Socio-economic and Environmental Impacts 1996-2008. P.G.
Economics Ltd, Dorchester, UK. http://www.pgeconomics.co.uk/pdf/2010-global-gm-crop-impact-study-final-April-
2010.pdf.
[4] Chakrabarti, S. K., Mandaokar, A. D., Kumar, P. A. and Sharma, R. P. (1998) - Efficacy of lepidopteran specific delta-
endotoxin to Bacillus thuringiensis against H. armigera. J. Invertebr. Pathol., 72: 336-337.
[5] Gujar, G. T, Kalia, V., Bunker, G. K. and Dhurua, S. (2010) - Impact of different levels of non-Bt cotton refuges on pest
populations, bollworm damage, and Bt cotton production. J. Asia-Pac. Ent. 13: 249253.
[6] James, C. (2010) - Global Status of Commercialized Biotech/GM Crops: 2010. ISAAA Brief No. 42. ISAAA: Ithaca, NY.
[7] Kranthi, K.R. Naidu, S., Dhawad, C.S., Tatwawadi, A., Mate, K., Patil, E., Bharose, A. A., Behere, G. T., Wadaskar, R. M.
and Kranthi, S. (2005) - Temporal and intra-plant variability of Cry1Ac expression in Bt cotton and its influence on the
survival of the cotton bollworm, H. armigera (Hbner) (Noctuidae: Lepidoptera), Curr. Sci. 89: 291-298.
[8] Padaria, R. N. Singh, B., Sivaramane, N., Naik, Y. K., Modi, R. and Surya, S. (2009) - A Logit Analysis of Bt Cotton
Adoption and Assessment of Farmers Training Need. Ind, Res. J. Ext. Edu. 9 (2): 39-45
[9] Regev, A., Keller, M., Strizhov, N., Sneh, B., Prudovsky, E., Chet, I., Ginzberg, I., Koncz-Kalman, Z., Koncz, C., Schell, J.
and Zilberstein, A. (1996) - Synergistic activity of a Bacillus thuringiensis delta-endotoxin and a bacterial endochitinase
against Spodoptera littoralis larvae. Appl. Environ. Microbiol., 62 (10): 35813586.
[10] Singh, P. K., Kumar, M., Chaturvedi, C. P., Yadav, D. and Tuli, R. (2004) - Development of a hybrid delta-endotoxin and
its expression in tobacco and cotton for control of a polyphagous pest S. litura. Transgenic Res., 13 (5): 397-410.
[11] Subramanian, A. amd Qaim, M. (2009) - Village-wide Effects of Agricultural Biotechnology: The Case of Bt Cotton in
India. World Development, 37 (1): 256-267.
[12] Tindall, K. V., Willrich Siebert, M., Leonard, B. R., All, J. and Haile, F. J. (2009) - Efficacy of Cry1Ac:Cry1F proteins in
cotton leaf tissue against fall armyworm, beet armyworm, and soybean looper (Lepidoptera: Noctuidae). J. Econ. Entomol.,
102 (4):1497-1505.
Influence of Weather Parameters on Population of

Mealybug, Phenacoccus solenopsis and its Natural
Enemies on Bt Cotton
B.V. Patil, S.G. Hanchinal, M. Bheemanna and A.C. Hosamani
Main Agricultural Research Station, University of Agricultural Sciences, Raichur-584102
E-mail: shanchinal@gmail.com
AbstractSeasonal incidence of mealybug, Phenacoccus solenopsis (Tinsley) on cotton was studied at main
agricultural research station, UAS, Raichur during 2008-09 and 2009-10 cropping seasons. Mealy bug incidence was
started in the month of September (40
th
standard week) during both the seasons. Initially mealy bug population was
low and gradually increased as the crop stage advanced. Steep increase of mealybug population was observed after
January and reached peak in the month of March. Population was varied from 66.28 to 146.64 and 12.32 to 122.64 per
10 cm apical shoot during 2008-09 and 2009-10 seasons, respectively. In general predator population was low through
out the season in both the years. Hymenopteran parasitoids activity was started in the month of October and it was
ranged between 0.52 to 22.12 per cent during 2008-09 and 0.08 to 34.64 per cent during 2009-10. Parasitoid cocoons
were maximum in the month of March which recorded 22.12 and 34.64 per cent during 2008-09 and 2009-10 seasons,
respectively. Maximum temperature was positively correlated and significant where as other parameters were
negatively correlated.
INTRODUCTION
Mealybugs (Hemiptera: Pseudococcidae) are the soft bodied insects found feeding on variety of plants. In
the recent past various mealybug species have been recorded on many field crops, orchards, vegetables
and wild plants due to certain abiotic changes in climate and environment. Two species of mealybugs ie
Phenacoccus solenopsis and Maconellicoccus hirsutus (Green) damaged the cotton crop in 47 locations
in nine cotton growing states (Nagarare et el., 2009). Recently, mealy bug has taken upper hand among
sucking pests in cotton in the states like Punjab, Rajastan, Gujarat and Maharashtra in North India and
moderate incidence in Central and South India. Among two species of mealy bug, Phenacoccus
solenopsis was the dominant species damaged the cotton crop in Punjab. Mealy bug has been recorded on
most of the cotton growing areas in Karnataka, especially incidence was more in the isolated patches of
Raichur and Bellary districts (Hanchinal et al., 2009 ). It has been also recorded on many alternative
hosts which includes sunflower, vegetables, weeds, ornamentals etc., Saini et al.(2009) and Deshmukh et
al.(2009)
Mealy bugs feed on all parts of a plant, particularly on growing tips or on leaves that join stems or
along leaf veins. The crawlers disperse from the ovisac by way of walking, wind, or ants. The nymphs
feed and develop into adults in approximately 30 days. The insect has a life cycle of 24 to 30 days. The
female mealy bug produces 10-15 generations per year in colonies of 500-600 eggs (Tanwar et al., 2007).
The investigation was carried out at the main agricultural research station, UAS, Raichur during 2008-09
and 2009-10 cropping seasons. Population dynamics of mealybug was recorded on Bt cotton (BG-II,
NCS-145). One acre of unprotected cotton plot was maintained at MARS, Raichur. Observations were
recorded weekly interval on 20 randomly selected plants. Mealybugs were recorded from 10cm apical
shoot length from September to March during both the seasons. Natural enemies like predators and
parasitoids were also recorded per plant. Correlation was worked out between seasonal fluctuation of
mealy bug, predators and parasitoid cocoons and percent emergence of parasitoids in the laboratory.
Correlation was also worked out between weather parameters and natural enemies of mealybug during
2008-09 and 2009-10 cropping period.
31
Mealybug incidence varied from 0.50 to 180.42 per 10 cm apical shoot from 38
th
to 14
th
meteorological
week in main agricultural research station, Raichur during 2008-09 cropping season(Table.1). During
2009-10 cropping period population varied between lowest of 0.14 in October first week to highest of
184.32 mealybugs per 10 cm apical shoot during March third week. Mealybug infestation started
appearing in the month of September and gradually increased as crop growth advanced during both the
seasons. Mealybug population was 0.50 and 0.14 per 10cm apical shoot in the 38
th
and 40
th

meteorological week during 2008-09 and 2009-10 seasons, respectively and progressive increase in the
population was recorded throughout the season. Population reached to 115.42/10 cm apical shoot in the
third week of January and there after population increased suddenly and attained 180.42/10cm apical
shoot in the 7
th
meteorological week during 2008-09 season. Similarly during 2009-10 season peak
activity observed between February to March (64.68 to 184.32/10 cm apical shoot).Later on infestation of
mealybug declined gradually at end of the cropping period in both the seasons (Fig.1). Overall mealybug
population was low in the second season and it was due to increased activity of parasitoids.
In general predator population was low during the cropping season. Maximum population of
coccinellids, chrysoperla and spiders were 0.14, 0.13 and 0.16 per plant during 2008-09 season and 0.30,
0.27 and 0.32 per plant during 2009-2010 season, respectively. Per cent parasitoid cocoons ranged
between 0.52 to 20.02 and 008 to 34.64 during 2008-09 and 2009-10 seasons, respectively. Parasitoid
activity started in the month of October and reached to peak during February and March months in both
the seasons. Highest per cent parasitoid cocoons of 22.12 and 34.64 on cotton plants were recorded
during 8
th
and 12
th
meteorological week during 2008-09 and 2009-10 cropping period, respectively. Peak
activity of parasitoid cocoons were coincidence with the higher population of mealybug during same
period in both the seasons.
Among five different hymenopteran parasitoids and a dipteran parasitoid, Aenasius bambawalei
Hayat was the dominant species. Similarly Pinjarkar et al (2009) reported that A. bambawalei was the
only species recorded on P. solenopsis in Maharashtra. Correlation coefficients were calculated between
field infestation of mealy bug, coccinellids, chrysoperla, Spiders, percent parasitoid cocoons and weather
parameters.
The impact of weather parameters has revealed that maximum temperature showed a positive and
significant correlation with the mealybug population (0.821), while others showed significantly negative
correlation. Presence of predators with respect to weather parameters were non significant. Minimum
temperature and number of rainy days showed significantly negative correlation (-0.537 and -0.419) with
coccinellids and relative humidity was positively correlated (0.377) with chrysoperla predators. However,
presence of parasitoid cocoons on mealybug infested cotton plants showed significantly positive
correlation (0.734) with maximum temperature while other parameters showed significantly negative
correlation (Table 3). Present results also corroborated with (Dhawan et al., 2009) who reported that
positive correlation with the maximum temperature and negative correlation among the relative humidity
and rainfall in case of mealybug in Punjab.
The relationship of mealybug population and its natural enemies like coccinellids, Chrysoperla,
spiders, parasitoid cocoons and parasitoid emergence was studied using correlation coefficients. Presence
of predators were non significant, however positive correlation was observed with coccinellids (0.184)
while Chrysoperla (-0.244) and spiders (-0.129) were negatively correlated. Mealybug population was
significantly influenced by the parasitoids and their emergence in laboratory. Parasitoids cocoons (0.973)
were highly significant and showed positive correlation. Correlation of P. solenopsis population with
parasitoid emergence in the laboratory was significant and positively correlated with all the five
parasitoids (Table 4).

Influence of Weather Parameters on Population of Mealybug, Phenacoccus solenopsis and its Natural Enemies 195
The partial regression co-efficient of the mealybugs, parasitoid cocoons on plants and parasitoids
emergence in laboratory were found to be highly significant. However, the partial regression coefficients
of weather factors on predators population was non significant. Regression equations calculated for
maximum temperature were Y = -455.83 + 15.07, Y = -68.48 + 2.32 and Y = -21.23 + 0.74 with R
2
value
of 0.79, 0.71 and 0.76 for mealybugs, parasitoid cocoons and parasitoid emergence, respectively. It
indicated that about 79.30, 71.10 and 76.90 per cent of population fluctuation depends on weather
parameters. The multiple regression equation fitted with A. bambawalei and weather parameter to predict
the mealybug population (Y) was:
Y= -164.055X
1
+ 6.853X
2
-2.615X
3
+ 0.047X
4
+ 1.489X
5
- 0.198X
6
-0.302 X
7
+ 3.895X
8

with a R
2
value of 0.865 at main agricultural research station, Raichur. Where X
1,
X
2,
X
3,
X
4,
X
5,
X
6,
X
7
and X
8
denote maximum temperature, minimum temperature, rainfall, rainy days,
relative humidity maximum, relative humidity minimum and parasitoids, respectively
TABLE 1: SEASONAL INCIDENCE OF MEALYBUGS, PREDATORS AND PARASITOIDS ON COTTON (NCS-145 BT) UNDER IRRIGATED ECOSYSTEM DURING 2008-09
Months ISD
Week
Mealybugs*/ 10 cm
Apical Shoot
Predators Per Plant Per Cent Parasitoid Cocoons
Per Plant
Coccinellids Chrysoperla Spiders
Sep 1-7 36 0.00 0.02 0.00 0.04 0.00
Sep 8-14 37 0.00 0.00 0.04 0.00 0.00
Sep 15-21 38 0.50 0.02 0.02 0.00 0.00
S ep 22-28 39 0.52 0.00 0.00 0.02 0.00
Sep 29-5 40 0.61 0.02 0.00 0.02 0.00
Oct 6-12 41 0.65 0.04 0.00 0.02 0.00
Oct 13-19 42 0.72 0.06 0.00 0.04 0.00
Oct 20-26 43 0.84 0.02 0.00 0.06 0.00
Oct 27-2 44 0.86 0.00 0.00 0.02 0.52
Nov 3-9 45 2.16 0.02 0.04 0.04 1.28
Nov 10-16 46 6.24 0.00 0.00 0.00 1.32
Nov 17-23 47 6.58 0.00 0.02 0.00 2.12
Nov 24-30 48 8.24 0.00 0.00 0.16 3.62
Dec 1-7 49 10.64 0.04 0.12 0.04 4.54
Dec 8-14 50 13.21 0.02 0.13 0.02 6.24
Dec 15-21 51 16.58 0.08 0.04 0.08 7.60
Dec 22-28 52 22.42 0.02 0.00 0.04 9.21
Dec 29-4 1 36.52 0.00 0.00 0.00 9.44
Jan 5 -11 2 66.28 0.12 0.00 0.04 9.82
Jan 12-18 3 85.34 0.14 0.02 0.00 10.25
Jan 19-25 4 115.42 0.00 0.00 0.02 12.62
Jan 26 -1 5 154.62 0.10 0.00 0.00 13.45
Feb 2 -8 6 142.16 0.00 0.00 0.02 15.62
Feb 9 -15 7 180.42 0.08 0.00 0.00 20.65
Feb 16 -22 8 158.44 0.00 0.00 0.04 22.12
Feb 23 -1 9 162.24 0.02 0.00 0.00 20.02
Mar 2 - 8 10 154.50 0.00 0.00 0.02 16.42
Mar 9-15 11 152.26 0.00 0.00 0.00 12.64
Mar 16-22 12 150.65 0.00 0.00 0.02 10.20
Mar 23-29 13 148.64 0.00 0.00 0.00 4.62
March 30-5 14 146.64 0.00 0.00 0.00 3.26
Mean 54.03 0.02 0.01 0.02 6.04
S.D. 68.43 0.04 0.03 0.03 6.90
* Mean of 20 plants
S.D = standard deviation

TABLE 2: SEASONAL INCIDENCE OF MEALY BUGS, PREDATORS AND PARASITOIDS ON COTTON (NCS-145 BT) UNDER IRRIGATED ECOSYSTEM DURING 2009-10 SEASON

Months ISD Week Number of Mealybugs Per 10
Cm Apical Shoot Length
Number of Predators Per Plant* Per Cent Parasitoid
Cocoons Per Plant
Coccinellids Chrysoperla Spiders
Sep 10-16 37 0.00 0.00 0.02 0.05 0.00
Sep 17-23 38 0.00 0.00 0.04 0.02 0.00
Sep 24-30 39 0.00 0.00 0.02 0.15 0.00
Oct 1-7 40 0.14 0.00 0.12 0.02 0.08
Oct 8-14 41 0.33 0.02 0.05 0.30 0.12
Oct 15-21 42 0.41 0.03 0.12 0.02 0.18
Oct 22-28 43 0.52 0.04 0.06 0.25 0.21
Oct 29-4 44 2.53 0.02 0.02 0.06 0.23
Nov 5-11 45 0.22 0.03 0.14 0.32 0.08
Nov 12-18 46 0.45 0.01 0.30 0.04 0.06
Nov 19-25 47 0.62 0.01 0.12 0.25 0.36
Nov 26-2 48 0.66 0.08 0.02 0.00 0.92
Dec 3-9 49 1.22 0.12 0.04 0.16 1.15
Dec 10-16 50 1.64 0.07 0.12 0.04 3.55
Dec 17-23 51 3.27 0.05 0.25 0.12 4.23
Dec 24-31 52 6.24 0.06 0.27 0.28 5.11
Jan1-7 1 12.32 0.02 0.02 0.24 7.25
Jan 8-14 2 16.42 0.04 0.04 0.27 8.44
Jan 15-21 3 28.25 0.14 0.07 0.15 8.82
Jan 22-28 4 35.37 0.12 0.02 0.24 9.27
Jan 29-4 5 45.42 0.15 0.05 0.25 12.45
Feb 5-11 6 64.68 0.22 0.02 0.32 13.84
Feb 12-18 7 82.13 0.30 0.12 0.22 15.08
Feb 19-25 8 88.55 0.04 0.27 0.30 15.66
Feb 26-4 9 108.13 0.02 0.00 0.04 18.12
Mar 5-11 10 122.21 0.04 0.00 0.12 22.42
Mar 12-18 11 184.32 0.05 0.00 0.02 26.56
Mar 19-25 12 154.20 0.03 0.00 0.00 34.64
Mar 26-1 13 122.64 0.00 0.00 0.00 32.20
Mean 33.84 0.05 0.07 0.14 7.54
S.D. 52.71 0.07 0.09 0.12 10.04
S.D = standard deviation
* Mean of 20 plants.
TABLE 3: CORRELATION CO-EFFICIENT BETWEEN WEATHER PARAMETERS AND SEASONAL FLUCTUATION OF MEALYBUGS, PREDATORS AND PARASITOIDS ON BT COTTON
Parameters Mealy Bugs Coccinellids Chrysoperla Spiders Per Cent Parasitoid Cocoons Per Plant
Maximum Temperature (
o
C) 0.821** -0.021 -0.280 -0.280 0.734**
Minimum Temperature (
o
C) 0.147 -0.537** -0.07 -0.393* 0.017
Rainfall (mm) -0.237 -0.261 0.058 -0.247 -0.277
No. of Rainy days -0.402* -0.419* -0.047 -0.311 -0.465**
Relative Humidity-I -0.736** 0.235 0.377* 0.331 -0.686**
Relative Humidity-II -0.688** -0.064 0.088 0.109 -0.677**
* Differs significantly (P = 0.05)
**Differs significantly (P = 0.01)
TABLE 4: CORRELATION CO-EFFICIENT BETWEEN SEASONAL FLUCTUATION OF MEALY BUGS, PREDATORS AND PARASITOIDS
Parameters Mealybugs
Coccinellid predators 0.184
Chrysoperla predators - 0.244
Spiders - 0.129
Per cent parasitoid cocoons 0.973 **
Per cent parasitoid emergence in laboratory
Aenasius bambawalei 0.923**
Promuscidea unfasciativentris 0.840**
Homalotylus eytelweinii 0.899**
Prochiloneurus pulchellus 0.882**
Anagyrus dactylopii 0.820**
* Differs significantly (P = 0.05)
** Differs significantly (P = 0.01)
Influence of Weather Parameters on Population of Mealybug, Phenacoccus solenopsis and its Natural Enemies 197

Fig. 1: Average Population of Mealybug and Natural Enemies on Bt Cotton during 2008-09 and 2009-10 Seasons
REFERENCES
[1] Deshmukh, A. J. Vennila, S., Pinjarakar, D.B, Ghodki, B.S. and Kranthi, K.R, 2009, Host range of mealybug, Phenacoccus
solenopsis Tinsley in cotton +pigeon pea cropping system of central India. : In proceedings of National symposium on Bt-
cotton: Opportunities and Prospectus,CICR,Nagpur,November 17-19,pp150
[2] Dhavan, A. K, Kamaldeep, S,A and Sarika, S, 2009. Distribution of mealybug, Phenacoccus solenopsis Tinsley in cotton
with relation to weather factors in south-Western districts of Punjab. J.ent.Res.,33(1):59-63
[3] Hanchinal, S,G, Patil, B,V, Bheemanna, M, Hosamani, A.C and Sharanbassappa, 2009. Incidence of mealy bug on cotton in
Tungbhadra project area: In proceedings of Dr. Leslie C. Coleman memorial national symposium on plant protection, 4 - 6
December 2008, University of Agricultural Sciences, GKVK, Bangalore 560 065
[4] Hanchinal, S,G, Patil, B,V, Bheemanna, M,and.Hosamani, A.C,2009, Incidence of mealybug Phenacoccus solenopsis
Tinsley and its natural enemies on cotton in Karnataka: In proceedings of National symposium on Bt-cotton:Opportunities
and Prospectus,CICR,Nagpur,November 17-19,pp150
[5] Nagrare,V.S, Kranthi,S, Biradar,V.K, Zade,N.N, Sangode,V, Kakde,G,Shukla,R.M, Shivare,D, Khadi,B.M, and
Kranthi,K.R,2009.Widespread infestation of the exotic mealybug species, Phenacoccus solenopsis (Tinsley) (Hemiptera:
Pseudococcidae) on cotton in India,Bulletin of Entomological Research: P.1-5
[6] Pinjarakar,D.B,Vennila,S,Ramamuthy,V.V,Kranthi,K.R,Ghodki,B.S and Deshmukh,A.J, 2009, Diversity and abundance of
Hymenopteran parasitoids of mealybugs in rainfed cotton: In proceedings of National symposium on Bt-
cotton:Opportunities and Prospectus,CICR,Nagpur,November 17-19,pp150
[7] Saini,R.K,Palaram, Sharma,S.S and Rohilla,H.R, mealybug Phenacoccus solenopsis Tinsley and its survival in cotton
ecosystem in Haryana: In proceedings of National symposium on Bt-cotton: Opportunities and Prospectus, CICR, Nagpur,
November 17-19,pp150
[8] Tanwar, R.K, Jeyakumar, P and Monga,D., 2007, Mealybugs and their management.Tech.Bull.No.19.NCIPM, New Delhi
(India)
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0.18
36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 1 2 3 4 5 6 7 8 9 10 11 12 13 14
Standard weeks
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e
a
l
y
b
u
g
s
/
1
0

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a
p
i
c
a
l

s
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40
60
80
100
120
140
160
180
200
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coccinellids chrysoperla spiders %parasitoid cocoons mealybug
Insecticide Induced Resurgence of Mealybug,

Phenacoccus solenopsis Tinsley in Cotton
Rishi Kumar
1
, Dinesh Swami
1
, Vijender Pal
1
and K.R. Kranthi
2

1
Central Institute for Cotton Research, Regional Station, Sirsa, Haryana-125055,
2
Central Institute for Cotton Research, Nagpur
E-mail: rishipareek70@yahoo.co.in
AbstractThe knowledge relating to risk associated with the insecticide as regards resurgence is of paramount
importance, the influence of the commonly used insecticides alone and in combination was studied for their role in
resurgence of mealybug population on cotton cultivar RCH134 during 2008 and 2009.The insecticides i.e.
cypermethrin, monocrotophos, acephate, ethion, profenophos, spinosad, cypermethrin + monocrotophos, cypermethrin
+ acephate, cypermethrin + ethion, cypermethrin + profenophos were applied 11 time at 10 day intervals in mealybug
infested fields spreading throughout the cotton season at their recommended dosages. On the basis of two year
cumulative data 14.53 per cent resurgence in mealybug population was recorded due to Spinosad. No resurgence in
mealybug population during the year 2008 after 1
st
and II
nd
spray was observed but 0 .69 to 11.24% resurgence was
recorded after 3
rd
, 4
th
, 6
th
, 7
th
, 8
th
, 9
th
, 10
th
and 11
th
spray after 7
th
day of each spray application. During the 2009,
4.53 to 43.53 % resurgence in mealy bug due to spinosad was recorded after 2
nd
, 3
rd
, 4
th
, 5
th
, 6
th
, 7
th
, 8
th
, 9
th
, 10
th
and
11
th
spray after 7
th
day of each spray application. The studies conducted under polyhouse conditions also revealed
12.92 % resurgence in mealybug population due to spinosad after 5 sprays at weekly intervals.
Among the other insecticides applied, 10.89 to 21.92% resurgence due to cypermethrin during the 2009 was
recorded after 5
th
, 6
th
, 7
th
, 8
th
, 9
th
, 10
th
, and 11
th
spray after 7
th
day of each spray application. 12.12 to 29.08 %
resurgence due to monocrotophos was recorded after 6
th
, 7
th
, 8
th
, 9
th
and 11
th
spray after 7
th
day of each spray
application. As per the record, though the infestation of mealybug was more during 2008 but the resurgence in
mealybug population due to insecticides was more during 2009. The reason for the resurgence like biochemical changes
in plant, changes in insect reproduction physiology or ecological changes could not be traced at this stage which has to
be confirmed through more planned and systemic studies.
INTRODUCTION
Resurgence was defined as an increase in target arthropod pest species abundance to a level which
exceeds that of control or untreated population following the application of insecticides (Hardin et al.,
1995). Insecticides-induced outbreaks of insects have been reported as early as 1951 in walnut (Bartlett
and Ewart, 1951). Ripper (1956) listed more than 50 species of phytophagous arthropods whose
population has resurged with wide spread use of insecticides. After the introduction of Bt cotton, the
insecticidal usage in cotton was for sucking pests only. Sucking pests especially, whitefly, jassid, thrips
and mealy bug have become quite serious from seedling stage. The mealybug species Phenacoccus
solenopsis Tinsley, an exotic species appeared in 2006 in cotton growing zones of country (Nagrare et al
2009) and insecticide application became an integral part of its management. The wide spread use of
chemical pesticide is known to induce the population resurgence of pests (Gu et at., 1996 and Yin et al.,
2008). Mealy bug is amenable to control with insecticides, but repeated application often results in
problems such as induction of resurgence and development of resistance. Insecticides may affect the
physiology of target insect pest directly through stimulation of growth and reproduction or indirectly
through alteration in the nutritional quality of host plant that leads to favorable biological effects on
target pests, such as shortening of development period, increased survival, increased feeding rate and
reproduction or other related changes in behavior of the insects (Ravindhran and Xavier, 1997 and Singh
et. al., 2008). As large numbers of insecticides alone and in combinations are repeatedly used for the
management of mealy bug, the present study was therefore, undertaken to determine the influence of
these insecticides in relation to resurgence.
32
Insecticide Induced Resurgence of Mealybug, Phenacoccus solenopsis Tinsley in Cotton 199
The experiment was conducted at Experimental area of Central Institute for Cotton Research, Regional
Station; Sirsa to study the effect of most frequently used insecticides alone and in combination by the
farmers on the resurgence of mealybug by using RCH-134Bt, the most popular Bt cotton hybrid of the
North Zone of India. The experiment was sown on 18.05.2008 and 17.05.2009.The experiment was
conducted in three replications with a plot size of 9.6 x 7.0 M in randomized block design (RBD), with
spacing of 100 x 60cm. These insecticides, 11 treatments (Cypermethrin, Monocrotophos, Acephate,
Ethion, Profenophos, Spinosad, Cypermethrin+Monocrotophos, Cypermethrin+Acephate,
Cypermethrin+Ethion, Cypermethrin+Profenophos and No spray) and 11 repeated applications were
applied at 10 days interval (For 2 years) .The observations on mealybug were recorded one day before
application of insecticides (pre-treatment) and one and seven days after application of insecticides (post-
treatment). The observations were recorded from 5 randomly selected and tagged plants per plot
(observed over time).
The studies conducted in field were again confirmed through Polyhouse experiment, where Cotton
was planted in pots. A single pot containing one plant was considered as one replication so 5 pots were
taken for each treatment. The polyhouse studies were conducted on Spinosad, Profenophos, Fipronil and
control. The potted cotton plant of one month age were artificially infested with the mealybugs and after
the establishment of mealybug when the population reached to the II
nd
grade injury level, the spray
application of each insecticide at recommended dose were applied through Baby Hand Sprayer at 10 days
intervals. The data on mealybug population was recorded as pre spray and 1 and 7

day after application of
insecticides. The % resurgence in population was calculated as per the method used for field experiments.
The data was compiled and the % resurgence was calculated on the basis of formula suggested by
Henderson and Tilton (1955) and slightly modified by Jayaraj and Reghupathy,1987 and finally approved
through All India Cotton Improvement Project, Coimbatore, Tamilnadu (Anonymous, 1993)
Resurgence (%) = Ts X C
F
- 1 X 100
Cs T
F
Where
Ts - Subsequent observations

in treated plot (Post treatment)
Cs - Subsequent observations

in control plot (Post treatment)
T
F
First observation in a treated plot (Pre-treatment)
C
F -
First observation in a control plot (Pre-treatment)
On the basis of pooled data of two years and 11 sprays applied at 10 days intervals, 14.53 per cent
resurgence in mealybug population was recorded due to application of Spinosad (Table-1).
Besides the cumulative effect the individual spray application also indicated resurgence in mealybug
population and during the year 2008 (Table - 3) after 1
st
and II
nd
spray no resurgence in mealybug
population was recorded due to Spinosad but the induced resurgence due to spinosad in mealybug
population was recorded after 3
rd
spray (Nil and 5.93%), 4
th
spray (0.69% and 20.90%), 6
th
spray (0.57%
and Nil), 7
th
th
th
spray (0.81% and 11.24%), 10
th
spray
(Nil and 7.25%) and 11
th
spray (Nil and 5.35%) at 1

and 7 day of spray application. Some inconsistent
resurgence in mealybug population after 10
th
spray of Acephate (8.21%) and after 9
th
spray of Ethion
(5.29%) was reported at 7 days of spray application during 2008.Among the combinations, 2.53%
resurgence due to Cypermethrin+Ethion after 9
th
spray application and 5.62% resurgence due to
Cypermethrin+Profenophos after 2nd spray application, in mealybug population was reported at 7 days
of spray application.
During 2009, resurgence due to Spinosad application (Table - 4) in mealy bug population was
recorded after 2
nd
spray (Nil & 4.53%), 3
rd
spray (11.44% & 13.88%), 4
th
spray (12.21% & 15.22%), 5
th

(14.20% & 27.72 %), 6
th
spray ( 14.12% & 36.47%), 7
th
spray (18.06% & 30.24%), 8
th
spray (16.23% &
33.05%) 9
th
spray (33.73% & 43.53%), 10
th
spray (Nil & 13.69%) and 11
th
spray (11.18 & 21.15 % ) at
1
st
and 7
th
day of spray applications.Inconsistent resurgence due to cypermethrin application during the
2009 was recorded after 5
th
spray (Nil & 10.89%), 6
th
spray (Nil & 15.87%), 7
th
spray (17.56% &
19.99%), 8
th
spray (12.21% & 18.23%), 9
th
spray (Nil & 21.92%), 10
th
spray (Nil & 10.72%), and 11
th

spray application (9.79% & 12.56%) at 1 and 7

day of spray applications.
Due to monocrotophos in 2009 after 6
th
spray (Nil & 12.12%), 7
th
spray (16.27% & 29.08%), 8
th
spray (16.29% & 19.57%), 9
th
spray (Nil & 28.00%) and 11
th
spray (14.28% & 25.70 %) resurgence was
recorded after 1 and 7 days of spray applications. Unlike to 2008, in 2009 resurgence in mealybug
population was not recorded due to the application of combinations of insecticides.
Comparatively more resurgence in mealybug population was recorded during 2008 than to 2009. The
studies conducted under polyhouse conditions also revealed 9.71 and 12.92 % resurgence in mealybug
population after 5 sprays applied at weekly intervals (Table - 2).The resurgence recorded other than
spinosad was inconsistent as it was not recorded after each spray during both the years.
As the mealybug appeared in cotton in 2006 from Gujarat (Kuchh) to Punjab (Bathinda) and it
multiplied and spread very rapidly across India. This situation led to confusion among farmers and an era
of usage of unwanted, unrecommended insecticides started. Farmers were spraying against the pest
whatsoever was available in market. Many insecticides which were totally out of use in cotton crop
started again. The purpose of the experiment was to study whether any of these commonly used
insecticides are responsible for resurgence in mealybug population though few were giving good results.
There are large numbers of reports and findings on the resurgence of mealybug due to synthetic
pyrethroids like the adverse effect of insecticide treatments on mealy bug population density and pest
status is characterized by the fact that the cryptic nature of mealy bug renders the use of many of the
insecticides ineffective, and the frequent use of non-selective insecticide to be responsible for the
outbreak of mealy bugs (Browning, 1994 and Smith et al., 1997). The phenomenon of resurgence has
recently been reviewed and discussed by Hardin et al., (1995). The resurgence of Japanese mealybug
against Cypermethrin was reported by Morishita, 2005. Weires (1984) reported the resurgence of
Comstock mealy bug, Pseudococcus comstoki (Kuwani) on apple treated with flucythrinate. Mendel. et.
al. (1994) reported the outbreak of mealy bug due to the use of the pyriproxifen for controlling the
Aonidiella aurantii. Rosen (1974) reported the outbreak of Planococcus citri and Pseudococcus
longispinus because of the frequent use of organophosphate and carbamates. The outbreaks of mealybug
on grape vine in part of Karnataka after insecticide sprayed were also recorded by Manjunath (1985). The
spinosad triggered resurgence in mealybug population reported in present studies could be due to direct
or indirect effects which need careful and systematic studies. In majority of insecticides induced
resurgence, the elimination of natural enemies due to insecticides is considered as an important factor
but, the spinosad was recorded safer to the potential parasitoid, Aenasius bambawalei Hayat (Rishi et al,
2009) but monocrotophos was recorded as most toxic for it and in present studies the activity of the
parasitoid Aenasius bambawalei was not affected as per visual observations.. The confirmatory studies
conducted in polyhouse also indicate towards changes in plant or insect physiology as there was no
parasitoid activity observed under polyhouse condition.
The resurgence studies conducted through periodical application of most commonly used insecticides
in cotton pest management resulted into resurgence in mealybug population. The resurgence recorded in
many insecticides alone and in combination was not consistent. However, the resurgence recorded in
mealybug population in spinosad treated plots was observed after each spray application. The resurgence
recorded in mealybug population during 2008 was less as compared to 2009. The exact mechanism for
spinosad triggered resurgence in mealybug population has to found out.
Insecticide Induced Resurgence of Mealybug, Phenacoccus solenopsis Tinsley in Cotton 201
TABLE 1: CUMULATIVE* POPULATION OF MEALY BUG (ON 5 CM PORTION OF STEM) AND PER CENT RESURGENCE DUE TO INSECTICIDES
Treatment Cumulative Population (on the Basis of 11 Spray) Resurgence (%)
Pre-Treatment After 1 Day After 7 Day After 1 Day After 7 Day
Cypermethrin 5.205 4.39 4.84 -14.94 -2.125
Monocrotophos 4.65 3.78 4.14 -23.07 -9.03
Acephate 4.255 3.25 3.36 -32.16 -29.05
Ethion 5.31 4.26 4.17 -29.49 -15.765
Profenophos 2.705 1.06 1.495 -60.65 -41.285
Spinosad 6.075 5.96 6.99 -6.72 14.53
Cyp+Mono 4.495 3.53 3.8 -29.84 -21.38
Cyp+Acephate 4.8 3.63 3.605 -24.57 -24.635
Cyp+Ethion 5.305 4.155 4.345 -31.215 -23.295
Cyp+Profenophos 4.29 3.125 3.495 -31.585 -20.105
No spray 5.03 5.19 5.25 0.0 0.0
(Pooled data of 11 sprays applied during 2008 and 2009); -ve values indicate no resurgence
TABLE 2: INSECTICIDE INDUCED RESURGENCE IN MEALY BUG POPULATION UNDER POLY HOUSE CONDITION DURING 2010
Treatments Per cent Resurgence
1
st
Spray 2
nd
Spray 3
rd
Spray 4
th
Spray 5
th
Spray Cumulative Population
1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS
Profenophos -59.18 -96.11 -100.00 -100.00 -100.00 -100.00 -100.00 -100.00 -100.00 -100.00 -55.72 -95.67
Spinosad 44.30 6.71 -56.74 11.32 47.88 14.39 8.43 12.19 33.91 21.42 9.71 12.92
Control 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00
-ve values indicate no resurgence
TABLE 3: PER CENT RESURGENCE IN MEALY BUG POPULATION DUE TO INSECTICIDE DURING 2008
Treatmens Per cent Resurgence
1
st
Spray 2
nd
Spray 3
rd
Spray 4
th
Spray 5
th
Spray 6
th
Spray 7
th
Spray 8
th
Spray 9
th
Spray 10
th
Spray 11
th
Spray
1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS
Cypermethrin 0.0 0.0 -33.33 -24.16 -23.05 -24.32 -42.52 -9.62 -10.69 -3.08 -12.77 -12.90 -36.47 -16.92 -17.21 -3.95 -0.54 -6.96 -31.12 -18.07 -0.67 -21.83
Monocrotophos 0.0 0.0 -33.33 -18.75 -44.51 -47.94 -72.11 -49.96 -1.23 -32.00 -19.70 -15.72 -3.08 -8.02 -12.14 -7.83 -21.70 -18.27 -25.29 -10.21 -29.24 -29.53
Acephate 0.0 0.0 -30.76 -25.00 -2.75 -56.85 -73.68 -45.45 -1.89 -56.55 -7.74 -3.03 -17.50 -14.70 -20.73 -12.46 -10.91 -24.49 -23.92 8.21 -36.19 -29.03
Ethion 0.0 0.0 -60.60 -3.97 -19.48 -16.19 -26.98 -19.17 -1.33 -49.99 -12.60 -0.39 -4.37 -12.66 -32.63 -19.32 -21.51 5.29 -11.03 -39.59 -24.81 -44.42
Profenophos 0.0 0.0 -50.00 -18.75 -61.66 -24.10 -84.83 -70.76 -54.30 -40.85 -74.52 -45.29 -54.84 -19.84 -63.22 -50.39 -32.27 -52.25 -52.21 -32.08 -60.07 -8.67
Spinosad 0.0 0.0 -21.21 -18.75 -7.07 5.93 0.69 20.90 -2.67 -14.90 0.57 -13.02 -20.65 0.53 -28.16 4.00 0.81 11.24 -20.05 7.25 -5.84 5.35
Cyp+Mono 0.0 0.0 -45.45 -11.36 -64.34 -29.39 -22.94 -19.14 -4.37 -38.97 -14.25 -12.96 -19.14 -3.04 -37.25 -15.58 -21.14 2.53 -14.95 -5.00 -0.63 -31.53
Cyp+Acephate 0.0 0.0 -21.21 -1.51 -7.27 -54.00 -32.17 -4.24 -1.62 -67.39 -22.96 -24.94 -21.53 -10.57 -19.77 -7.78 -50.53 -27.78 -25.66 -19.99 -17.91 -17.15
Cyp+Ethion 0.0 0.0 -21.42 -11.01 -31.31 -25.25 -59.65 -31.70 -0.12 -62.50 -15.46 -6.91 -20.52 -4.22 -34.63 -9.33 -2.59 2.53 -0.69 -25.38 -15.90 -20.62
Cyp+Profeno 0.0 0.0 -50.00 5.62 -8.73 -55.64 -18.60 3.09 -7.22 -51.07 -26.13 -17.42 -11.56 -0.18 -32.63 -18.55 -35.71 -25.20 -17.54 -13.71 -25.52 -3.77
No spray 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0
-ve values indicate no resurgence
TABLE 4: PER CENT RESURGENCE IN MEALY BUG POPULATION DUE TO INSECTICIDE DURING 2009
Treat Per Cent Resurgence
1
st
Spray 2
nd
Spray 3
rd
Spray 4
th
Spray 5
th
Spray 6
th
Spray 7
th
Spray 8
th
Spray 9
th
Spray 10
th
Spray 11
th
Spray
1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS 1DAS 7DAS
Cyper -93.48 -65.57 -23.70 -33.69 -9.70 -8.13 -27.12 -65.43 -53.97 10.89 -44.50 15.87 17.56 19.99 12.21 18.23 -18.49 21.92 -4.83 10.72 9.79 12.56
Mono -93.36 -70.63 -30.06 -37.11 -9.35 -5.17 -29.49 -48.42 -11.55 -38.05 -41.90 12.12 16.27 29.08 16.29 19.57 -54.17 28.00 -23.74 -15.65 14.28 25.70
Acephate -97.38 -89.77 -34.45 -26.57 -8.56 -12.84 -38.00 -53.23 -33.08 -18.88 -34.70 -31.58 -15.45 -31.51 -36.39 -44.66 -64.94 -38.03 -31.30 -14.42 -40.87 -29.11
Ethion -97.38 -95.08 -30.40 -23.94 -16.41 -24.54 -17.23 -59.88 -10.58 -9.92 -36.97 -36.76 -14.86 -14.29 -65.84 -53.20 -73.25 -39.64 -23.86 -6.19 -26.03 -19.34
Profen -94.99 -56.13 -40.84 -61.57 -20.69 -27.07 -73.44 -69.27 -57.35 -49.60 -57.03 -32.14 -63.61 -53.99 -69.71 -41.53 -80.02 -49.76 -71.04 -42.42 -83.73 -83.24
Spinosad -79.67 -51.75 -22.86 4.53 11.44 13.88 12.21 15.22 14.20 27.72 14.12 36.47 18.06 30.24 16.23 33.05 33.73 43.53 -17.28 13.69 11.18 21.15
Cyp+Mon -83.60 -64.36 -34.50 -22.51 -22.53 -14.86 -6.44 -9.26 -9.59 -11.60 -13.14 18.39 -14.52 -8.03 -17.75 -34.48 -58.08 -9.92 -55.80 -33.36 -17.06 -4.29
Cyp+Acep -92.72 -90.75 -48.14 -30.65 -15.49 -19.59 -20.33 -12.53 -29.31 -22.56 -34.03 -43.40 -20.22 -22.86 -68.45 -29.63 -53.01 -19.38 -16.14 -9.52 -18.34 -25.35
Cyp+Ethio -83.29 -62.88 -40.41 -44.28 -3.79 -7.91 -32.66 -18.44 -21.67 -34.85 -36.91 -10.17 -28.50 -30.87 -87.80 -27.59 -62.12 -29.68 -27.91 -10.17 -9.00 -2.25
Cyp+Profe -81.27 -34.47 -34.25 -36.04 -3.11 -3.31 -74.12 -47.22 -48.69 -41.19 -48.18 -10.81 -36.45 -38.55 -44.24 -16.36 -68.29 -37.78 -17.53 -8.62 -61.84 -37.98
No spray 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0
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Species Diversity, Pestiferous Nature, Bionomics and

Management of Mirid Bugs and Flower Bud Maggots:
the New Key Pests of Bt Cottons
S. Udikeri
1
, S. Kranthi
2
, K.R. Kranthi
2
, N. Vandal
1
, A. Hallad
1

S.B. Patil
1
and B.M. Khadi
1
Agricultural Research Station.Dharwad-580007 Karnataka India
2
CICR,Nagpur-440010 Maharashtra India
e-mail: ssudikeri@gmail.com
AbstractUnforeseen insect pests have emerged as potential threats in India and elsewhere due to large scale
cultivation of Bt cottons. Upto 69 % reduction in usage of pesticides has been achieved and reliance on synthetic
pyrethroids, broad spectrum highly toxic organophosphates has been almost ceased, paving way for dominance of
emerging pests. Hence, since 2007 field and laboratory investigations are in progress under TMC to unearth
potential threats from emerging pests to cotton cultivation. Three different species of mirid bugs (Miridae: Hemiptera)
viz., Creontiades biseratense (Distant), Compylomma livida (Reuter) and Hyalopeplus linefer (Walker) have been
found infesting cotton recently. Creontiades biseratense has become major production constraint presently in South
India and Maharastra. They are number one pests in Karnataka state presently. Adults and nymphs suck the sap
from base of square and tiny bolls leading to heavy shedding of these fruiting structures. Feeding on matured bolls
leads to parrot beaking and improper opening. It has five nymphal stages that span 14 days and adult longevity of 13
and 21 days respectively for males and females. No Bt transgenic cotton cultivar or event could show appreciable
resistance to mirid bugs in cultivar association studies. The estimated minimum avoidable yield loss is 20.6 %.
Acephate 75 SP @ 700 gai/ha appeared to be promising chemical control option which limited the population to 14
mirids / 25 squares with maximum seed cotton yield of 2664 Kg/ha. Another mirid tea mosquito Helopeltis bradyi
(Waterhouse) has been noticed for the first time in interspecific Bt cotton hybrids causing 85 % yield loss through
severe boll and square damage. This paper also presents a new report on flower bud maggot Dasineura gossypii
Fletcher (Cecidomyiidae: Diptera) which appeared as potential pest for first time in the history of cotton entomology.
Currently > 90 % fruiting body damage has been recorded in largely cultivated Bt cotton cultivars viz Kanaka and
Neeraj. The reproductive biology of this pest spreads over 12 -13 days spending all stages in squares or flower buds
leading to heavy loss. Bioefficacy of Malathion 50 EC @ 2.0 ml / lit was better over the rest with 4.3 damaged buds /
plant limiting avoidable yield loss to 480 kg/ha or 24 per cent.
INTRODUCTION
Since commercialization of Bt transgenic genotypes in 2002 there is miraculous increase in area and
production of cotton in India. During 2009/10 the area under cotton was 103.29 lakh ha being highest in
the world. With 295 lakh bales production the country remained second largest producer of seed cotton
globally. The contribution from Bt cultivars is nearly 90 % which occupied 8.4 m ha area. Upto 69 %
reduction in usage of pesticides has been achieved through Bt transgenic cottons. The reliance on
synthetic pyrethroids and highly toxic broad spectrum organophosphate insecticides has ceased. Further
commercialization of Bollgard-II (2006) cottons expressing Cry1Ac and Cry2Ab together paved way for
dominance of emerging pests in cotton. By 2010 six different Bt transgenic events have been approved
for commercial cultivation in India. Hence there is mosaic of different hybrids and Bt genes offering
varied level of resistance /susceptibility to bollworms and sucking pests. Emerging pests have become
significant issue in sustainability of Bt cottons in many countries. The incidence, spread and chemical
control exercised over mealy bugs Phenacoccus solenopsis Tinsley (Pseudococcidae: Hemiptera)
recently in India stands as an example for altered insect pest scenario. In South India and Maharastra the
mirid bugs (often referred as true bugs) are now creating havoc. Creontiades biseratense (Distant) since
its first appearance in 2005 (Patil et al 2005) is rampant these days. The present paper deals with status of
mirid bugs and flower bud maggots as emerging pests of Bt of cotton in India.
33
Diversity of mirid bugs was assessed through regular survey in farmers fields in Karnataka state. The
association of cultivars with mirid bugs was assessed by growing different Bt cottons (and non Bt as
check) in two replicated randomized block design. The number of cultivars included varied in different
years but were tested for two consecutive seasons. Each genotype was grown in six rows at spacing of 90
cm X 60 cm. Standard package of practices were followed except that it include any pesticide sprays.
Avoidable loss was assessed by protecting the crop twice in the peak mirid bug incidence period with
different concentrations of most effective chemical Acephate 75 SP to create differential population. A
similar study was conducted involving different insecticides and biorationales to assess best management
option. In this study, two rounds of treatment applications were maintained and were carried out between
2007 to 2011 under rain fed conditions at Agricultural Research Station, Dharwad (Karnataka: India),
which is situated between 15 07' N latitude and 76 06' E longitude with an altitude of 678 m above
MSL in the northern transitional region (zone-8) of Karnataka (India). The average rainfall of ARS,
Dharwad is 733.8 mm. The mirid species under consideration was C. biseratense only for all these
experiments as a dominant and regularly occurring species. The information from other states covered (if
any) is based on personal communications with cotton entomologist under TMC /AICCIP projects.
The bionomics and oviposition pattern of C.biseratense discussed here is based on laboratory study
conducted during 2009. The details are available in the publication Udikeri et al (2010). The informations
on another mirid threat Helopeltis bradyi (Waterhouse) and a new pest flower bud maggots Dasineura
gossypii (Fletcher) are based on survey and strict vigilance over emerging pests in the state. The
bionomics of flower bud maggot has been studied in laboratory (on Bt cotton plants) at ARS, Dharwad
2010. The avoidable loss -cultivar association studies were carried out in Haveri district (Karnataka)
Species Complex, Distribution of Mirid Bugs and Severity in India
Three species of mirid bugs are found infesting cotton in India. None of these have been historically
referred as key pests of cotton. It is only after widespread cultivation of Bt cottons that mirid bugs have
assumed key status. Three different species of mirid bugs (Miridae: Hemiptera) viz., Creontiades
biseratense (Distant), Campylomma livida (Reuter) and Hyalopeplus linefer (Walker) were infesting
cotton since 2005. Unlike widespread distribution of mealy bug Phenacoccous solenopsis Tinsley
(Nagrare et al., 2009) mirid bugs have been restricted to Central and South India. The dominant species
is Creontiades biseratense (Distant). It is most potential, widespread and also becoming a production
constraint every year (Plate 1 and 2). In Karnataka Creontiades biseratense mirid bugs are the major pest
as in TamilNadu, Maharastra and Andhra Pradesh. Campylomma livida (Reuter) is dominant species in
Maharastra, however it is noticed in Karnataka also. Hyalopeplus linefer (Walker) is seen in Karnataka
and Maharastra, but, not as regular pest. Its status is negligible (Udikeri et al., 2008).
Identifying Characters
The commonly occurring key mirid bug Creontiades biseratense appears in different colour morphs,
brown and green being common. Black and red morphs are also often noticed. Such green and brown
mirids are common Australia and US also. But they are colour morphs. In recent years, two species of
mirid bugs were recorded in Australia, of which Green mirid (Creontiades dilutus) considered as a
significant pest and brown mirid (C. pacificus) are quite common in pulse crops (Khan, 2004). Adult of
Indian cotton mirid bug Creontiades biseratense are brown in color with dark brown T-shaped band on
pronotum. Nymphs are greenish in color with dark brown wing pads. The characters of Megacoelum
biserratense reported long from Thailand (Hormachan et al., 1998) are similar to the Creontiades
biseratense. Both could same also. Taxonomic revision is essential in this regard. Major identifying
character of Hyalopeplus linefer is the presence of brownish parallel streaks on the pronotum. Distinct
color morphs have been noticed in H. lineifer also. Nymphs of this species are creamish yellow in color
with long antenna and wing pad. Both of these species are larger in size than Compylomma livida. The
wing margins in these species are fringed and eyes are diaptic.

Species Diversity, Pestiferous Nature, Bionomics and Management of Mirid Bugs and Flower Bud Maggots 205
Nature of Damage
Both adults and nymphs damage developing flower buds/ squares and tender bolls. One to two days old
bolls with dried petals intact provide a good habitat to the insects for feeding and sheltering. The
characteristic symptoms of feeding on the flower bud shows oozing out of yellow fluid from the buds and
staining of this yellow fluid on the inner surface of the bracts. Infested tender bolls have number of black
patches on all sides of the outer surface of boll rind. It was observed shedding of most of the damaged
squares and tender bolls. Usually the adults are swift fliers and brown in color. Nymphs are small with
yellowish green abdomen and fast moving when disturbed. Feeding on matured bolls leads to parrot
beaking and improper opening. It was estimated that damage leading to shedding of tender bolls for two
day period revealed a loss of seed cotton yield. The symptoms of infestation and impact of mirid bugs
observed in India do not deviate from the pestiferous cotton mirids described elsewhere (Udikeri et al.,
2008).
Cultivar Association
A total of 100 Bt and four conventional cotton hybrids were evaluated to assess host plant resistance to
mirid bug C biseratense. Conducted over three seasons none could show resistance to mirid bugs. Thus
mirid attack was not genotype dependent, however slightly higher numbers on interspecific biotech
hybrids were observed. There was no significant difference among BG and BG-II, Non Bt and Bt hybrids
also. Two cultivars viz., Kanak (MRC 7351) and Neeraj (MRC 7201) always demonstrated higher
incidence of mirid bugs. These two hybrids have been affected in farmers fields too (Table 1.)
TABLE 1: BT COTTON GENOTYPES WITH COMPARATIVELY LOW INCIDENCE OF MIRID BUGS
Cultivar Group and Bt Event Genotypes with Less than Average 28.08 Bugs per 25 Squares
HxB BG-I NCHB 945,RCH-708
HxB BG-II Nil
HxB Non Bt hybrids DCH-32 N Bt,
HxH BG-I NCS-145, BN Bt, RCH-2Bt, RCH-20Bt, NCS-954,NCS-929,Jk-ISHWAR,JK-DURGA,
CHIRANJEEVI Bt, AKKA, DRUVA, NCS-907
HxH BG-II NCS 145 BG-II, KCH 15, TULSI-9, TULSI 45, SP 1037, CHIRANJEEVI, JAI-BG-II,
AKKA-BG-II, RCH-2, NCS-207
HxH Non Bt hybrids NCH 145, DHH-11 N Bt,
Susceptible genotype KANAKA, NIRAJ
Assessment of Avoidable Loss
Two applications of Acephate 75 SP @ 1400 g ai /ha at peak incidence offered maximum protection
against mirid bugs. The estimated loss was 449 Kg /ha or 20.6% over zero protection. This was on an
average true with either BG or BG-II cultivars under moderate insect pressure. This could also be more if
incidence is more or avoidable loss may increase with extended spray schedule. However two sprays
were ideal considering the pest and damage potential nature (Figure 1).

Fig. 1: Avoidable Yield Loss Due to Mirid Bug Incidence
g y g
0
10
20
30
40
50
60
70
175 350 700 1400 0
Different dosages of Acephate 75 SP (gai/ha)
M
i
r
i
d

b
u
g
s
/
2
5
s
q
u
a
r
e
s
0
100
200
300
400
500
600
A
v
o
i
d
a
b
l
e

y
i
e
l
d

l
o
s
s
(
k
g
/
h
a
)
Mirid bugs/25 squares Avaoidable loss (kg/ha)
Management of Mirid Bugs
Among various treatments, Acephate 75 SP @ 1.0 g/l was found be most significant treatment in
reducing mirid infestation. Acephate could limit the mirid incidence to 13.87 bugs/25 squares against 46
bug in untreated control leading to maximum recorded seed cotton yield 2664Kg /ha. It was followed by
Acetamiprid 20 SP and Imidacloprid 200SL. In Australia, trials have shown that reduced rate of
Indoxacarb or Fipronil combined with salt rendered effective control of mirid bugs (Khan, 2003). Fungal
pathogens viz., Verticillium, Metahrrhizium and Beauveria were not promising (Figure 2).

Fig. 2: Management of Mirid Bugs Through Insecticides and Boirationals
Bionomics of Mirid Bug C. biseratense and Oviposition Pattern
The females of mirid bugs C. biseratense preferred to lay eggs in petiole where in 855 eggs were traced.
There are five instars in the life history extending the total life cycle upto 39-41 days. The details are
available in Udikeri et al.(2010) (Table 2).
TABLE 2: BIOLOGICAL PARAMETERS AND MORPHOMETRIC MEASUREMENTS OF MIRID BUG C. BISERATENSE
Stage MeanSD Length (mm) MeanSD Breadth (mm) MeanSD
Egg (Incubation period) Days 5.601.1 1.559.5 0.456.9
Nymphal Duration (Days)
I instar 2.690.36 1.150.14 0.292.7
II instar 2.760.18 2.90.13 0.860.15
III instar 2.830.21 3.30.20 1.12.4
IV instar 2.890.24 4.41.27 2.71.6
V instar 2.830.20 5.50.26 2.60.25
Total Nymphal Period (Days) 141.19 - -
Pre mating period 1.750.42 - -
Mating period 3.600.51 - -
Pre oviposition 2.510.24 - -
Oviposition 9.511.2 - -
Post oviposition 8.220.64 - -
Fecundity (Number of eggs) 15.01.2 - -
Adult Longevity (Days)
Male 13.202.61 7.00.15 3.09.8
Female 21.03.50 9.00.1 3.40.15
Adult Life Cycle (Days)
Male 41.136.1 - -
Female 39.945.8 - -

0
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Management of Flower Bud Maggots
The bio efficacy of Malthion 50EC @ 2.0 ml/l was found to be most significant treatment in reducing
flower bud maggots infestation with 4.3 damaged buds / plant and maximum seed cotton (2475 kg /ha)
yield. It accounted for an avoidable yield loss of 480 kg/ha or 24 per cent. The next best chemical was
Profenophos 50 EC @ 2 ml/ lit with 24.0% avoidable loss (Figure 3).

Fig. 3: Management of Flower Bud Maggots (Midge) and Avoidable Loss in Bt Cotton
New Report of Tea Mosquito or Guava Kajji Bug as Pest of Bt cotton
Helopeltis bryadi (Waterhouse) a Hemipteran pest has been found damaging the Bt cottons severely. It is
a common pest on Guava / Cashew/ tea etc. and is called kajji bug as it causes black lesions on the almost
all leaves and young shoots, squares and bolls. However, it appears rarely on cotton crop. It has been
reported for the first time on DCH-32 during 1996 in Honnali of Davangere district. Later in 2002, on
the same genotype it was noticed in H. D. Kote taluka of Mysore district. The most severe damage of this
pest insect was noticed on Bt cotton in Hosalli village of Uttar Kannada district. Affected genotypes are
interspecific biotech hybrids, MRC -6918 and RCH-708 (Udikeri et al., 2011).
Both nymph and adults suck the sap from leaves and young shoots, squares and bolls. As a result of it
leaves get rolled at the edge with brown central black lessons particularly near the main veins. Cankers
like lesions develop on the lower green bolls. Linear scars with white papery layer appearance in tender
shoots. Retarded growth leads to gradual dying of shoots. Mature bolls bear blackish brown circular pits
towards tip of the bolls. Rotting of boll takes place due to entry of rainwater and such infested dried up
bolls / squares remain intact in plant (Plate 3 and 4).
Flower Bud Maggot or Gall Midge of Cotton: A New Report
During 2009 severe incidence of gall midge Dasineura gossypii Fletcher, 1914 (Cecidomyiidae: Diptera)
was seen in a farmers field at Hesarur (Taluk: Savanur District: Haveri). The crop was sown in first
week June and by August incidence took severe proportions. Later in Kakol, Konanatambagi etc villages
(Haveri District) also it has been noticed in severe form. In the research farm (ARS, Dharwad) it has been
noticed in negligible proportions earlier, but, since couple of years its incidence is increasing. In Raichur,
Bellary, Belgaum, Haveri, Gulburga and Dharwad district also the pest has been recorded during survey
but, in low proportions (figure 4).
0
2
4
6
8
10
12
14
16
Acephate 75 SP-
1.0g
Acephate 75 SP +
DDVP 76 EC-
1.0g+0.25ml
Fipronil 5% SC-
1.0ml
Malathian 50EC-
2.0ml
Prof enophos 50
EC-2.0ml
Untreated control-
0
D
a
m
a
g
e

(
%
)
0
50
100
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A
v
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a
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l
o
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s

(
k
g
/
h
a
)
Avoidable loss (kg/ha) Flower bud damge (%)

Fig. 4: Per Cent Square Damage of Flower Bud Maggot or Gall Midge in Different Bt Cotton
As the pest remains inside the flower and multiplies quickly, it was difficult to manage this pest by
the farmers even with 3-4 sprays. The insecticides used viz Imidacloprid (Confidor 200 SL),
Oxydemeton methyl (Metasystox 25 EC), Monocrotophos (Monocil 40 SL), Acephate (Starthene 75 SP),
Neem oil etc could not render satisfactory results. When these insecticides are used along with DDVP
(Nuvan 78 EC) as tank mix (@ 0.25 ml/l) the spread of incidence was checked to certain extent. In all the
locations where the pest has caused wide spread damage the cultivar was MRC-7351 BG-II (Kanaka)
only.
History of the Pest/ Previous Reports
Dasineura gossypii was described by Professor T. B. Fletcher in 1940 in Pusa, Bihar based on collections
from cotton flower buds (Fletcher, 1916). From Tamil Nadu also (Coimbatore) it has been reported
earlier as a pest on cotton as referred to as floral bud maggot (Ayyar 1932). Thus it could find a place in
Cotton in India: A Monograph Vol. II: 1960 (Dastur et al., 1960). After the initial reports, no
single reference about its appearance as a major/ minor pest is available. This is the first report of
Dasineura gossypii as a major pest in India.
Nature of Damage
Maggots feed on anthers and stamenal column leading to degradation / decaying. Three to fifteen
maggots are observed in a flower bud. The infested flower buds fail to grow properly and they will not
open as the petals as well as tissue inside dries. Thus comprehensive symptoms give a picture of flower
drying through organ degradation and death. The pupation take place inside dried flowers itself.
0
10
20
30
40
50
60
70
80
90
100
1
Bt Cotton cultivars
S
q
u
a
r
e

d
a
m
g
e

(
%
)
Kanaka
Neeraj
MRC-7383
Surpass
Miracle
Mallika Bt
RCH-2 Bt
Bunny Bt
Tulsi-9 Bg-I
Jai Bt
MRC-6918
MRC-7918
Chiranjeevi Bg-I
In general square formed will not turn into a boll, due to death at flowering stage. Tissue drying
and/or death unevenly lead to twisted or contorted stamenal column/ anthers. Compared to normal boll
persistent stamina column is seen in affected ones but it needs confirmation. In some cases, where
fertilization is not affected, the boll formation is affected. The size remains smaller. Tissue degradation is
prominent on boll rind also. The bolls will not reach normal size and no proper filling with fibre is seen
(plate 5 & 6).
Bionomics of Dasineura gossypii and Oviposition Pattern
The females of Dasineura gossypii preferred to lay eggs in square tips where in 42-45 eggs were traced.
There are three instars in the life history and pupation takes 4 to 5 days on bracts extending total life
cycle upto 10 -13 days. Further studies are under progress.
Large scale cultivation of bollworm resistant Bt transgenic cottons suppressed the dreaded pests
successfully world wide. Significant reduction in usage of insecticide especially broad spectrum
organophosphates and pyrethroids gave scope for emergence of new pests especially the sap feeders.
Mirid bugs either prevailing hitherto or newer ones have assumed key status warranting couple of sprays
during reproductive phase. Increased incidence of mirids in cotton may give rise to host range expansion
as well as enhanced damage in alternate hosts of these pests. Key strategies and integrated approaches are
essential for sustainable use of Bt technology.
ACKNOWLEDGEMENT
The funds received from Indian Council of Agricultural Research (ICAR) New Delhi in addition to the
technical support accorded by Central Institute for Cotton Research (CICR) Nagpur through TMC MM-I
3.1 project Emerging and key pests: their characterization, taxonomy, genetic diversity and control is
gratefully acknowledged.
Dr. C.A. Viraktamath. UAS. Bengaluru and Dr R.M.Sharma. ZSI. Jabalpur are acknowledged for
taxonomic services.
REFERENCES
[1] Ayyar, J. V. R. (1932)- Insects affecting the cotton plant in India. Madras Agril Dept Bulletin 28: 1-28.
[2] Dastur, R. H., Asana, R. D., Sawhney, K., Sikka, S. M., Vasudeva, R. S., Quadiruddin, K. and Roa V. P., and Sethi, B. L.
(1960)- Indian Central Cotton Committee., Bombay
[3] Hormachan, P. A.,Wongpiyasatid and Piyapuntawanon (1998)- New record of Megacoelum biseratense (Heteroptera:
Hemiptera) in Thailand. Proc. KUDRI Res. Conf., 20-23.
[4] Khan, M. (2003)-Salt mixtures for mirid management. The Australian Cotton Grower 24: 10.
[5] Khan, M., Kelly, D., Hickman, M., Mensah, R., Brier, H., and Wilson, L. (2004)- Mirid ecology in Australian
cotton.www.csiro.org
[6] Nagrare, V. S., Kranthi, S., Biradar, V. K., Zade, N. N, Sangode, V., Kakde, G., Shukla, R.M., Shivare, D., Khadi, B. M.,
Kranthi, K. R. (2009)- Widespread infestation of the exotic mealybug species, Phenacoccus solenopsis (Tinsley)
(Hemiptera: Pseudococcidae), on cotton in India. Bull. Entomol. Res. 99: 537-41.
[7] Patil, B. V., Bheemanna, M., Patil, S. B., Udikeri, S. S., and Hosaman, I. (2006)- A record of mirid bug Creontiades
biseratense (Distant) on cotton from Karnataka., India. Insect Environ. 11: 176-77.
[8] Udikeri, S. S. (2006)- Evaluation of new generation Bt cotton genotypes, Sustainability of Cry protein expression,
computation of ETL, Effect on aphid predators and development of IPM module for Bt Cotton under rainfed conditions. Ph.
D. Thesis, Univ. Agric. Sci., Dharwad, Karnataka (India).
[9] Udikeri, S. S. (2008)- Mirid Menace: An emerging potential sucking pest problem in cotton. The ICAC recorder Vol-
XXVI No.4
[10] Udikeri, S. S., Kranthi, K. R., Patil, S. B., Modagi, S.A and Vandal, N.B. (2010)- Bionomics of mirid bug Creontiades
biseratense (Distant) and oviposition pattern in Bt cotton, Karnataka J. Agri. Sci. 23: 153-156.
[11] Udikeri, S.S., Kranthi, K.R., Patil, S.B. and Khadi, B.M. (2011)- Emerging pests of Bt cotton and dynamics of insect pests
in different events of Bt cotton. Paper presented in 5th Asian Cotton Research and Development Network Meeting, Lahore,
Pakistan 23-25th February 2011. (www.icac.org)
Influence of Spatial Cropping Patterns of Cotton

Cultivation on Population Dynamics of Mirid Bug,
Creontiades biseratense (Distant)
B. Dhara Jothi
1
, T. Sonai Rajan
1
, V.S. Nagrare
2
, M. Amutha
1
,
Rishi Kumar
3
and T. Surulivelu
1

1
Central Institute for Cotton Research, Regional Station, Coimbatore, India
2
Central Institute for Cotton Research (CICR), P. B. No. 2., Shankar Nagar P. O., Nagpur, India
3
Central Institute for Cotton Research (CICR), Regional Station, Sirsa, Haryana, India
e-mail: dhara56@yahoo.co.in
AbstractMirid bug Creontiades biseratense (Distant) has been reported as an emerging pest on Bt cotton from the
States of Tamil Nadu and Karnataka. Nymphs and adults of C. biseratense cause damage to squares, flowers and
tender bolls leading to the affected parts gradually turning to yellow followed by shriveling and premature dropping.
Such damage results in significant reduction in seed cotton yield. Spatial cropping patterns with different adjacent
crops of cotton fields are expected to exert influence on the population dynamics of mirids. Present study assessed the
population dynamics of C. biseratense across different spatial cropping patterns of cotton viz., cotton surrounded by
non-target crop (Tomato) and cotton with intercrop (cowpea) during (2008-09) and additional adjacency cropping
patterns viz., cotton adjacent to weedy road and cotton adjacent to fallow weeds during 2009-10 with cotton as sole
crop as control during both the years were studied under farmers field conditions at Coimbatore, Tamil Nadu.
Influence of the spatial cropping patterns on nymphs, adults, natural enemies and the damage potential of the pest on
squares and bolls were documented throughout the season, and compared. During 2008-09, nymphal population
varied significantly with the cropping system between 37
th
standard meteorological week (SMW) and 41
st
SMW
wherein minimum population was recorded in cotton + cow pea (0.85 - 2.90/plant) consistently. Among the five
cropping patterns during 2009-10, cotton adjacent to weedy road recorded the maximum seasonal mean population of
nymphs besides for two periods. The adult population recorded across different cropping patterns was of the order:
cotton + cowpea (0.80-1.20/plant) > cotton surrounded by tomato (1.05-1.70/plant) > cotton as sole crop > cotton
adjacent to weedy road > cotton adjacent to fallow fields. Significant difference in square damage was recorded during
second fortnights of September, November and December during 2008-09. Square damage recorded was minimum in
cotton + cowpea (8.40-12.76%) andcotton + tomato (10.62-14.86%) and maximum in cotton as sole crop (14.67-
30.02%). During 2009-10, though cotton adjacent to weedy road recorded maximum square damage initially, cotton
surrounded by tomato recorded maximum damage in the later stages of the crop. Cotton + cowpea cropping system
recorded minimum boll damage (9.06 - 18.61%) as compared to other patterns of cotton cultivation. Cotton
surrounded by tomato recorded maximum Square and boll damage. Significant influence of cropping patterns on the
spider population was also recorded. The results indicated that cotton + cowpea intercrop with weed free surroundings
reduced the mirid bug population, and hence could form an important component of IPM package for the management
of the pest.
INTRODUCTION
Genetically engineered cotton crop that express endotoxin (Cry protein) from Bacillus thuringiensis
(Bt) is mainly toxic to the bollworms, (American bollworm, Spotted and Spiny bollworm and Pink
bollworm) semiloopers and hairy caterpillars in India (Kranthi, 2009). Reports (Qaim and Zibberman,
2003; Barwale et al., 2004, Bennet et al., 2004 & Morse et al., 2005) showed that yields increased
substantially by adopting Bt cotton and farmers in India were able to reduce insecticides use by at least 2
3 applications. Reduced insecticide application coupled with selective use of insecticides have allowed
minor pests to survive and emerge as important ones and the changed pest complex comprises of mirids,
aphids, whiteflies, thrips and mealybugs (www.crdc.com.au). In China, mirid bug have historically been
considered occasional (or) minor pests in most crops, occurring at relatively low population levels and
only sporadically pests management intervention (Lu and Wu, 2008). The mirids (true bugs) comprise a
large and diverse insect family miridae belonging to the order Hemiptera. These are small terrestrial
insects, usually oval shaped (or) elongate measuring less than 12 mm in length (en.wikipedia.org). Most
34
Influence of Spatial Cropping Patterns of Cotton Cultivation on Population Dynamics of Mirid Bug 211
of the mirids are agricultural pests with a wide host range of sunflower, safflower, pigeon pea, lucern,
legumes, maize, sorghum, bajra (Khan et al., 2004). The cotton mirid bug, Creontiades biseratense
(Distant) is an emerging insect pest on Bt cotton in Karnataka and Tamil Nadu, India causing heavy
shedding of squares and bolls which lead to significant reduction in seed cotton yield (Patil et al., 2006,
Surulivelu and Dharajothi, 2007, Ravi, 2007 and Udikeri et al., 2009). Mirid bugs can easily attain
outbreak levels; switch host crops (or) experience geographic spread due to their environmental
adaptability (Lu & Wu, 2008 and Ting, 1963 & 64), high population growth rate (Lu & Wu, 2008 and
Ting, 1963) and strong dispersal capacity. Detailed studies on the influence of spatial cropping pattern of
cotton cultivation on population dynamics of mirid bug, C. biseratense were conducted in Coimbatore
region and presented in this paper.
To study the influence of spatial cropping patterns of cotton cultivation on population dynamics of cotton
mirid bug, C. biseratense studies were conducted at Coimbatore under farmers field conditions during
the winter cotton season of 2008 09 and 2009 10. The village farms are located 30 Kms away from
Coimbatore. During 2008 09, three fields with cotton as sole crop, cotton with cow pea as intercrop and
cotton surrounded by non-target crop (tomato) and during 2009 10 apart from the above three adjacent
cropping patterns (cotton adjacent to weedy road and cotton adjacent to fallow land) were selected as
study fields. Cotton as sole crop was kept as control in both the years. Periodical observations were
recorded at weekly intervals on the population dynamics of mirid bug - nymphs, adults, natural enemies
and damage caused by the mirids on squares and bolls. All the data were analyzed statistically and
interpretations were drawn.
Dynamics of nymphs and adults of C. biseratense were recorded under 3 spatial cropping patterns of
cotton during 2008-09. Nymphal population appeared during September II fortnight to December II
fortnight. Within the observation period nymphal population was significantly varied from September to
October I fortnight, minimum population of nymphs were recorded in cotton with cowpea (0.85 to 2.90)
followed by cotton surrounded by tomato (2.40 to 3.75) and cotton as sole crop 2.60 to 3.0 (except
October I fortnight). However overall seasonal mean of the nymphs indicated that there was no
significant difference among the three cropping patterns (Table 1& Fig.1.a). Adult population recorded in
all the cropping systems vary significantly from September I fortnight to November II fortnight. Initially
minimum population of adult was recorded on cotton as sole crop (1.45 to 5.0) from September I
fortnight to October I fortnight. However, influence of cotton with cow pea cropping pattern was
recorded with the minimum population of adults varied from 0.6 to 3.45 followed by cotton surrounded
by tomato (0.75 to 4.25) and cotton as sole crop (1.25 to 5.50) from October II fortnight to November II
fortnight. Overall mean population of adults recorded were significantly low in cotton with cowpea as
intercrop followed by cotton surrounded by tomato and cotton as sole crop (Table 1& Fig. 1b.).
TABLE 1: INFLUENCE OF SPATIAL CROPPING PATTERNS OF COTTON CULTIVATION ON POPULATION DYNAMICS OF MIRID BUG, CREONTIADES BISERATENSE
S.
No
Cropping System Population & Damage/ 20 Plants
Nymph Adult Spider Square Damage (%) Boll Damage (%)
08 - 09 09 - 10 08 - 09 09 - 10 08 - 09 09 - 10 08 - 09 09 - 10 08 - 09 09 - 10
1. Cotton as sole crop 2.32 0.86 3.14 0.42 1.03 0.36 18.19 12.04 28.06 13.95
2. Cotton surrounded by Tomato 2.77 1.14 1.95 0.75 0.97 0.43 14.58 11.98 28.97 19.05
3. Cotton with cowpea 2.63 1.04 1.32 0.59 1.05 0.30 9.95 9.31 20.60 18.75
4. Cotton adjacent to weedy road - 1.29 - 1.88 - 0.53 - 12.51 - 16.58
5. Cotton adjacent to fallow land - 0.89 - 0.99 - 0.40 - 6.91 - 17.58
SEd 0.11 0.07 0.06 0.07 0.05 0.06 1.04 1.79 1.36 1.70
CD (P = 0.05) N.S 0.14 0.12 0.14 N.S 0.12 2.11 N.S 2.76 N.S.

Fig. 1: Influence of Cropping Pattern of Cotton on Dynamics of Mirid Bug during 2008-09
a.NymphalPopulation
0
1
2
3
4
5
6
7
SepIfort SepII fort Oct Ifort Oct IIfort NovIfort NovIIfort DecIfort DecIIfort
Standard weeks
N
y
m
p
h
2
0
p
l
a
n
t
s
CotonasSol e crop CottonSurroundedbytomato Cottonwi thCowpea
b.AdultPopulation
0
1
2
3
4
5
6
SepI fort SepII fort Oct Ifort Oct IIfort NovIfort NovIIfort DecIfort DecIIfort
Standard weeks
A
d
u
l
t
2
0
p
l
a
n
t
s
c.Squaredamage
0
5
10
15
20
25
30
35
SepI fort SepII fort Oct Ifort Oct IIfort NovIfort NovIIfort DecIfort DecIIfort
Standardweeks
S
q
u
a
r
e
d
a
m
a
g
e
2
0
p
l
a
n
t
s
d.Bolldamage
0
5
10
15
20
25
30
35
SepIIfort Oct Ifort Oct IIfort NovIfort NovIIfort DecIfort DecIIfort
Standardweeks
B
o
l
l
d
a
m
a
g
e
2
0
p
l
a
n
t
s
e.Spiderpopulation
0
0.5
1
1.5
2
2.5
SepIfort SepIIfort OctIfort OctIIfort NovIfort NovIIfort DecIfort DecIIfort
Standardweek
P
o
p
u
l
a
t
i
o
n
/
2
0
p
l
a
n
t
s
CotonasSol ecrop CottonSurroundedbytomato Cottonwi thCowpea

Fig. 2: Influence of Cropping Pattern of Cotton on Dynamics of Mirid Bug during 2009-10
a.NymphPopulation
0
0.5
1
1.5
2
2.5
3
3.5
4
Oct Ifort Oct IIfort NovIfort NovIIfort DecIfort DecIIfort DecIIfort
Standardweeks
P
o
p
u
l
a
t
i
o
n
/
2
0
P
l
a
n
t
s
CotonasSol ecrop Cottonsuroundedtomato Cottonwi thcowpea
Cotonadjacent toweedy road Cottonadjacenttofal l owl and
b.Adultpopulation
0
1
2
3
4
5
6
7
Standard weeks
P
o
p
u
l
a
t
i
o
n
/
2
0
P
l
a
n
t
s
Cotonadjacent toweedy road Cottonadjacent tofal l owl and
c.SquareDamage
0
5
10
15
20
25
30
35
Oct Ifort OctIIfort NovIfort NovIIfort DecIfort DecIIfort DecIIfort
Standardweeks
S
q
u
a
r
e
d
a
m
a
g
e
/
2
0
p
l
a
n
t
s
CotonasSol ecrop Cottonsuroundedtomato
Cottonwi thcowpea Cotonadjacenttoweedy road
Cottonadjacenttofal l owl and
d.Bolldamage
0
5
10
15
20
25
30
35
40
45
Standardweeks
B
o
l
l
d
a
m
a
g
e
/
2
0
p
l
a
n
t
s
Cotonadjacenttoweedy road Cottonadjacenttofal l owl and
e.SpiderPopulation
0
0.2
0.4
0.6
0.8
1
1.2
1.4
OctIfort OctIIfort NovIfort NovIIfort DecIfort DecIIfort DecIIfort
Standardweeks
P
o
p
u
l
a
t
i
o
n
/
2
0
p
l
a
n
t
s
CotonasSol ecrop Cottonsuroundedtomato
Cottonwi thcowpea Cotonadjacenttoweedyroad
Cottonadjacenttofal l owl and
Significant difference in square damage was observed during II fortnight of September, November
and December. Square damage was recorded as minimum in the cotton + cowpea (8.40-12.76) and cotton
+ tomato (10.62-14.86) and maximum in cotton as sole crop (14.67-30.02). No significant difference was
recorded during September I fortnight, October I & II fortnight, November II fortnight on the square
damage (Table 1& Fig 1c.). Mean square damage was minimum in cotton with cowpea as intercrop
followed by cotton surrounded by tomato and maximum square damage was recorded in cotton as sole
crop.
Significant difference in the per cent of boll damage was observed within the cropping patterns
during II fortnights of September, October, November and December. Cotton + cowpea cropping system
recorded minimum per cent of boll damage (9.06 - 18.61) as compared to other cropping patterns. No
significant difference in the per cent of boll damage was recorded during October I fortnight and
December II fortnight within the cropping system (Table 1 & Fig 1d.). Seasonal mean damage within the
cropping pattern also indicated minimum per cent of boll damage in cotton with cowpea as intercrop. No
significant difference in the spider population within the cropping patterns, however numerically higher
number of spiders was recorded in cotton with cow pea (Table 1 & Fig. 1e.).
During 2009 - 10 the influence of single cropping system on the nymphal population was not
observed consistently. The population varied significantly within the 5 cropping patterns only during
October I & II fortnight, November II fortnight and December I fortnight. On October I fortnight cotton
adjacent to weed road influenced the population of nymphs of the pest by recording highest population of
3.65 / 20 squares. During October II fortnight cotton adjacent to fallow weeds and sole crop of cotton
cropping system influenced the nymphal population by recording 1.85 and 2.95 / 20 squares respectively.
On November II fortnight cotton with cow pea and cotton surrounded by tomato recorded highest
population of 0.95 and 1.40 nymphs / 20 squares respectively (Table 1 & Fig 2a.).
Seasonal mean population was highest in cotton adjacent to weedy road & cotton surrounded by
tomato and the significantly minimum population was recorded in cotton as sole crop, cotton adjacent to
fallow land and cotton + cowpea and were on par with each other.
Significantly higher and lower number of adults was recorded in cotton adjacent to weedy road and
cotton as sole crop, respectively. Populations on all other cropping patterns were on par with each other.
Adult population of mirid bug was highly influenced by the cropping systems. During I and II fortnight
October and November cotton adjacent to weedy road and cotton adjacent to fallow weeds recorded
significantly highest level of adult population (Table 1 and Fig 2b.).
Square damage was significantly different within the cropping pattern during October I fortnight to
December II fortnight. Cotton surrounded by weedy road and cotton surrounded by tomato recorded
higher percentage of square damage during I and II fortnights of October, November and December
(Table 1 and Fig 2c). Significant difference in the per cent of boll damage was recorded during II
fortnight of November and December. Among the cropping pattern cotton surrounded by tomato and
cotton surrounded by weedy road recorded higher per cent of infestation consistently for two weeks
(Table 1 and Fig 2 d).
Overall spider numbers were minimum however, within the available population among the cropping
pattern, cotton adjacent to weedy road recorded higher mean population of spiders (Table 1 and Fig 2 e).
In general mirid population level was minimum during the observation period. Population of
nymphs, adults, square and boll damage observed during 2008-09 was higher than 2009-10. However,
with a mean population ranging from 2.32-2.77 / 20 plants 0.86-1.29 / 20 plants during 2008-09 and
2009-10 respectively caused % of square and boll damage ranging from 9.95 - 18.19, 6.91 - 12.51 and
20.60 - 28.97 and 13.95 -19.05 during 2008-09 and 2009-10 respectively (Table 1). Square and boll
damage were correspondingly higher during the availability of peak nymph and adult population.
Planting crops that are more attractive to mirids close to cotton crops may reduce population in
cotton. Lucerne has been shown to be an excellent trap crop for mirids, (www.cotton.crc.org.au). In the
present study influence of cropping patterns were studied for the level of mirid nymphs and adult
population level. Results indicated that cowpea intercropped in cotton recorded minimum population of
mirid nymphs, adults than in other cropping pattern. This is in agreement with Altieri, 1987 and
Venugopal Rao (1995) who has reported that polyculture, intercropping provides a favourable habitat for
the buildup of natural enemies which makes less convenient to establish crop pests. According to the
present study cotton intercropped with pulses (Cowpea) registered minimum damage in squares and bolls
as compared to other cropping patterns. As per the present study, cotton surrounded by tomato and cotton
surrounded by weedy road recorded higher level of infestation. Mensah and Khan, (1997) reported the
movement of Creontiades dilutus adults into adjacent weeds. The present study indicated that cotton +
cowpea (as intercrop) and weed free surroundings will reduce the mirid bug population, and form an
important component of IPM package for the management of sucking pests including the mirid bugs.
ACKNOWLEDGEMENT
We acknowledge the financial assistance by World Bank through Indian Council of Agricultural
Research, New Delhi to carry out the present study as a part of National Agricultural Innovation Project
(NAIP/DSS/C 2046) at Central Institute for Cotton Research, Regional Station, Coimbatore.
REFERENCES
[1] Altieri, M.A. 1987. The Scientific basis of alternative agriculture. Agro. Ecology, Boulder / London: Crestview/ITP. pp.
[2] Barwale, R.B., V.R., Gadwal, U. Zehr, and B. Zehr, 2004. Prospects for Btcotton technology in India.
Ag Bioforum 7: 23 26.
[3] Bennet, R.M., Y., Ismael, U. Kambhampatti, and S. Morse. 2004. Economic impact of genetically modified cotton in India.
AgbioForum 7: 96 100.
[4] http:// www.cotton.crc.org.au
[5] http://en.wikipedia.org/wiki/miridae
[6] Khan, M., D. Kelly, M. Hickman, R. Mensah, H. Brier and L. Wilson. 2004. Mirid ecology in Australia. CRS Research
Rev. 15.
[7] Kranthi, K.R. and D.A. Russell. 2009. Changing trends in cotton pest management. R. Peshin, A. k. Dhawan (eds.),
Integrated Pest Management: Innovation Development Process. Pp. 499 541.
[8] Lu Y.H. and K.M. Wu, Biology and control of cotton mirids, (Golden shield Press, Beijing, 2008).
[9] Mensah, R.K. and M. Khan. 1997. Use of Medicago sativa (L.) interplantings / trap crops in the management of the green
mirid Creontiades dilutus (Stal) in commercial cotton in Australia. Int. Journ of pests Mgt. 43 (3): 197 202.
[10] Morse, S., R.M. Bennett, and Y. Ismael, 2005. Genetically modified insect resistance in cotton: Some economic impacts in
India. Crop Protection 24(5): 433 440.
[11] Patil, B.V., M., Bheemanna, S.B. Patil, S.S. Udikeri, and A. Hosamani, 2006. Record of mirid bug, Creontiades biseratense
(Distant) on cotton from Karnataka, India, Insect Environ. 11: 176 177.
[12] Ravi, P.R., 2007. Bio-ecology, loss estimation and management of Mirid bug Creontiades biseratense (Distant)
(Hemiptera: Miridae) on Bt cotton. M.Sc., (Agri) Thesis, Univ. Agric. Sci., Dharwad (India).
[13] Surulivelu, T. and B. Dhara Jothi. 2007. Mirid bug, Creontiades biseratense (Distant) damage on cotton in Coimbatore,
http://www.cicr.gov.in.
[14] Ting, Y.Q., Acta Phytophyl. Sin. 2, 285 (1963)
[15] Ting, Y.Q., Acta Entomol. Sin. 13, 298 (1964)
[16] Udikeri, S.S., S.B. Patil, H.M. Shaila, G.S. Guruprasad, S.S. Patil, K.R. Kranthi, and B.M. Khadi. 2009. Mirid menace a
potential emerging sucking pest problem in cotton.
[17] http://www.icac.org.
[18] Venugopal Rao, N. 1995. Bio-ecology and management of H. armigera in the cotton ecosystem of Andhra Pradesh.
Bulletin of APAU, R. Nagar, Hyderabad.
[19] Qaim, M. and D. Zibberman, 2003. Yield effects of genetically modified crops in developing countries. Science 299: 900
902.
Determination of Economic Injury Level for

Defoliator Spodoptera litura (Fab.) on Bt Cotton
M. Bheemanna, S. Hanchinal, A.K. Hosamani and R. Chowdary
University of Agricultural Sciences, Raichur, India
e-mail: bheemuent@rediffmail.com
AbstractCotton defoliator Spodoptera litura (Fab.) is considered to be one of the important pests after introduction
of Bt cotton. The damage potential and economic injury level (EIL) for S. litura larvae on Bt cotton (NCS 145 BG) at
different days after sowing were worked out. The EIL was worked out for two years during 2008-09 and 2009-10
seasons. Economic injury level was 2.64, 3.47 and 4.27 larvae per plant at 90,120 and 135 days after sowing,
respectively during 2008-09. Similarly during 2009-10, EIL was 1.68, 2.44, 2.54 and 3.59 larvae per plant at 90, 105,
120 and 135 DAS, respectively.
INTRODUCTION
The knowledge of economic threshold level (ETL) helps in determining whether an insect is to be
classified as a pest or not. Without a doubt, economic decision levels are the key stones of insect pest
management programmes and they are indispensable because they indicate the course of action to be
taken in any given pest situation (Pedigo,1991). The ideas expressed by Pierce(1934) with regard to the
assessment of insect damage and the initiation of control measures become one incentive for
development of the concept of economic injury level (EIL). In later years, it was Stern et. al (1959) who
formally proposed the concept of economic threshold level as the number of insects(density or intensity)
when management action should be taken to prevent the increasing pest population from reaching
economic injury level.
The effective management of most of the pests in cotton depends on the use of chemical insecticides,
but its use should be judicious, need based and based on ETL (Economic threshold level) concepts. The
earlier concept of pest control arising cent per cent elimination of pests from the agricultural ecosystem
was replaced by the term Pest Management concept, the established and accurate determination of
action thresholds of any pest species is a prerequisite. Considering these points the present investigation
was undertaken to assess the economic injury level (EIL) of S. litura infesting Bt cotton.
The calculation of EIL for S.litura was done based on the economic losses in yield resulting from insect
pest and the cost involved in plant protection against the particular pest. An experiment in randomized
block design (RBD) was laid out during 2008-09 and 2009-10 under field conditions at Main
Agricultural Research Station, UAS, Raichur with eight treatments and three replications using the
popular Bt cotton hybrid (NCS-145 BG).
The treatments were
T1- Complete protection
T2- Releasing 5 larvae/plant
35
Determination of Economic Injury Level for Defoliator Spodoptera litura (Fab.) on Bt Cotton 217
For each treatment, six plants were maintained in each plot, such three replications were maintained
for each treatment. All the plots were covered with fine nylon mesh to prevent infestation from outside.
Second instar larvae of S .litura obtained from laboratory culture were released on cotton plants at 90,120
and 135 days after sowing during 2008-09 and 90,105,120 and 135 days after sowing during 2009-10
season. Different blocks were maintained for releasing the larvae at different days after sowing to assess
the yield loss. The cages were so designed that they did not allow free passage of larvae. The bottom
edges of the cages were inserted into the soil on all the sides so as to check the escape or entry of larvae.
Nylon net cages were erected on bamboo sticks fixed in four corners.
The larvae were released thrice during 2008-09 and four times during 2009-10 season. The total
weight of seed cotton lint from all the treatments was worked out by correlation co-efficients and
regression equations. Yield data were converted into q/ha. Yield loss from different treatments was
derived by delinting the yield in the respective treatment from the yield in control (where no larvae were
released).The value of yield loss was determined at the wholesale market price of cotton prevailing at
Raichur just after harvest during two seasons.
The EIL was computed based on the procedure given by Stone and Pedigo (1972) and modified by
Ogmana and Pedigo(1974) using the following formula

Gainthreshold=
Management cost (Rs/ha)
Market value of cotton (Rs/q)

Management cost was calculated for the insecticide Buprofezin @ 1ml/lt (Rs 2500/lt) along with
application cost including the labour charges. Two sprays were required to keep the crop free from
infestation.
The correlation co efficient r between the variables, population level of the pest reduction in cotton
yield per hectare were worked out using the following formula.
r=
N.xyxy
(Nx (x) )(Ny (y) )
2 2 2 2

Where,
N is the total number of observations
0X is the population levels of larvae/plant
0Y is the reduction in grain yield
Regression equation of y = a + bx form was deduced where a is the intercept, b is the yield
reduction per larva. Thus b is the per plant reduction in yield by increasing five larva. Substituting the
values in the above formula EIL was computed.
There was significant difference in leaf damage between treatments, which received varied number of
larvae and the plants which received no larvae during 2008-09 (Table 1).The per cent leaf damage ranged
from zero in completely protected (T1) plants to 46.44 where 35 larvae per plant where released (T8) at
90 DAS (Days after sowing). The plants which received 5, 10 and 15 larvae per plant were on par with
each other and differed significantly with treatment which received 20,25,30 and 35 larvae per plant. The
treatments which received 20 and 25 larvae per plant were on par with each other and differed
significantly with treatments which received 30 and 35 larvae per plant.

TABLE 1: PER CENT LEAF DAMAGE AND SEED COTTON YIELD DUE TO SPODOPTERA LITURA ON BT COTTON DURING 2008-09
No. of Larvae
Released Per Plant
Per Cent Leaf Damage Seed Cotton Yield (q/ha) Per Cent Reduction
over Control
90 DAS 120 DAS 135 DAS 90 DAS 120 DAS 135 DAS 90 DAS 120 DAS 135 DAS
Complete Protection 0.00
a
(0.00) 0.00
a
(0.00) 0.00
a
(0.00) 32.14
a
31.33
a
31.88
a
0.00 0.00 0.00
5 5.62
b
(13.37) 6.84
b
(14.20) 16.44
b
(23.82) 30.48
b
29.24
b
30.24
a
5.16 6.67 5.23
10 9.48
c
(17.79) 8.24
bc
(16.60) 18.84
b
(25.70) 29.12
c
29.08
b
30.08
a
9.40 7.18 5.75
15 12.44
c
(20.56 10.24
c
(18.29) 22.34
c
(28.17) 28.44
cd
28.84
b
30.11
ab
11.51 7.95 5.65
20 24.36
d
(29.52) 22.24
d
(28.10) 24.56
d
(29.70) 28.04
cd
28.46
b
29.48
ab
12.76 9.16 7.66
25 28.50
d
(32.26) 28.42
e
(32.21) 28..36
e
(32.17) 27.52
d
27.84
bc
29.22
ab
14.37 11.14 8.49
30 35.64
e
(36.65) 32.12
e
(34.51) 32.36
f
(34.66) 27.48
d
27.44
bc
28.48
ab
14.50 12.42 10.85
35 46.44
f
(42.96) 38.44
f
(38.31) 36.33
g
(37.06) 26.12
e
26.58
c
28.12
b
18.73 15.16 12.00
S Em 1.38 0.99 0.75 0.36 0.48 0.65
CD 4.19 3.04 2.29 1.08 1.46 1.96
Figures in the same column with similar alphabets do not differ significantly at P=0.05 by DMRT
TABLE 2: PER CENT LEAF DAMAGE AND SEED COTTON YIELD DUE TO SPODOPTERA LITURA ON BT COTTON DURING 2009-10
No. of
Larvae
Released
Per Plant
Per Cent Leaf Damage Seed Cotton Yield (q/ha) Per Cent Reduction Over Control
90 DAS 105 DAS 120 DAS 135 DAS 90DAS 105DAS 120DAS 135DAS 90DAS 105DAS 120DAS 135DAS
Complete
Protection
0.00
a
(0.00) 0.00
a
(0.00) 0.00
a
(0.00) 0.00
a
(0.00) 32.08
a
36.32
a
36.88
a
36.84
a
0.00 0.00 0.00 0.00
5 0.00
a
(0.00) 14.64
b
(22.41) 24.36
b
(29.57) 26.44
b
(30.91) 30.24
b
36.21
a
36.24
a
36.58
ab
5.74 0.30 1.74 0.71
10 0.00
a
(0.00) 18.24
c
(25.22) 25.64
b
(30.42) 28.84
c
(32.46) 30.12
b
36.13
a
36.84
a
36.12
ab
6.11 0.52 0.11 1.95
15 0.00
a
(0.00) 18.25
c
(25.20) 30.31
c
(33.40) 32.34
d
(34.64) 29.52
bc
36.04
a
36.12
a
36.00
ab
7.98 0.77 2.06 2.28
20 5.34
b
(13.40) 26.88
d
(31.20) 32.58
d
(34.81) 34.56
e
(35.99) 28.36
c
35.58
a
35.88
ab
34.64
ab
11.60 2.04 2.71 5.97
25 8.45
c
(16.90) 30.22
e
(33.22) 37.34
e
(37.67) 38.36
f
(38.26) 27.48
cd
34.21
b
34.89
b
34.12
ab
14.34 5.81 5.40 7.38
30 8.22
c
(16.70) 36.52
f
(37.16) 40.46
f
(39.50) 42.36
g
(40.60) 26.12
d
32.21
c
32.85
c
33.42
ab
18.58 11.32 10.93 9.28
35 10.11
c
(18.50) 42.44
g
(46.39) 46.55
g
(43.02) 52.33
h
(46.34) 23.44
e
30.11
d
30.45
d
33.02
b
26.93 17.10 17.43 10.37
S Em 0.78 0.47 0.32 0.42 1.44 0.10 0.38 1.22
CD 2.36 1.44 0.98 1.30 4.34 0.31 1.14 3.58
DAS: Days after Sowing
Values in parenthesis are arcsin transformed values
Figures in the same column with similar alphabets do not differ significantly at P=0.05 by DM
The correlation between the number of larvae per plant and per cent leaf damage was positive and
significant (r = 0.99) seed cotton yield at 90 days after sowing plot was ranged between 26.12 to 32.14
q/ha with yield reduction over control varied from 5.16 to 18.73 per cent. The correlation between the
number of larvae per plant and seed cotton yield was negative and significant (r = - 0.96), where as
correlation between the larval population per plant and reduction in cotton yield over control was positive
and significant (r = 0.96). From the regression equation yield reduction per larva (b) was 0.42 and gain
threshold was 1.11, hence EIL was 2.64 larvae/plant at 90 DAS.
Similarly larvae released at 120 days after sowing, per cent leaf damage was varied from zero in
complete protection plant to 38.44 in 35 larvae released plot. There was no significant difference between
the treatments which received 5, 10 and 15 larvae per plant and these treatments significantly differed
from rest of the treatments. Seed cotton yield was ranged between 26.58 to 31.33 q/ha with yield
reduction over control varied from 6.67 to 15.16 per cent. There was no significant difference in the seed
cotton yield of different treatments except complete protection.
The correlation between the number of larvae per plant and the percent leaf damage was positive and
significant (r = 0.98) where as correlation between number of larvae and seed cotton yield was negative
and significant (r = - 0.95). Correlation between the number of larvae released and percent reduction of
yield over control was positive and significant (r = 0.93). From the regression equation yield reduction
per larva was 0.32 and gain threshold was 1.11, hence EIL was 3.47 larvae/plant at 120 days after
sowing.
Percent leaf damage ranged from zero in complete protection plot to 36.33 in the treatment which
received 35 larvae per plant at 135 DAS plots. There was no significant yield reduction among different
treatments and it ranged between 28.12 to 31.88 q/ha with yield reduction over control that varied from
5.23 to 12.00 per cent. Correlation between the number of larvae released and per cent leaf damage and
Determination of Economic Injury Level for Defoliator Spodoptera litura (Fab.) on Bt Cotton 219
percent yield reduction over control was positive and significant (r = 0.95), where as correlation between
number of larvae and seed cotton yield was negative and significant (r = -0.85). Regression equation
indicated that yield reduction per larvae (b) was 0.26, gain threshold was 1.11 and hence EIL was 4.27
larvae per plant at 135 DAS.
During 2009-10 season, EIL was worked out for different days after sowing viz, 90,105,120 and 135
days after sowing.
Per cent leaf damage was ranged between zero to 10.11 at 90 DAS with seed cotton yield varied from
23.44 to 32.08 q/ha. The correlation between the number of larvae, per cent leaf damage and percent
yield reduction over control was positive and significant (r = 0.93 and r = 0.97), where as correlation
between number of larvae and seed cotton yield was negative and significant (r = -0.97). From the
regression equation yield reduction per larvae (b) was 0.62 and gain threshold was 1.04, hence EIL was
1.68 larvae per plant at 90 DAS.
Per cent leaf damage differed significantly with each other among different treatments at 105,120 and
135 days after sowing with per cent ranging from zero to 42.44, zero to 46.55 and zero to 52.33,
respectively. There was no significant difference in the yield among various treatments which received
5,10,15,20 and 25 larvae/plant but differed significantly from the treatments which received 30 and 35
larvae per plant at 105 and 120 DAS.
There was no significant difference in the seed cotton yield among the different treatments at 135
DAS. The correlations between number of larvae and seed cotton yield were negative and significant at
105,120 and 135 with r value of - 0.88, - 0.88 and 0.98 respectively. Correlation between number of
larvae and per cent yield reduction over control was positive and significant at 90,105,120 and 135 DAS
with r value of 0.97, 0.86, 0.86 and 0.85, respectively. From the regression equation yield reduction per
larvae (b) was 0.43, 0.41 and 0.29 with gain threshold value of 1.04 at 105,120 and 135 days after sowing
respectively EIL was 2.44, 2.54 and 3.59 larvae per plant at 105,120 and 135 DAS, respectively.
Similar studies were made by Miranda et al (2007) who developed Economic injury level
of Spodoptera frugiperda in Brazilian cotton.
ACKNOWLEDGEMENT
This study is an output of the Technology Mission on Cotton MMI (2007-2012) funded by the Ministry
of Agriculture with technical support from CICR, Nagpur.
REFERENCES
[1] Miranda, J.E, Barbosa, K.A and Nascimento, S. (2007)- Economic injury level of Spodoptera frugiperda in
Brazilian cotton crops. www.backupfly.com
[2] Ogunlana, M.O. and Pedigo, L. P. (1974)- Economic injury levels of potato leaf hopper on soybeans in Iowa. J. Econ. Ent.
67: 29- 32.
[3] Pedigo, L. P. (1991)- Entomology and Pest Management, Macmillan Publishing Company, New York. pp. 107 119.
[4] Pierce, W. D. (1934)- At what point does insect attack becomes damage. Entomol. News. 45: 1-4.
[5] Stern, V. NiL, R. F. Smith, R. Vanden Bosch, and KS. Hagen. (1959)- The integrated control concept, Htlgarda 29: 81-101.
[6] Stone, J.D. and Pedigo, L. P. (1972)- Development of economic injury level of the green clover worm on soybean in Iowa.
J. Econ. Ent. 65: 197-201.
Development of Metapopulation Approach

for Landscape-level Lygus hesperus
Management in Texas
M.N. Parajulee, R.B. Shrestha, W.O. Mcspadden and S.C. Carroll
Texas A&M System AgriLife Research, 1102 East FM 1294, Lubbock, Texas 79403, USA
AbstractInsect source-sink dynamics are vital to ecologically intensive pest management. Maintaining sink plant
hosts, or trap crops, and destroying alternate hosts or breeding places adjacent to the field crop are effective pest
management strategies for some arthropods. However, determining whether a host acts as a source or a sink is
challenging, especially when the pest species is highly mobile and polyphagous. The western tarnished plant bug,
Lygus hesperus, is highly polyphagous, and can utilize >300 hosts. Its presence has been documented in 26 roadside
weed hosts in the Texas High Plains. Previous studies demonstrated that Lygus hesperus prefer alfalfa over cotton and
several alternate weed hosts. A four-year project involved surveying and sampling for Lygus hesperus in the
agricultural landscapes of several sub-regions of the southwestern United States, including the Texas High Plains. In
Texas, geographic information of the landscape vegetation complex was compiled from a 150-km radius in the Texas
High Plains. In one study, fifty irrigated cotton fields representing the crop diversity within this region were sampled
via sweep-net for 10 weeks. This effort also included sampling of up to six non-cotton insect habitats within a 3-km
radius of each field. Seasonal average Lygus hesperus abundance data were regressed with 27 field characteristics
(variables), including habitat-specific land cover, distance between focal cotton fields and non-cotton habitats,
longitude, latitude, elevation, habitat heterogeneity index, and several environmental/ecological variables. Significant
variables were selected using a stepwise regression at 15% probability rate. A 10-parameter linear model explained
93% of the variation in the data. Major parameters contributing significantly to variation in Lygus hesperus
abundance in cotton were corn and sunflower acreages, focal cotton field distances from several non-cotton hosts, and
habitat heterogeneity index. In addition, field marking and capture (FMC) studies were conducted using protein
markers and enzyme-linked immunosorbent assays to characterize Lygus hesperus intercrop movement behavior. The
FMC approach can be used to study the effects of various crop management practices on L. hesperus intercrop
movement and can potentially be applied to other pests and cropping systems.
INTRODUCTION
Cotton [Gossypium hirsutum (L.)] is grown in 17 states in the U.S. with coverage exceeding 9 million
harvested acres. U.S. cotton farmers harvest ~16.7 million bales of cotton annually, valued at $8 billion
(@$1$ per lb) (NASS, 2010 to 2006, last five year average data). Texas is the largest cotton-producing
state in the U.S. In Texas, on an average 5.4 million acres of cotton are planted and 7.3 million bales of
cotton are produced (valued at $3.5 billion) in 2010 (Williams, 2011). In fact, approximately 50% of U.S.
cotton is produced in Texas, while 65% of Texas cotton is produced in the Texas High Plains. Thus,
cotton produced in the Texas High Plains accounts for 32% of total U.S. cotton production.
Texas High Plains cotton production was a low-insecticide use system until boll weevil populations
arrived in early 1990s. A significant shift in arthropod pest management approach was expected after
eradication programs eliminated the boll weevil from the Texas High Plains. Increased adoption of
transgenic cotton cultivars conferring tolerance to lepidopteran pests reduced the insecticide load in
cotton insect management. Despite this reduction, increased public concerns for environmental safety and
questions regarding the sustainability of current crop protection practices have placed a premium on the
development of integrated pest management (IPM) approaches.
The IPM approaches have been widely used to manage cotton pests. These approaches include
biological, cultural, chemical, and plant resistance methods (Frisbie et al. 1989). Integration of these
approaches has been hindered due to a lack of understanding of the biology and ecology of the target
pests, often resulting in the unilateral reliance on insecticides. The ecological effects of widespread
insecticide use have often fueled other pest problems, environmental pollution, and affected non-target
36
Development of Metapopulation Approach for Landscape-level Lygus hesperus Management in Texas 221
organisms. Recognition of the ecological changes and problems associated with reliance on insecticides
fostered the development of IPM concepts which are now well-established in all cotton-production
regions of the U.S. (Luttrell 1994).
The western tarnished plant bug, Lygus hesperus (Knight), is highly polyphagous, and can survive on
a broad range of hosts (Day 1996). Its presence has been reported in 26 unique roadside weed hosts in the
Texas High Plains (Parajulee et al. 2003). Previous studies have demonstrated that Lygus hesperus prefer
alfalfa over cotton and several weed hosts (Sevacherian and Stern 1974, Barman et al. 2010). These data
suggest that the insect source-sink dynamics could be a valuable component of L. hesperus management
strategy.
Maintaining sink plant hosts, or trap crops, and destroying source plant hosts (alternate hosts or
breeding sites) adjacent to the field crop are effective strategies in managing many pest populations.
Determining whether a particular host acts as a source or a sink is challenging, especially when the pest
species is highly mobile and polyphagous. Lygus hesperus sub-populations in the crop field are
continuously interacting with sub-population in nearby host habitats through intermigration. These strong
interactions represent an important opportunity for exploitation in an integrated pest management system,
particularly, since even if all Lygus hesperus are removed from a specific field, re-colonization pressure
due to these interactions will continue. In fact, landscape-level management of metapopulation dynamics
can be a sustainable, economical, and environmentally conscious approach of pest management.
Previous studies have indicated that Lygus hesperus move from alfalfa and other weed hosts into
cotton (Sevacherian and Stern 1975, Fleischer et al. 1988). Lygus hesperus management decisions in
cotton might be greatly affected by advancing the understanding of host availability and Lygus hesperus
source-sink dynamics. For example, Lygus hesperus population dispersal from alfalfa to adjacent cotton
might be increased by government-enforced mowing of rural roadside weed hosts such as volunteer
alfalfa. Researchers in California demonstrated that strip-cutting commercial alfalfa mitigates Lygus
hesperus dispersal to cotton (Mueller et al. 2005). Similarly, an area-wide Lygus hesperus management
project in Mississippi, funded by U.S. Department of Agriculture, demonstrated that roadside weed
management is an effective means of minimizing tarnished plant bugs, Lygus lineolaris (Palisot de
Beauvois), in adjacent cotton (Snodgrass et al. 2000). L. hesperus intercrop movement behavior has not
been fully characterized in cotton-alfalfa systems in the Texas High Plains. Doing so could improve L.
hesperus management strategies in the region and the utility of such strategy could be expanded to other
cotton producing regions of the United States.
Development of a sustainable and environmentally conscious Lygus hesperus management approach
requires a deeper understanding of metapopulation dynamics. Information on the Lygus hesperus
metapopulation dynamics is lacking in current scientific literature. Therefore, the goals of this research
are to 1) identify the Lygus hesperus sub-populations in Texas High Plains via habitat survey, 2)
characterize the metapopulation dynamics through landscape-level studies, and 3) validate Lygus
hesperus sub-population interactions (intermigration) in a field marking and intercrop movement study.
Lygus hesperus Habitat Survey
A four-year (2002-2005) survey of cotton and non-cotton hosts was conducted to examine the role of
non-cotton hosts in supporting Lygus hesperus populations in cotton in the Texas High Plains. An area
wide survey of prevalent non-cotton hosts was conducted in April (pre-season survey) and in late
July/early August (to coincide with peak cotton blooming/fruiting) in each of the twenty-five counties
comprising the Plains Cotton Growers, Inc. (PCG) service area in the Texas High Plains. At four
locations in each county, two 100-sweep samples were collected per host habitat plant species using
standard 15 sweep-nets, resulting in 800 sweeps per habitat per county. In each of the twenty-five PCG
service area counties, cotton was sampled in late July/early August at a rate of approximately 500 sweeps
per county.
Landscape-Level Study of Lygus hesperus
A two year (2008-2009) survey of cotton and non-cotton hosts was conducted to examine the effects of
host habitat spatial distribution and cotton surrounding landscape composition heterogeneity on Lygus
hesperus abundance in Texas High Plains cotton. During each years cotton growing season, fifty
irrigated cotton fields were selected from a six-county (~5,400 mile
2
) (2008) or seven-county (~6,300
mile
2
) (2009) area near Lubbock, Texas. These selected focal field and their surrounding habitats were
used for a late-season cartographic study and classification of agricultural fields and non-cotton host
habitats within a 3-km radius and weekly sweep-net sampling. Of the fifty selected cotton focal fields,
ten were chosen for more detailed sampling work, to consist of weekly sampling of six non-cotton host
habitat patches within a 3-km buffer zone. Sweep-net samples were placed in cold storage and later
processed to determine insect abundances.
Late-season host habitat classification and ground-truthing of the agricultural landscape surrounding
cotton focal fields to a buffer radius of 3 km were performed with the aid of GPS technology and in
conjunction with manual cartographic techniques. The accuracy of the resulting habitat maps were
subsequently validated through meticulous matching and association with high-resolution USDA-FSA-
AFPO NAIP digitally orthorectified aerial imagery, as well as USDA-NASS cropland data layers. A
high-throughput geographical information system (GIS), ESRI ArcGIS Desktop, was used for data
storage and processing.
Lygus hesperus Intercrop Movement Study
A two-year (2008-2009) experiment was conducted at the Texas AgriLife Research farm located near
Lubbock, Texas to assess bidirectional Lygus hesperus intercrop movement in a cotton-alfalfa system.
Lygus hesperus intercrop movement was determined by field-marking Lygus hesperus insects in alfalfa
and adjacent cotton using two different protein markers, capturing the insects via keep it simple KIS
vacuum sampling, and then detecting protein markers using indirect ELISA. Field-marking and Lygus
hesperus sampling in alfalfa and cotton was initiated at the 7-8 true leaf cotton stage. Weekly spray
applications of 10% egg white (EW) marker solution (185 L/ha) in alfalfa and 10% non-fat dairy milk
(NFDM) marker solution (185 L/ha) in cotton were made from cotton squaring to cotton boll maturation.
KIS sampling was conducted following a 24-hour post-spray foraging period. Indirect ELISA was used
to detect Lygus hesperus protein marker acquisition. Based on indirect ELISA results, Lygus hesperus
samples were classified into four categories: residents, immigrants, visitors, and roamers. Lygus hesperus
testing positive for a protein marker applied in the opposing host were categorized as immigrants. Lygus
hesperus testing positive only for the protein marker applied to the capture source host were categorized
as residents. Lygus hesperus testing negative for both protein markers were recorded as visitors. Lygus
hesperus testing positive for both protein markers, evidence of protein acquisition in both hosts, were
recorded as roamers.
External immigration (visitors, for example) was eliminated from movement calculations due to
irrelevance. For analytical purposes, 24-hour unidirectional and bidirectional movement were combined
into a final Lygus hesperus intercrop movement indicator, Total Lygus hesperus Influx, for each host.
Thus, for each host, immigrants + roamers = Total Lygus hesperus Influx. Data were analyzed by
ANOVA using PROC MIXED, SAS 9.2. Means were separated using LSMEANS with 0.1 alpha levels.
The relationship between Lygus hesperus abundance in cotton and the number of immigrants from alfalfa
was evaluated via correlation and regression analyses of two-year data. The relationship between alfalfa
Lygus hesperus immigrants and roamers was also determined via correlation and regression analyses.
Non-cotton Host Habitats of Lygus hesperus
Throughout surveying, Lygus hesperus bugs were collected from 23 host plants (Table 1). Among non-
cotton host plants, wild mustards (flixweed, tumble mustard, black mustard, and London rocket)
supported the highest number of Lygus hesperus adults and nymphs, but these hosts senesced well before
cotton was available as a suitable host for Lygus hesperus (Table 1). In most of the High Plains region,
London rocket was available as a Lygus hesperus host as early as in late January, whereas no other
apparent host plants were available for Lygus hesperus until late February. Therefore, it appears that
London rocket is responsible for supporting early-emerging Lygus hesperus (emerging from
overwintering quarters) in the northern region of the High Plains.
TABLE 1: NUMBER OF 100-SWEEP SAMPLES AND SEASONAL AVERAGE ABUNDANCE (NUMBER/100 SWEEPS) OF LYGUS HESPERUS BUGS IN COTTON AND 22 NON-COTTON HOSTS
IN THE TEXAS HIGH PLAINS (ALL 25 COUNTIES AVERAGED), 2003.
Host Common Name Scientific Name Number of Samples Adult Nymph Total
Flixweed Descurainia sophia 144 170.2 50.6 220.8
Tumble mustard Sisymbrium altissimum 13 105.1 51.6 156.7
Black mustard Brassica nigra 15 102.2 20.6 122.8
London rocket Sisymbrium irio 41 96.2 24.2 120.4
Yellow sweet clover Melilotus officinalis 18 79.4 12.4 91.8
Alfalfa Medicago sativa 222 45.8 10.3 56.1
Curly dock Rumex crispus 3 47 1.7 48.7
Russian thistle Salsola iberica 71 39.6 3.9 43.5
Pigweed Amaranthus spp. 76 27.5 7.1 34.6
Redstem filaree Erodium cicutarium 5 14 4.6 18.6
Prairie sunflower Helianthus petiolaris 4 10 1.8 11.8
Scarlet gaura Gaura coccinea 4 9.5 0.5 10
Wooly leaf bursage Ambrosia grayi 36 9.4 0.5 9.9
Texas blueweed Helianthus ciliaris 49 7.6 1.3 8.9
Kochia Kochia scoparia 30 7 0.4 7.4
Huisache daisy Amblyolepis setigera 3 7.3 0 7.3
Gumweed Grindelia squarrosa 5 3.2 1.8 5
Ragweed Ambrosia artemisiifolia 26 3.5 0 3.5
Silverleaf nightshade Solanum elaeagnifolium 45 2.8 0.3 3.1
Blue mustard Chorispora tenella 21 2.8 0.1 2.9
Cotton Gossypium hirsutum 143 2.5 0.3 2.8
Wavy gaura Gaura sinuata 3 1.7 0 1.7
Wild sunflower Helianthus annuus 20 0.5 0.1 0.6
In the northern region, five (Hale County) to 12 (Lubbock County) non-cotton host plants were
observed to bridge the sequence between non-cotton host plants and cotton during the cotton squaring
stage. However, in the southern region, alfalfa and Russian thistle were the only non-cotton hosts that
bridged the host sequence with cotton during cotton fruiting season. Among the weed hosts surveyed,
samples collected from huisache daisy, ragweed, and wavy gaura did not have any Lygus hesperus
nymphs, indicating that these weed hosts may not be suitable for Lygus hesperus reproduction. Higher
numbers of Lygus hesperus nymphs (>10 nymphs per 100 sweeps) were found in flixweed, tumble
mustard, black mustard, London rocket, yellow sweet clover, and alfalfa. Identification of reproductively
suitable host habitats for Lygus hesperus would later help in determining sub-populations and estimating
the total Lygus hesperus population in Texas High Plains landscape.
Effect of Landscape Characteristics on Lygus hesperus Populations in Cotton
Twenty-five landscape variables or characteristics were calculated for each focal field surveyed in 2008
and 2009. The landscape characters analyzed were Latitude, Longitude, Corn Area (ha), Mixed Weeds
Area (ha), Non-Habitat Area (ha), Playa Area (ha), Habitat Heterogeneity (Shannons H), Alfalfa Area
(ha), Sorghum Area (ha), Alfalfa Distance (m), Cotton Distance (m), Cotton Area (ha), Conservation
Research Program grass or CRP Area (ha), Mixed Weeds Distance (m), Non-Habitat Distance (m),
Sunflower Distance (m), Urban Area (ha), Excess Area (ha), Sunflower Area (ha), Corn Distance (m),
Urban Distance (m), CRP Distance (m), Playa Distance (m), and Sorghum Distance (m). Distances were
measured between the closest boundaries of two habitat patches.
Of twenty-five landscape variables, only six (Latitude, Corn Area, Mixed Weeds Area, Non-Habitat
Area, Playa Area and Habitat Heterogeneity) were found to be significantly positively correlated with the
number of Lygus hesperus collected from cotton focal fields (Table 2, Fig. 1). Five landscape variables
(Cotton Area, CRP Area, Mixed Weeds Distance, Non-Habitat Distance, and Sunflower Patch Distance)
were significantly negatively correlated with Lygus hesperus abundance in focal field cotton.
TABLE 2: RELATIONSHIPS OF AVERAGE NUMBER OF LYGUS HESPERUS IN COTTON FIELD (PER 1000 SWEEPS) WITH VARIOUS HABITAT ABUNDANCE AND PROXIMITY VARIABLES:
THE PEARSON CORRELATION ANALYSIS.
Variables Correlation Significance
Latitude + s
Corn Area (ha) + s
Mixed Weed Area (ha) + s
Non-habitat Area (ha) + s
Playa Area (ha) + s
Habitat heterogeneity (Shannons H) + s
Alfalfa Area (ha) + ns
Longitude + ns
Sorghum Area (ha) + ns
Alfalfa Distance (m) + ns
Cotton Distance (m) + ns
Cotton Area (ha) - s
CRP Area (ha) - s
Mixed Weed Distance (m) - s
Non-habitat Distance (m) - s
Sunflower Distance (m) - s
Urban Area (ha) - ns
Excess Area (ha) - ns
Sunflower Area (ha) - ns
Corn Distance (m) - ns
Urban D Distance (m) - ns
CRP Distance (m) - ns
Playa Distance (m) - ns
Sorghum Distance (m) - ns
At alpha=0.1, s=significant, ns=non-significant correlation.

Fig. 1: Color-coded 2009 THP Corn Hectares (Per Focal Field Buffer Zone) Versus Seasonal Average Lygus Abundances Per Cotton Focal Field
(Indicated by Red Columns). 2009 Corn Crop Coverage in the THP is Shown in the Background.
The season-long cotton survey showed that the Lygus hesperus metapopulation is divided amongst
many habitats and geographically specific sub-populations. Cotton growers are primarily concerned with
managing Lygus hesperus sub-populations present in their cotton fields. Understanding metapopulation
dynamics among these sub-populations is essential for successful management of Lygus hesperus in
cotton. Promoting those landscape characteristics showing negative relationships with Lygus hesperus
sub-populations in cotton and minimizing those showing positive relationships with Lygus hesperus in
cotton is necessary for landscape-level Lygus hesperus metapopulation dynamics management.
A stepwise regression analysis of all twenty-five landscape variables predicting Lygus hesperus
abundance in focal field cotton showed that only nine variables were useful in a predictive model (Table
3). The multivariate regression model developed based on two years of focal field survey data was
significant, with a 93% model R
2
(Fig. 2). Model validation and optimization is yet to be performed, but
landscape metapopulation management based upon a strong predictive model is necessary to map
potential risks for Lygus hesperus infestation and outbreak. Such a risk map will help in management
decision-making and in directing the overall landscape-level pest management effort.
TABLE 3: MULTIVARIATE LINEAR MODEL VARIABLES AND PARAMETERS PREDICTING AVERAGE LYGUS HESPERUS DENSITY IN FOCAL FIELD COTTON BASED ON VARIOUS LANDSCAPE PARAMETERS.
Variables Parameter F Value Pr > F Partial R
2
Model R
2

Intercept 162.2713 10.94 0.01 - -
Corn area 0.0105 48.43 <.0001 0.36 0.36
Sunflower area 0.1189 47.81 <.0001 0.15 0.52
Playa distance -0.0126 30.70 0.00 0.08 0.60
Mixed weed distance 0.0134 32.23 0.00 0.07 0.66
Corn distance 0.0070 33.97 0.00 0.10 0.77
CRP distance -0.0101 19.48 0.00 0.05 0.82
Urban distance 0.0043 11.42 0.01 0.04 0.86
Longitude -0.0002 10.44 0.01 0.04 0.90
Habitat heterogeneity -2.2409 5.12 0.05 0.03 0.93

Fig. 2: Model Prediction of Average Lygus Abundance in Focal Fields
Fig. 3: Weekly Average Lygus Abundance in Cotton and Alfalfa and Net Lygus Intercrop Movement Between Alfalfa and Adjacent Cotton, Lubbock, Texas,
2008 (left) and 2009 (right). Note: 1 ha = 833.3 KIS Samples, the Negative Value Suggest the Opposite Direction of Lygus Movement (i.e., from Alfalfa
Into Cotton). The Similar Alphabet on the Top or Bottom of the bar Among the Sampling Weeks Within a Specific Host and Year are Not Significantly
Different at Alpha=0.1 When Means Were Separated by LSMEAN.
A study investigating physical tracking of Lygus hesperus intermigration dynamics between or
among major habitats was conducted to validate this model and further characterize Lygus hesperus
metapopulation dynamics.
Intercrop Movement Dynamics of Lygus hesperus
Influx into Cotton
Analysis of variance of Lygus hesperus influx into cotton (year and host combined) revealed significant
differences in the pattern of Lygus hesperus influx into cotton among the sampling weeks (df = 6, 24; F =
5.2; P = <0.0015). There were significant interactions between Lygus hesperus intercrop movement with
week and year (df = 6, 24; F = 3.74; P = <0.0091). Lygus hesperus influx to a cotton field from nearby
alfalfa was very low when cotton was in its vegetative growth stage (0-40 DAP, days after planting, prior
to July). During the first five weeks following cotton planting, Lygus hesperus was not detected in cotton.
Once cotton began squaring (40-45 DAP), Lygus hesperus began moving into cotton from alfalfa
therefore obviously alfalfa was a source of Lygus hesperus to nearby cotton during the cotton squaring
stage.
In 2008, Lygus hesperus influx into cotton was highest at 83 DAP (fifth sampling week), followed by
97, 62, 76, 47, 69, and 104 DAP. In 2009, Lygus hesperus influx into cotton was highest at 56, 82, and 90
DAP (fifth sampling week), followed by 107, 65, 42, and 56 DAP. Peak Lygus hesperus influx into
cotton in both years occurred during the second week of August (during the fifth sampling week), when
cotton was in full bloom. Lygus hesperus influx was low in all weeks prior to this peak (during squaring).
The first peak of Lygus hesperus influx in cotton in 2009 occurred at 56 DAP or during the third
sampling week. It is unanticipated to have detected peak influx during squaring, as this is not considered
to be the most favorable cotton stage with respect to Lygus hesperus. High influx during squaring in 2009
might be accounted for by temporarily reduced alfalfa quality due to poor irrigation timing as a result of
an irrigation system backlog on the research farm and concurrently high temperatures and low relative
humidity.
Influx into Alfalfa
In order to understand the effect of cotton Lygus hesperus influx on a Lygus hesperus population in
cotton, and to understand the potential for cotton injury due to Lygus hesperus, Lygus hesperus retention
in cotton, rather than influx, is more important. For the purposes of this study, Lygus hesperus retention
can be calculated by subtracting outflow from inflow. Inflow can be estimated via sampling, but
estimating outflow is difficult. Assuming the cotton-alfalfa system as closed, Lygus hesperus outflow
from cotton can be estimated by sampling alfalfa and determining the quantity of influx into adjacent
alfalfa. In this study, Lygus hesperus influx in cotton and alfalfa was quantified for each sampling week.
Analysis of variance of Lygus hesperus influx in alfalfa (both years and both hosts combined) revealed
significant differences in the patterns of Lygus hesperus intercrop movement among the sampling weeks
(df = 6, 24; F = 10.65; P = <0.0001). Week and year interacted significantly (df = 6, 24; F = 10.78; P =
<0.0001) in terms of Lygus hesperus intercrop movement. In 2008, Lygus hesperus influx into alfalfa
from nearby cotton was near-zero when cotton was in its vegetative growth stage (0-40 DAP, prior to
July). Lygus hesperus was not detected in cotton during the first five weeks following cotton planting.
Once cotton began squaring (40-45 DAP), Lygus hesperus began moving between alfalfa and cotton.
Net Movement
Net Lygus hesperus intercrop movement between cotton and alfalfa was calculated by subtracting cotton
Total Lygus hesperus Influx (EW-marked Lygus hesperus captured in cotton) from cotton Total Lygus
hesperus Out flux (NFDM-marked Lygus hesperus captured in alfalfa). Positive net movement values
indicate net Lygus hesperus gains in cotton. Likewise, negative net movement values indicate net Lygus
hesperus losses. Lygus hesperus net movement data have the potential to indicate the timing of host
source-sink dynamics information which may be of value in making pest management decisions.
Lygus hesperus net movement into cotton varied significantly (df = 1, 2; F = 230.41; P = 0.0043)
between 2008 and 2009. Year and phenological stage affected Lygus hesperus net movement
significantly (df = 2, 32; F = 9.57; P = 0.0006). In both 2008 and 2009, Lygus hesperus net movement
was negative during cotton squaring, indicating net outflow from cotton. Net outflow peaked during the
second week of July, at 49-62 DAP in 2008 and 2009 (Fig. 3). During this period, few Lygus hesperus
(<1 Lygus hesperus /12 m
2
) were retained in cotton, and most Lygus hesperus visiting cotton moved back
to alfalfa. In 2008, as cotton grew older, and cotton squares continued to grow and blooming began,
Lygus hesperus net movement, with respect to cotton, gradually increased from negative toward zero, and
became positive as cotton approached full bloom (76 DAP). Thereafter, Lygus hesperus net movement
remained positive in cotton, indicating an increased cotton capability to retain more Lygus hesperus
having moved from alfalfa. In 2008, average Lygus hesperus net movement into cotton was significantly
higher (df = 2, 16; F = 3.64; P = 0.05) during cotton blooming (821.9 Lygus hesperus per ha or 0.99
Lygus hesperus/12 m
2
influx) and boll maturation (358.2 Lygus hesperus per ha or 0.43 Lygus
hesperus/12 m
2
influx) than during squaring (1,458 Lygus hesperus per ha or 1.75 Lygus hesperus/12 m
2

out flux). The Lygus hesperus net movement pattern in 2009 differed from that in 2008.
In 2009, Lygus hesperus net movement in cotton never became positive during the cotton growing
season. This indicates that alfalfa remained more attractive than cotton throughout the cotton growing
season. Unexpectedly, Lygus hesperus density in cotton increased continually, and cotton was able to
retain some Lygus hesperus migrants from alfalfa, even when Lygus hesperus net movement was
negative in cotton. It is somewhat puzzling to have observed Lygus hesperus net movement favoring
alfalfa while simultaneously observing increases in EW-marked Lygus hesperus retention and population
in cotton. This phenomenon indicates that calculation of Lygus hesperus net movement did not account
for actual Lygus hesperus population changes due to intercrop movement between cotton and alfalfa.
Although differences in crop structure, combined with the chosen sampling method, may have led to
overestimation of Lygus hesperus densities in alfalfa, it is likely that naturally higher Lygus hesperus
densities in alfalfa and naturally high Lygus hesperus intercrop movement between cotton and alfalfa
may have contributed to said overestimation. Many Lygus hesperus from alfalfa may have visited cotton,
but most returned to alfalfa. Only a few actually settled in cotton. Each time a large number of Lygus
hesperus move from alfalfa to cotton, a few Lygus hesperus may remain and settle, which explains the
steady, gradual Lygus hesperus population increase in cotton. However, since most returned to alfalfa,
Lygus hesperus net movement calculations indicated high Lygus hesperus influx into alfalfa.
Because Lygus hesperus is highly mobile, and moves quite freely back and forth between hosts in the
open agroecosystem, currently available knowledge and technology are insufficient to accurately
quantify net movement mathematically, as opposed to in a hypothetical closed ecosystem with only
unidirectional movement.
CONCLUSION AND FUTURE RESEARCH
Lygus hesperus
Sub-populations in agricultural field crops and host habitats continuously interact, and these interactions
represent an excellent opportunity for exploitation in Lygus hesperus metapopulation management. This
is particularly true, given that even if all Lygus hesperus are removed from a specific crop field, Lygus
hesperus source populations residing in nearby habitats will continue to exert considerable re-
colonization pressure and pose an infestation risk. Managing pests at the landscape-level via intelligent
exploitation of metapopulation dynamics may prove to be sustainable, economical, and environmentally
conscious tool for use in conjunction with other methods in an integrated pest management system. In
fact, development of elegant, environmentally conscious pest management approaches requires a deeper
understanding of metapopulation dynamics. Further detailed investigations of Lygus hesperus
metapopulation dynamics in the Texas High Plains is necessary for continued development of landscape-
level, sustainable, integrated approaches to L. hesperus management.
ACKNOWLEDGMENT
This project was partially funded by Cotton Incorporated Core Program, USDA CSREES RAMP,
International Cotton Research Center, and Plains Cotton Growers, Inc.
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Survival of Pink Bollworm, Pectinophora gossypiella

(Saunders) in Bt and Non Bt Cotton In Normal
and Late Sowing With A Special Emphasize
to Avoid Population Pressure
S. Mohan and S. Nandini
Department of Agricultural Entomology, Centre for plant protection studies,
Tamil Nadu Agricultural University, Coimbatore3
INTRODUCTION
Cotton Gossypium hirsutum L., is one of the principal commercial crops playing a key role in economic,
social, and political affairs of the country. India is an important grower of cotton on a global scale. In
India, cotton is cultivated in an area of 110 lakh ha with an average productivity of 503 kg/ha.
(Anonymous, 2011).
The insect pests spectrum of cotton is quite complex and as many as 1326 species of insect pests
have been reported on this crop throughout the world. Among the bollworms, the pink bollworm
assumed major pest status in recent past (Ghosh, 2001). World over, Pink bollworm Pectinophora
gossypiella (Saunders) has become economically the most destructive pest of cotton and has known to
cause 2.8 to 61.9 per cent loss in seed cotton yield, 2.1 to 47.10 per cent loss in oil content and 10.70 to
59.20 per cent loss in normal opening of bolls (Patil, 2003).
In India Bt cotton has gained considerable importance in recent years. The area under Bt cotton
reached 7.6 million hectares in 2008-09 constituting nearly 81% of the total cotton area in India. As a
result, the production also reached 4.9 million tons (Anonymous, 2009). Bt cotton (Bollgard) offers
high level of resistance against cotton bollworm complex ie., Helicoverpa armigera (Hubner), Earias
vittella (Fabricius) and Pectinophora gossypiella (Saunders) both under laboratory as well as field
conditions (Kranthi and Kranthi., 2004).
However recent report of ineffectiveness of Bollgard (Cry1Ac) to pink boll worm in certain parts of
Gujarat (The Hindu, Dated 6.3.2010) made it necessary to investigate the pink bollworm incidence in Bt
cotton. Hence, studies were carried out under the All India Coordinated Cotton Improvement Project
(ICAR) regarding the incidence of pink bollworm in Bt and Non Bt cotton at the Cotton farm of Tamil
Nadu Agricultural University during winter season, 2010-11.
The Bt cotton hybrid, Bt Bunny (Cry1Ac) was sown in one acre along with a non-Bt cotton, MCU-5
in another one acre. Sowing was done on 13
th
August 2010 with a spacing of 90 cm x 60 cm by following
all standard agronomical practices.
The observation on rosette flowers due to pink bollworm infestation was made, starting from 70 DAS
and continued upto 130 DAS at weekly intervals, on twenty five plants selected randomly and tagged in
both Bt and Non Bt cotton fields.
Collections of twenty five green bolls were made from fields where Bt and Non Bt were sown during
125 and 140 DAS. Collected bolls were taken to the laboratory and each green boll was cut opened along
with ridges of the locules with help of sharp cutter carefully and the damage was estimated by counting
the number of live PBW larvae in each boll. At the time of cotton picking, 50 fully opened bolls were
sampled randomly from each field and the total number locules and damaged locules due to PBW larva
were counted.
37
Pink Bollworm Damage in Bt Cotton and Non Cotton
The first incidence of rosette flowers was observed when the crop was 100 days old in Non Bt (MCU 5).
The rosette flowers observed in MCU 5 ranged from 13 in numbers/100 flowers. First to third instar
larvae were found in the rosette flowers. No rosette flower was observed in Bt Bunny.
TABLE 1: OBSERVATION ON ROSETTE FLOWERPINK BOLLWORM DAMAGE

Date of Collection
Non Bt field (MCU 5)
Total Flowers Observed No. of Rosette flowers No. of Larvae Collected Instar of Larvae
I II III IV
13/11/2010 100 0 0 0 0 0 0
24/11/2010 100 1 0 0 0 0 0
09/12/2010 100 2 1 0 1 0 0
17/12/2010 100 5 3 1 1 0 0
03/01/2011 100 3 2 0 1 1 0
Green boll damage in MCU 5 ranged from 4-8%. The first incidence of pink bollworm in MCU 5
occurred on 125 DAS. The stage of the larvae observed was 2 to 4
th
instar in MCU 5. No bolls were
infested by PBW larvae in Bt Bunny.
Bt field (Bt Bunny)
Total Flowers Observed No. of Rosette Flowers No. of Larvae Collected
Instar of Larvae
I II III IV
100 0 0 0 0 0 0
100 0 0 0 0 0 0
100 0 0 0 0 0 0
100 0 0 0 0 0 0
100 0 0 0 0 0 0
TABLE 2: OBSERVATION ON GREEN BOLL DAMAGE AND LOCULE DAMAGE

Date of Collection
Non Bt Field (MCU 5)
Total Green Bolls
Observed
No. of Damaged
GB
No. of Locules
Damaged
No of Larvae
Collected
Instar of Larvae
I II III IV
13/12/2010 25 0 0 0 0 0 0 0
18/12/2010 25 0 0 0 0 0 0 0
03/01/2011 25 1 1 1 0 1 0 0
23/01/2011 25 2 1 2 0 0 1 1
In MCU 5, the open boll damage (at harvest) (135 DAS) and locule damage were observed. The open
boll damage due to pink bollworm ranged from 8-20% and locule damage ranged from 1-3%. The larval
stage observed was 3 and 4
th
instar.
Bt Field (Bt Bunny)
Total Green Bolls
Observed
No. of Damaged GB
No. of Locules
Damaged
No of Larvae Collected
Instar of Larvae
I II III IV
25 0 0 0 0 0 0 0
25 0 0 0 0 0 0 0
25 0 0 0 0 0 0 0
25 0 0 0 0 0 0 0
TABLE 3: OBSERVATION ON LOCULE DAMAGE (AT HARVEST)

Date of Collection
Non Bt field (MCU 5)
Total Open Bolls
Observed
No. of Damaged
OB
No. of Locules
Damaged
No of Larvae
Collected
Instar of Larvae
I II III IV
27/12/2010 25 2 1 0 0 0 0 0
13/01/2011 25 4 2 2 0 0 1 0
23/01/2011 25 3 2 1 0 0 0 1
6/02/2011 25 5 3 2 0 0 0 1
Bt field (Bt Bunny)
Total Open Bolls
Observed
No. of Damaged OB
No. of Locules
Damaged
No of Larvae Collected
Instar of Larvae
I II III IV
25 0 0 0 0 0 0 0
25 0 0 0 0 0 0 0
25 0 0 0 0 0 0 0
25 0 0 0 0 0 0 0
Survival of Pink Bollworm, Pectinophora gossypiella (Saunders) in Bt and Non Bt Cotton In Normal and Late Sowing 231
No pink bollworm damage was observed in Bt Bunny during this period also. These findings are in
agreement with the reports of Udikeri et al. (2003) and Surulivelu et al. (2004) who recorded
significantly very less locule damage in Bt cotton hybrids over non Bt cotton hybrid.
The present finding is important in the sense that even in Bollgard (Cry1Ac) we did not observe
any pink bollworm damage. Currently, we have Bollgard II (Cry1Ac +Cry2Ab) under cultivation in
majority of the cotton growing areas of India. As even the Bt Bunny with the single gene (Bollgard)
had no pink bollworm incidence, we assume that the Bollgard II with Cry IAc and Cry2Ab can do still
better without any resistance problem as it has double gene. However careful and continuous monitoring
of field incidence is a must for evolving better management strategy of pink bollworm in cotton.
REFERENCES
[1] Anonymous. (2011) - Annual report. All India Coordinated Cotton Improvement Project, Hisar, Haryana.
[2] Anonymous. (2009) - A Status report on Bt cotton. Asia-Pacific Consortium on Agricultural Biotechnology and National
Research Centre on Plant Biotechnology, 2009. New Delhi.
[3] Ghosh, S. K., (2001) - G. M. Crops: Rationally irresistible, Curr. Sci., 6: 655-660.
[4] Kranthi, K. R. and Kranthi, N. R., (2004) - Modelling adaptability of the cotton bollworm, Helicoverpa armigera (Hubner)
to Bt cotton in India. Curr. Sci., 87: 1096-1107.
[5] Patil, S. B., (2003) - Studies on management of cotton pink bollworm Pectionophora gossypiella (Saunders) (Lepidoptera:
Gelechiidae). Ph. D. Thesis, Univ. Agric. Sci., Dharwad (India).
[6] Surulivelu, T., Sumathi, E., Mathirajan, V. G. and Rajendran, T. P., (2004) - Temporal distribution of pink bollworm in Bt
cotton hybrids. In: Int. Symp. Strat. Sust Cotton Prodn, - A Global Vision Vol. 3. Crop Protection, 23-25 November 2004,
UAS, Dharwad, Karnataka (India), pp. 86-88.
[7] Udikeri, S. S., Patil, S. B., Nadaf, A. M. and Khadi. (2003) - Performance of Bt-cotton genotypes under unprotected
conditions. Proc. World Cotton Res. Con., - III. 9-13 March, 2003, Cape Town, South Africa: 1282-1286.
Dynamics of Biotypes b and q of Bemisia tabaci

in Cotton Fields and Their Relevance
to Insecticide Resistance
A.R. Horowitz
1
, H. Breslauer
1
, M. Rippa
1
, S. Kontsedalov
2
, M. Ghanim
2
,
P. Weintraub
1
and I. Ishaaya
2
1
Department of Entomology, Agricultural Research Organization,
Gilat Research Center, M.P. Negev, 85280
2
The Volcani Center, Bet Dagan 50250, Israel
E-mail: hrami@volcani.agri.gov.il
AbstractAn extensive survey for identifying Bemisia tabaci biotypes and monitoring insecticide resistance was
conducted from 2003 to 2010 in cotton fields from several locations in Israel. Two biotypes of B. tabaci, B and Q, were
identified; and some differences in the biotype dynamics were recorded from different areas. From 2003 to 2007 in
northern Israel, a higher proportion of the B biotype was consistently found in the early season. However, by the end of
the season a definite rise of the Q biotype was recorded, ranging from 60 to 100%, along with high resistance to the
insect growth regulator (IGR) pyriproxyfen and to the neonicotinoid insecticides. The Q biotype was predominant
throughout the season in fields located in the east-central part of Israel, with high resistance to pyriproxyfen; on the
other hand, in cotton fields located in southern Israel, the B was the widespread biotype during the entire season with
no significant resistance to pyriproxyfen.
From 2008 to date, we identified a significant shift in the biotype dynamics: the B biotype is currently
predominating in all cotton fields, reaching up to 90% or more. Concurrently, resistance to pyriproxyfen and
neonicotinoids has reduced considerably. The possible reasons for the change in the dynamics of B. tabaci biotypes are
discussed.
INTRODUCTION
The whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a key pest in many agricultural
crops including vegetables, ornamentals and field crops (Byrne and Bellows 1991, Oliveira et al. 2001,
Stansly and Naranjo 2010). It directly damages the plants by feeding on phloem sap, and excretes
honeydew on leaves and fruit. The sticky, sugary surface forms a substrate for the growth of black sooty
mold that stains the cotton and covers the leaves; thus, impairing photosynthesis. The resulting stickiness
and discoloration greatly reduce the value of agricultural crops such as ornamentals, vegetables and
cotton. In cotton, the honeydew may cause fiber stickiness that interferes with the spinning process in the
textile mills, and greatly reduces the product's value (Hequet et al. 2007). Bemisia tabaci is a vector of
several important families of plant viruses (Jones 2003, Hogenhout et al. 2008) including cotton (e.g. the
cotton leaf curl geminivirus (CLCuV).
Bemisia tabaci is known for its genetic diversity, which is expressed in a complex of biotypes
(Brown et al. 1995, Perring 2001, De Barro et al. 2005) or, as recently suggested, a complex of distinct
cryptic species (Xu et al. 2010, De Barro et al. 2011). The biotypes are largely differentiated based on
biochemical or molecular polymorphism, and differ in their characteristics such as host plant range, the
capacity to cause plant disorders, attraction by natural enemies, expression of resistance, and plant virus-
transmission capabilities (e.g. Bedford et al. 1994, Brown et al. 1995, Sanchez-Campos et al. 1999,
Perring 2001, Horowitz et al. 2005). Recent reports have suggested that the floral composition of
bacterial symbionts might be specific to certain biotypes (Gottlieb et al. 2006, Chiel et al. 2007) and
might confer upon them resistance to insecticides (Kontsedalov et al. 2008). The most widespread
biotype, B, was identified in the late 1980s (Costa and Brown 1991, Costa et al. 1993), following
extensive outbreaks of B. tabaci in the southwestern USA, and has a worldwide distribution. An
additional common biotype, Q, which probably originated in the Iberian Peninsula (Guirao et al. 1997),
has since spread globally (Horowitz et al. 2003, Boykin et al. 2007, Chu et al. 2010).
38
Dynamics of Biotypes b and q of Bemisia tabaci in Cotton Fields and Their Relevance to Insecticide Resistance 233
An extensive survey for identifying B. tabaci biotypes along with monitoring insecticide resistance
was conducted from 2003 to 2011 in cotton fields in several locations of Israel. The main objective of
this paper is to summarize the results of this survey and discuss the shift in biotype composition and
resistance status that has occurred during recent cotton seasons.
METHODS AND MATERIALS
Collections of Whiteflies
Collections of whiteflies from commercial cotton fields in Israel were conducted from 2003 to 2011
throughout six regions (Fig. 1) to characterize the distributions of the B and Q biotypes along with
resistance to the new insecticides pyriproxyfen (IGR) and neonicotinoids (e.g. acetamiprid and
thiamethoxam). In most years, each field was sampled twice per season, once in late June and another in
early September. The exception was in 2005 and 2006, where fields were sampled every month from
June to October. Whiteflies were sampled randomly from plants in each field using an aspirator.
Samplers performed a random walk diagonally through the center of each field; beginning 50-100 meters
inside the field edge and walking towards the center, until a total of 80-100 whiteflies were sampled. The
samples were kept in vials with 90% ethanol at -20C until biotypes were identified using DNA markers
(Khasdan et al. 2005). A total of 15-20 individuals per sample were used for molecular analyses. The
average proportion of the B and Q was calculated across the fields in each region. In case of high
population of whiteflies, a large amount of adults was taken for resistance monitoring bioassays.

Fig. 1: Collection Sites of Bemisia Tabaci Populations in Israel During the 2003 2011 Cotton Growing Seasons. The Locations: 1- Western Galilee;
2 Carmel Coast; 3 Central Israel; 4 Ayalon Valley; 5 South District; 6 Western Negev.

Egypt
D
e
a
d

S
e
a
M
e
d
i
t
e
r
r
a
n
e
a
n

S
e
a
Jordan
Lebanon
Syria
25 km 25 km
Elat
Beer-Sheva
Gaza
Tel-Aviv
Jerusalem
Jericho
Haifa
Nazareth
Egypt
D
e
a
d

S
e
a
M
e
d
i
t
e
r
r
a
n
e
a
n

S
e
a
Jordan
Lebanon
Syria
25 km 25 km
Elat
Beer-Sheva
Gaza
Tel-Aviv
Jerusalem
Jericho
Haifa
Nazareth

Fig. 2: Proportion of B and Q Biotypes of Bemisia Tabaci Collected in the Ayalon Valley During the 2003 2011 Cotton Growing Seasons

Fig. 3: Proportion of B and Q Biotypes of Bemisia Tabaci Collected in the Carmel Coast During the 2003 2011 Cotton Growing Seasons

Fig. 4: Log Concentration-Response Curves (on a Probit Scale) for the Effect of Pyriproxyfen on Bemisia Tabaci Populations Collected
from 2003 to 2010 from Cotton Fields in the Ayalon Valley, Israel
2003 2004 2005 2006 2007 2008 2009 2010 2011
Year
0
20
40
60
80
100
P
r
o
p
o
r
t
i
o
n

B

b
i
o
t
y
p
e

(
%
)
Early Season Late Season
Ayalon Valley
2003 2004 2005 2006 2007 2008 2009 2010 2011
Year
0
20
40
60
80
100
P
r
o
p
o
r
t
i
o
n

B

b
i
o
t
y
p
e

(
%
)
Early Season Late Season
Carmel Coast
0.0001 0.001 0.01 0.1 1 10 100 1000
Concentration, ppm
1
5
10
30
50
70
90
95
99
E
g
g

m
o
r
t
a
l
i
t
y
,

%
S
2003
2006
2007
2010
Ayalon Valley

Fig. 5: Log Concentration-Response Curves (on a Probit Scale) for the Effect of Acetamiprid on Bemisia Tabaci Populations Collected
from 2003 to 2010 from Cotton Fields in the Ayalon Valley, Israel
Resistance bioassays
Insecticides
The following insecticides were used for bioassay procedures. Pyriproxyfen, commercial name Tiger,
was formulated as 10% EC (emulsion concentrate) (Sumitomo Co. Tokyo, Japan). Acetamiprid,
commercial name Mospilan, was formulated as 200 g [AI]/ Kg SP (soluble powder9, (Nippon Soda Co.,
Tokyo, Japan): both were obtained from Agan Chemical Company, Ashdod, Israel. Thiamethoxam,
commercial name Actara, was formulated as 25 250 g [AI]/ Kg WG (water-dispersible granules),
(Syngenta AG, Basel, Switzerland, obtained from C.T.S., Hod Hasharon, Israel).
Bioassay Procedure
Cotton seedlings (15-20 cm tall with two true leaves) were dipped in aqueous concentrations of
insecticide, or in deionized water (untreated control), and then allowing the plant to air-dry for 2 h.
Fifteen B. tabaci females, confined in clip-on leaf cages, were exposed to treated cotton seedlings for 48
h and kept under standard laboratory conditions of 26 2C, 60% RH, and a photoperiod of 14:10 (L:
D). For acetamiprid and thiamethoxam, female adult mortality was determined; for pyriproxyfen, females
were removed and fecundity was recorded. Egg viability (egg hatch suppression) was determined eight
days after treatment (Ishaaya and Horowitz, 1992). Each bioassay was done with at least four
concentrations, each with five replicates, and repeated three times on different days. Mortality curves
along with LC values were then determined.
Data Analysis
Probit analyses of the concentration-dependent mortality data were made using POLO-PC (LeOra
Software, 1987) after correction with Abbotts (1925) formula. Failure of 95% C.L. (confidence limits) to
overlap at a particular lethal concentration indicated a significant difference.
Figures 2 and 3 show the changes in the ratios of the biotypes B and Q from 2003 until 2011 cotton
growing seasons in two representative locations, Ayalon Valley (east-central Israel) and Carmel Coast
(northern Israel), respectively. In the Ayalon Valley, the Q biotype was the predominant until 2007, and
0.1 1 10 100 1000
Concentration, ppm
1
5
10
30
50
70
90
95
99
A
d
u
l
t

M
o
r
t
a
l
i
t
y
,

%
S
2003
2005
2007
2010
Ayalon Valley
acetamiprid
approximately 20% biotype B was sampled in 2005 2007 during the early seasons. However, in 2009
2011 there was a drastic shift where almost no Q biotypes were sampled in the cotton fields (Fig. 2). In
the Carmel Coast, more B biotypes were sampled than in the Ayalon Valley 2003 2008 seasons (no
cotton was grown in this area in 2009). In 2003 2006, around 50% of biotype B was sampled in the
early seasons but in the late seasons, the Q was more widespread. However during 2010 2011 the B
was the predominant biotype, as was observed in the Ayalon Valley (Fig. 3). During recent years, similar
pattern was found in other open field-crops; however, in protected crops where chemical regime is
common, the Q was the predominant biotype (Kontsedalov et al. in press).
In addition to the biotyping of whiteflies, in some seasons, resistance to insecticides was monitored
especially in the two aforementioned locations, Ayalon Valley and Carmel Coast. In Fig. 4 the dynamic
of resistance to pyriproxyfen in the Ayalon Valley is demonstrated in some of the late seasons from 2003
2010 as compared with the susceptible standard strain. In the 2003, 2006 and 2007 seasons, high
resistance was found ranging from 300 to 1000-fold. As resistance to pyriproxyfen has been detected
since the 1990s, very high resistance in B. tabaci was found in both locations (Horowitz et al., 2002,
Horowitz et al. 2005). However, in 2010, susceptibility to pyriproxyfen was almost restored to a level of
the standard strain (Fig. 4). The resistance to acetamiprid was less decisive (about 4-6 fold); however,
high cross-resistance was detected to thiamethoxam, an insecticide that has never been applied in cotton
in this area (Horowitz et al., 2004).
Horowitz et al. (2005) have demonstrated that the B biotype was more competitive than the Q
biotype under untreated conditions; however, repeated applications of either pyriproxyfen or a
neonicotinoid would encourage the Q biotype and depress the B biotype. Crowder et al. (2010) assumed
that the main reason for the high competitiveness trait of B biotype is its ability to copulate more
effectively than the Q biotype, resulting in a faster population growth, and consequently an exclusion of
the Q biotype.
There could be some explanations for the shift in the biotype composition. Firstly, reduction in
acreage of cotton fields during recent years along with a decrease in insecticide use, especially
pyriproxyfen, for controlling the whitefly. This decrease resulted in less selection for the Q biotype along
with the fact that the B biotype without insecticide selection is more competitive than the Q. Nonetheless,
as was mentioned, in greenhouses with many chemical treatments the Q was the predominant
(Kontsedalov et al. in press). We assume that applications of the IGR pyriproxyfen are the main cause for
the establishment of the Q biotype in some cotton fields. However, other reasons could result from
encouragement of a specific biotype such as different regional climate (Mahadav et al. 2009,
Kontsedalov et al. in press) and differences in agroecology and topography among growing regions
(Castle et al. 2010). Another possibility to the high levels of the B biotype is the increase in resistance to
insecticides. In general, no resistance to pyriproxyfen was detected in the B biotype in Israel (Horowitz et
al. 2002, 2005), although the B biotype was found to be resistant to the same insecticide in the US
(Crowder et al. 2010). However, high resistance to neonicotinoids in the B biotype from Israel was
reported (Rauch and Nauen, 2003). Our results from the Ayalon Valley in 2010 did not confirm
resistance to pyriproxyfen in an Israeli-B biotype but more monitoring is required to validate such
resistance.
A practical implementation resulted from our research is the possible reuse of pyriproxyfen in areas
where the B is the predominant; however, it should be accompanied with strict biotyping determination
before and after a pyriproxyfen application to prevent any field failure.
ACKNOWLEDGMENT
The authors gratefully acknowledge the Chief Scientist of the Ministry of Agriculture and the Israeli
Cotton Board for their partial support of the research. This paper is contribution No.507/2011, from the
Agricultural Research Organization, the Volcani Center, Bet Dagan, Israel.
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Gambit of IPM for Insect Resistant

Transgenic Cotton
N.V.V.S. Durga Prasad, G.M.V. Prasad Rao and V. Chenga Reddy

Insect Pest Management, Regional Agricultural Research Station, Lam, ANGRAU, Guntur
AbstractField experiments were conducted at Regional Agricultural Research station, Lam, Guntur to study the
performance of Transgenic Bt cotton BG, BG II and non Bt cotton of Mallika hybrid under IPM, conventional control
(CC) and unprotected conditions (check) for two consecutive seasons. The incidence of whiteflies was significantly low
in IPM block of all the hybrids compared to conventional control, while the incidence of thrips, aphids and jassids were
marginally high under IPM module. The larval incidence and fruiting bodies damage of American and pink bollworms
were significantly high in IPM module compared to conventional control. The activity of natural enemies under
ecofriendly IPM in all the hybrids were more compared to conventional control and check without pesticidal
application. Though there were slight difference in yield levels the cost benefit ratio was high from IPM module (1:1.55
in BG, 1:1.73 in BG II and 1:1.35 in NBt) compared to conventional control (1:1.34 in BG, 1:1.45 in BG II and 1:1.23
in NBt) due to low cost of plant protection. The untreated check module without any protection to pest complex in Bt
cotton system recorded low yield and cost benefit ratio which may not be a viable system. Hence, Bt cotton hybrids
should be viewed as a foundation on which Integrated Pest Management (IPM) has to be built up to combat the pest
problems for sustainable transgenic Bt cotton cultivation.
INTRODUCTION
Cotton is one of the most important commercial crop of Andhra Pradesh and insect pest complex is one
of the major constraint for cotton cultivation. Over dependence and indiscriminate use of insecticides led
to the control failures of the cotton pests due to development of resistance in the major pests and
resurgence of minor pests besides deleterious effect on the bio control agents paving the way for the
development of IPM. There are number of reports on successful IPM programmes against different pests
on major crops in different states of our country. IPM helps in preservation of natural biodiversity apart
from reducing the cost of plant protection by using environmentally safer methods of pest management.
The rapid changes witnessed in the biotechnology resulted in the development of genetically modified
crops which evolved as effective alternate tools for pest management. The genetically modified
transgenic Bt cotton (BG) was commercially available in India from 2002 and stacked BG II hybrids
from 2006 which found favor with farmers with considerable increase in area of Bt cotton in the country.
Though the Bt cotton hybrids are resistant to bollworms, there are reports regarding the high incidence of
sucking pests in Bt cotton hybrids (Cui and Xia, 2000; Barwale et al., 2002) and varied trends on the
occurrence of natural enemies in Bt cotton hybrids. Transgenic Bt cotton with potential to combat certain
insect pests should become a major component in IPM. Therefore it is most important to evaluate the
impact of present IPM strategies visa a vis conventional insecticide dependent control and under
completely unprotected conditions on Transgenic BG and BGII cotton to formulate suitable IPM
module for Bt cotton towards sustainable cultivation. Hence, the present study was aimed to assess the
advantage of IPM module for insect resistant transgenic Bt cotton under different modules.
The experiment was conducted at Regional Agricultural Research station (RARS), Lam, Guntur, Andhra
Pradesh, India during kharif, 2009-10 and 2010-11 under rainfed conditions in black cotton soils. Mallika
BG, Mallika BG II and non Bt cotton hybrids
(NBt hybrid) were grown in three modules IPM, conventional control (CC) and unprotected
conditions (Check) in three replications. All the hybrids were sowm during IInd fortnight of July in both
39
the seasons at a spacing of 105 cm X 60 cm. The agronomic practices and fertilizer application were
taken up similarly as per the recommendations in all the blocks except plant protection measures.
IPM MODULE
The following IPM practices were followed
Seed treatment with imidacloprid @ 5g/kg seed
Stem application with monocrotophos @1:4 ratio at 30,45 and imidacloprid @ 1:20 ratio at 60
DAS
Intercropping with green gram
Growing jowar as border crop
Erection of bird perches
Monitoring of bollworms through pheromone traps
Yellow sticky traps for whitefly
Use of botanical insecticides, 5 % NSKE and neemoil
Need based application of insecticides based on ETL
CONVENTIONAL CONTROL BLOCKS (CC)
Insecticidal interventions were taken up for every 10 to 15 days in all Bt and non Bt hybrids under
conventional control.
Unprotected Block (Check)
Bt and Non Bt cotton hybrids were kept under completely unprotected conditions without any pest
management interventions.
The incidence of sucking pests, bollworms, fruiting body damage due to bollworms and the
occurrence of natural enemies were recorded at weekly interval from each block from 25 randomly
selected plants. Sucking pests such as aphids, leafhoppers, thrips and whiteflies were recorded from three
leaves, each one from top, middle and bottom canopies of the plant. While for the bollworms damage to
fruiting parts and natural enemies were recorded from whole plant. Cost of cultivation and yields were
recorded from different control modules and cost benefit ratios were calculated. The incidence of
different pests, damage and cost of cultivation were given as seasonal mean for both the seasons. The
data was subjected to suitable transformations and analyzed through ANOVA using MSTATC
The pooled data over the two seasons showed that the seasonal mean incidence of whitefly was
significantly low under IPM module (2.35 in BG, 3.03 in BG II and 3.43 in NBt per 3 leaves) compared
to conventional control (CC) (5.52 in BG, 4.46 in BG II and 4.82 in NBt per 3 leaves) and check (2.70 in
BG I, 4.25 in BG II and 3.70 in NBt per 3 leaves), IPM practices played a major role in suppressing
whitefly population. The population of thrips, aphids and jassids (1.34, 1.62 and 2.19/3 leaves,
respectively in BG; 1.65, 2.24 and 1.85/3 leaves, respectively in BG II; 1.33, 1.81 and 2.10/3 leaves,
respectively in NBt) were low under conventional control when compared to population levels in IPM
(2.21, 2.76 and 3.45/3 leaves, respectively in BG; 2.29, 3.36 and 2.93/3 leaves, respectively in BG II;
2.77, 2.53 and 3.62/3 leaves, respectively in NBt) and check (1.60, 9.44 and 5.91/3 leaves, respectively
in BG; 1.38, 7.24 and 5.20/3 leaves, respectively in BG II; 1.48, 7.99 and 5.48/3 leaves, respectively in
NBt), which indicates that the incidence of thrips, aphids and jassids were reduced by frequent
insecticidal interventions effectively than IPM practices (Table 1). But the incidence of whiteflies was
high in conventional control block compared to IPM and check which clearly indicates the resurgence of
whiteflies due to frequent insecticidal sprayings. In general, the population of all the sucking pests was
high in check block compared to IPM and conventional control modules except whiteflies. The lower
Gambit of IPM for Insect Resistant Transgenic Cotton 241
incidence of sucking pests under conventional control can be attributed to insecticidal interventions at
frequent intervals, but resulted in resurgence of whiteflies. While adoption of IPM practices such as seed
treatment, stem application, intercropping with green gram and spraying with 5%NSKE reduced the
population of sucking pests under IPM without any insecticidal interventions. The efficacy of
imidacloprid at 5, 7.5 and 10 g /kg seed as seed dresser was proved effective earlier by many workers in
controlling the populations of leafhoppers, aphids, thrips and whiteflies up to 30-50 days after sowing
(Gupta et al., 1998; Satpute et al., 2001; Kannan et al., 2004). Rama Rao et al. (1998) reported that stem
application with imidacloprid (200 SL) at 1:20 dilution at 20, 40 and 60 DAS was highly effective in
controlling aphids, leafhoppers and mealy bugs in cotton. Venkatesan et al. (1987) observed the lower
incidence of leafhoppers on cotton when intercropped with green gram and black gram, while Rao and
Chari (1992) reported that the cotton crop bordered by sorghum showed significantly lower aleyrodid
populations. Sohi et al. (2004) reported that the incidence of leafhoppers and whiteflies per leaf were low
in IPM fields compared to that of non IPM fields.
TABLE 1: INFLUENCE OF TRANSGENIC BT COTTON AND NON BT COTTON ON PEST COMPLEX IN DIFFERENT MODULES
Pests/damage
BG I Hybrid BG II Hybrid Non Bt Hybrid (NBt)
F test C.D.
IPM CC Check IPM CC Check IPM CC Check
Thrips/3 leaves 2.21(1.79) 1.34(1.53) 1.60(1.61) 2.29(1.81) 1.65(1.63) 1.38(1.54) 2.77(1.94) 1.33(1.53) 1.48(1.57) SIG 0.06
Aphids/3 leaves 2.76(1.94) 1.62(1.62) 9.44(3.23) 3.36(2.01) 2.24(1.80) 7.24(2.87) 2.53(1.88) 1.81(1.68) 7.99(3.00) SIG 0.10
Jassids/3 leaves 3.45(2.11) 2.19(1.79) 5.91(2.63) 2.93(1.98) 1.85(1.69) 5.20(2.49) 3.62(2.15) 2.10(1.76) 5.48(2.54) SIG 0.09
Whiteflies/3 leaves 2.35(1.83) 5.52(2.55) 2.70(1.92) 3.03(2.01) 4.46(2.34) 4.25(2.29) 3.43(2.11) 4.82(2.41) 3.70(2.17) SIG 0.11
Predators/plant 0.14(1.07) 0.08(1.04) 0.12(1.06) 0.16(1.08) 0.24(1.12 0.16(1.08) 0.16(1.08) 0.08(1.04) 0.18(1.09) SIG 0.02
Helicoverpa
larva/plant
0.03(1.01) 0.00(1.00) 0.01(1.01) 0.00(1.00) 0.00(1.00) 0.01(1.01) 0.18(1.09) 0.09(1.04) 0.24(1.12) SIG 0.013
Pink bollworm
larvae/10 bolls
0.00(1.00) 0.00(1.00) 0.00(1.00) 0.00(1.00) 0.00(1.00) 0.00(1.00) 0.08(1.04) 0.11(1.06) 0.16(1.08) SIG 0.006
Fruiting bodies
(squares) damage%
0.91[5.46] 0.00[0.00] 0.35[3.39] 0.44[3.81] 0.00[0.00] 0.10[1.80] 3.64[10.99] 1.03[5.83] 3.44[10.68] SIG 0.47
Boll damage% 0.07[1.45] 0.00[0.00] 0.05[1.32] 0.00[0.00] 0.00[0.00] 0.00[0.00] 1.11[6.04] 0.19[2.50] 1.10[6.02] SIG 0.44
Locule damage- green
bolls % (PBW)
0.00[0.00] 0.00[0.00] 0.00[0.00] 0.00[0.00] 0.00[0.00] 0.00[0.00] 0.60[4.41] 0.90[5.43] 2.05[8.23] SIG 0.49
Figures in ( ) parentheses are x+1 transformed values
Figures in [ ] parentheses are Arcsin transformed values
TABLE 2: PLANT PROTECTION AND ECONOMICS OF TRANSGENIC BT COTTON AND NON BT COTTON IN DIFFERENT MODULES
Particulars
BG I Hybrid BG II Hybrid Non Bt Hybrid (NBt) F test C.D.
IPM CC Check IPM CC Check IPM CC Check
Number of Sprays
a)Stem application 3 0 0 3 0 0 3 0 0
b)Sucking pests 2 7 0 2 7 0 2 7 0
c) Helicoverpa 0 1 0 0 1 0 1 3 0
d) Spodoptera 1 2 0 0 1 0 1 2 0
e)Pink bollworm 0 0 0 0 0 0 0 0 0
Total 6 9 0 5 8 0 7 11 0
Plant protection cost (Rs/ha) 6515 13490 0 4550 11500 0 8175 15835 0
Yield (q/ha) 19.62 20.93 12.55 20.19 21.45 13.20 17.56 20.10 10.55 SIG 1.89
Gross income (Rs/ha) 70600 75280 45972 72325 77110 48202 36930 72384 39440
Cost of cultivation (Rs/ha) 45940 56115 37375 42725 53375 37625 47600 58960 36375
Net profit (Rs/ha) 24660 19165 8597 29600 23735 10577 16330 13424 3065
Cost benefit ratio 1:1.55 1:1.34 1:1.25 1:1.73 1:1.45 1:1.31 1:1.35 1:1.23 1: 1.09
The larval population was nil in IPM due to toxic effect of Bt protein present in cotton plant, where
as in Conventional control of BG II and BG hybrids due to the toxic effect of Bt protein and insecticides.
In non Bt hybrids very low incidence of 0.18, 0.09 and 0.24 larvae/plant were recorded in IPM,
conventional control and check modules, respectively and never reached ETL (Table 1). The higher
population of natural enemies and spraying of NSKE 5 % may reduced Helicoverpa armigera (Hub.)
larval population by suppression of the egg load under IPM, while it can be attributed to insecticidal
sprays under conventional control. Ramteke et al. (2002) reported that NSKE (5 %) and neem oil (300
ppm) were more economical in suppressing the population of H.armigera in cotton. Sohi et al. (2004)
reported that the larval incidence of H.armigera per plant, E.vittella and P.gossypiella in intact green
fruiting bodies were low in IPM fields compared to that of non IPM fields.
The per cent square (i.e. 0.81 in BG hybrid, 0.44 in BG II hybrid and 3.64 in NBt hybrid) and boll
damages (i.e. 0.07 in BG hybrid and 1.11 in NBt hybrid) were high in IPM module compared to
conventional control module which can be attributed to insecticidal interventions, while boll damage was
nil in IPM block of BG II hybrid which is attributed to the presence of double gene might be given full
protection against bollworms. In contrast, Kulkarni et al. (2004) reported that the mean boll damage was
12.16 % in Bt cotton with IPM against 14.58 % in Bt cotton with insecticide spraying. Bambawale et al.
(2004) reported significant reduction in boll damage by the bollworms in Bt cotton with IPM which was
11.5 % as against 29.4 % in conventional control.
The larval incidence of pink bollworm through destructive sampling showed that the larval
population was nil in both IPM and conventional control modules of both BG and BG II hybrids with no
locule damage in green bolls, whereas in NBt hybrid per cent locule damage (0.60) was low in IPM
block compared to conventional control (0.90) and check (2.05) due to pink bollworm and the number of
pink bollworm larvae/10 bolls was 0.08, 0.11 and 0.16 under IPM, conventional control and check,
respectively. (Table 1). Earlier, Sohi et al., (2004) reported that the larval incidence of P.gossypiella in
intact green fruiting bodies were low in IPM fields compared to that of non IPM fields.
The occurrence of natural enemies was high under IPM (0.14, 0.16 and 0.16/plant in BG, BG II and
NBt hybrids, respectively) and check blocks (0.12, 0.16 and 0.18 /plant in BG, BG II and NBt hybrids,
respectively) in all the modules, while it was low under conventional control (0.08, 0.24 and 0.08 /plant
in BG, BG II and NBt hybrids, respectively) in all the modules except in BG II cotton hybrid(Table 1).
The populations of natural enemies were conserved under IPM due to growing of intercrop, application
of safer chemicals like NSKE, avoidance of early season sprays by adopting seed treatment and stem
application. The increased incidence of natural enemies due to seed treatment (Kannan et al., 2004),
intercropping with greengram or black gram (Venkatesan et al., 1987) were well documented. Kulkarni
et al. (2004) reported that the mean population of natural enemies was significantly higher i.e. 9.93/ plant
in Bt cotton IPM when compared to 7.87/ plant in Bt cotton with insecticidal sprayings. The population
of coccinellids and chrysopids increased by 45.5 and 38.7 per cent, respectively in Bt cotton IPM plots
over control (Cui and Xia, 1998).
The seed cotton yield was slightly high from conventional control compared to IPM and checks in
all the modules. But, the cost benefit ratio was favourable from IPM (1:1.55, 1:173 and 1:1.35 in in BG,
BG II and NBt hybrids, respectively) compared to conventional control (1:1.34, 1:1.45 and 1:1.23 in BG,
BG II and NBt hybrids, respectively) and check (1:1.25, 1:1.31 and 1:1.09 in BG, BG II and NBt hybrids,
respectively) (Table.2). Though the yield was high, the net profit and cost benefit ratio were low from
conventional control modules due to higher plant protection cost than IPM and check. The total number
of interventions in BG, BG II and NBt hybrids were only 6, 5 and 7, respectively with low cost
techniques like stem application and spraying of botanical insecticides lead to low plant protection cost of
Rs. 6515, 4550 and 8175/ha in BG, BG II and NBt hybrids, respectively under IPM as against 9,8 and 11
insecticidal interventions with Rs. 13490, 11500 and 15835/ha under conventional control in all the three
hybrids, which resulted in low net profit though the yield was high in conventional control. The present
results clearly indicated that the use of insecticides indiscriminately lead to the reduction in net profits
with low cost benefit ratio and adverse effect on natural enemies. The present findings are in conformity
with Sohi et al. (2004) who reported that there was 53.3 % reduction in insecticide use in IPM fields with
12.2 number of insecticide sprays as against 18.7 sprays in non IPM fields which resulted in higher
economic returns. Kumar (2006) reported that growing of Bt cotton reduced the cost of bollworm control
upto 60 % with corresponding saving from plant protection cost and increase in seed cotton yield. Rao et
al. (1995) reported that the seed cotton yield from IPM plots was high which resulted in a higher cost
benefit ratio (1:5.3) in comparison with conventional farming practice (1:2.5). Hence transgenic Bt cotton
with inbuilt resistance to bollworms under IPM umbrella can combat pest problems for sustainable
cotton cultivation.

Gambit of IPM for Insect Resistant Transgenic Cotton 243
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R.K.Yadav), 28-29 Jan,2006, CCS HAU, Hisar, pp. 32-35.
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Cotton Pest Management Programmes using

Threshold-Based Interventions Developed
by CIRAD and its Partners in SubSaharan
African Countries
Silvie P.J.
1
, Adegnika M.A.
2
, Akantetou K.P.
3
, Ayeva B.
3
, Bonni G.
4
, Brevault, T.
5
Gautier
C.
6
, Hma O.
7
Houndete T.A.
8
, Ochou G.
9
, Prudent P.
6

Renou A.
10
and Togola M.
11
1
CIRAD, UPR Systmes de culture annuels, Montpellier, France
2
INRAB/CRA-CF, Cotonou, Benin,
3
ITRA/ CRA-SH, Anie, Togo
4
INRAB/CRA-CF, Parakou, Benin,
5
CIRAD, UPR Systmes de culture annuels, Montpellier, France
6
CIRAD, UPR Systmes de culture annuels, Maroua, Cameroon
7
INERA, Bobo-Dioulasso, Burkina Faso,
8
INRAB/CRA-CF, Bohicon, Benin
9
CNRA, Abidjan, Cte DIvoire,
6
CIRAD, UPR Systmes de culture annuels, Maroua, Cameroon
10
CIRAD, UPR Systmes de culture annuels, Bamako, Mali,
11
IER, Sikasso, Mali
AbstractIn the late 1980s, after a long period during which insecticides were sprayed at preset dates to control
cotton pests and their damage, some French-speaking countries in sub-Saharan areas decided to disseminate a more
sustainable crop protection approach among smallholders: targeted staggered control (LEC, for Lutte tage cible).
According to this approach, decisions on some insecticide sprays were made on the basis of infestation levels or the
extent of crop damage induced by major pests: Aphis gossypii aphids, Haritalodes (= Syllepte) derogata leaf-eating
caterpillars, and more generally Helicoverpa armigera, Diparopsis watersi, Earias insulana and E. biplaga bollworms.
Polyphagotarsonemus latus mites were sometimes included on this list. Due to changes in cotton production
conditions over the past 10 years, especially the development of pyrethroid resistance in H. armigera, and depending
on the country, this programme has been abandoned, preserved or replaced by other programmes. The strict use of
thresholds in Mali was thus taken to be a logical follow-up to LEC, which is still widely implemented. A targeted
threshold-based programme was developed in Togo. Cameroon abandoned LEC and opted for a sequential plan for
individual decision (SPID) programme (LOIC, for Lutte aprs observation individuelle des chenilles), based on
control after sequential sampling of bollworms. In Benin, LEC has been presented in two forms, i.e. complete and
partial, tailored to two regions delineated according to the extent of damage of some bollworms that live inside cotton
bolls, i.e. Pectinophora gossypiella, Thaumatotibia (= Cryptophlebia) leucotreta. In Ivory Coast, where these
Lepidopteran pests are also present, the use of treatment thresholds is limited to the beginning of the cotton crop cycle.
On the contrary, in Burkina Faso, thresholds are used after the first two calendar sprayings. The present article fully
describes these new crop protection programmes, sampling methods and associated intervention thresholds, in addition
to the advantages and constraints associated with their adoption.
INTRODUCTION
The concept of Integrated Pest Management remains widely recommended in crop protection
programmes against pests. Before any decision is taken to apply a toxic product, the use of action
thresholds is an essential step (Stern et al., 1959). In cotton production, the use of action thresholds
(called thresholds in the rest of this article), linked to the level of pests or damage, has been adopted in
numerous countries where the growth of this crop is conducted in a mechanical way over large acreages,
with the aim of reducing the quantity of pesticide use (the undesirable effects of pesticides are known)
and, as a result, reducing production costs.
At the end of the 1980s in the French-speaking sub-Saharan Africa region, cotton producing
countries developed various forms of integrated protection programmes for cotton which were called
Lutte tage cible (LEC) or targeted staggered control (TSC). A basic programme, sometimes seen
as a protection or safety net, is recommended according to a calendar preset in the conventional
standard protection programme (usually five or six sprays at fortnightly intervals), with the application of
40
Cotton Pest Management Programmes using Threshold-Based Interventions Developed by CIRAD and its Partners 245
insecticide doses reduced by half. Additional treatments or doses are carried out when the action
thresholds are reached. Amongst the diversified fauna of Arthropods which are present on this crop
(Vaissayre, 1994) certain groups of pests have been taken into account more specifically for the
application of these thresholds. Therefore observation has mainly focused on aphids (Aphis gossypii),
leaf-eating caterpillars such as Haritalodes (= Syllepte) derogata, Anomis flava, or Spodoptera littoralis
and caterpillars described as carpophagous such as Helicoverpa armigera, Diparopsis watersi, Earias
insulana and E.biplaga, which feed on fruiting organs and are visible on the outside of them (i.e.
exocarpic). In some countries, such as Togo and Benin for example, the presence of carpophagous
caterpillars which live inside green bolls (endocarpic) has led to the definition of two phytosanitary
regions (a northern zone and a southern zone). In fact, populations of these species are higher in the
southern regions where they cause significant damage to cotton and maize. The species concerned are
Thaumatotibia (= Cryptophlebia) leucotreta, which is also present in Ivory Coast (Ochou et al.,
1998a,b), and the Lepidoptera Mussidia nigrivenella, a pest best known in maize but sometimes reported
in significant numbers in southern Togo (Silvie, 1990).
A first summary detailing protection programmes using thresholds was published by Silvie et al.
(2001). Since this time, the context in which cotton is produced in sub-Saharan Africa has changed.
Major events have occurred locally, such as the privatization of cotton networks, leading to the division
of operations and reorganisation of the production chain. In Burkina Faso, for example, the production
zones have been consigned to three different entities, of which Faso Coton is part. In the Cte dIvoire,
the Ivoire Coton group manages the north-west of the country. This geographical carving up of cotton
production has led to a fragmentation which does not always make it easy to interpret production at the
national scale, particularly when it comes to agricultural statistics. There is also a possible threat to pest
management, caused by the definition of different phytosanitary protection strategies from one
geographical zone to another within the same country. Regarding pest management, the ways cotton
crops are protected have changed since the evolution of resistance to pyrethroids in the cotton bollworm
H. armigera was highlighted in countries such as the Cte dIvoire (Martin et al., 2002, 2005; Vaissayre
et al., 1998), Benin (Djihinto et al., 2009) and, later, Cameroon (Achaleke & Brvault, 2010 ; Brvault
et al., 2008). On the other hand, the recent highlighting of resistance in Bemisia tabaci (Houndt et al.,
2010) does not appear to have changed the pest management programmes applied. Finally, another recent
change has been the introduction of genetically modified cotton varieties carrying the cry1Ac and cry2Ab
genes, which come from the bacterium Bacillus thuringiensis (Bt). Thanks to the production of
insecticidal proteins, these cotton plants, known as Bt cotton, are resistant to the major Lepidopteran
pests (Hma et al., 2008; Traore et al., 2008). Following these changes and according to the country,
LEC programmes have been abandoned, modified or replaced by other types of protection programme
which still use thresholds to decide when to apply insecticides.
The recently published summaries on cotton pest management (Peshin et al., 2009 ; Kranthi &
Kranthi, 2010) rarely mention the experiences of French-speaking Africa in this area (van Huis, 2009 ;
Kranthi & Russell, 2009). It seems useful therefore, 10 years on, to update our knowledge on the use of
thresholds in cotton pest management programmes in French-speaking sub-Saharan Africa.
Published knowledge on the current status of cotton protection programmes varies according to the
country. As an example, the latest information on the protection of cotton crops in Chad and Central
African Republic dates back to data published by Nibouche et al. (2003a). For these authors, the
existence of various protection programmes, offered simultaneously to farmers, and changes in
application techniques (from one to 10 liters of water plus insecticide applied per hectare), appear to be
the cause for withdrawing the LEC programmes in these countries.
PAST AND CURRENT STATUS OF THRESHOLD-BASED PROGRAMMES IN WEST AFRICAN COUNTRIES
LEC programmes are based on a basic calendar programme. This calendar is the same as the
conventional programme, comprising five or six sprays at fortnightly intervals starting from the 45
th
day
after emergence (early squaring), but with insecticide doses reduced by half.
246
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TABLE 3: COMPARISON OF MARGINS (IN CFA (1)/HA) CALCULATED FOR VARIOUS COTTON PROTECTION PROGRAMMES IN FRENCH-SPEAKING SUB-SAHARAN AFRICA
Margin and Country Conventional' Programme Programmes using Thresholds (LEC - LOIC) Reference
MARI
(2)

Burkina Faso 41 243 68 206
COMPACI et al., 2010 Cameroon 51 340 79 630
Ivory Coast 33 300 107 300
Net margin
Benin Weak guidance LEC
Mathess et al., 2005
North 33 682 46 391
North-central -11 431 20 468
Centre -64 384 14 026
South -91 806
(1)
cfa = Franc of the African Financial Community (1 = 655.96 cfa)
(2)
MARI = Margin After Repayment of Inputs
Faced with the importance of resistance, Togo, that abandoned the LEC programme in 2000, adopted
a reinforced programme. Special scouting for H. armigera is systematically conducted six days after the
five treatments (northern region) or six treatments (southern region) of the conventional calendar
programme.
Between 2000 and 2002 in Ivory Coast, the Ivoire Coton company adopted a programme that used
thresholds only during the first protection window (Tab. 2). From 2009, the same company has been
using this strategy within the scope of the COMPACI project (Ochou and Amon, 2010). The
programme uses thresholds only This covers the first two application dates in the calendar programme
(conventional). Following this, the calendar programme is followed. The explanation for this is the
presence of endocarpic caterpillars, which are difficult to control with insecticide applications.
In contrast, for its crops grown in Burkina Faso, the Faso Coton company has decided to employ a
programme based on thresholds but only after two first treatments in a predefined calendar programme
(Leynaert, 2010).
Mali followed the logic set in motion through its adoption of an initial LEC programme and by 2008
had implemented a programme of treatments decided on sensu stricto thresholds (SST). With this
programme, there is no longer a basic programme of insecticide applications. The decision to spray is
taken only after the passing of defined thresholds, which remain the same as those in the LEC
programme (Tab. 1). However, it is clearly defined for economic reasons that the interval between two
consecutive treatments must be no less than 14 days. In 2008, Mali implemented 14,000 hectares of
cotton land with LEC programme, 52,000 hectares with the SST programme, while the preset calendar or
conventional programme was performed on 112,000 hectares. Therefore, programmes using thresholds
represented 37% of the area devoted to cotton production.
In Benin, two forms of the programme adapted to the two regions, LEC complete (north) and LEC
partial (south), have evolved during the development of the Projet dAmlioration et de Diversification
des Systmes dExploitation (improvement and diversification of growing systems project) (Prudent et
al.. 2006). A total area of 17,000 hectares was managed under LEC in 2009, representing nearly 12% of
Benins total cotton area.
In 2006, after studies by Nibouche et al. (2003b) and Beyo et al. (2004), Cameroon opted for a
protection programme called LOIC (Lutte aprs Observation Individuelle des Chenilles) or SPID
(Sequential Plan for Individual Decision). This programme uses sequential sampling on individual
bollworms. It therefore does not include the counting of leaf-eating caterpillars and aphids or their
damage. This simplified observation method for pests was adopted by 2,195 growers in 2009,
representing 1,170 hectares of cotton crops (less than 1% of the total surface area).
There has been a marked diversification in the cotton protection programmes using thresholds over
the past 10 years. They are more fully detailed below.

Sampling of Plants, Pests and Decision-Making Procedures
The sampling procedures are very similar in each country, with the exception of Cameroon which has
opted for sequential sampling.
For all the countries, observation field by field and the decision to spray (or not to spray) concern
only the field under observation. Treatment by crop block, which used to be conducted in Cameroon, is
no longer possible everywhere because of the presence of other crops in these blocks.
The number of plants observed in Cameroon varies according to the results obtained as scouting in
the field continues. A maximum of 25 plants is analyzed per area of 0.25 hectares. For the other
countries, between 25 and 40 plants are observed. They are chosen individually at random (therefore not
next to each other in the same row), in general following one or two diagonals. Their observation is
completed and damage or pest presence is noted, at a minimum, for bollworms (H. armigera, D. watersi,
Earias spp.), leaf-eating caterpillars (H. derogata, A. flava, S. littoralis) and the aphid A. gossypii. The
threshold levels leading to the decision to spray vary according to the country (Tab. 1 and 2). For
example, the threshold of three caterpillars of H. armigera per 30 observed cotton plants is used in Togo
to decide if treatment is needed against this pest (Tab. 1). This threshold is 5 larvae for 40 observed
plants in Benin. In other countries, all the exocarpic larvae are considered together. Mite
(Polyphagotarsonemus latus) damage is also taken into account in Ivory Coast and Benin (Tab. 1).
Leafhoppers (Cicadellidae) are taken into account only in the Cte dIvoire and the Bemisia tabaci
whitefly in Burkina Faso (Tab.2). In the latter country, following the introduction of Bt cotton plants,
there are no specific thresholds used for bugs (Pentatomidae, Pyrrhocoridae or Miridae) which represent
an important group in West African growing systems (Poutouli et al., 2011).
For easier record keeping in the field, various types of pegboards have been created for LEC
programmes in each country. An original pegboard was specially created for the LOIC programme in
Cameroon, which uses the sequential sampling technique (Beyo et al., 2004).
Active Ingredients, Dosage and Spraying Technique
The first protection window (W1) no longer includes pyrethroids. The two treatments in the first window
(Tab. 2) are designed to control the first caterpillars of H. armigera. The active ingredients are used in
conventional programme doses, sometimes described as full doses. In some countries, such as
Cameroon and Mali, this family of insecticides is not recommended in the third protection window either
(Tab. 2). Following this, in Burkina Faso, there is no reduction in the dose and the products used are
binary associations of pyrethroids (against caterpillars) and organophosphorous insecticides aimed at
aphids, whiteflies and mites (Leynaert, comm. pers.) or neocicotinoid. In Malis LEC programme, from
the second protection window products based on the binary association of pyrethroids-
organophosphorous insecticides are applied in half doses (compared to the full dose). Similarly, in
Benin, the doses of the last four applications are reduced. They are conducted using a combination of
cypermethrin and triazophos (SHERPHOS
370 or 320 EC depending on the region, north or south). For

additional applications, based on thresholds, and according to the country, simple or combined products
are used. In Benin, if the mite threshold is exceeded, triazophos (HOSTATHION
400) should be applied

at the rate of 200ml/ha (in the north) or 300ml/ha (in the south). For bollworms, aphid and H. armigera
thresholds, the products used can be respectively CYPERCAL
87.5 EC (cypermethrin) at 200ml/ha,

GAZELLE
200 SL (acetamiprid) at 40ml/ha and LASER
480 SC (spinosad) at 100ml/ha.

In Togo, the principle of alternating active ingredients is followed. In the northern area, for example,
the active ingredient used for the reinforced treatment against H. armigera is different to that used for
the first two treatments in the calendar. However, the choice of active ingredients beyond pyrethroids
remains limited. Endosulfan, which was originally used, was prohibited in 2009 for toxicological
reasons. Spinosad, an active ingredient recognized as being efficient for the control of H. armigera, is not
generally used because of its cost. Products such as indoxacarb (AVAUNT
150 SC) or
organophosphorous ingredients such as profenofos (CALFOS
720 EC) are available and used alone

(Tab. 3). In Burkina Faso, profenofos has been used at only 500g/ha rather than 720g/ha (used in Togo).
Emamectin benzoate could also be used on its own at 9.5g/ha in the next future (Leynaert, comm. pers.).
In the case of Cameroon (Prudent, comm.. pers.), there are no thresholds used to control aphids or
whiteflies. The treatment decision is taken by the producer and the agent who supports him when they
judge that the infestation is too large. A neonicotinoid, acetamiprid, is applied at 10g/ha against aphids or
20g/ha against whiteflies.
The widespread spraying technique is to use a volume of 10 litres of fluid per hectare (Very Low
Volume). The operator passes through every third row (0.80m between rows) with a sprayer which has a
rotary disc from, for example, the Berthoud company (model C5-10
, equipped with a green nozzle, in

Togo) or MicronSprayer (from MicroUlva
with a black nozzle, in Togo, or Ulva Plus
, in Mali and
Cameroon). These devices have a reservoir which is carried on the back of sufficient capacity (10 litres).
This technique uses the wind to help spread the toxic mixture (water plus insecticide) which always
carries an increased risk of contamination for the operator, especially when the wind changes direction
during spraying.
ADVANTAGES OF THRESHOLD-BASED PROGRAMMES
In French sub-Saharan Africa the main advantages expected for protection programmes using thresholds
(threshold programmes) are economic, combining a reduction in the insecticide use, number of
applications or quantity and increase of yields.
Insecticide Reduction
When pest pressure is low, programmes using sensu stricto thresholds, such as in Mali and Cameroon,
lead to a reduction in the number of treatments. For example, in Cameroon the LOIC programme, tested
over 2,000 hectares in 2006 and 2007, led to a reduction in the number of sprays in five of the 17 villages
that were monitored (Brvault et al., 2009). In 2008, the farmers who adopted this programme carried out
less insecticide treatments than other growers and their average cottonseed yields were no lower
(Bertrand et al., 2010).
But the application of thresholds does not always lead to a reduction of all insecticide used. In Togo
in 1995, before the implementation of the current reinforced programme, 50% savings in cypermethrin
were noted by Ayeva and Agossou (2000) with, however, additional use of chlorpyrifos-ethyl to control
infestations of the leaf-eating caterpillar H. derogata.
Yield Increase
When reducing the number of applications and the volume of insecticides used, there is often an increase
in average yields in fields which are protected by threshold programmes. This increase in yield, which
could in part be due to closer monitoring in the fields, is dependant on the weather and agronomic
conditions. In Mali in 2010, an average yield of 833kg of seed cotton per hectare was achieved with the
application of sensu stricto application thresholds, whereas it was 1,051kg two years earlier. In
Cameroon in 2009, a comparison of the conventional and LOIC programmes on 266 producers fields
showed a gain of 259kg/ha of cottonseed in favour of the latter programme (Gautier et al., 2010).

Economic Balance
Reducing the quantity of insecticides sprayed leads to economic savings. Ayeva and Agossou (2000), in
a comparison of the same 20 fields in Togo in 1995, identified a monetary saving of 30% in the costs
when using LEC programmes compared to the conventional programme. In Cameroon, Gauthier et al.
(2010) identified an additional cost of 1335cfa/ha (= 2.0) with the conventional spray programme.
Calculating margins is an analysis tool often used for comparing crop protection programmes. Thus
COMPACI et al. (2010) have calculated the margins after repayment of inputs (MARI) in the case of
calendar-based (conventional), threshold and LEC programmes (Tab. 4).
In Benin, the Matthess et al. (2005) study concerned three programmes actually used in the country,
conventional, organic cotton and LEC, and two programmes defined by extrapolation, Bt cotton and
fair trade cotton. As in the case of MARI, the net margin determined by these authors was in favour of
LEC programmes. The profitability of the LEC programmes in Benin is also confirmed by Prudent et al.
(2006).
Ecological Impact and Human Health
Lower pollution of watercourses and air and greater conservation of the fauna which regulates pest
populations are ecological advantages which have not been measured in a quantitative way. The use of
indicators of environmental quality is an aspect which merits further investigation. The risk of
contamination to the person applying the treatment has not generally been measured. With the possible
increase in the number of passages when a threshold is reached at the end of seven days, it could be
thought that the risk of contact with insecticides would increase.
MAJOR CONSTRAINTS
Transfer of Knowledge
The first constraints met are those linked to the dissemination of any innovation among smallholders.
Communicating in local languages concepts such as the management of insecticide resistance and
thresholds may present translation problems (Tourneux, 2003). The coexistence of several protection
programmes within the same country, or a new parameter to consider, and several production chains
(organic cotton, fair trade cotton) can further complicate the task of disseminating these programmes to
small growers.
The major constraint is the lack of knowledge of observers on the biology and ecology of Arthropods
(Sinzongan et al., 2004), on sampling techniques and on spotting and identifying problems associated to
pests and their damage, plant diseases and mineral deficiencies. Besides field diagnostic tools such as
pegboards, audio-visual tools and booklets identifying problems are generally available to personnel who
are trained. Beneficial insects (natural enemies of damage-causing insect pests) are also detailed.
Observers are members of farmers associations. Some, such as those in Mali, are qualified neo-literates
and are part of a programme to eliminate illiteracy.
Knowledge about pesticides and spraying equipment is sometimes deficient (Sougnabe et al., 2010),
including the risks relating to their use and guidelines for protecting the user, linked with the problem of
interpreting symbolic messages such as the warning pictograms on bottles (Tourneux, 1993). The option
of creating rural schools, proposed by Ochou et al. (1998a,c), or Champs-Ecoles des Producteurs (CEP),
equivalent to Farmer Field Schools (FFS), offering participative training, have been developed in the sub-
regional GIPD programme (Gestion Intgre de la Production et des Dprdateurs, integrated
management of production and pests) in Mali, Benin, Burkina Faso and Senegal (COS-Coton, 2011). But
this approach is considered as too expensive (Treen & Burgstaller, 2003).

Compliance with Threshold Principles
Independent of the adoption of threshold protection programmes, the economic crisis of the cotton chains
in cotton-producing countries have led to a reduction in the acreages dedicated to cotton growing over the
past few years. Investigations carried out within farmers associations have shown that official
recommendations are not strictly applied, with frequent under-dosing of plant protection products for
economic reasons (Sinzongan et al., 2004) or the inappropriate use of insecticides on other crops such as
cowpea (Vigna unguiculata) and tomato (Sougnabe et al., 2010). Given this context, the increase in the
number of treatments reported in Benin by Williamson et al. (2008) appears difficult to interpret.
Strict compliance with recommendations is difficult to obtain for several reasons. Prudent et al.
(2007) have shown, for example, in Benin that planters who have learned LEC techniques find it difficult
to remember the methods a few years after training. The complexity of the observations which have to be
carried out have been mentioned by Sinzogan et al. (2004). Another constraint is the necessity of
conducting weekly observations. Finally, instances of under-dosage, or non-application of insecticide,
despite a threshold being reached, are sometimes seen. The opposite is true too, with cases reported of
treatments being carried out even if the defined threshold has not been reached.
Economic Benefits
At the producer level, the job of observation in the field may be given to paid observers. In Togo, for
example, where an observer carried out the job in 10 fields in 1995, the payment was 100 cfa (= 0.15)
per observation and per field. The payment for this service, when carried out by a third party like this, is a
limitation very often mentioned by owner-producers. Another cost mentioned by producers is for the
management of the insecticides that have not been used for threshold treatments. This cost has sometimes
been included in the purchase price of products destined for producers wanting to apply LEC
programmes, but this discriminatory policy has triggered complaints from the producers involved.
Packaging in 15 litre containers is a handicap in Cameroon because each drum opened and not
completely used must still be paid for.
There are also economic constraints to be considered at the organisational level. The cost of
cascade or stepped training is never mentioned. In the programme offered in the Cte dIvoire, for
example, National Research (CNRA) has to train 205 extension officers, who in turn train 500 producer-
instructors who in turn train 1,500 producers. As a result of this, by 2012 it is forecast that in three years
1,500 producer-instructors and 500 LEC producers will have been trained (Ochou and Amon, 2010).
And yet this training represents a major effort and investment for a number of extension staff (Bertrand
et al., 2010).
CONCLUSION
In French-speaking sub-Saharan Africa, the current situation for cotton protection programmes using
action thresholds reveals a great variation from one country to another. All the same, their development
over significant acreages in some countries provides a measure of the interest shown in this type of
programme by both producers and the organisations which provide them with technical and financial
assistance. The multiplicity of the programmes offered is sometimes, but not always, linked to an
ecological reality. For example, in those regions with endocarpic Lepidopteran species (southern Benin,
Togo and the Cte dIvoire) it is more difficult to envisage the application of thresholds. For cost
reasons, there is no general monitoring of adult populations of Thaumatotibia leucotreta or Pectinophora
gossypiella with sexual pheromones, and producers are reluctant to destroy green cotton bolls to evaluate
the presence of these pests. Furthermore, the rosetted bloom damage caused by P. gossypiella does not
allow an action threshold to be established. The situation for these pests therefore remains unchanged
since the studies presented by Vaissayre (1994).

An increase in this diversity of protection programmes is a reasonable perspective, linked with new
projects under development financed by external institutions (for example, COMPACI, Cotton made in
Africa, Better Cotton Initiative). The GIPD programme, which is under development in Mali, is the only
project until now which seeks to take natural enemies into consideration through the calculation of target
pest/natural enemy ratios, such as those offered in Australia, albeit in a very different context. Producers
knowledge of these beneficial aids to crop production is often limited (Prudent et al., 2007) and a special
effort will have to be made in terms of training.
A simplification of the numerous existing threshold-based protection programmes, logically oriented
by an ecological and a regional analysis would probably be more satisfactory for a better diffusion among
smallholders, and consequently, for a reduction of costs. It will need the development of a network
involving researchers, growers, and all the actors of the production chain. For a large scale monitoring of
the impact of these new programmes, the contribution of producers will be essential.
A future challenge will be posed to pest management in countries which adopt or will adopt
transgenic resistant cotton to Lepidoptera. Until now, only Burkina Faso has very recently grown
genetically modified (GM) cotton over large acreages. According to the available information (Leynaert,
2010, COS-Coton, 2011), the first four treatments in the conventional programme were eliminated,
while the two applications at the end of the cycle were maintained, to control biting and sucking insects.
Research is continuing to evaluate the impact on non-targeted fauna, particularly bugs, and, with National
Research, adjustments are being made to the protection programme. In this context, definition of
thresholds for likely-emerging pests, as Mirids or Pentatomids bugs observed in other countries for
example, will be very useful.
Thus, the challenge is to develop a more theoretical approach for a better definition of threshold -
than the empirical one applied in many cases and at the same time, to succeed by a participative way of
extension and field application of the threshold, with a clear message, a good management of inputs and a
collective evaluation of the economical benefits of threshold-based programmes.
ACKNOWLEDGEMENT
The authors would like to thank the cotton companies who were willing to provide the agricultural
statistics used in this article and in particular Mr Paul Asfom (Sodecoton, Cameroon), as well as Mr Marc
Leynaert (Faso Coton, Burkina Faso) for the technical information pertinent to this country.
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Can Natural Refuges Delay Insect Resistance

to Bt Cotton
Brvault Thierry
1,2
, Nibouche Samuel
3
, Achaleke Joseph
4
and Carrire Yves
2

1
CIRAD, UPR 102, F-34398 Montpellier, France
2
Department of Entomology, University of Arizona, Tucson, AZ, USA
3
CIRAD, UMR PVBMT, F-97410 Saint-Pierre, La Runion, France
4
IRAD, PRASAC-ARDESAC, Garoua, Cameroon
AbstractNon-cotton host plants without Bacillus thuringiensis (Bt) toxins can provide refuges that delay
resistance to Bt cotton in polyphagous insect pests. It has proven difficult, however, to determine the effective
contribution of such refuges and their role in delaying resistance evolution. Here we used biogeochemical markers to
quantify movement of Helicoverpa armigera moths from non-cotton hosts to cotton fields throughout the cropping
season, in three agricultural landscapes of the West African cotton belt (Cameroon) where Bt cotton was absent. We
show that the contribution of non-cotton hosts as a source of moths was spatially and temporally variable, but at least
equivalent to a 7.5% sprayed refuge of non-Bt cotton. Simulation models incorporating H. armigera biological
parameters, however, indicate that planting non-Bt cotton refuges may be needed to significantly delay resistance to
cotton producing the toxins Cry1Ac and Cry2Ab. Specifically, when the concentration of one toxin (here Cry1Ac)
declined seasonally, resistance to Bt cotton occurred rapidly when refuges of non-Bt cotton were rare, because
resistance was essentially driven by one toxin (here Cry2Ab). The use of biogeochemical markers to quantify insect
movement can provide a valuable tool to evaluate the role of non-cotton refuges in delaying the evolution of H.
armigera resistance to Bt cotton.
Cotton is widely grown in West Africa, where it helps sustain millions of resource-poor farmers and
rural communities. Transgenic cotton producing the Bacillus thuringiensis (Bt) toxins Cry1Ac and
Cry2Ab was recently introduced to Burkina Faso (1) to increase agricultural profitability. Such Bt cotton
is called pyramid because it produces two distinct Bt toxins active against some pest species (2-5).
Management of insect resistance to Bt crops requires production of abundant susceptible individuals in
refuges of non-Bt host plants that disperse and mate with the rare resistant survivors in Bt fields (2-5).
Because the most important insect pest of cotton in West Africa, Helicoverpa armigera, is polyphagous
and highly mobile (6, 7), non-cotton host plants could reduce the reliance on refuges of non-Bt cotton to
delay resistance. While some studies have evaluated production of H. armigera by non-cotton host plants
elsewhere (8-11), movement of moths from non-cotton hosts to cotton fields has never been quantified in
space and time. Nevertheless, it is often assumed that cotton refuges are not required to delay H.
armigera resistance to Cry1Ac/Cry2Ab cotton in agroecosystems where small fields of diversified crops
and patches of non-cultivated hosts are close together (10), such as in West Africa.
We used biogeochemical markers to measure movement of H. armigera moths from non-cotton hosts
to cotton fields in Cameroon (in the West African cotton belt). A total of 18 moth collections were taken
from pheromone traps in cotton fields from June to November 2006 at three locations (Guider, Djalingo,
and Tchollir). Larval host plants were identified by analyzing moths abdomens for gossypol (a
phytochemical present in cotton) and wings for stable carbon isotope ratio. We categorized plants as
cotton, non-cotton C3 plants (e.g., weeds such as Cleome spp. and Hyptis sp.), and C4 plants (e.g., corn).
Most moths trapped early in the growing season (June-July) had signatures of C3 and C4 non-cotton
plants (Fig. 1a-c). The few gossypol-positive moths detected at that time likely originated from
overwintering pupae and possibly from early-planted cotton or cotton left in fields from the previous
growing season. When the first moth generation emerged from cotton (August), most moths had
signatures of C3 and C4 non-cotton plants (Fig. 1a-c). The contribution of non-cotton refuges to the pool
of moths trapped in cotton fields decreased during the second (September) and third (October)
generations, particularly at Djalingo, and to a lesser extent Tchollir and Guider. At cotton harvest
(November), most moths originated from non-cotton C3 plants at Djalingo and Tchollir, whereas moths
41
from cotton still contributed significantly to the pool of moths at Guider (Fig. 1a-c), where cotton is
usually harvested a few weeks later.

Fig. 1: (a-c). Moths Trapped in Cotton Fields (%) that Originated from Non-cotton Host Plants. Remaining Moths (100 % Indicated by bar) Originated
from Cotton. Moths were Trapped at Three Locations (Guider, Djalingo, and Tchollir) in Cameroon in 2006.
(d) Typical Sequence of Helicoverpa Armigera Host Plants in the West Africa Cotton Belt Throughout the Cropping Season.
Curves Represent Temporal Occurrence and Relative Area of Host plants.
We used a two-locus population genetics model incorporating empirical estimates of H. armigera
biological parameters to evaluate how movement from non-cotton refuges may affect the evolution of
resistance to Cry1Ac/Cry2Ab cotton at each of the three locations. The model considered the seasonal
decline in mortality of a strain resistant to Cry2Ab on Cry1Ac/Cry2Ab cotton (12), which paralleled the
decline in Cry1Ac concentration generally observed in Bt cotton during the course of the growing season
(5, 13). Such reduction in mortality of Cry2Ab-resistant insects on Cry1Ac/Cry2Ab cotton invalidates
one of the fundamental assumptions of the pyramid strategy, i.e., the killing of insects resistant to one
toxin by the other toxin, and thus could accelerate resistance evolution (2-5, 14).

Seasonal declines in Cry1Ac-induced mortality and more stable Cry2Ab-induced mortality
necessarily generates stronger selection for resistance to Cry2Ab than Cry1Ac. Simulations showed that
the evolution of resistance was primarily driven by Cry2Ab resistance alleles, as the initial resistance
allele frequency and the dominance of Cry1Ac resistance had little effect. Among-site variability affected
the role of non-cotton refuges in delaying resistance evolution (Fig. 2a,b). With partially recessive
resistance to Cry2Ab (D
LC
= 0.1) and initial resistance allele frequency of 0.0033 to Cry2Ab, non-cotton
refuges delayed resistance 32 years at Guider, 16 years at Tchollir, and 8 years at Djalingo (Fig. 2a).
With partially recessive resistance to Cry2Ab (D
LC
= 0.1) and higher initial resistance allele frequency of
0.033 to Cry2Ab, however, resistance evolution was faster and non-cotton refuges delayed resistance 17
years at Guider, 9 years at Tchollir, and 6 years at Djalingo (Fig. 2b). With higher dominance of
Cry2Ab resistance (D
LC
= 0.3 or 0.5), sprayed refuges of 20% non-Bt cotton in addition to non-cotton
refuges delayed resistance 8 years at Guider, 11 years at Tchollire, and 8 years at Djalingo (Fig. 2b).
In a worst-case scenario with an initial resistance frequency of 0.033 and semi-dominant resistance to
Cry2Ab (D
LC
= 0.5), sprayed refuges of 50% non-Bt cotton delayed resistance 15 years at Guider, 8 years
at Tchollir, and 6 years at Djalingo (Fig. 2b).
Can Natural Refuges Delay Insect Resistance to Bt Cotton 257

Fig. 2: Effect of the Abundance of Sprayed Refuges of Non-Bt cotton (%) on the Evolution of Helicoverpa Armigera Resistance to Cry1Ac/Cry2Ab Cotton
at three Locations (Guider, Djalingo, and Tchollir) in Cameroon. For Cry2Ab, the Initial Resistance Allele Frequency Was 0.0033 (a) or 0.033 (b), and
Resistance was Partially Recessive (D
LC
= 0.1, Dashed line) or semi-Dominant (D
LC
= 0.5, Solid Line). For Cry1Ac, the Initial Resistance Allele Frequency
was 0.0003 and Resistance was Partially Recessive (D
LC
= 0.3). The Criterion for Resistance Evolution was >20% Survival on Bt Cotton
Our seasonal assessment of H. armigera movement shows that non-cotton refuges were equivalent to
7.5% non-Bt cotton refuges treated with insecticides throughout the cotton-growing season (Fig. 1b).
Despite the important but temporally and regionally variable moth contribution from non-cotton hosts to
putative Bt cotton fields, our modeling results show low efficacy of the pyramid strategy when the
concentration of Cry1Ac declines during the growing season, resistance to Cry2Ab is non-recessive, and
only non-cotton refuges are available. Under the first two conditions, refuges of non-Bt cotton would be
needed to significantly delay resistance, unless high sustained movement from non-cotton refuges to
cotton fields occurs during the growing season (e.g., Guider), or long-range migration is more important
northward than southward. More generally, we demonstrate that biogeochemical markers provide a basis
to evaluate the role of a variety of refuges in delaying the evolution of resistance to Bt crops in
polyphagous insect pests. Such markers will be useful to assess the role of non-cotton hosts in delaying
H. armigera resistance to Bt in Burkina Faso and other West African countries that may adopt Bt cotton.
REFERENCES
[1] C. James, ISAAA Briefs 39, 129-132 (2008).
[2] B.E. Tabashnik et al., Nat. Biotechnol. 26, 199-202 (2008).
[3] B.E Tabashnik. et al., J. Econ. Entomol. 102, 2011-2025 (2009).
[4] F. Gould, Annu. Rev. Entomol. 43, 701-726 (1998).
[5] A.M. Showalter et al., J. Insect Sci. 9, 1-39 (2009).
[6] T. Brvault et al., Bull. Entomol. Res. 98, 565-573 (2008).
[7] J.M. Vassal et al., Comm. Appl. Biol. Sci., 73, 433-437 (2008).
[8] W.M. Green et al., Afr. Entomol. 11, 21-29 (2003).
[9] K.C. Ravi et al., Environ. Entomol. 34, 59-69 (2005).
[10] K.M. Wu, Y.Y. Guo, Annu. Rev. Entomol. 50, 31-52 (2005).
[11] G.H. Baker, C.R. Tann, G.P. Fitt, Aust. J. Agr. Res. 59, 723-732 (2008).
[12] R.J. Mahon, K.M. Olsen, J. Econ. Entomol. 102, 708-716 (2009).
[13] K.R. Kranthi et al., Curr. Sci. 89, 291-298 (2005).
[14] F. Gould et al., J. Econ. Entomol. 99, 2091-2099 (2006).
Can Tomato be a Potential Host Plant

for Pink Bollworm
N. Ariela
1
, S. Harpaz Liora
2
, R. Mario
3
, S.

Roee
4
and H.A. Rami
3

1
Israel Cotton Board, P.O.B. 384 Herzlia B' 46103 Israel;
2
Northern R & D, P.O. Box 831, Kiryat Shmona11016, Israel
3
Department of Entomology, Agricultural Research Organization,
Gilat Research Center, M.P. Negev, 85280, Israel
4
Department of Entomology, The Robert H. Smith Faculty of Agriculture, Food and Environment,
The Hebrew University of Jerusalem P.O.Box12, Rehovot76100, Israel
E-mail: ariela@cotton.co.il
AbstractThe pink bollworm (PBW), Pectinophora gossypiella (Saunders) is a major pest of cotton in Israel.
Recently, processed tomatoes growers from northern Israel have reported that, suspected PBW larvae were found
inside tomato fruits in the field. Since tomato (Solanum lycopersicum) has not been recorded as a host plant for the
PBW, a laboratory study was conducted to find out whether PBW neonates can penetrate tomato fruits and complete a
whole life cycle on them. PBW eggs were placed on tomato fruits; thereafter, some neonates penetrated the fruits and
succeeded to complete a whole life cycle in tomato fruit. In another experiment, tomato plants were placed in net cages
and PBW adults were introduced into the cages. Females laid eggs on the tomato plants and a few larvae developed in
the fruits. These findings shed new light on the understanding of PBW host range and have implications on area wide
IPM programs.
INTRODUCTION
The pink bollworm (PBW), Pectinophora gossypiella (Saunders) is the major pest of cotton in Israel; and
mating disruption is a very common practice in all cotton fields (Kehat and Dunkelblum 1993, Kehat et
al. 1998). During the past ten years, the pest has spread all over the country, causing a real threat to
cotton growth in Israel. PBW is found mainly on cotton, although a few larvae were noticed also on
other Malvaceae species such as Hibiscus sp. and Okra Recently processed tomatoes growers from
northern Israel have reported that suspected PBW larvae were found inside tomato fruits in the field. As
tomato (Solanum lycopersicum) has not been recorded as a host plant for the PBWs, (Shiller et al. 1962),
a laboratory study was conducted to find out whether PBW neonates can penetrate tomato fruits and
complete on them a whole life cycle.
Thirty ripe tomato fruits with their vines were put into plastic cups along with twenty PBW eggs placed
on each fruit. The fruits were held under standard laboratory conditions of 272C, 50% humidity and
photoperiod of 14:10 hours light: dark conditions. Every few days the fruits were checked for larva
penetration.
In another experiment, three tomato plants, each from a different variety, (Brigade, 5811, 9780) were
put in net cages.
60 males and 60 females were introduced to each cage. The cages were placed on tables in outdoor
conditions (summer).
After 14 days, the fruits were checked for PBW eggs and entrance holes; then, the fruits were cut into
slices to find PBW larvae.

42
Can Tomato be a Potential Host Plant for Pink Bollworm 259
At the first experiment, tiny holes were found on the upper part of the tomato fruit underneath the sepal.
Later on, PBW larvae were found inside the fruits feeding on flesh and seeds (figure 1).

Fig. 1: PBW Larvae Inside a Tomato Fruit
Two weeks later, exit holes and damage to fruits were detected and larvae dropped down and pupated
(figure 2, 3). Adults that emerged from the pupae mated normally and laid fertile eggs. In conclusion, the
PBW has succeeded to complete a whole life cycle in tomato fruit.

Fig. 2: Exit Hole of PBW

Fig. 3: Damage Made by PBW

At the second experiment with tomato plants, we found entrance holes and larvae only in the variety
9780. Penetrations of PBW larvae were detected inside just two fruits out of 25 red and green fruits that
were on the plants.
The results showed that PBW not only could develop in tomato fruits but females might lay eggs
spontaneously on tomato plants.
These findings shed new light on the understanding of PBW host range and have implications on area
wide IPM programs.
Further choice experiments will be planned to learn whether PBW females would select and lay eggs
on tomato fruits in the presence of cotton plants.
REFERENCES
[1] Kehat, M, and Dunkelblum, E. 1993. Sex pheromones: Achievements in monitoring and mating disruption of cotton pests
in Israel. Arch. Insect Biochem. Physiol. 22: 425-431.
[2] Kehat, M., Anshelevich, L, Gordon, D., Harel, M. and Dunkelblum, D. 1998. Evaluation of Shin-Etsu twist-tie rope
dispensers by the mating table technique for disrupting mating of the cotton bollworm, Helicoverpa armigera (Lepidoptera:
Noctuidae), and the pink bollworm, Pectinophora gossypiella (Lepidoptera: Gelechiidae). Bull. Entomol. Res. 88: 141-148.
[3] Shiller, I., L. W. Noble, and L.C. Fife. 1962. Host plants of pink bollworm. J. Entomol. 55: 67-70.
Impact of IRM Strategies on Bt Cotton

in Andhra Pradesh
T.V.K. Singh, N.V.V.S.D. Prasad, S. Sharma and S. Dayakar
Acharya N.G.Ranga Agricultural University, Rajendranagar, Hyderabad, India
E-mail: tvksingh@yahoo.com
AbstractCotton is extensively cultivated in entire Andhra Pradesh, which is one of the important agrarian states in
India under diverse farming situations with high inputs. Cotton in highly vulnerable to pest attack and insect pests
cause losses up to 87% in seed cotton yield (Taley et al. 1988). Among insect pests aphids [Aphis gossypii (Glover)],
Leaf hopper [Amrasca biguttula biguttula (Jshida)], whiteflies [Bemisia tabaci (Genn.)], thrips [Thrips tabaci (Linde)]
and boll worm complex viz., Gram caterpillar [Helicoverpa armigera (Hub.)], Tobacco caterpillar [Spodoptera litura
(Boisd.)] and Pink bollworm [Pectinophora gossypiella (Saund)] are considered to be the major constraints. Excessive
and indiscriminate use of insecticides in cotton has led to problems of insecticide resistance, pest resurgence,
accumulation of harmful residues and toxicity to non-target organisms. This has prompted the necessity for the
development of strategies for judicious management of insecticides and a window based insecticide resistance
management (IRM) strategies on cotton was implemented in three districts of Andhra Pradesh viz., Guntur,
Khammam and Kurnool. The strategies blend all crop production practices to incorporate proper and low use of
insecticides. Natural enemy populations are least disturbed and, different groups of chemicals are alternated.
The dissemination of insecticide Resistance Management (IRM) strategies at village level by way of trainings and
field visits prompted the adaptation of strategies by farmers for managing cotton pest complex on Bt cotton. To
disseminate IRM strategies, a total of 165 villages were adopted in three districts from 2008 to 2011 along with 75
villages that were selected as non IRM villages for comparing the impact of IRM strategies. The adoption of IRM
strategies led to reduction in pest incidence in IRM villages. Boll worm incidence was very less in IRM and non IRM
villages. The population of sucking pests was less in IRM villages than non IRM villages.
The strategic positioning of insecticides coupled with ecofriendly technologies led to abundance of natural enemies
in cotton ecosystem in IRM fields, while the incidence of these insects was lower in non IRM fields due to insecticidal
sprays. Impact of adoption of IRM strategies resulted in the reduction in insecticidal sprays (28.84%) in IRM villages
over non IRM villages. Cotton yield was higher in IRM adopted village (26.67 qt/ha) compared to non IRM villages
(22.46 qt/ha). Net profit per/ha was more in IRM villages than non IRM villages. Farmers, by adopting IRM
strategies realized higher net returns by saving in plant protection cost due to less number of insecticidal sprays and
increased seed cotton yield.
INTRODUCTION
Cotton popularly known as white gold is the most important commercial crop in India and plays a vital
role in agricultural, industrial, social and monetary affairs of the country. Area wise, India ranks first in
global scenario (about 33 per cent of the world cotton area) but with regard to production, it is ranked
next to China, which is the top producer (AICCIP,2011). The production increased from a meager 2.8
million bales (170 kg lint/bale) in 1947-48 to a high of 17.6 million bales in 1996-97 and a record of 31.5
million bales was recorded during 2007-08(AICCIP, 2008). During 2009-10, it was grown on an area of
10.3 million hectares with the production of 29.5 million bales and average lint yield of 486 kg/ha.
Among cotton growing states, Gujarat leads in production with 9.8 million bales followed by
Maharashtra (6.3 million bales) and Andhra Pradesh (5.2 million bales). However, the productivity of
cotton in India is still far less than other cotton growing countries of the world, viz., Australia (1579
kg/ha) Brazil (1480 kg/ha), China (1301kg/ha), USA (943 kg/ha), Uzbekistan (775 kg/ha) and Pakistan
(1579kg/ha) (AICCIP, 2011).
The insect pests are one of the major constraints in achieving optimum yield potential. Cotton crop
harbored 1326 species of insects from sowing to maturity in different cotton growing areas of the world
(Hargreaves, 1948) and 162 species have been reported on the crop in India. Among these, 9 are of
utmost importance inflicting significant losses in yield. The monetary value of yield losses due to insect
pests has been estimated to be Rs. 33, 9660/- million annually (Dhaliwal et al., 2010).
43
Before the introduction of Bt cotton, cotton growers were mainly using the synthetic insecticides to
combat the pests. As a result, bollworms, developed resistance to almost all major classes of pesticides.
Development of transgenic cotton resulted in an immense increase in seed cotton yield and reduction in
insecticidal sprays (Barwale et al., 2004) and it helped the farmers to manage the population of H.
armigera, the most important pest causing about 31.0 per cent loss in non-transgenic cotton (Grover and
Pental, 2003). Keeping in view the above facts, the present study on impact adoption of insecticide
resistance management (IRM) strategies in Bt cotton was planned to manage insect pests below
economic threshold level (ETL), reduction in number of sprays, and increase the cotton yield by
disseminating the IRM strategies in the adopted villages in Andhra Pradesh.
TABLE 1: DETAILS OF VILLAGES UNDER IRM AND NON-IRM IN ANDHRA PRADESH DURING 2010-11
Year Guntur Khammam Kurnool

No. of IRM
Villages
No. of Non-IRM
Villages
No. of IRM
Villages
No. of Non-IRM
Villages
No. of IRM
Villages
No. of Non-IRM
Villages
2008-09 15 2 15 15 60 15
2009-10 15 5 15 15 15 5
2010-11 10 5 10 3 10 10
TOTAL 40 12 40 33 85 30
Total IRM Villages = 165
Total Non-IRM Villages = 75
IRM module developed by CICR for Bt Cotton Pest Management was implemented for three consecutive
years during 2008 to 2011 and evaluated in 165 villages in three districts viz., Guntur, Khammam and
Kurnool districts of Andhra Pradesh. Seventy five villages were also selected as non IRM villages for
comparing the impact of IRM strategies(Table-1). A total of 6245 farmers (Table-2) followed IRM
strategies in an area of 12898.54ha during 2008 to 2011. Recommended package of practices of
ANGRAU was followed (ANGRAU Panchangam, 2011). In the beginning of every year, farmers were
educated about the IRM strategies. Various techniques, field days and field visits were conducted for
demonstrating IRM strategies in IRM adopted villages. Literature in local language pertaining to
agronomic practices, insect pests, economic threshold levels and their management strategies were
distributed to farmers.
TABLE 2: DETAILS OF BENEFICIARY FARMERS AND AREA COVERED IN ANDHRA PRADESH DURING
Year Guntur Khammam Kurnool

No. of IRM
Beneficiary
Farmers
Area
under
IRM (ha)
Area under
Non-IRM
(ha)
No. of IRM
Beneficiary
Farmers
Area
under IRM
(ha)
Area under
Non-IRM
(ha)
No. of IRM
Beneficiary
Farmers
Area
under
IRM (ha)
Area under
Non-IRM
(ha)
2008-09 737 2252.50 159.10 819 1110.50 62.10 1600 3343.56 536.20
2009-10 653 1605.20 306.20 787 1393.20 1008.00 385 565.40 128.94
2010-11 420 1026.20 360.00 544 1175.20 501.80 300 426.78 32.00
TOTAL 1810 4883.90 825.30 2150 3678.90 1571.90 2285 4335.74 697.14
IRM strategies which were implemented are as follows
Early Sucking Pests: No Foliar Spray (Till 60 Das)
Cultivation of sucking pest tolerant genotypes (Bt or non BT)
Intercropping with cowpea, soyabean and blackgram
Eradication of weeds in and around the cotton fields
Avoidance of chlorothalonil and organophospate sprays for sucking pest control
Stem application or soil application of dimethoate or acephate at 30-40 DAS and 50-60 DAS for
control of thrips, mirid bugs, mealybugs and other sucking pests
Neem oil 2.5 lit/ha mixed with 0.1% Nirma washing soap powder

Impact of IRM Strategies on Bt Cotton in Andhra Pradesh 263
Biological and Biopesticide Window (61-90 Das) Initial Bollworm Infestation
Verticillium lecanii to be used for sucking pest control especially for the control of mealy bugs
Cryptolaemus montrouzieri as inoculative releases on weeds or fruit crops adjacent to cotton
fields
Use of HaNPV on Bt cotton at 50% bollworm infested plants followed by the application of 5%
NSKE a week later
Not to spray against minor lepidopteran insects such as cotton leaf folder and cotton semilooper
Trichogramma can be used on non-Bt genotypes at 70-80 DAS
Not to spray Bt formulation on Bt cotton to avoid further selection pressure
Insecticide Window (91-120 Das)
Use of spinosad or emamectin benzoate on only non-Bt cotton at ETLs of 50% infested plants.
Avoid these insecticides on Bt cotton
Use of indoxacarb only once only on non-Bt cotton for control at ETLs of 90-100% plants
showing flared up squares
Use organophospate or carbamates only once either on Bt cotton or non-Bt cotton as effective
larvicides for control of bollworms at ETLs of 90-100% plants
Pink Bollworm Window (>120 Das) Pyrethroids
ETL based spray: Eight pink bollworm moths per trap per night for 3 consecutive nights. The
application of thiodicarb as late season sprays would be effective for pink bollworm
management.
Pyrethorid resistance in H.armigera is generally high, but pyrethroids are very effective against
pink and spotted bollworms and are ideally suited for the late season window.
The data pertaining to cultivation of different hybrids, sowing time, different agronomic practices
adopted along with the yeasrs by the individual IRM farmers was recorded and pooled.
Pests incidence: The population of leafhoppers, whiteflies, thrips, mealy bugs, tobacco caterpillar and
pink boll worm remained below the ETL in all the IRM adopted villages and was significantly less than
Non IRM villages (Table-3).
TABLE 3: OCCURRENCE OF INSECT PESTS AND NATURAL ENEMIES IN ANDHRA PRADESH PROJECT VILLAGES
Insects Villages Guntur* Khammam* Kurnool*
Leafhopper / 3 leaves
IRM 1.43 1.62 1.33
Non-IRM 2.12 1.94 2.41
Whiteflies /3 leaves
IRM 2.01 1.64 1.87
Non IRM 2.94 2.54 2.48
Thrips / 3 leaves
IRM 0.62 0.50 0.41
Non-IRM 0.83 0.76 1.14
Mealy bugs / 2.5 cm
IRM 1.14 0.25 0.11
Non IRM 1.45 0.34 0.19
Tobacco caterpillar/plant
IRM 0.07 0.22 0.55
Non-IRM 0.23 0.53 0.62
Pink Bollworm/plant
IRM 0.12 0.02 0.09
Non-IRM 0.22 0.11 0.23
Natural enemies/plant
IRM 0.68 0.61 0.63
Non-IRM 0.54 0.30 0.20
*Mean of 3 years for the entire occurrence of insecticidal pest
The strategies positioning of insecticides coupled with ecofriendly technologies lead to abundance of
natural enemies in cotton ecosystem in IRM fields, while the incidence of these insects was lower in non
IRM fields due to insecticidal sprays (Table-3).
Impact of IRM strategies: The IRM strategies disseminated in IRM adopted villages on no. of sprays,
cost of sprays, cotton yield, gross income and net profit is presented in Table-4. The mean no. of sprays
for pests was 3.74 in IRM villages and 5.41 in non IRM villages. The mean cost of sprays was higher in
non-IRM villages (Rs.3230) as compared to IRM villages (Rs. 2244).
TABLE 4: IMPACT OF IRM TECHNOLOGY IN ANDHRA PRADESH
Attributes IRM Villages IRM Villages
No.of Sprays 3.74 5.41
Cost of sprays (Rs) 2244 3230
Cotton yield (qt/ha) 26.67 22.46
Gross income (Rs/ha) 65264 54420
Net profit (Rs/ha) 41351 34640
The yield was also higher in villages where IRM strategies were adopted (26.67 qt/ha) over non-IRM
villages (22.46 qt/ha). The grass income and net profit was more in IRM villages.
The present findings are in conformity with the results of Rajak et al., (1997) and Kranthi et
al.,(2000) who reported reduction in pesticide consumption in IRM adopted villages and increase in
yields.
ACKNOWLEDGEMENT
This work was funded by the Ministry of Agriculture under the Technology Mission on Cotton II through
DOCD with technical support in a network mode from Director, CICR, Nagpur.
REFERENCES
[1] AICCIP (2008)-All India Coordinated Cotton Improvement Project. Annual Report 2009-10, Central Institute of Cotton
Research, Regional station, Coimbatore. pp 3-5.
[2] AICCIP. (2011)- All India Coordinated Cotton Improvement Project. Annual Report 2010-11, Central Institute of Cotton
Research, Regional station, Coimbatore. pp 3-5.
[3] ANGRAU (2011)-Vyavasaya Panchangam ANGRAU, Hyd.
[4] Barwale, R.B., Godwal, V.R., Usha, Z. and Zehr, B (2004) - Prospect for Bt cotton technology is India. AgbioForm.7: 23-6.
[5] Dhaliwal, G.S., Jindal, V. and Dhawan, A.K. (2010) - Insect pest problems and crop losses: changing trends. Indian J.
Ecology 37: 1-7.
[6] Grover, A. and Pental, D. (2003) - Breeding objectives and requirements for producing transgenic for the major field crops
of India. Curr. Sci. 84: 310-20.
[7] Hargreaves, H. (1948) - List of recorded cotton insects of the world. Pp50. Commonwealth Institute of Entomology,
London.
[8] Kranthi, K.R., Banerjee, S.K. and Russell, D. (2000) - IRM strategies for sustainable cotton pest management in India.
Pestology 24: 58-67.
[9] Rajak, R.L.; Diwaker, M.C. and Mishra, M.P. (1997) - National IPM program in India. Pesticide Information 23: 23-26.
Efforts to Mitigate Stickiness Problem in Sudan

A. Abdelatif and E. Babiker

Agricultural Research Corporation, Sudan
AbstractStickiness is one of the limiting factors for cotton production and marketing in many countries and obliged
cotton grower, worldwide, to sell their sticky cotton at lower prices. In the Sudan, research programs were carried out
by Agricultural Research Corporation (ARC), addressing the causes and control measures in an attempt to find a
remedy for the problem. Of these efforts manipulating the morphological and physiological characters of the cotton
plant in such away to reduce the whitefly population and allow for easy biological, chemical and cultural control,
resulted in developing very promising lines. In addition, identification of the type of sugar causing cotton stickiness
and the establishment of reliable methods for grading cotton stickiness were developed. Stickiness research project was
endorsed and financed by the Common Fund for Commodities (CFC) during 1997-2001. The objectives of the project
were to develop a methodology to separate sticky from non-sticky cotton. Another objective of the project was the
determination of threshold levels of stickiness for spinning under different conditions to enable the utilization of sticky
cotton in spinning process.
The study revealed considerable variability in stickiness levels among the cotton production areas, and also,
considerably low levels of stickiness were observed in some schemes. Cultural practices were needed where a long term
improvement of stickiness free production were observed.
INTRODUCTION
Cotton Gossypium is the major natural textile fibre crop worldwide. In Sudan, cotton has been grown
for centuries. The cotton plant is indigenous and a number of its wild relatives (members of the genus
Gossypium) existed in various parts of the country.
Commercial growing of the crop, however, started in 1867. However, the big jump was in 1926,
which marked the official start of functioning of the Gezira Scheme. Likewise, large production has,
since the beginning, been backed by a strong research program The Agricultural Research Corporation
(ARC) has an intensive program to develop new varieties, increase yield and improve quality to meet the
recent demand of consumers. The bulk of the production is exported as raw fibre (90%) in a highly
competitive world market. During the last three decades Sudan cotton faced strong competition in the
world market. Sudan cotton suffered mainly from low yields and low quality due to contamination.
Major activities of the research program addressed the yield and fibre quality problems. In recent years,
however, contamination issues started to acquire their fair share in the research strategies. The main
objective of this paper is to focus on efforts to mitigate stickiness in cotton, grown in the Sudan.
STICKINESS PROBLEM
Stickiness was observed in Sudan since the early 1960s, but was sporadic at that time and of little
importance. During the 1980s the phenomenon became worldwide thus affecting marketability of Sudan
cotton. It caused substantial economic losses to the cotton producers, worldwide, and obliged them to sell
their sticky cotton at lower prices. In case of the Sudanese cotton the discount prices ranged from 5-30%.
(Fadlalla, 1998).
Stickiness of cotton lint was found to be caused by honey-dew excreted by the two insects whitefly
and aphids (Gameel 1969).However, other factors causing stickiness contamination has been reported in
the literature of these, broken seeds, immature fibres as well as sugary substance of cotton plant which
may directly or indirectly affect cotton lint at later processes. (Kalifa1980,Watson 2000).
NATIONAL RESEARCH PROGRAM
Intensive research was carried out regarding stickiness of cotton in Sudan. First, a scientific research
committee was established in 1967 with the objective of investigating the nature and the origin of
44
substance causing stickiness, then followed by the National Research Committee on Cotton severed
programs (Ali and Khalifa 1982, Khalifa 2001) were launched to address stickness. The programs
included:
Type of sugars causing cotton stickiness.
Quick methods for grading cotton stickiness
Ginning efficiency, and the spinning performance.
Integrated pest management (IPM package)
Breeding of cotton varieties tolerant to whitefly infestation.
These research programs continued and in a very short time main results by researcher were revealed.
Ali and Khalifa (1980) found that the sugar deposits mainly consist of fructose, glucose and mannose.
The whitefly excretions contained two additional unidentified components X and Y which were absent in
aphid secretions. They also reported that the sugar deposits causing stickiness were mainly the excretions
of whitefly; those of the aphids ranking second.
Results of the chemical method correlate very well with the results of the mini-card (Ali and Khalifa
1980). This method was modified to suit commercial application by shortening the test period, as well as
reducing the amount of the chemical used (Ali 1998). It was also reported that, the sticky cotton may
decrease the output of the roller gin to about 5-7 kg/gin/hour, compared to 25-30 kg/gin/hour for the
clean cotton (Khalifa and Gameel. 1983). It was found that the distribution of honeydew within the same
plant was variable. The level of cotton stickiness was higher in lint collected from bottom and middle of
the plant compared with the top (Khalifa 1982). Whitefly usually prefers humid, warm and shady
conditions, as well as protection from wind. (Gameel 1982). It was also found that the medium staple
cotton (Acala) showed higher stickiness level compared to extra long staple cotton (Barakat). This is
mainly because hirsutum (Acala) varieties are hairy and bushy, and hence more susceptible to whitefly
infestation (Khalifa 1982). It was found that a single adult could produce excretion that can cover 38
mm
2
of leaf surface in one day (Gameel 1968).
The whitefly has a wide range of host plants and cotton is normally planted in Sudan during the
period July- August. When the other host plants start to dry up white flies migrate to cotton and start
breeding rapidly during September-November. They have about 10-12 generations per growing season
(Khalifa 1982). Also, distribution of honey-dew within the same plant was variable. A long term program
was conducted (early 1980s) to breed for tolerant and resistant cultivars to whitefly infestation. Its main
objective was to manipulate the morphological and physiological characters of the cotton plant in such a
way so as to reduce the whitefly population and allow for easy biological, chemical and cultural control
(Okra shape high gossypol content). Despite research effort the problem of stickness in Sudan cotton
persisted.
GLOBAL RESEARCH PROGRAM
Research programs addressing the causes and control measures were carried out by ARC. During 1998-
2000, a stickiness research project financed by the Common Fund for Commodities (CFC) was executed
with the objectives of developing an objective methodology (rather than the current subjective methods
in use) to separate sticky from non-sticky cotton in order that the non-sticky part could be sold at due
price. The partners of this project were the Sudan Cotton Company (SCC) and the Agricultural Research
Corporation (ARC) in Sudan, the Institut Franais du Textile et de lHabillement (IFTH) and the Centre
de Coopration Internationale de Recherche Agronomique pour le Dveloppement (CIRAD) in France.
The methodology was developed and, in addition, the study revealed considerable variability in
stickiness levels among the cotton production areas, and considerably low levels of stickiness were
observed in some schemes.
Efforts to Mitigate Stickiness Problem in Sudan 267
.
Source: Gourlot et al -2011: ITMF continuation survey (Gourlot et al., 2011) indicated that few stickness at a problem
with Sudaness cotton.
Fig. 1: Mentioned Stickiness Problems for Sudan Production (in % of Answers), ITMF Cotton Contamination Surveys
REFERENCES
[1] Ali, N. A. and H. Khalifa (1980). Development of methods to measurecotton stickiness. Cot.Fib.Trop.xxv,4, 311-313.
[2] Fadlalla, A.S. (1998), Summary of project Rationale, Objectives and
[3] Execution. Annual Report, cotton Stickiness Project.
[4] Khalifa, H. (1980). Cotton stickiness. Paper presented before the
[5] Constituent Assembly of the International Committee for Cotton Testing, Bremen- Germany.
[6] Khalifa, H. (1982) Variation of cotton stickiness and methods of sampling Proc. of International Committee for Cotton
Testing Conference.Bremen-Germany.
[7] Khalifa, H. and Gameel. O.I. (1982). Breeding cotton varieties resistant to
[8] Whitfly (Bemisa tabaci:Genn).Symposium on cotton production and marketing. Khartoum..Sudan.pp.9.
[9] Khalifa, H.(1982). The control of cotton stickiness through breeding resistant cotton (Bemisa tabaci:Genn). Proc. Of
workshop of Advisory Group Meeting on the use of Nuclear Techniques for the improvement of oil seeds and other
industrial crops. Proc.IAEA/FOA p233-240, Dakar- Senegal.
[10] Gourlot J.-P.
)
, Abdin M. A.
)
and Latif A., Abdalla A. (2011) long term benefit of a CFC/ICAC project global improvement
of the situition
0
10
20
30
40
50
60
70
80
90
100
1989 1991 1993 1995 1997 1999 2001 2003 2005 2007 2009 2011 2013
Year
Present Status of Mealy Bug Phenacoccus solenopsis

(Tinsley) on Cotton and Other Plants
in Sindh (Pakistan)
Khuhro S.N.
1
, A.M. Kalroo
1
and R. Mahmood
2

1
Central Cotton Research Institute Sakrand67210 SindhPakistan
2
CABI South Asia, RawalpindiPakistan
AbstractIn Pakistan mealy bug Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) was recorded first
time in 2005 on cotton and other plants. The survey was carried out in different districts of Sindh to know the status
of the mealy bug on cotton and other plants. The pest was widely sprayed in the surveyed areas attacking a number of
plants including cotton. Mean maximum population (mealy bug 2
nd
& 3
rd
instars and adults/shoot) was recorded in
the districts Shaheed Benazirabad 46.93 followed by Ghotki district (38.88), Sukkur, (32.17), Naushahro Feroze
(32.07), Khairpur (29.67), and Dadu district (14.69). Mealy bug was recorded on 22 plants in Shaheed Benazirabad
district. On unsprayed cotton (95%) mealy bugs were found parasitized by Aenasius bambawalei Hayat, followed by
(92%), on Abutilon indicum (91%), okra (87%), datura, (86%) , china rose (80%) on egg plant, and on tomato (77%)
during 2010. However, mealy bugs parasitized by Aenasius bambawalei very low in 2011 due to indiscriminate use of
pesticides and appearance of hyper parasitoid. Different insecticides were also tested for controlling mealy bug on
cotton. Maximum mortality of the mealy bug recorded in plots treated with Movento 20 SC (95.2%), followed by
Movento energy 480 SC (94.8%), Confidor 50 SC+ Ultra (93.3%), Profenofos 50 EC (92.69%), Confidor 70 WG
(92.40%), Fyfanon 57 EC (91.1%), Bono 20 SC (89.60), and Malatox 57 EC (84.65%) up to one week of spray. The
meteorological data revealed that mealy bug was more common in the field at temperatures in the range of 30.5-
39.5C.
Keywords: Phenacoccus solenopsis, Aenasius bambawalei, Parasitism and population
INTRODUCTION
Cotton, Gossypium hirsutum L., is the most important fiber crop of Pakistan. It is used in textile as well
as oil industries and earns foreign exchange through export in shape of raw cotton, cotton yarn, cloth,
garments and other products. It makes about 80% of national edible oil production (Agha, 1994). It
engages millions of employees in the farms and factories. It provides edible oil, animal feed, fiber, and
fuel to a large proportion of the urban and rural populations. It supplies raw material for about 1200
ginning units, 180 spinning units, about 470 textile mills, and 50 vegetable oil mills operating in the
country. It is also a major export item from the crop sector because it directly or indirectly contributes
about 66 percent to Pakistans export earnings (Government of Pakistan, 1995). Unfortunately, the crop
was severely attacked by many sucking and chewing insect pests including cotton mealy bug. Mealy
bugs have recently become abundant on cotton in Pakistan. These soft bodied insects belong to family
Pseudococcidae of order Hemiptera. About 5000 species of mealy bugs have been reported from 246
families of plants throughout the world. Among these, 56 species have been reported from 15 genera of
family Malvaceae, including cotton and many other plants of economic importance (Ben-Dov, 1994).
Mealy bugs were never been reported from cotton in Pakistan until 2005 when for the first time
Phenacoccus solenopsis Tinsley was recorded from Vehari-Punjab. This insect alone was held
responsible for the loss of 0.2 million bales (bale weighs 375 lbs or 170 kg) in 2007 in Pakistan
(Muhammad, 2007). Mahmood et al (2011) reviewed its world distribution. According to them it is a
new world species and has recently entered a number of countries in Asia and Australia. They reported
that this insect is widespread on the plains of Pakistan. Results of present studies carried out in different
districts of Sindh on its distribution and abundance are reported in this paper.

45
Present Status of Mealy Bug Phenacoccus solenopsis (Tinsley) on Cotton and Other Plants in Sindh (Pakistan) 269
Mealy Bug Population on Cotton and other Hosts
Regular survey of mealy bug was under taken to record the phenology and host range in different
areas/districts of upper Sindh including Shaheed Benazirabad, Naushahro Feroze, Dadu, Khairpur,
Sukkur and Ghotki from May to December 2010. Five terminal shoots each measuring 15cm long were
taken at random one each from the four corners and in the center of the of the cotton field. Samples of
mealy bugs collected from fields were kept in jars (laboratory (at 26 2
o
C temperature and 75.5% R.H).
Samples were kept in Petri-dishes for a week for parasitoids emergence, in the lab 2
0
C temperature and
75.5% R.H. Counts were made of healthy and mummified 2
nd
, 3
rd
instars and adult mealy bug individuals
from the samples. The observations on cotton were made from May to December 2010. Samples of same
size were also taken from other plants where the mealy bug was found. Similar experiments were
improved upon in 2011.
Efficacy of Insecticides for Controlling Mealy Bug on Cotton Crop
Eight insecticides were tested for the efficacy against cotton mealy bug. The crop was sown on 27-05-
2010 and crop was sprayed on 03-08-2010. The trial was conducted at CCRI-Sakrand Farm in
Randomized Complete Block Design (RCBD) with four replications. Plot size was kept at 30 x 40.
Spray were initiated when the mealy bug population increased. The control plot was kept unsprayed for
comparison of the pest population.
Phenology
During observations the mealy bug was found breeding profusely on cotton and other plants in May-
December. It seems to breed almost throughout the year.
Population Trends of the Mealy Bug on Cotton
The results showed that the mealy bug infestation started initially after germination of cotton plants. The
minimum infestation was in June, 2010 and maximum in September was recorded at all areas surveyed
(Table-1). The mealy bug infestations were comparatively higher at Shaheed Benazirabad and Ghotki,
compared with Khairpur, Naushahro Feroze, Sukkur and Dadu districts of Sind province (Table-1).
TABLE: 1. POPULATION OF MEALY BUG IN DIFFERENT DISTRICTS OF SINDH-IN 2010

Mealy bug Numbers Per Terminal Shoot in Following Districts
Shaheed Benazirabad Naushahro Feroze Dadu Khairpur Sukkur Ghotki
Jun. 10.14 5.11 0.78 0.14 4.45 0.22
Jul. 16.33 14.11 2.36 3.43 13.78 13.50
Aug. 23.70 17.10 7.10 19.30 16.8 10.90
Sept. 84.30 8.54 18.0 60.80 24.10 133.33
Oct. 113.00 98.30 22.45 67.30 65.80 72.20
Nov. 50.00 76.00 39.75 53.67 86.50 39.00
Dec. 31.00 5.29 12.33 3.00 13.75 3.00
Mean 46.93 32.07 14.69 29.67 32.17 38.88
Mealy bug on plants other than cotton
The mealy bug was recorded from more than 22 plants however it was consistently found on egg
plant, tomato, Abutilon indicum, okra, hollyhock and china rose. At the peak period of its population in
September, it was found most abundant on cotton, followed by china rose, Abutilon indicum, okra,
eggplant, tomato and hollyhock (Table-2). Commonality of the mealy bug on different plants has been
reported by Arif, et al. (2009) who reported the inadence of mealy bug Phenacoccus solenopsis on about
154 plants but was most abundant on cotton.
TABLE 2: MEALY BUG POPULATIONS ON DIFFERENT PLANTS HOSTS IN SINDH IN SEPTEMBER, 2010
English/ Local Name Technical Names Mean Mealy Bug Infestation/ Shoot
1. Egg plant Solanum melongena 45.12
2. Tomato Lycopersicon esculentum 23.41
3. Abutilon Abutilon indicum 65.74
4. Cotton Gossypum hirsutum 84.64
5. Datura Datura alba 24.46
6. Okra Abelmoschus esculentus 77.13
7. Hollyhock Alcea setosa 12.71
8. China Rose Hibiscus rosa-sinensis 69.23
Natural Enemies of the Mealy Bug
Since mealy bug appearance was recorded during 2005 in Pakistan only insecticides have been tried to
control the mealy bug on cotton. Natural enemies did not have much role in controlling the mealy bug.
Mahmood et al. (2011) developed techniques of conserving predators and parasitoids in field conditions
and successfully bred millions of parasitoids and predators using plant debris (mealy bug infested drying
twigs and leaves). They reported a number of predators associated with the mealy bug in 2006-2007,
however, parasitoid Aenasius bambawalei Hayat was first time reported during 2008 from Tando Jam
Sindh-Pakistan (Solangi and Mahmood, 2011). This parasitoid spreads fast and keeps the mealy bug
under control. In sprayed cotton fields though parasitoid was rare it was most common on unsprayed
cotton fields and helped keep the pest under control (Table- 3). The parasitoid was not only common in
cotton but also was common on other plants and most of the mealy bugs were found parasitized. In 2011
the parasitoids population was less than 2010 (Table-4).
The main reason of low population of parasitoid is the adverse effect of large scale use of pesticides
in cotton and vegetables. Moreover a hyper parasitoid Promuscidea unfasciativentris Girault has
appeared thereby impacting parasitoid population. As a result of decline in population of the parasitoid
the mealy bug population has increased severely.
TABLE 3: PARASITISM OF AENASIUS BAMBAWALEI ON DIFFERENT HOST PLANTS AT SHAHEED BENAZIRABAD DISTRICT IN AUGUST 2010
English/Local Name Technical Names Parasitism Percent
1. Eggplant Solanum melongena 80
2. Tomato Lycopersicon esculentum 77
3. Abutilon Abutilon indicum 92
4. Cotton Gossypum hirsutum 95
5. Datura Datura alba 87
6. Okra Abelmoschus esculentus 91
7. China Rose Hibiscus rosa-sinensis 86
Efficacy of Insecticides for Controlling Mealy bug on Cotton Crop
TABLE 5: EFFICACY OF INSECTICIDES FOR CONTROLLING MEALY BUG AT CCRI-SAKRAND DURING AUGUST 2010
Treatment Dose/ acre (ml/g) Post-Treatment Average
Population/Shoot
Mortality (%)
48 hours 72 hours 1 week 48 hours 72 hours 1 week
Bono 20 SL 125 ml 26.07 17.41 13.17 76.89 85.70 89.60
Malatox 57 EC 750 ml 31.12 22.01 19.45 72.41 81.90 84.65
Profenofos 50 EC 500 ml 18.31 11.21 9.26 83.76 90.78 92.69
Confidor 20 SC 250 ml 17.0 8.0 5.0 83.5 93.5 95.2
Fyfanon 57 EC 500 ml 33.0 21.0 10.0 66.6 82.9 91.1
Confidor 20 SL+ ultra 250 ml 34.0 13.0 7.0 65.8 89.8 93.3
Confidor 70 WG 140 gm 25.0 12.0 7.0 74.6 88.1 92.4
Movento energy 480 SC 150 + 250 ml 30.0 10.0 5.0 70.2 91.6 94.8
Control - 112.81 121.62 126.74 - - -

Present Status of Mealy Bug Phenacoccus solenopsis (Tinsley) on Cotton and Other Plants in Sindh (Pakistan) 271
Results given in Table-5 indicate that, Movento 20 SC gave maximum (95.2%) mortality followed
by Movento Energy 480 SC, Confidor 50 SC+Ultrs, Profenofos 50 EC, Confidor 70 WG, Fyfanon 57
EC, Bono 20 SC and Malatox 57 EC up to one week of spray. Similes results were reported by (Aheer,
et al. 2009) who also mentioned that all tested insecticides proved significantly effective against mealy
bug up to 7 days after treatment.
METEOROLOGY DATA
The meteorology data was recorded during the survey of cotton mealy bug at CCRI-Sakrand Farm. The
results showed that the mealy bug built up its population when the temperature 29
0
C and was maximum
in the temperature range of 30.5-39.5
0
C and decreased at temperatures below 29
0
C. (Table 2,3&6).
TABLE 6: METEOROLOGICAL DATA OF 2010 SEASON RECORDED AT CENTRAL COTTON RESEARCH INSTITUTE, SAKRAND, SINDH-PAKISTAN
Month Mean Mealy Bug
Population/ Shoot at
Shaheed Benazirabad
Average
Maximum Temp.
and Range
o
C
Average
Minimum Temp.
and Range (
o
C)
Mean relative
Humidity and
Range (%)
Rainfall
(mm)
Jun. 10.14 40.6(29.0-45.0) 27.4 (25.0-31.0) 57.8(47.0-91.5) 45.2
Jul. 16.33 38.6(35.0-43.0) 28.1(24.5-29.0) 65.7(56.0-90.5) 136.2
Aug. 23.70 35.7 (30.0-38.0) 27.2 (24.5-29.0) 73.8(63.5-90.5) 72.0
Sep. 84.30 34.3 (33.5-38.0) 25.1(21.5-30.0) 75.6(56.0-93.5) 50.0
Oct. 113.00 35.7 (30.5-39.5) 21.9(18.0-25.0) 55.9(41.7-66.0) -
Nov. 50.00 29.1(24.0-33.0) 14.9 (8.5-20.5) 52.7(41.3-69.0) -
Dec. 31.00 23.2(20.0-25.0) 7.5(4.0-10.0) 55.3(42.0-72.7) -
ACKNOWLEDGEMENT
We acknowledge the financial assistance by former Ministry of Food and Agriculture, Government of
Pakistan through PSDP, to carry out the present studies under the project Biological control of major
cotton pests including mealy bug in Pakistan Sakrand Component. We specially thank Mr. Arshad
Ahmed, Vice President and Dr. Tasawar Malik, Ex-Director Research, Pakistan Central Cotton
Committee (PCCC) and Dr. Ibad Badar Siddiqi, Project Director BCMCP for their consistent support in
conducting research.
REFERENCES
[1] Agha, H. K. 1994. Crop Production. Published by Pakistan Book Foundation, Islamabad Pp.6.
[2] Aheer, G. M. Riaz Ahmad; Amjad Ali . 2009. Efficacy of different insecticides against cotton mealybug, Phenacoccus
solani Ferris. Journal of Agricultural Research (Lahore) Vol. 47 No. 1 pp. 47-52
[3] Arif M.I, Wazir S, Rafiq M, Ghaffar A, and Mahmood R. 2011. (Incidence of Aenasius bambawalei Hayat on mealybug
Phenacoccus solenopsis Tinsley and its hyperparasite, Promuscidea unfasciativentris Girault at Multan).
http://www.icac.org/tis/regional_networks/asian_network/meeting_5/documents/papers /PapArifMI-et_al.pdf
[4] Arif, M.I., M. Rafiq, and A. Ghaffar, 2009. Host plants of cotton mealy bug (Phenacoccus solenopsis): A new menace to
cotton agro ecosystem of Punjab, Pakistan. International Journal of Agriculture and Biology 11: 163-167.
[5] Ben-Dov, Y. 1994. A systematic catalogue of the mealy bugs of the world (Insecta: Homoptera: Coccoidea:
Pseeudococcidae and Putoidae). Intercept Ltd., Andover, P: 686.
[6] Mahmood, R, M. N. Aslam, G. S. Solangi and A. Samad. 2011. Historical Perspective and achievements in biological
management of cotton mealy bug Phenacoccus solenopsis Tinsley in Pakistan. 5th Meeting Asian Cotton Research and
Development Network, held during February 23-25. Lahore, pp. 1-17. Online at:
[7] http://www.icac.org/tis/regional_networks/asian_network/meeting_5/documents/papers/MahmoodR.pdf
[8] Muhammad, A. 2007. Mealy bug: Cotton Crops Worst Catastrophe. Centre for Agro-Informatics Research (CAIR),
Pakistan. Available on-line at http://agroict.org/pdf_news/Mealybug.pdf accessed Jul.2008 (verified 27 May 2009).
[9] Government of Pakistan, 1995. Economic Survey, 1994-95. Islamabad: Finance Division, Economic Advisers Wing.
[10] Solangi G. S. and R. Mahmood. 2011. Biology, host specificity and population trends of Aenasius bambawalei Hayat and
its role in controlling mealy bug Phencoccus solenopsis Tinsley at Tandojam Sindh. 5th Meeting Asian Cotton Research
and Development Network held on February 23-25. Lahore, pp. 1-7. Online at:
[11] http://www.icac.org/tis/regional_networks/asian_network/meeting_5/documents/papers/PapSolangiGS-et_al.pdf
Changing Scenario of Cotton Diseases

in IndiaThe Challenge Ahead
D. Monga
1
, K.R. Kranthi
2
, N. Gopalakrishnan
3
and C.D. Mayee
4

1
Central Institute for Cotton Research (CICR), Regional Station, Sirsa
2
C.I.C.R. Nagpur,
3
Rishi Bhawan, New Delhi
4
Agriculture Scientists Recruitment Board, New Delhi
AbstractThe cotton disease scenario has shown a continuous change during the past sixty four years since
independence. When mainly indigenous diploid cottons were being grown in fifties, Fusarium wilt, root rot, seedling
blight, anthracnose and grey mildew were the major problems. With the large scale cultivation of tetraploid upland
cotton (Gossypium hirsutum), bacterial blight became the major disease to which indigenous cottons were highly
resistant. After the introduction of Bt cotton hybrids during 2002 onwards and continuous increase in area under
these hybrids to around 85% of total cotton area till date, the disease scenario has also shown some change. The grey
mildew, once a serious problem for diploid cottons especially in central India has now become a major problem in Bt
cotton hybrids. Grey mildew (percent disease intensity) in central zone was recorded on Bt cotton hybrids during
2010-11 in Maharashtra in the irrigated areas of Vidarbha region (9.2 to 20.4 % & Nanded-6.5 to 27.2%). In south
zone it was severe in two states i.e. Karnataka (5.0-30.0%), and Andhra Pradesh (28.9-46.4) during the season.
Among other important diseases on Bt hybrids, Bacterial blight was reported as important disease in central zone in
Maharashtra ( Vidarbha- 8.3 to 22.2 %; Nanded 2.2 to 15.7 %) and in south zone in Karnataka (5.0-15.0 %) and
Andhra Pradesh ( 8.0-47.6%). Alternaria blight was observed serious during 2010-11 season in Gujrats Saurashtra
area (2.0-15.0%) and Maharashtras Rahuri (10.2-35.8%) and Nanded (5.0-21.5%) and in south zone states ie
Karnataka (5.0-30.0%), Andhra Pradesh (10.0-54.6%) and Tamil Nadu from 12.6 to 38.8%. (Anonymous 2011).
Fusarium wilt has become less important as upland cotton now occupying 85% area is immune to Indian race of
the pathogen. Verticillium wilt which appeared in Tamil Nadu remained restricted mainly to that state only.
In north India, the leaf curl disease caused by gemini virus and transmitted by white fly Bemisia tabaci has
become a threat to cotton cultivation due to development of new recombinant strains and introduction of a number of
susceptible Bt cotton hybrids in north zone. A disease identified as Tobacco Streak Virus (Ilar virus) transmitted by
thrips was observed in the transgenic cotton growing region of Southern Maharashtra and Andhra Pradesh. (Sharma
et al, 2007). Avoidable losses due to important diseases like cotton leaf curl virus, (53.6% ),bacterial leaf
blight(20.6%), Alternaria leaf spot (26.6%), grey mildew( 29.2%)and Myrothecium leaf spot ( 29.1%) have been
documented. Newer chemicals like propiconazole, captan+hexaconazole, tetraconazole and strobilurin compounds
(fungicides) and copper hydroxide (bactericides) have been successfully tested for the management of foliar disease of
cotton. Strategies for the integrated management of diseases causing losses in terms of yield and quality need to be
redefined.
INTRODUCTION
Cotton is an important crop for the sustainable economy of India and livelihood of the Indian farming
community. It is cultivated in 11.0 M hectares in the country. India accounts for about 32% of the global
cotton area and contributes to 21% of the global cotton produce, currently ranking second after China.
The production increased from a meager 2.3 M bales (170 kg lint/bale) in 1947-48 to an all time highest
record of 31.5 M bales during 2007-08. Cotton provides employment and sustenance to a population of
nearly 42 M people, who are involved directly or indirectly in cotton production, processing, textiles and
related activities. India has the unique distinction of being the only country in the world to cultivate all
four cultivable Gossypium species, Gossypium arboreum and G.herbaceum (Asian cotton), G.
barbadense (Egyptian cotton) and G. hirsutum (American upland cotton) besides hybrid cotton.
Approximately 65% of Indias cotton is produced under rainfed conditions and 35% on irrigated lands.
Cotton is cultivated in three distinct agro-ecological regions (north, central and south) of the country. The
northern zone is almost totally irrigated, while the percentage of irrigated area is much lower in the
central (23%) and southern zones (40%). Cotton crop is particularly sensitive to a number of biotic and
abiotic stresses and the disease problems are also distinct to some extant in agro-ecological regions
46
Changing Scenario of Cotton Diseases in IndiaThe Challenge Ahead 273
referred above. A number of diseases are prevalent on cotton crop in one part of the country or another.
Under north zone, cotton leaf curl virus and root rot diseases are the major problems whereas grey
mildew, bacterial blight and Alternaria blight are severe in one or the other region in central and southern
zone.
In the present review paper, the historical background and present status of cotton diseases in India has
been presented. The information gathered on seed borne diseases, soil borne diseases and emerging
diseases is highlighted. Among emerging diseases, cotton leaf curl virus, grey mildew and fungal foliar
diseases are covered. An attempt is made to present this information in the context of Bt cotton hybrids
presently grown in the country. This is followed by information on changing disease scenario of
important diseases with major emphasis on cotton leaf curl virus disease and development of new
recombinants during recent years, preparation of disease maps and epidemiological studies and their
implications on disease scenario. Economic losses due to diseases and new molecules for disease
management are described in subsequent sections.
A number of diseases are prevalent on cotton crop in one part of the country or another (Table 1).
TABLE 1: MAJOR COTTON DISEASES IN INDIA AND EMERGING SCENARIO
Disease Causal Agent Remark
Seed Borne and Foliar Diseases
Cotton leaf curl Gemini virus North zone (Potential threat)
Grey mildew Ramularia areola Central & South zone (Emerging)
Bacterial blight Xanthomonas axanopodis pv malvacearum Maharashtra, Gujrat, Karnataka
Alternaria leaf spot Alternaria macrospora Maharashtra, Gujrat, Karnataka
Myrothecium leaf spot Myrothecium roridum Madhya Pardesh
Leaf Rust Phakopsora gossypii Kanataka, Andhra Pardesh (Emerging)
Cercospora leaf spots Cercospora gossypina Andhra Pardesh (Minor)
Helminthosporium leaf spots Helminthosporium gosyypii Andhra Pardesh (Minor)
Anthracnose Colletototricum capsici South zone (Minor)
Tobacco streak virus Ilar virus Andhra Pardesh(Emerging)
Soil Borne Diseases
Wilt Fusarium oxysporum fsp.vasinfectum Restricted to diploids
Root rot Rhizoctonia solani, R. bataticola Scattered
Verticillium wilt Verticillium dahliae Tamil Nadu, Karnataka
HISTORICAL BACKGROUND AND PRESENT STATUS OF COTTON DISEASES IN INDIA
The cotton disease scenario has shown a continuous change during the past sixty four years. Initially,
mainly indigenous diploid(Gossypium arboreum & G herbaceum)cottons were being grown in fifties and
the Fusarium wilt, root rot, seedling blight, anthracnose and grey mildew were the major problems. With
the large scale cultivation of tetraploid upland cotton (G hirsutum), bacterial blight became the major
problem to which indigenous cottons were highly resistant. The susceptibility of American cottons is
attributed to their not having been exposed to the disease till its introduction into the Americas in post
Columbian times. Fusarium wilt became less important as upland cotton (Bt cotton hybrids) now
occupying above 85 percent area is immune to Indian race of the pathogen. Verticillium wilt which
appeared in Tamil Nadu remained restricted mainly to South zone only. The grey mildew, once a serious
problem for diploid cottons especially in central India with the continued cultivation and imposed
selection pressure got adopted to tetraploid cotton and their hybrids as well. Presently, it is a problem in
central and south India in Bt-cotton hybrids. Alternaria blight and Myrothecium leaf spots are prevalent
everywhere but are severe in the states of Karnataka and Madhya Pardesh, respectively. Other diseases
such as Cercospora and Helminthosporium leaf spots are sporadic only. Rust, although a minor disease
may assume significance in southern states in near future. In north India, the cotton leaf curl virus disease
(CLCuD) caused by a Gemini virus and transmitted by whitefly, Bemisia tabaci has become a major
threat to cotton cultivation since its appearance in 1993.
Grey mildew (percent disease intensity) in central zone was recorded on Bt cotton hybrids during
2010-11 in Maharashtra in the irrigated areas of Vidarbha region (9.2 to 20.4 % & Nanded-6.5 to 27.2%).
In south zone it was severe in two states ie Karnataka (5.0-30.0%), and Andhra Pradesh (28.9-46.4)
during the season. Among other important diseases on Bt hybrids, Bacterial blight was reported as
important disease in central zone in Maharashtra (Vidarbha - 8.3 to 22.2 %; Nanded 2.2 to 15.7 %) and in
south zone in Karnataka (5.0-15.0 %) and Andhra Pradesh (8.0-47.6%). Alternaria blight was observed
serious during 2010-11 season in Gujrats Saurashtra area (2.0-15.0%) and Maharashtras Rahuri (10.2-
35.8%) & Nanded (5.0-21.5%) and in south zone states ie Karnataka (5.0-30.0%), Andhra Pradesh (10.0-
54.6%) and Tamil Nadu from 12.6 to 38.8%. (Anonymous 2010). A disease identified as Tobacco Streak
Virus (Ilar virus) transmitted by thrips was observed in the transgenic cotton growing region of Southern
Maharashtra and Andhra Pradesh. (Sharma et al, 2007). During the surveys conducted around Guntur
(Andhra Pradesh) from September 2010 to January 2011 Tobacco Streak Virus disease incidence on
different Bt cotton hybrids varied from 1.0 to 43.7%. Maximum incidence was recorded during
September in four months old crop. The disease did not appear to cause significant losses at present
(Anonymous,2011).
The prevailing disease problems can be broadly divided into (i) Seed borne diseases (ii) soil borne
diseases and (iii) Emerging diseases.
Seed Borne Diseases
Studies on seed transmission of cotton diseases conducted at central institute for cotton research, Nagpur
(1983-1998) have indicated that leaf and boll spot pathogen Alternaria macrospora and the anthracnose
pathogen Colletotricum capsici could become deep seated (embryo borne) and seed transmitted in diploid
cottons (Gossypium arboreum, G herbaceum) varieties and hybrids. The bacterial blight caused by
bacterium Xanthomonas axanopodis pv. malvacearum was found seed transmitted mainly in tetraploid
cotton (G hirsutum, G barbadense) varieties and hybrids. The black boll rot fungus Botrydiplodia
theobromae and the stem break/root rot pathogen Macrophomina phaseolina were recorded seed
transmitted both in diploid and tetraploid varieties and hybrids (Mukewar and Kairon,2001).
Myrothecium blight has also been shown as seed borne in nature ( Srinivasan, 1994).
Soil Borne Diseases
Root Rot caused by Rhizoctonia solani and R. bataticola and wilt caused by Fusarium oxysporum f.sp.
vasinfectum are the two major soil borne fungal disease problems in India. Another soil borne disease
Verticillium wilt has been observed in some areas of Tamil Nadu and Karnataka. The root rot disease is
serious in northern India and detailed studies on various aspects have been undertaken (Monga and
Raj,1994; Monga and Raj,1994a; Monga, 1995 ; Monga and Raj, 1996; Monga and Raj,1996 b; Monga,
1997; Monga and Raj,2000 ; Monga, 2001 ; Monga and Raj, 2003 ; Monga, et. al., 2004a). The disease
affects both the hirsutum (American cotton) and arboreum (Desi) cotton species, being more serious on
desi cottons (Monga and Raj, 1994a). The wilt disease caused by Fusarium oxysporum f sp. vasinfectum
appears at any stage of plant development and affects only Desi cottons. Symptoms of Verticillium wilt
depend on the cultivar, virulence of the fungal isolate, development stage of the plant and environmental
conditions especially temperature.
EMERGING DISEASES
Cotton Leaf Curl Virus Disease
Cotton leaf curl virus disease caused by whitefly (Bemisia tabaci) transmitted Gemini virus with single
stranded circular DNA was observed during 1993 around the border areas in Rajasthan and Punjab. The
disease in a short span of 4-5 years spread in the entire north zone as the G hirsutum varieties like F-846,
RST-9 and HS-6 being grown in the region at that time were highly susceptible to this disease. The
initiation of disease is characterized by small vein thickening (SVT) type symptoms on young upper
leaves of plants. Upward/downward leaf curling followed by formation of cup shaped leaf laminar out
growth of veinal tissue on the abaxial side of the leaves is other important symptom. In severe cases
reduction of internodal length leading to stunting and reduced flowering/fruiting is also noted.
TABLE 2: PROMISING CLCUD TOLERANT VARIETIES/ HYBRIDS/ BT HYBRIDS IN NORTH ZONE
Name of Variety/ Hybrid Source
H-1226, H-1117(Varieties), HHH-223, HHH-287(Hybrid) CCS Haryana Agricultural University, Hisar
F-1861,LH 2076(Varieties), LHH-144 (Hybrid ) Punjab Agricultural University, Ludhiana
RS-875, RS-810,RS-2013 Rajasthan Agricultural University, (Bikaner) Sriganganagar
Shresth (CSSH 198), Kalyan (CSHH-238), Simran (CSHH 243) Central Institute for Cotton Research, Regional Station,
Sirsa
MRC 7361, MRC 6025, MRC 7031 BG II, MRC-7017 BG II,
MRC-6304, SP-7007, SP-7010,SWCH 4711, BIOSEED 6488,
BIOSEED 6488 BG-II, BIOSEED 6317, BIOSEED 2113,
BIOSEED 6588 BGII, PCH 877, BIOSEED 6588, ANKUR
3028, SHAKTI -9, VBCH 1008, VBCH 1534, VBCH 1518
BGII, NCEH 31, NCEH 6, JKCH-1, RCH 605, RCH 569 BG
II, NCS 855 BGII, NCS 905, VICH-307, VICH-309 BGII, PCH
401,
Private Sector
TABLE 3: COTTON LEAF CURL VIRUS DISEASE HOSTS REPORTED FROM INDIA
Name of Host Type of Test Reference
Sida sps, Abutilon Indicum, Hibiscus rosa
sinensis, Althea rosea
Based on visual symptoms Singh et al.,1994
Phaseolus vulgaris, Capsicum annum, Nicotiana
tabacum, Lycopersicum esculentum
Transmission studies and ELISA Nateshan et al.,1996
Abelmoschus esculentus, Althea rosea, Physalis
floridana, Nocotiana benthamiana, Phaseolus
vulgaris
Transmission studies Radhakrishnan et al., 2001
Althea rosea, Sida sps., Ageratum sps., Hibiscus
rosa sinensis
DNA-A probe hybridization Sharma, 2002
Tribulus terrestris, Cucumis sps. CLCuRv-CPgene and DNA beta
amplification
Sivalingam et al., 2004
Chorchorus acutangularis, Melilotus indica,
Ageratum conyzoides
DNA-A & DNA beta probe
hybridization
Radhakrishnan et al., 2004

Nicotiana tabacum, Lycopersicum esculentum,
Zinnia elegans, Mentha arvensis, Capsicum sps,
Hibiscus rosa sinensis, Abelmoschus esculentus,
Sida alba
PCR using CP primer

Kang et al., 2004

Sida sps., Achyranthus sps., Clearodendron sps. CP gene amplification Monga et al., 2005
Convolvulus arvensis, Capsicum sps., Pathenium
sps., Solanum nigrum, Digeria arvensis, Lantana
camara, Achyranthus aspera, Chenopodium album,
Spinacea sps., Xanthium strumarium
CP gene amplification

Monga et al., 2011b
A vigorous exercise was then taken up by the state agricultural universities and institutions under
Indian Council of Agricultural Research (ICAR) in the region to work out strategies for its management.
Molecular diagnostic tools for detection of virus were developed.(Chakrabarty et al., 2005). The disease
could be managed by development of resistant varieties/hybrids (Table 2), control of its vector whitefly
and eradication of weeds (Table 3) harboring cotton leaf curl virus disease (Narula et al., 1999 and
Monga et al., 2001). The disease was brought under control and the damage caused by it was
considerably reduced. The disease till date is restricted to northern cotton growing zone.
The Bt cotton hybrids were introduced in north zone in 2005 by the private sector initially with six
hybrids approved by Genetic Engineering Approval Committee. Subsequently, however, a large number
of hybrids were permitted for cultivation with in a span of five years and amongst them a number of
hybrids were observed to be highly susceptible to cotton leaf curl virus disease. As a result, the incidence
of disease increased and it became an emerging problem after the introduction of susceptible Bt cotton
hybrids in north zone. Yield loss estimation was studied in Bt cotton hybrids from 2008-09 to 2010-11
based on percent disease index (PDI) ranging from 5% to 60% and disease severity grades from I to IV.
Based on PDI seed cotton yield reduction ranged from 0.08-59.5% at 5-60 PDI. Seed cotton yield
reduction ranged from 7.2-80.1% at severity grades of one to four in different hybrids (Monga
et al., 2011b). At present, Bt cotton hybrids tolerant to cotton leaf curl virus disease have been identified
under Technology Mission of Cotton project and being advocated to farmers (Table 2).Experiments
conducted under AICCIP (2009-11) have shown average losses of 53.6% in some Bt cotton hybrids.
Grey Mildew
The disease caused by Ramularia areola is characterized by irregular, angular, pale, translucent spots
measuring 1-10 mm in size surrounded by veinlets. The disease appears on the older leaves usually when
the plants are reaching maturity. A frosty or mildew growth consisting of conidiophores of the fungus
appear first on the under surface and subsequently on the upper surface of affected leaves. As the
infection progresses leaves become yellowish brown and fall off prematurely. The incidence of grey
mildew is assuming a serious position in central and southern zone. Majority of released Bt hybrids fall
in moderately susceptible to highly susceptible category (Hosagoudar et al., 2008).
Foliar Spots
The bacterial blight caused by Xamthomonas axanopodis pv malvacearum with four distinct phases of
the disease (seedling phase, angular leaf spot and vein blight phase, black arm phase and boll rot phase)
used to cause considerable losses till 90s. Gossypium barbadense is more severely affected than G.
hirsutum. Resistant genes and occurrence of physiological races of the pathogen were described in detail.
Sources of resistance available in G hirsutum include 101-102B, BJA-592, Reba B-50, P14-T-128, HG-9,
Tamcot-CAMD-E, TxBonham, BJR 734, C-1412, Badnawar-1 and Khandwa-2 and have been used
extensively to develop resistant varieties/hybrids ( Srinivasan, 1994). The primary symptoms due to
Alternaria macrospora on leaves are small pale to brown round or circular spots (0.5-3.0 mm diameter)
showing concentric rings with cracked centre. These spots coalesce to form larger lesions (1 cm
diameter). Severe infection may lead to considerable defoliation. Stem cankers are formed in severe cases
and the infection may even reach bolls. Natural infection of seeds or seed inoculation results in disease
on cotyledons. The characteristic symptoms caused by Myrothecium roridum are the appearance of
circular or oval light ash coloured spots with violet to reddish brown margin. Fruiting bodies
(Sporodochia) are produced in concentric rings and protrude from lower as well as upper surface of
leaves. Under severe conditions, the lint gets strained to yellow or light brown. The rust caused by
Phakospora gossypii (Arth) Hirat. F. occurs sporadically in Tamil Nadu, Andhra Pardesh and Karnataka
during December-March. Its early appearance has potential to cause considerable loss by decreasing the
photosynthetic area and heavy defoliation. The pathogen initially affects the older leaves and then
spreads to the younger ones. Only the uredial stage of the rust occurs in India. Uredial sori appear on the
leaves as small (1-3 mm) pinkish brown spots which may coalesce to form larger patches. The uredia are
oval to circular on the pedicels and branches and the urediospores are exposed on rupture of the
epidermis. The early incidence of rust was noted during the last two seasons in Karnataka and Andhra
Pradesh (Anonymous, 2010 and 2011).
CHANGING DISEASE SCENARIO
When the incidence of important diseases in varieties/hybrids/Bt cotton hybrids was studied across the
country for the past three years, it was noted that CLCuD, Alternaria blight and Grey mildew showed an
increasing trend where as bacterial blight incidence did not show any trend but varied between 17.2 -
36.7% over locations (Table 4).

TABLE 4: INCIDENCE OF DISEASES (PDI) IN SCREENING NURSERIES
Disease Location Bt Hybrid/ Hybrid/ Variety 2008-09 2009-10 2010-11
Percent Disease Index
CLCuD Faridkot (N.Z.) F-846 0 29 96.2
Sriganganagar (N.Z.) RST-9 35.96 100 100
Bacterial blight Akola(C.Z.) RCH-2 26.3 30.42 22.08
Dharwad (S.Z.) RCH-2 36.73 27.08 17.2*
Surat (C.Z.) G. Cot hybrid-10 35.5 25.13 34.5*
Alternaria blight Rahuri (C.Z.) LRA-5166 28.66 29.3 36.6
Dharwad (S.Z.) RCH-2 21.28 32.67 39.5
Grey mildew Dharwad (S.Z.) RCH-2 11.53 18.75 32.2
N.Z.-North zone, C.Z.-Central zone, S.Z.-South zone
*Bunny Bt at Dharwad and G. Cot hybrid 12 at Surat was tested during 2010-11
The Bt-cotton hybrids have shown higher incidence of fungal foliar spots including grey mildews and
bacterial leaf blight diseases. A survey carried out by Hosagoudar et al. (2008) in eight districts in north
Karnataka on Bt cotton hybrids revealed higher incidence of grey mildew (5-40%), Alternaria blight
(5.0-35%) and bacterial blight (5.0-25%). In another study conducted in central zone, the bacterial blight
disease increased progressively and reached its peak in RCH-2 Bt cotton hybrid exhibiting 46.7%
incidence with intensity of 20.0% compared to 45.0% incidence and 18.7% intensity in susceptible
variety LRA-5166. Maximum temperature and sunshine hours exhibited positive and significant
correlation with disease incidence. Maximum temperature of 32.5
0
C and relative humidity above 81
percent with sunshine hours 7.9 contributed to rapid spread and development of this disease (Ingole et
al., 2008).
In northern cotton growing zone of the country consisting of about 15 lakh ha area, Leaf curl virus
disease is one of the most significant but highly complex disease of cotton caused by the whitefly
transmitted Geminivirus. Since the outbreak of the disease in 1994 in Sri Ganganagar region of Rajasthan
in north India, the disease established endemically in the entire North West Indian states of Punjab,
Haryana and Rajasthan causing moderate to severe losses. The cotton leaf curl virus (CLCuV) emerged
with renewed aggressiveness during the crop season of 2009-10, when some of the hitherto resistant
genotypes and hybrids succumbed to its onslaught. Regular monitoring of CLCuV affected cotton is done
to characterize variability in symptoms, diversity of sequences within the associated isolates and
variability in disease pattern, if any. Four distinct symptom types were documented viz, leaf curl with
prominent enations, severe leaf curl without prominent enation, upward and downward curling of leaves.
Sequences of DNA-A and beta DNA components of the isolates associated with different symptoms
showed existence of significant variation and recombination with other strains of CLCuV. Sequence
identity matrix and RDP analysis of DNA-A and beta DNA components of six virus isolates analyzed
over a period of four years from 2006 showed sequence homology and recombination among several
isolates from India and Pakistan. Isolate G6-DC, isolated from cotton cv. RS2013, with compromised
resistance and severe leaf curl isolate S2 analyzed during 2009-10, showed close resemblance to several
CLCuV isolates from Pakistan. DNA-A component of G6-DC had major recombination events with two
Pak strains, besides other Indian strains while S2 isolate showed major recombination with three Pakistan
strains. Accumulation of recombination events over the years coupled with favorable environmental
conditions appeared to have knocked down the resistance of cotton ( Chakrabarty et al.,2010).
After the appearance of cotton leaf curl virus disease in a severe form during 2009-10 crop season in
some areas of north zone, district level disease development maps were prepared and it was noted that in
Punjab, out of nine cotton growing districts, the disease was very severe (PDI >50%) in Ferozepur
followed by severe (25-50%) in Muktsar and Faridkot and moderate (5-25%) in Moga, Bhatinda, Sangrur
and Mansa districts. In Patiala, it was low (1.1-5%) whereas It was observed in traces (0-1.0%) in
Ludhiana. However, during 2010-11 season the disease was observed to be in severe form from moderate
during 2009-10 in Sangrur and Mansa district also indicating increased severity. Similarly in Haryana the
disease was observed in traces in the major cotton growing districts of Sirsa, Fatehabad, Hisar and Jind
whereas it was not observed in other districts like Rohtak, Bhiwani, Jhajjar, Mahindergarh and Rewari
during 2009-10. However during 2010-11 season the disease was quite widespread in Haryana and was
found to be moderate in Sirsa, Fatehabad and Hisar followed by low in Bhiwani and traces in Rohtak
districts. In Rajasthan during both the years, the disease was moderate in Sriganganagar district and low
in Hanumangarh (Monga et al., 2011a).
In recent years epidemiological studies have thrown light on the weather factors associated with
disease development and its progress. Step wise multiple regression analysis revealed that weather
parameters altogether accounted for 53.0 86.7% significant variation of cotton leaf curl virus disease
during 1999-2005 at Regional Research Station, Faridkot in the state of Punjab. Minimum temperature
alone contributed 70.2% negative significant variation whereas minimum relative humidity contributed
86.7% positive significant variation. Two week lag weather parameters played significant role in
appearance of the disease over the years (Singh et al.,2010).
In another study (1999-2009) conducted at CICR Regional Station Sirsa, the multiple regression
equation of current weekly progress of disease( per cent incidence) during 27 31 Met weeks was tried
with thirty independent variables (six weather factors for current as well as four prior/lag weeks) using
stepwise regression. The value of coefficient of determination ( R
2
) was found to be 0.8230. Minimum
temperature of current and one lag week and sunshine of three lag weeks were significantly negatively
correlated and contributed to 47% variation whereas morning RH of three lag weeks, evening RH of
current week & four lag weeks, rain fall of current and three lag weeks were significantly positively
correlated and contributed to 35% variation.(Monga et al.,2011c).
ECONOMIC LOSSES CAUSED DUE TO DISEASES
Alternaria leaf spots can cause loss upto 26.6% based on results (2006-07 to 2008-09) of study
conducted in central India at Rahuri and south zone locations at Guntur and Dharwad. Five sprays of
Propiconazole (0.1%) at 35, 50, 65, 80, and 95 DAS decreased percent disease index (PDI) from 31.6 to
20.8% thereby reducing this yield loss due to Alternaria leaf spots in variety LRA-5166 ( Anonymous,
2009). In case of grey mildew disease also, a reduction of loss due to grey mildew disease up to 29.2%
with the application of five sprays of carbendazim (35,50,65,80 and 95 days after sowing) in Bt cotton
hybrid Bunny was demonstrated based on a study (2008-09 to 2010-11) conducted across central and
south zone, (Dharwad, Guntur and Nanded ). PDI showed reduction to 8.1 as compared to 20.9 in control
(Anonymous,2011).
In another important fungal disease, Myrothecium leaf spot, a reduction of loss up to 29.1% with the
application of five fungicidal sprays of Propiconazole (@ 0.1%) at an interval of 35, 50, 65, 80 and 90
DAS in variety JK-4 was observed on the basis of trial in central zone at Khandwa (2007-08 to 2009-10).
Percent disease index (PDI) showed reduction to 7.4 as compared to 22.5 in control (Anonymous,2010).
Losses to the tune of 33.8% with 0.1% propiconazole spray at 35,50,65,80 and 95 days after sowing due
to Helminthosporium leaf spot disease could be avoided in cotton variety LRA-5166 based on (2007-08
&2008-09) studies carried out at Guntur in South zone ( Bhattiprolu,2010).
Reduction of losses due to bacterial leaf blight up to 20.6% with the application of five sprays at
35,50,65,80 and 95 days after sowing of Copper oxychloride (0.2-0.3%) and Streptocyclin (100-500ppm)
on the basis of two year trials (2009-10 &2010-11) in central zone at Surat and Akola and south zone at
Dharwad and Guntur were also noted (Anonymous 2010 and 2011).
NEW MOLECULES FOR DISEASE MANAGEMENT
Tetraconazole 11.6 % w/w 900 ml/ha showed effective control ( percent disease index 9.9 compared to
28.1 in control) of Alternaria leaf blight (tested at Arupkotai, Junagarh, Rahuri, Nanded during 2009-10
&2010-11) and led to 30.5% increase of seed cotton yield over control. Kresoxim methyl (Ergon 44.3%
at 500ml/ha) when tested against foliar pathogens( Alternaria blight, myrothecium leaf spot and grey
mildew) at seven locations showed significant reduction of percent disease index. (Anonymous 2010
and 2011).
The fungicide captan+hexaconazole (Taqat @500g/ha) tested at Coimbatore, Junagarh, Faridkot,
Guntur and Dharwad during 2007-08 &2008-09 significantly reduced fungal foliar leaf spots (
Alternaria, Myrothecium, Grey mildew, Helminthosporium and Cercospora leaf spots) with an increase
in seed cotton yied of 12% over control(Anonymous 2008 &2009). Taqat at at 500g/ha was economical
in managing fungal leaf spot diseases at Guntur with benefit cost ratio of 1.42 (
Bhattiprolu,2010).Evaluation of copper hydroxide ( Dharwad, Surat, Akola, Khandwa, Nanded and
Rahuri) during 2007-08 &2008-09 revealed significant reduction of bacterial blight and Alternaria spots
at 1500g/ha with maximum increase of seed cotton yield of 20.8% over control.(Anonymous 2008 and
2009).
CONCLUSION
Cotton leaf curl virus disease, an important problem presently restricted to north zone need to be dealt
more seriously in the context of changed scenario leading to the development of recombinants and
breakdown of resistance. The new sources of resistance should be identified from available germplasm.
The introgression of resistance from other available sources is another option. The work on development
of transgenics using RNAi technology is in progress and shall go a long way in development of
resistance against this important viral disease. Other components of integrated disease management
strategy like cultural practices including weed management and vector control using innovative methods
need to be pursued vigorously to obtain a holistic approach. Certain other diseases like Alternaria,
Bacterial blight and grey mildew showing significant appearance in few areas at present shall need better
management options including nanotechnology. A vigil is required on the emerging problems like
tobacco streak virus and their likelihood to cause losses and minor disease like rust becoming major due
to early appearance in south zone. Another important aspect will be to focus on the disease development
and progress vis-a-vis climate changes to understand disease epidemiology and plan management
strategies.
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th
meeting of Asian Cotton Research and Development Network (Full paper at
ICAC website) held at Lahore from February 23
rd
to 25
th
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th
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rd
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th
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International Gemini Virus Symposium, July 24-28, 2001, John Innes Centre, Norwich, Norfolk, U. K., p53.
[30] Radhakrishnan, S., Malathi, V.G. and Varma, A. (2004) -Biological characterization of an isolate of cotton leaf curl
Rajasthan virus from northern India and identification of sources of resistance - Indian Phytopathol., 57: 174-180.
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dissemination in the field - Ph.D. thesis, Department of Plant Pathology, College of Agriculture, CCS HAU, Hisar.
[32] Sharma, O. P., Bambawale, O. M., Datar, V. V., Chattannavar, S. N., Jain, R. K., and Singh Amerika (2007)- Diseases and
disorders of cotton in changing scenario. NCIPM Technical Bulletin. Pp.20.
[33] Singh, Daljeet, Singh, Pritpal, Gill, J. S. and Brar, J. S. (2010) Weather based prediction model for forecasting cotton leaf
curl disease in American cotton - Indian Phytopathol., 63: 87-90
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leaf curl virus disease in Punjab - J. Insect Sci., 7: 194-198.
[35] Sivalingam, P. N., Padmalatha, K. V., Mandal, B., Monga, D., Ajmera, B. D. and Malathi, V.G. (2004)-Detection of
begomoviruses in weeds and crop plants in and around cotton field surveillance: Disease forecasting and management held
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CIRCOT, Bombay, p 311.
Emerging and Key Insect Pests on Bt Cotton

Their Identification, Taxonomy,
Genetic Diversity and Management
S. Kranthi
1
, K.R. Kranthi
1
, Rishi Kumar
2
, Dharajothi
3
, S.S. Udikeri
4
,
G.M.V. Prasad Rao
5
, P.R. Zanwar
6
, V.N. Nagrare
1
, C.B. Naik
1
, V. Singh
7

V.V. Ramamurthy
8
and D. Monga
2

1
Crop Protection Division, Central Institute for Cotton Research, Nagpur
2
Central Institute for Cotton Research, Regional Station, Sirsa
3
Central Institute for Cotton Research, Regional Station, Nagpur
4
Agriculture Research Station, UAS, Dharwad
5
ANGRAU, Lam farm Guntur
6
Cotton Research Station, Marathwada Agricultural University, Nanded
7
Regional Agricultural Research Station, Sriganganagar, Rajasthan
8
Entomology Division, Indian Agricultural Research Institute, New Delhi
AbstractTechnology Mission on Cotton in India has proved to be successful in the planning, implementation,
execution and monitoring of research projects in a stipulated time with a focused approach. Emerging and key insect
pests on Bt cottontheir identification, genetic diversity and management is one of the projects that addressed the
changing pest problems in different regions through strategic research. Mealybugs (Phenacoccus solenopsis,
Paracoccus marginatus), mirids (Creontiades biseratense, Campylomma livida,Hyalopeplus linefer ) , flower bud
maggots (Dasineura gossypii), safflower caterpillar (Perigea capensis) , Tea mosquito bug (Helopeltis bryadi) were
emerging insect pests while leaf hoppers (Empoasca devastans), whiteflies (Bemisia sp), pink bollworm (Pectinophora
gossypiella) and the armyworm (Spodoptera spp.) were the key pests on Bt cotton. Incidence and damage caused by
these pests varied across regions and Bt genotypes being cultivated. Timely taxonomic identification of the mealy bug,
P. solenopsis and subsequent molecular study to suggest its narrow genetic diversity led to the development of
meaningful management strategies to limit its spread. Studies on the mt COI region of the key pest E. devastans
revealed that leaf hopper populations on cotton although morphologically and taxonomically similar were genetically
distinct from leaf hoppers of South and Central India. Implications on pest management in light of this finding are
presented. Flower bug maggots that were hitherto not reported on cotton were found to cause extensive damage in
parts of Karnataka. The life cycle of D. gossypii was elucidated to identify vulnerable stages in its life cycle that can be
exploited for pest management. Two botanical formulations Mealy Kill 50EC (against sucking pests) and Mealy Quit
(against mealybugs) were identified, developed and validated in multilocation trials. Entomofungi were evaluated for
their efficacy in sucking pest management.
INTRODUCTION
India accounts for about 32% of the global cotton area and contributes to 21% of the global cotton
production after China. The production increased from a meager 2.3 M bales in 1947-48 to 17.6 M bales
in 1996-97 to an all time highest record of 31.5 M bales during 2010-2011. Prior to 2002, cotton
production in the country was plagued by bollworm that was a major limiting factor in obtaining the full
yield potential of a genotype. This was coupled with the use of genotypes with low yield potential per se.
With the introduction of Bt cotton, bollworms have been effectively controlled thus minimizing yield
losses. The biggest gain from the technology was in the form of reduced insecticide usage from 46% in
2001 to less than 26% after 2006 and to a further 21% in 2009-10 and 2010-11. The reduction in
insecticide usage in India from Rs. 7180 M in 2004 for cotton lepidopteran caterpillars to Rs.1100M with
only Rs.230M for the control of American bollworm in 2010 is the spectacular effect of Bt cotton (Vision
2030).
While effectively controlling the American bollworm widespread cultivation of Bt cotton has
resulted in emerging pest problems some of which are discussed below.
47
Emerging Pests
Observations were recorded at weekly intervals on Bt cotton from 25 DAS to 120 DAS in Sirsa
(Haryana), Nagpur, (Maharashtra) Coimbatore (Tamil Nadu), Guntur (Andhra Pradesh) and Dharwad
(Karnataka) in farmers fields cultivating Bt hybrids. Those insects apart from the known insect pests of
cotton that appeared in large numbers were recorded. Damage was also recorded.
Feeding of Perigea Capensis under Laboratory Conditions
Field collected larvae on Bunny Bt were collected from Nanded, Yavatmal and were reared to F1 on non
Bt cotton terminal leaves. F1 neonates (30 larvae per event in 3 replicates) were provided with one rupee
sized terminal leaves of different Cry events under no choice conditions and the leaves were changed
each day for a period of 7 days. Mortality was recorded every day and larval weights were recorded at the
end of the bioassay period.
Genetic Diversity of Mealy Bug, Phenacoccus Solenopsis and Cotton Leaf Hopper, Empoasca Devastans
Field collected mealy bugs from 49 locations were preserved in absolute ethanol and brought to the lab
for further studies in 2007. In 2008 mealy bug samples were collected from cotton fields reared to F1 on
potato sprouts and F1 females were used for molecular diversity studies.
Field collected leaf hopper nymphs from cotton at peak vegetative stage were collected in ethanol
from all cotton growing locations of the country with samples representing 3 fields in a village in turn
covering 3 districts of a state. Nine states were covered during the course of study.
DNA was isolated from individual insect samples and PCR COI specific primers were designed to
amplify the COI region of the mitochondrial genome of leaf hopper. Mealy bug PCR amplicons were
generated using primers designed specifically to amplify 18s and 28s rDNA. Using annealing
temperatures 50.8
o
C PCR amplicons of the CO1 region of leaf hopper were generated. The annealing
temperature used to generate PCR amplicons of mealy bugs was 58
o
C. Amplicons were subjected to
double pass analysis and the resulting sequences were aligned and phylogenetic tree was drawn using
MEGA4 (Tamura et. al., 2007).
Formulation of Mealy Kill 50 EC
Using products of insect induced signal transduction pathway limonene was extracted from citrus peel
using the cold press method and evaluated in insect bioassays against aphids, jassids whiteflies and mealy
bugs. Commercially available synthetic analogues of limonene with 98% purity was evaluated under no
choice conditions using log dose probit concentrations as diet incorporation, topical application and leaf
dip methods of bioassays and the LC
50s
were worked out (Finney, 1971). Soap nut powder was used as
emulsifier.
Emerging Pests
From the table it is evident that Mealy bugs were the dominant emerging pest on Bt in Haryana,
Maharashtra, Gujarat and Guntur while mirid bugs were dominant in Haveri and Belgaum districts of
Dharwad in Karnataka. Mealy bugs and mirid bugs were seen as emerging pests in Tamil Nadu (Table 1).
Species Composition of Emerging Pests
Phenacoccus solenopsis was the dominant species of mealy bugs found across the country during its first
year of incidence (2007-08). Subsequently Paracoccus marginatus emerged as a pest on cotton and other
crops of Tamil Nadu in 2009 since its first report in 2007 (Table 2).
Emerging and Key Insect Pests on Bt CottonTheir Identification, Taxonomy, Genetic Diversity and Management 283
TABLE 1: INCIDENCE OF EMERGING PESTS
S. No. State Locations Emerging
pest
Incidence* Damage
1 Haryana Odhan, SIrsa, Kaleriwal,
Dhabwali and Baraguda
Mealy bug 5-44% Grade 3: 1.66-19%
Grade 4: 1.33-19%
2 Maharashtra Nanded Mealy bug 33.70** NR
3 Gujarat Surat Mealy bug - Grade 4 in August.
Grade 2 till Jan
4 Andhra
Pradesh
Guntur Mealy bug 5.76- 35.35 % 1.71-4
5 Karnataka Haveri Belgaum Mirid bugs 43.85 bugs/25 squares -
6 Tamil Nadu Coimbatore Mealy bug 55-83.1% 1.0-1.22
Mirid bugs 16-85.1 bugs/100 squares -
* Refers to number of plants harboring mealy bugs and causing more than Grade 1 damage.
**Refers to number of mealy bugs on 2.5 cm stem length.
TABLE 2: DIFFERENCES BETWEEN THE 2 SPECIES OF MEALY BUGS
Phenacoccus Solenopsis Paracoccus Marginatus
Body quite large (5mm) with dark dorso sub medial bare
spots on inter segmental areas of thorax and abdomen, these
areas forming 1 pair of dark longitudinal lines on dorsum,
with 18 pairs of lateral wax filaments, posterior pair longer up
to 1/4
th
inch length of body.
The adult female is yellow covered with white waxy coating
and measures approximately 2.2 mm in length and 1.4mm
wide. A series of short waxy caudal filaments less than 1/4
th

the length of the body exist around the margin.
Live mealy bug colonies (P. solenopsis) collected across the country from 37 locations were
subjected to DNA isolation and PCR using 18S and 28S rDNA primers, elongation factor 1alpha and
elongation factor 1 Beta primers. PCR amplicons of approximately 350bp and 700bp were sequenced
using double pass analysis in 140 samples representing 3-4 samples per location. All the colonies
belonged to a single haplotype reflecting narrow genetic diversity. This information was important in
devising simple management strategies to be applied uniformly across the country.
Mirid Bugs
Three species have been found to cause damage of varying intensities on cotton- Creontiaedes
biseratense (Distant), Hyalopeplus lineifer (Walker), Campyloma livida (Reuters). Mirids feed on tender
shoots, squares and cause excessive shedding of flowers, small squares and parrot beaking of bolls. They
occur in large numbers moving rapidly on the plant and often miss the eye. Mirid bugs are reported to
cause maximum damage in Haveri and Belgaum districts of Karnataka with up to 2 mirids per square in
the months of October and November. Mirids were reported to cause an avoidable loss of 290Kg/Ha in
Dharwad. Yield loss due to mirids in Nagpur of Central India ranged from 25-30%. Avoidance of broad
spectrum insecticides seems to have assisted in their establishment as emerging pests of cotton. Also,
introduction of new genotypes hitherto unknown into the ecosystem seems to have encouraged the
occurrence of new pests.
Safflower caterpillar, Perigea capensis were collected as late instar larvae from Bt cotton leaves in
Vidarbha, Hingoli and Buldana. It was recorded for the first time along with Spodoptera in cotton fields
adjoining soybean in early vegetative stage. It does not feed significantly on Bt cotton leaves (BG and
BG II) in the lab as neonates die at the end of 7 days. Even though they survive a poor weight gain of
neonate larvae on non Bt cotton leaves was observed (<10mg in a 7 day period).
The adult moth is grayish brown in colour and looks like the pink bollworm moth. It has a pre-
oviposition period of 3 days. Eggs are laid in clusters and are covered with rough hair. Egg period is 3-5
days. Neonate larvae are white in colour and translucent and very active. Full grown larvae are light
green, fleshy with prominent yellow bands across the larval segments. Full grown larvae look like
Helicoverpa with larval period between 14-17 days and a pupal period of 4-5 days.

Genetic Diversity of Mealy Bugs and its Implication for Mealy Bug Management
PCR amplicons of approximately 350 bp and 700 bp were sequenced using double pass analysis in 140
samples representing 3-4 samples per location. All of them had the identical sequence except one sample
of the three from Sriganaganagar which we believe may be a different species. The cotton mealy bug was
identified as Phenacoccus solenopsis (taxonomically) without any genetic diversity (molecular analysis)
throughout the country
PCR Amplification of Mealy Bug DNA From different
location with 28S- A,F & R Primers

PCR Amplification of Mealy Bug DNA From different
location with 18S F & R Primers

1
st
Row:- Lane 1-3: Sirsa; Lane 4-6: Hissar; Lane 7-9:
Abohar; Lane 10-12: Fatehabad; Lane 13-15:
Shriganganagar; Lane 16-18: Hanumangarh; Lane 19-21:
Amravati; Lane 22-24: Jalna.
2
nd
Row:- Lane 1-3: Latur; Lane 4-6: Akola; Lane 7-9:
Hingoli; Lane 10-12: Washim; Lane 13-15:
Yavatmal; Lane 16-18: Nagpur; Lane 19-21: Nanded; Lane
22-24: Parbhani.
1
st
Row:- Lane 1-3: Sirsa; Lane 4-6: Hissar; Lane 7-9: Abohar;
Lane 10-12: Fatehabad; Lane 13-15:
Shriganganagar; Lane 16-18: Hanumangarh; Lane 19-21:
Amravati; Lane 22-24: Jalna.
2
nd
Row:- Lane 1-3: Latur; Lane 4-6: Akola; Lane 7-9:
Hingoli; Lane 10-12: Washim;
Lane 13-15: Yavatmal; Lane 16-18: Nagpur; Lane 19-21:
Nanded; Lane 22-24: Parbhani.
Fig. 1: PCR Amplicons of Mealy Bugs using 18s and 28s rDNA Specific Primers
Management strategies were devised based on the following basic information and the article by
Kranthi et. al., (2011) is recommended for further reading. Pigeon pea, maize and bajra are least
preferred by the mealy bugs. Mealy bugs survive on weeds during the season and also during off-season.
Aenasius bambawalei is the most effective parasitoid. The predatory beetles Cryptolaemus montrouzieri,
Brumus suturalis and Scymnus spp. are prominent in the ecosystems in India and Pakistan. The
entomopathogenic fungi, Metarrhizium anisopliae, Beauveria bassiana, Verticillium lecanii and
Fusarium pallidoroseum are effective in infecting mealybugs. Botanical mixtures containing neem oil,
citrus peel extracts and fish oil rosin were found to be effective in controlling the mealybugs. The insect
growth regulator, Buprofezin is effective in control. Insecticides such as Malathion and Acephate, which
are considered by the WHO as only slightly hazardous (WHO III category) can be used as soil
application near the root zone. All the populations collected in India were highly homogenous, indicating
scant genetic diversity in India. This implied that a common pest management strategy could be adopted
across the country.

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South India
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50
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eco-friendly
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ndia.
ealy bugs),
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ective as it
agents. The
urce of the
ul for pest
nsecticidal
as use of a
0 per acre.
It is not only specific to cotton but can be used on any crop for aphid, jassid and whitefly management.
Mealy Kill 50EC formulation was supplied to 9 AICCIP centres but was tested at 4 centres namely,
Raichur, TNAU, Sirsa and Faridkot, essentially against mealy bugs. It was tested at 20ml/L in north India
and 10ml/L in South India. It offered 34% reduction when sprayed once at Sirsa and was on par with
other bio-pesticides such as V. lecanii, M. anisopliae and B. bassiana. It was superior to the bio-
pesticides tested at Faridkot. There were no significant differences in yield in the insecticide treated plots
and Mealy Kill treated plots in Faridkot. In Raichur and TNAU the reduction in mealy bugs observed due
to Mealy Kill was 90% that was on par with the insecticidal check chlorpyriphos both in terms of pest
control and yield. Mealy Kill was superior to the other bio-pesticides tested, each, sprayed twice, at these
centres in terms of mealy bug control and yield.
ACKNOWLEDGEMENT
The funding for this work, received from TMC MMI from Ministry of Agriculture, is gratefully
acknowledged.
REFERENCES
[1] Vision 2030, CICR (2011). Compiled by K.R. Kranthi, M.V. Venugopalan and M.S. Yadav. Indian Council of Agricultural
Research, New Delhi.
[2] Tamura K, Dudley J, Nei M & Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software
version 4.0. Molecular Biology and Evolution 24: 1596-1599.
[3] Finney, D.J. (1971). Probit Analysis, third ed. Cambridge University Press, Cambridge.
[4] Kranthi, K.R., V. Nagrare, S. Vennila and S.Kranthi (2011). Package of practices for mealy bug management on cotton.
ICAC, 29 (1): 13-16.
Efficacy of Triazoles in Management

of Major Fungal Foliar Diseases of Cotton
A.S. Ashtaputre, N.S. Chattannavar, S. Patil, Rajesh
N.K. Pawar and G.N. Hosagoudar
University of Agricultural Sciences, Dharwad, Agricultural Research Station,
Dharwad Farm, Dharwad580007, Karnataka India
E-mail: sudheendra67@gmail.com
AbstractGrey mildew and Alternaria blight are the major fungal foliar diseases in northern region of Karnataka
and two year study was conducted to know the efficacy of triazoles against these major fugal foliar diseases of cotton,
grey mildew caused by Ramularia areola Atk. and Alternaria blight caused by Alternaria macrospore Zimm, during
kharif 2009 and 2010

under rainfed situation at Agricultural Research Station, Dharwad. The experiment was laid out
in replicated trial of randomised block with ten treatments. The study revealed that all the triazoles under study were
found to be effective in control of major foliar diseases, which in turn reflected in more yield. Among these triazoles,
Percent disease index(PDI) of Penconazole for Alternaria blight(AB) (6.10 PDI) and grey mildew(GM) (10.30 PDI)
followed by Hexaconazole (AB 8.20, GM 11.0 PDI), Difenconazole (AB 7.10, GM 11.10 PDI) and Tridemefan (AB
11.3, GM 13.5 PDI), reduced the disease severity of both the diseases effectively and also enhanced the yield. But three
sprays of Hexaconazole (0.1%) were more useful not only in reducing the cost of protection but also gave higher
benefits (B:C ratio 9.63) as compared to other treatments and can be used for the management of major fungal foliar
diseases of cotton. Hexaconazole can be recommended as one of the components in integrated disease management of
cotton as it showed the best result in the control of both diseases with higher cost benefit ratio and increased
yield(14.3 q/ha).
INTRODUCTION
Cotton, The White Gold enjoys a pre-eminent status among all cash crops in the country and is the
principal raw material for a flourishing textile industry. India now produces around 290.00 lakh bales of
cotton ranging from short staple to extra long staple from an area of 93.73 lakh hectares with productivity
of 526 kg per hectare (Anonymous, 2009). In Karnataka, the area under cotton cultivation is 3.90 lakh
hectares with a production of 9.00 lakh bales and an average productivity of 392 kg per hectare
(Anonymous, 2009). Cotton is known to suffer from number of diseases caused by fungal, bacterial and
viral origins. There is now more relative importance for different diseases may be air borne like grey
mildew, Alternaria leaf spot, Myrothecium leaf spot, bacterial blight, rust, cotton leaf curl virus (white fly
transmitted) or soil borne like seedling rots, Rhizoctonia root rot, Verticillium wilts and even some times
Sclerotium rolfsii affecting cotton across India. Only the type of the disease and its virulence differs with
different agro climatic regions. These changes may be due to change over from the cultivation of
Asiatic (G. herbeceum and G. arboreum) to American cottons (G. hirsutum) and hybrids. Most of them,
even though high yielding, are susceptible to diseases. Only the type of the disease and its virulence
differs with different agro climatic regions. These changes may be due to change over from the
cultivation of Asiatic (G. herbeceum and G. arboreum) to American cottons (G. hirsutum) and hybrids.
Most of them, even though high yielding are susceptible to diseases (Shivankar and Wangikar, 1992,
Chattannavar et al, 2009). Among the fungal diseases grey mildew and Alternaria blight are the
predominant ones causing economic losses to the cotton crop in the country. Management of diseases is a
continuous process due to development of different resistant races of pathogens imposed by climatic
changes, chemicals or even resistance to old resistant cultivars.

48
A field experiment was conducted at Agriculturl Research Station (Cotton), Dharwad farm, Dharwad ,
Karnataka during Kharif 2009-10 and 2010-11 to evaluate the field bio-efficacy of triazole group of
fungicides against major fungal foliar diseases grey mildew caused by Ramularia areola Atk. and
Alternaria blight or Alternaria leaf spot caused by Alternaria macrospore Zimm were compared with
standard recommendation Carbendazim 50% WP foliar spray treatment @ 0.1%. The experiment was
planned in Randomised block Design and replicated thrice on Bt cotton hybrid Bunny Bt. The
individual treatment plot size was 6.0 x 5.4 m
2
with spacing of 90 x 60 cms. Normal recommended
cultural practices were adopted. Three sprays of all treatments were undertaken immediately after the
appearance of the disease at an interval of 12 days. The observations on percent disease index of
Alternaria blight and grey mildew were recorded 15 days after the last spray, on five randomly selected
plants in each treatment. In each treatment, ten plants were randomly selected and tagged. Three
branches were randomly tagged per plant and the intensity of Alternaria blight and grey mildew on all
the leaves of these tagged branches were graded by adopting 0 to 4 scale as given by Sheo Raj (1988).
Per Cent Disease Index (PDI)
The results obtained during 2009 with respect to Alternaria blight, revealed that, all the treatments were
significantly superior over untreated control. From the data, it is clear that, the treatments viz.,
Penconazole, Difenconazole, and Hexaconazole were found on par with each other with PDI of 5.80,
6.70 and 8.30 respectively and they were significantly superior to all other treatments followed by
Propiconazole, Mycobutanil, Tridemefon with PDI of 9.40, 10.30 and 11.40 respectively. The results
obtained during kharif, 2010 followed similar trend of results but in slightly higher intensity of incidence
of disease, as observed during kharif, 2009.
TABLE 1: EFFICACY OF TRIAZOLES AGAINST ALTERNARIA BLIGHT AND GREY MILDEW OF COTTON
Sl.
No
Treatments Alternaria blight
PDI
Pooled
mean
PDI
Grey mildew
PDI

Pooled
mean
PDI
Yield (q/ha)

Pooled
Yield
(q/ha)
B:C
2009-10 2010-11 2009-10 2010-11 2009-10 2010-11
T
1
Mycobutanil @
1gm/litre
10.30
(18.73)*
11.20
(19.53)
10.70
(19.13)
8.60 (17.05) 20.30(26.79) 13.90(21.92) 12.80 13.7 13.2 1.81
T
2
Hexaconazole
@ 1ml/litre
8.30
(16.67)
8.20
(16.59)
8.20
(16.63)
9.40(17.86) 12.70(20.90) 11.00(19.38) 14.30 14.8 14.5 9.63
T
3
Penconazole @
1ml/litre
5.80
(13.93)
6.40
(14.60)
6.10
(14.27)
6.70(15.04) 14.50(22.40) 10.30(18.72) 17.20 15.6 16.4 5.80
T
4
Propiconazole @
1ml/litre
9.40
(17.90)
9.30
(17.73)
9.40
(17.82)
9.50(17.93) 21.00(27.28) 14.80(22.60) 14.10 15.7 14.9 6.2
T
5
Difenconazole@
1ml/litre
6.70
(15.03)
7.40
(15.77)
7.10
(15.40)
8.60(17.00) 13.80(21.83) 11.10 (19.42) 14.80 15.5 15.2 3.27
T
6
Tridimefon @
1gm/litre
11.40
(19.70)
11.20
(19.57)
11.30
(19.63)
8.20(16.63) 19.80(26.40) 13.50(21.51) 14.10 15.3 14.7 3.12
T
7
Tridemorph @
1ml/litre
19.90
(26.50)
20.00
(26.58)
20.00
(26.54)
7.60(15.96) 23.50(28.98) 14.60(22.47) 13.60 14.2 13.9 4.83
T
8
Carbendazim @
1gm/litre
20.10
(25.67)
21.30
(27.50)
20.00
(26.58)
8.20(16.67) 21.30(27.50) 14.10(22.08) 13.20 13.9 13.5 5.77
T
9
Propineb @
3gm/litre
18.20
(25.27)
17.00
(24.35)
17.60
(24.81)
10.80(19.18) 23.00(28.65) 16.40(23.92) 12.90 14.2 13.6 3.65
T
10
Control 30.30
(33.37)
33.00
(35.08)
31.60
(34.22)
23.70(29.10) 34.60(36.03) 29.00(32.57) 11.20 12.7 11.9 -
SEm 1.187 1.068 0.748 1.192 1.645 0.989 0.351 0.389 0.219
CD at 5% 3.527 3.174 2.222 3.543 4.887 2.938 1.041 1.157 0.651
The pooled data (Table 1) of two years for Alternaria blight and grey mildew indicated that all the
treatments were significantly superior over untreated control. The triazoles under study were found to be
significantly effective in the management of the diseases. The least PDI was observed in Penconazole of
6.10 PDI and 10.30 PDI for Alternaria blight and grey mildew respectively followed by Difenconazole
Efficacy of Triazoles in Management of Major Fungal Foliar Diseases of Cotton 289
(7.10 PDI for A. blight and 11.10 PDI for grey mildew) and Hexaconazole (8.20 for A. blight and 11.00
PDI for grey mildew) which were on par with each other and significantly superior over rest of the
treatments followed by all other triazole group of fungicides under study.
Cotton Yield
The cotton yield was significantly superior in all the treatments as compared to untreated control. The
results indicated that, all the triazoles under study have showed higher yield. Next best treatments were
viz., Tridemorph (13.9 q/ha), Propineb (13.6 q/ha), Carbendazim (13.5 q/ha) and Mycobutanil (13.2
q/ha), on par with each other, but differed significantly with the untreated control.
The pooled data of two years depicted that, the triazole group of fungicides was found to be more
effective in enhancing the yields significantly (Table 1). Pooled maximum yield of both the years was
noticed in Penconazole (16.4 q/ha), which was significantly superior over all other treatments, followed
by Difenconazole (15.2 q/ha), Propiconazole (14.9 q/ha), Triadimefon (14.7 q/ha) and Hexaconazole
(14.5 q/ha) . The least yield was noticed in untreated control (11.9 q/ha).All the treatments were found to
be significantly differ with untreated control.
Benefit Cost Ratio (BCR)
From the pooled data of two years, it is evident that maximum B: C ratio was observed in Hexaconazole
(9.63) followed by Propiconazole (6.2) and Penconazole (5.80) (Table 1).
In the present investigation, it is evident that all triazoles under study were found to be effective in
control of the grey mildew and Alternaria blight disease, which in turn reflected in more cotton yield.
Among these triazoles, Penconazole followed by Hexaconazole and Difenconazole reduced the disease
severity of both the diseases effectively and also enhanced the yield. These findings are in accordance
with Khodke and Raut( 2009) who reported that these triazoles gave the effective control of grey mildew.
The benefit cost ratio is an important parameter for recommendation of any treatment for successful
control of plant disease. In the present study, though the treatments containing three sprays of
Penconazole, Hexaconazole, Difenconazole, Triadimefon and Propiconazole gave significant control of
both the diseases, maximum Cost Benefit ratio of 9.63 was realized in treatments containing three sprays
of Hexaconazole (0.1%) followed by Propiconazole (6.2) and Penconazole (5.80). This clearly indicated
that three sprays of Hexaconazole (0.1%) are more useful not only in reducing the cost of protection but
also gave higher benefits as compared to other treatments and can be recommended as one of the
components in integrated disease management of cotton. This is followed by Difenconazole and
Penconazole applications. Similar types of findings are observed by many workers (Khodke and Raut,
2009, Algarsamy and Tagarajan, 1986). Hence, spraying of Hexaconazole (0.1%) could be considered as
an effective management practice to manage major fungal foliar diseases.
REFERENCES
[1] Anonymous, 2009, Ann. Rep. of All India Co-ordinated Cotton Improvement Project, for 2008-09, Central Institute for
Cotton Research Regional Station, Coimbatore.
[2] Algarsamy, C. and Tagarajan, R., 1986, Efficacy of fungicides against grey mldew disease of Cotton. Madras agric. J.,73:
651-652
[3] Chattannavar, S. N., Hosagoudar, G. N., Ashtaputre, S. A. and Ammajamma, R., 2009, Evaluation of cotton genotypes for
grey mildew and Alternaria blight diseases. J. Cotton Res. Dev., 23(1) : 159-162.
[4] Khodke, S.W and Raut, B.T., 2009, Chemical management of grey mildew caused by Ramularia areola Atk. of diploid
cotton
[5] Sheo Raj, 1988, Grading for cotton disease, CICR, Nagpur. Bull., pp. 1-7.
[6] Shivankar, S. K. and Wangikar, P. D., 1992, Estimation of crop loss due to grey mildew disease of cotton caused by
Ramularia areola. Indian Phytopath. 45: 74-76.
Damage Caused in Cotton by Different Levels

of Ramulosis in Brazil
Alderi Emdio De Arajo
1
,
Alexandre Cunha De Barcellos Ferreira
2
and Camilo De Lelis Morello
2
1
Embrapa Algodo, CP: Campina Grande, PB, Brazil 174, CEP 58428095
2
Embrapa Algodo, Research Group of Cerrado, Embrapa Arroz e Feijo C.P. 179,
CEP 75375000, Santo Antnio de Gois, GO, Brazil
e-mail: alderi@cnpa.embrapa.br
AbstractThe ramulosis caused by Colletotrichum gossypii var. cephalosporioides is one of the most important
diseases of cotton in Brazil. The damage can range from 20 to 30% reaching 85% in severe cases. This study aimed to
assess the damage to cotton caused by different levels of disease severity. The experiment was carried out in state of
Gois, in the season of 2006. The treatments were five severity indexes based on the following descriptive key: 1- plant
without symptoms; 2-plants with necrotic spots in the young leaves; 3- necrotic spots in the leaves, shortening of
internodes and initial broom; 4- necrotic spots in the leaves, shortening of internodes, broom little developed and
height reduction; 5- necrotic spots in the leaves, shortening of internodes, broom very developed and height reduction.
The plants with 40 days old age were inoculated with a suspension of 2x10
5
conidia/ml of the pathogen and to assure
the occurrence of different levels of severity, the treatments with low scores of the key were sprayed with fungicides
according to disease development. The experimental design was in randomized blocks with 5 treatments and 4
repetitions. Were assessed the following variables: height of the plants, number of bolls, weight of the bolls and lint
production. The more important damage caused by disease were the reduction in the weight of the bolls and in the lint
production. To these variables the reduction was more than 70% when the severity of the disease was high. The
reduction in the plant height was higher when the disease severity achieved the 3 and 4 points of the key. Based on
these results we conclude that is very important the control of the disease in the initial stages until the point 2 of the
descriptive key to avoid significant damage to the fiber production
Keywords: disease, fungus, control, Colletotrichum
INTRODUCTION
The ramulosis, caused by Colletotrichum gossypii var. cephalosporioides, is one of the most important
diseases of cotton in Brazil. The main characteristic of this disease is the breaking of apical dominance,
which induces successive shoots, giving to the plant the appearance of a broom. The damage can range
from 20% to 30%, reaching 85% in severe cases. In Mato Grosso were reported damages of up to 80%
(Freire et al., 1997).
The environmental conditions more favorable to ramulosis are high rainfall, temperatures of 25 C to
30 C and relative humidity above 80% (Miranda; Suassuna, 2004; Silveira, 1965). In state of Mato
Grosso, the temperature range favorable for the development of the disease ranged from 20 C to 30 C
(Arajo, Farias, 2003), while in state of Minas Gerais the optimum temperature for the higher incidence
of disease was 18.3 C (Santos, 1993).
The first symptoms of ramulosis occur in young leaves and are characterized by circular necrotic
spots. Afterwards the tissue of these spots breaks up and detaches itself, resulting in star-shaped
perforations. The uneven growth of the tissue induces wrinkling of the leaf. Soon after the emergence of
the first leaf injury, death occurs from the apical meristem of the affected branch, halting its growth
stimulating the sprouting of lateral buds, which culminates in the formation of a cluster of branches with
short internodes and swollen, giving the plant the aspect of a broom (Arajo; Suassuna, 2003; Suassuna,
Coutinho, 2007).

49
Damage Caused in Cotton by Different Levels of Ramulosis in Brazil 291
The ramulosis can also affect the quality of fiber, like length, fineness, uniformity and micronaire and
the damage can cause the reduction in weight of bolls and in the percentage of fibers (Carvalho et al.,
1984).This study aimed to determine the damage caused to cotton by different levels of severity
ramulose.
The experiment was carried ou at the Experimental Station of Embrapa / GO Foundation in Santa Helena
de Gois, in the season of 2006. The cultivar used was BRS Ipe, whose seeds were treated with the
insecticide imidacloprid (270 g kg ai/100), and fungicides tolylfluanida (75 g kg ai/100) + pencycuron
(75 g ai/100 kg).
The treatments were of five levels of disease severity based on the descriptive key proposed by
Arajo et al. (2003): 1- plant without symptoms; 2-plants with necrotic spots in the young leaves; 3-
necrotic spots in the leaves, shortening of internodes and initial broom; 4- necrotic spots in the leaves,
shortening of internodes, broom little developed and height reduction; 5- necrotic spots in the leaves,
shortening of internodes, broom very developed and height reduction.
Infection of plants was obtained through inoculation of a suspension of 2x105conidia / ml of the
pathogen, 40 days after emergence. Plants with level 1 of the descriptive key were not inoculated. In
order to ensure the different levels of severity of the disease, plants with disease severity with levels 1
and 2 were sprayed with the fungicide propiconazole + trifloxystrobin(125 + 125 g / L a.i. / ha), based on
data obtained by monitoring the evolution of disease. The other levels of the descriptive key were
obtained through the systematic control of the development of the disease with spray with trifloxystrobin
+ propiconazole (125 +125 g / L a.i. / ha) when necessary in both cases. The first spray was made at the
levels 3 and 4, and one application was sufficient to maintain levels while level 5 was obtained without
spray.
The design was randomized blocks, with five treatments and four replicates, and the parcel had four
lines 5 m, considering how useful the two central rows. The evaluation was performed at 140 days after
emergence, having been employed the descriptive key proposed by Arajo et al. (2003). The measured
variables were: plant height, number of bolls / plant, boll weight and cotton lint production.
Based on the results shown in Table 1, it was observed that the disease has negatively affected all
variables. The largest damage were observed for boll weight and in production of cotton lint per plant.
For these variables, the decrease was greater than 70%, indicating that the ramulosis, in advanced stages,
causes severe damage to production. Although the differences in the number bolls per plant were not as
expressive, it is important to note that the weight the boll was reduced with the increase in disease
severity, reflecting directly in the production of cotton lint. The number of capsules suffered the greatest
reduction when there was increase in the severity of the disease, Based on the results shown in Table 1, it
was observed that the disease has negatively affected all variables. The largest damage were observed for
boll weight and in production of cotton lint per plant. For these variables, the decrease was greater than
70%, indicating that the ramulosis, in advanced stages, causes severe damage to production.
Although the differences in the number of bolls per plant were not as expressive, it is important to
note that the weight the boll was reduced with the increase in disease severity, reflecting directly in the
production of cotton lint. The number of capsules suffered the greatest reduction when there was increase
in the severity of the disease, as can be observed the data relating to notes 4 and 5 of the descriptive key.
This phenomenon is associated the fact that higher levels of severity of the ramulosis can induce an
increased production of branches vegetative rather than fruiting branches, what determines a reduction in
production boll. The reduction in plant size was observed more steeply from the notes 3 and 4 of the
descriptive key. From these levels disease severity, there were also further damage to production. Thus,
based the observations, there is the need for control the disease in the earliest stage, in view of the
irreversibility of damage since the early symptoms of broom, that begin to manifest when the plant shows
increased levels of severity recorded from the level 3 of the descriptive key.
These results corroborate those already obtained by Carvalho et al. (1984), in the state of
Pernambuco. However, it should be pay attention to the fact that climatic conditions of Gois state may
be more favorable to ramulosis, due to higher rainfall and regular rainfall usually recorded in the
Midwest of Brazil, with a view that C. gossypii var. cephalosporioides is widespread primarily by
splashing water, and the rain is a major agent of dispersal of inoculum. Therefore, monitoring of disease
in the state of Gois, in brazilian Midwest should be more systematic, given the more favorable
conditions the development of disease. Therefore, the use of resistant cultivars is recommended and the
permanent monitoring of farming, to prevent that the disease reach to rates of high severity and induce
significant damage to production.
The control measures that prevent the increase of inoculums such as the use of seed health and crop
rotation should be privileged. The chemical control should be implemented in the early stages and should
never exceed the level 2 of the descriptive key used for the evaluation of disease severity.
TABLE 1: PLANT HEIGHT (CM), NUMBER OF BOLLS, BOLL WEIGHT (G) AND COTTON LINT PRODUCTION / PLANT (G) OF COTTON FOR DIFFERENT LEVELS OF SEVERITY RAMULOSE.
SANTA HELENA DE GOIAS, BRAZIL, 2006.
Level of the Descriptive Key* Plant Height Number of Bolls Boll Weight Fiber Production
1 126,16 a 3,33a 32,51a 12,01a**
2 121,50ab 2,58ab 27,92ab 9,47b
3 111,34b 2,31ab 20,94bc 7,23b
4 85,41c 1,96b 14,54cd 5,51bc
5 69,8d 1,53b 8,48d 3,21c
VC 6,3 24,88 21,12 23,64
*Descriptive key: 1- plant without symptoms; 2-plants with necrotic spots in the young leaves; 3- necrotic spots in the
leaves, shortening of internodes and initial broom; 4- necrotic spots in the leaves, shortening of internodes, broom little
developed and height reduction; 5- necrotic spots in the leaves, shortening of internodes, broom very developed and height
reduction.
**Means followed by same letter vertically do not differ by Tukey test at 5% probability.
REFERENCES
[1] Arajo, A E., Suassuna, N. D., Farias, F. J. C., Freire, E. C. Escalas de Notas Para Avaliao de Doenas Foliares do
Algodoeiro. In: Congresso Brasileiro de Algodo, 4., 2003, Goinia. anais... campina grande. Embrapa Algodo, 2003, 1
cd-rom.
[2] Arajo, A. E.; Farias, F. J. C. Progress of witches broom disease of cotton in Mato Grosso State Brazil. In: World Cotton
Research Conference, 3., 2003, Cape Town, Anais... Cape Town, ICAC, p. 1428-1430.
[3] Carvalho, L. P.; Cavalcanti, F. B., Lima, E. F., Santos, E. V. Influncia da ramulose nas caractersticas de fibra do
algodoeiro. Fitopatologia Brasileira, v. 9, p. 593-598. 1984.
[4] Freire, E. C.; Soares, J. J.; Farias, F. J. C.; Arantes, E. M.; Andrade, F. P.; Paro, H.; Laca-Buendia, J. P. Cultura do
algodoeiro no estado de Mato Grosso. Campina Grande-PB: Embrapa Algodo, 1997, 65 p. (Embrapa Algodo. Circular
Tcnica 23).
[5] Miranda, J. E.; Suassuna, N. D. Guia de Identificao e controle das principais pragas e doenas do algodoeiro. Campina
Grande: Embrapa Algodo, 2004. 47 p. (Embrapa Algodo. Circular Tcnica, 76).
[6] Santos, G. R. Progresso da ramulose do algodoeiro e transmisso de Colletotrichum gossypii South var. cephalosporioides
Costa pelas sementes. 1993. 53 p. Dissertao (Mestrado em Fitopatologia) Universidade Federal de Viosa, Viosa, MG,
1993.
[7] Silveira, A. P. Fungos e bactrias. In: Instituto Brasileiro De Potassa. Cultura e adubao do algodoeiro. So Paulo, 1965.
p. 417- 419
Insecticidal Toxin Genes from Bacterial Symbiont of

Thermotolerant Isolate of Heterorhabditis indica,
Entomopathogenic Nematode
Nandini Gokte-Narkhedkar, Kanchan Bhanare, Prachi Nawkarkar,
Prashanth Chiliveri and K.R. Kranthi
Division of Crop Protection, Central Institute for Cotton Research, Nagpur
INTRODUCTION
In the last two to three decades use of chemical control for pest management has become less acceptable
as concerns about contamination of soil and water and deleterious effects on man and livestock have led
to restrictions on their use. This and development of resistance in insects against commonly used
chemicals has led to demand for development of alternates for pest management and biological control is
one such option. Entomopathogenic nematodes with their associated bacteria have been identified as
viable option for insect management and toxicity of EPN-bacterial system to insects is largely attributed
to toxins produced by bacterial symbiont . Considerable progress has been made in identification of toxin
genes from bacteria Photorhabdus and Xenorhabdus (Williamson and Kaya, 2003). Toxin genes from
EPN- bacterial system can be used as alternative to Bt toxins or can be used to pyramid multiple
resistance genes for broad range and effective resistance against insect pests. A thermophilic isolate of
EPN Heterorhabditis indica has been developed at CICR and bacterial isolate found associated with this
EPN was found to be very effective against sucking pests of cotton in field trials undertaken at CICR,
Nagpur, its regional station Sirsa and at Nanded. Therefore Bacteria isolated from thermophilic EPN
H.indica isolate were taken up for further characterization and identification of toxin genes.
Bacteria were isolated from juveniles of nematode H.indica on standard bacteriological media and
bacteria were taken up for molecular and biochemical characterization. (Bergys Manual). The
biochemical parameters taken up were Colony Morphology on Nutrient Agar,Gram Stain, Pigmentation,
Levan production, Methyl Red, Voges-Proskauer Test, starch hydrolysis, oxygen requirement, H2S
production, indole production, nitrate reduction, Urease test, ADH test, citrate, catalase, gelatinase,
motility, tyrosinase and Galactosidase tests. Carbohydrates fermentation studies for the bacterial isolate
were carried out for 21 carbohydrates.
For molecular characterization 16s ribosomal RNA sequence of bacterial isolate was
amplified using oligonuceotide primers (5GGA GAG TTA GAT CTT GGC TC3 sense and 5AAg
GAG GTG ATC CAG CCG CA3(Brunel et al., 1997). Samples amplified using 25l of reaction with
10mM of each primer, 0.1 g of DNA template, 12.5 l 2X PCR- master mix and distilled Water. PCR
conditions were same as Brunel et al., 1997 with amplification at 57
0
C. The sequence amplified was
around 1550 bp and it was cloned in pEMT vector for sequencing.
TOXIN ISOLATION
For isolation of toxins, the bacteria was cultured on LB broth for 48 hrs. on shaker. Extracellular and
intracellular fractions separated by centrifugation and sonication . Different fractions from the
extracellular and intracellular components of bacterium separated using columns, centrifugal devices and
gel filteration were bioassayed against 3
rd
instar larva of Helicoverpa armigera for insecticidal activity.
Protein content of different fractions was estimated and fractions were tested against 3
rd
instar larva of
Helicoverpa armigera for insecticidal activity.
50
DESIGNING OF PRIMERS FOR AMPLIFICATION OF TOXIN GENES
The primer pairs have been designed by identifying 8-10 amino acid stretch in protein that is rich in
amino acid codes by only one or more codons (Met, Trp, Phe, Cys, His, Lys, Asp, Gly, Gln, Tyr) and that
has no or few amino acids coded by six codon (Ser, Leu, Arg). Primers have also been designed by
aligning known toxin sequences from data bases.
F-5ACCGCCGAGTCCCTTGGCTA3,R-CGCTGCTGTCTGTGGAGCGTT
F-5CTTCGGCGCCATTCCCCGTT 3, R-GCGCTACTCTCGGCAGCAGG
F-5GCGGAGGATGGCCGCAAACT 3, R-CGTGCTGTGCTACCGCGTCA
F-5CTTCGGCGCCATTCCCCGTT 3,R-GCGCTACTCTCGGCAGCAGG
F-5CGGTGACGCCGCACAGTTCT3,R-TCTGTGCGACCGGAAACGGC
F -5 TACC AATA TGTTAATTG TGGAC 3, R R - 5 CCA TCA TTTCAC ATA ACCG 3
F-5 TTCG AATA CCAA TATG TTAA TTGTGGAC 3, R-5 CCA TCA TTTCAC ATA ACCG 3
F-5 ATTACCAATATGT TAATTGTGG 3, R - 5 TCATCATATATTTTATAATG
F -5 GGTCTAGAATGTAAAGGCAACAC-3'), R- 5'-GGAAGGACGGAAAGTGGAGA-3
F-(5'-ACCATACGCATCGGACAAAC-3'), R-5'-CGTAGCGGTTATTCACTCTTCT-3
F -TCAGACTGATGCCAAAGG, R - CCATCAATAGTTCCTGCC,
F -TCAGACTGATGCCAAAGG, R -CCATCAATAGTTCCTGCC
F-5 TACTTAGTTGAGCGGTCAGG, R - 5 GCCATGCTCAGTTACTGC
F-5 TACTTGCTCA GACATTTCTCTATGG 3,R TTATTTAATGGTGTAGCG 3
F 5ACCATACGCATCGGACAAAC-3, R 5CGTAGCGGTTATTCACTCTTCT-3
F- 5GGTCTAGAATGTAAAGGC-3, R -5GGAAGGACGGAAAGT 3
F- 5TACCACTGACAATACGTTTAT 3, R- 5CGGTTACTGACGATTGCTG3
F- 5 TCATGAAATACGTCCTAAGTGG 3, R- 5 AAA TATGT AAAACTATGGG GTTC3
F- 5 ACCTTAACTAATACAGACTTAG 3, R- 5 AA AGAAAAGAAATTTACGCGTG 3
F - 5 TGTAGTTACAAGAAAGAACC 3, R- 5 ATGTCTAAATACAAATTAAACC 3
F-5 CTTATACTATACTCAGGCAG 3, R- 5 ATTGCAAGATATTAATTACAAAG 3
Molecular Characterization of Bacterial Isolate

Fig. 1
The sequence amplified was around 1550 bp and it was cloned in pEMT vector for sequencing.
Plasmid DNA was isolated using Qiagen miniprep kit and sequenced. The sequences were blasted. The
sequence of bacterial isolate showed 96% similarity to Paenibacillus sp. As this bacterial isolate showed
toxicity to sucking insect pests, this bacterial isolate was also characterized for biochemical parameters.
Biochemical characterization of bacterial isolate associated with H. indica and identified as
Paenibacillus sp.
Insecticidal Toxin Genes from Bacterial Symbiont of Thermotolerant Isolate of Heterorhabditis indica 295
Test Result Test Result
Gram Stain Gram Positive. Methyl Red Test Negative.
Pigmentation No pigmentation. Voges-Proskauer Test Positive
Levan Production No levan Production. Gelatin Test Weakly positive
Urease Test Negative Oxygen requirement Facultatively anaerobic.
Tyrosinase Test Positive Production of H
2
S Gas. No H
2
S production was observed.
Citrate Test Positive Production of Indole. No Indole production was seen.
Catalase Test Positive Nitrate Reduction Negative
Amino Acid decarboxylase Negative Growth on Mc Conkey Agar Growth observed
Starch hydrolysis test Positive. Esculin Hydrolase Test Positive
Casein Hydrolysis Positive Arginine Hydrolase Test Negative
Motility Test Negative Oxidase Test Negative
Carbohydrate Fermentation test The Test strongly positive for Glucose, Fructose, Galactose, Maltose, Raffinose, Sucrose,
Salicin, Trehalose and weakly positive for adonitol.
The test is negative forArabinose, Cellobiose, Inositol, Inulin, Lactose, Mannose, Mannitol, Melibiose, Rhamnose,
Sorbitol, Xylose and Dulcitol. No gas production was observed.
Toxin Isolation
Different fractions from the extracellular and intracellular components separated using columns,
centrifugal devices and gel filteration were bioassayed against 3
rd
instar larva of Helicoverpa armigera
for insecticidal activity. Protein content of different fractions was estimated and found to range between
1.32 -1.68 mg/ml. The fractions were tested against 3
rd
instar larva of Helicoverpa armigera for
insecticidal activity.
TABLE 1: INSECTICIDAL EFFICACY OF DIFFERENT FRACTIONS AT 10 G
Fraction % Dead Intrahaemocoelic % Dead Oral
1KG 22 20
3KG 40 60
10KG 58 80
50kG 67 100
100KG 45 40
Control 10 0
1K-More than 1kDa, 3-more than 3kDa, 10, More than 1kDa, 50- More than 1kDa, 100 -More than
1kDa
Individual fractions at three different concentrations were (5, 10 and 15g) were injected into
haemocoel of 3
rd
instar H.armigera larvae. At l0 g difference in efficacy of different fractions was
evident and results are presented in Table 1. Control was maintained with physiological saline solution.
Observations on insect mortality after 24 hrs revealed that fraction 50 -100 kDa recorded more than 98%
mortality after 24 h while 10K fraction recorded 60% morality. In other fractions mortality was recorded
after 48 hrs only while in control there was nil mortality up to 48 hr. These fractions were also evaluated
for oral toxicity with H.armigera neonates. 50K fraction was also recorded to have oral toxicity.
50K fraction was run on native PAGE and individual bands were cut, eluted in buffer (140 mm NaCl,
2.7 mM KCl, 10mM Na
2
HPO
4
, 1.8 mMKH
2
PO
4
, pH 7.3) and analysed for insecticidal activity.

Fig. 2
The elutes of bands were applied to artificial diet for oral toxicity to Helicoverpa armigera neonates.
These were also injected in intrahaemoceolic for toxicity to H.armigera. LC
50
experiments were
conducted for 48 h with neonate larvae, and were replicated on three separate occasions with 12 larvae
per treatment. Growth inhibition studies were 72 h in duration and were repeated twice with 12
individuals per treatment. Mortality data from the LC
50
experiments was analyzed by Probit analysis.
Results indicate that two bands of approximately 950kDa had insecticidal effect. Lc50 for A band was
calculated at 0.1 g while Lc50 for B band was 0.12 g. At concentration of 0.18 g injected in
haemocoel mortality ranged between 89-87%. Oral toxicity to neonates of H. armigera was also
recorded. At 0.05 g oral toxicity to neonates was recorded with 78-85% mortality of neonates.
Evaluation of toxicity of these components against sucking pests is underway.
Rajagopal and Bhatnagar(2002) has isolated two protein complexes of approx. 1000kDa from
Photorhabdus luminescens subsp. akhurstii which were active against Spodoptera litura and Galleria
mellonella.
Amplification of Toxin Genes

Fig. 3
Insecticidal Toxin Genes from Bacterial Symbiont of Thermotolerant Isolate of Heterorhabditis indica 297
D6TcdB, Wg TcdA, D6TcdA2,Wg TcdA2,G1 TcdB, G1 TcdAB, G5 TcdA2, Photo TcdA, WgTcd
Ab could be amplified by using primers designed for amplification of toxin genes and standardization of
PCR conditions. These were cloned in pGEM-T vector and sequenced.
The sequences of PhotoTcdAB, TcdB were blasted. These were found to have 98% similarily with
Serine protease gene and phospholipase of Bacillus thuringiensis and B.cereus.
Amplification of Tcc Genes from Paenibacillus sp. is significant as this appears to be first reports of
a Paenibacillus species, strain, or protein having toxicity to lepidopterans. Furthermore, this may also
first known report of a Paenibacillus having toxin complex (TC)-like proteins controlling insects and like
pests. Genes from Photorhabdus encode large insecticidal toxin complexes which cause septicaemia in
insects. Arabidopsis thaliana plants expressing toxin A from Photorhabdus luminescens showed
considerable activity against lepidopteran insects and moderate activity against colepteran insects (Liu et
al, 2003). Identification and cloning of toxin genes from Paenibacillus would make available genes
effective against sucking pests. Further work on cloning of full length gene and their expression in
suitable vector is underway.
REFERENCES
[1] Brunel B., Givaudan A., Lanois A., Akhurst R.J. and Boemare N.E. (1997). Fast and accurate identification of Xenorhabdus
and Photorhabdus by restriction analysis of PCR amplification. Appl. Environ. Microbiol., 63:574-580.
[2] Liu, D., Burton, S., Glancy T., Li, Z.S., Hampton, R., Meade, T. and Merlo, D.J. (2003). Insect resistance conferred by 283
kDa Photorhabdus luminescens protein TcdA in Arabidopsis thaliana. Nat. Biotechnology 21: 1022-1028.
[3] Rajagopal, R. and Bhatnagar, R.K. (2002). Insecticidal toxic proteins produced by Photorhabdus luminescens akurstii, a
symbiont of Heterorhabdits indica. J.Nematol. 34 23-27.
[4] Williamson, V.M. and Kaya, H.K. (2003). Sequence of a symbiont. Nat. Biotechnology 21: 1294-1295.
Identification and Characterization of a Novel Source

of Resistance to Root-Knot Nematode in Cotton
Mota C. Fabiane, Giband Marc, Carneiro, D.G. Marina, Silva, H. Esdras, Furlanetto
Cleber, Nicole Michel, Barroso, A.V. Paulo and Carneiro and M.D.G. Regina
Research Scientist, Cirad, UMR AGAPEmbrapa Algodo, Rodovia Go462, Km 12, Zona Rural
75.375000 Santo Antnio de Gois, GoBrazil
E-mail: marc.giband@cirad.fr.
AbstractThe root-knot nematode (RKN) Meloidogyne incognita Kofoid and White 1919, Chitwood 1949 is a major
constraint in cotton (Gossypium hirsutum L.) production in numerous countries. Control of RKN has been hampered
by the lack of high-quality local varieties exhibiting high levels of resistance as well as the lack of options for crop
rotation. High levels of resistance occur in breeding lines, but this high level of resistance has not been readily
transferred to cultivated varieties. Resistance to RKN is also found in wild tetraploid cotton accessions that represent
valuable resources for novel genes/mechanisms to be used for cotton improvement.
In this work, accessions of Gossypium spp. were evaluated for resistance to RKN in greenhouse experiments.
Responses to infection by M. incognita varied among the tested accessions, ranging from highly susceptible to
resistant. Some accessions displayed a significant reduction in the nematode reproduction. Histological observations of
one of the highly resistant G. barbadense accession showed that resistance may occur through two-stage mechanism
involving a hypersensitive-like response.
The highly resistant accession was crossed with a susceptible one to generate F1 and F2 plants for further genetic
studies. Analysis of the response of these F1 and F2 plants to RKN inoculation indicated that resistance is recessive,
and controlled by at least one major gene. Analyses using molecular markers associated to known RKN resistance loci
showed that the allele(s) involved are different from those previously described.
The characterization of the genetics and of the defense mechanisms associated with this novel source of resistance
to RKN in cotton constituted a first step towards its use in crop improvement.
Keywords: Gossypium, cotton, root-knot nematode, host-plant resistance, hypersensitive response
INTRODUCTION
The root-knot nematode (RKN) [Meloidogyne incognita Kofoid and White 1919 (Chitwood 1949)] is a
major constraint in cotton (Gossypium hirsutum L.) production in a number of countries, causing direct
damages and increasing in the severity of other root diseases, including Fusarium wilt disease (Hyer et
al. 1979; Shepherd 1982; Jeffers and Roberts 1993). The importance of this pest has been increasing over
the years, and in some regions, it has become one of the major causes of yield reduction.
Resistant varieties not only help control the disease and maintaining crop productivity, but they also
help decrease nematode populations in the soil and protect following rotations (Williamson and Hussey
1996; Ogallo et al. 1999; Starr et al. 2007; Davis and Kemerait 2009). Control of RKN has been
hampered by the lack of high-quality locally-adapted varieties exhibiting high levels of resistance as well
as the lack of adequate options for crop rotation.
Search for high levels of RKN resistant in cotton germplasm has been undertaken over the years, in
both cultivated species as well as in wild relatives (Jenkins et al.1979; Shepherd 1983; Robinson and
Percival 1997). Despite these efforts, few accessions with a high level of resistance have been identified.
In a more recent study, Robinson et al. (2004) identified three accessions of G. hirsutum (TX-25, TX-
1828, and TX-1860) that showed resistance levels equivalent to that of Auburn 623 RNR. This elite
breeding line (Shepherd 1974a), that was selected from crossing between two moderately-resistant
accessions Clevewilt 6 and Wild Mexican Jack Jones (Shepherd 1974b), exhibits the highest level of
resistance to RKN known to date in cotton, and has been used to derive a number of breeding lines
(Shepherd et al. 1996). Nevertheless, the high level of resistance of Auburn 623 RNR and of its
derivatives (M-series) has not been transferred to superior cultivars. Only very recently were
51
Identification and Characterization of a Novel Source of Resistance to Root-Knot Nematode in Cotton 299
lines with high levels of resistance released (Davis et al. 2011; Starr et al. 2011). The cultivar Clevewilt 6
is also at the origin of the obsolete varieties Stoneville LA 887 (Jones et al. 1991) and Paymaster (Hartz)
1560, that were widely cultivated for their moderate levels of resistance to RKN, and of their sister lines
(La. RN 4-4, La. RN 909, La. RN 910, La. RN 1032) (Jones et al. 1988). To date, the only available
moderately RKN-resistant varieties with desirable agronomical and quality standards are Acala Nem X
(Oakley 1995) and Acala NemX H Y (Anonymous 2005), which have a restricted diffusion due to their
particular characteristics (Acala-type cotton).
Variability in virulence of RKN isolates on resistant cotton genotypes has been demonstrated
(Robinson and Percival 1997; Zhou et al. 2000). Furthermore, selection of isolates with increased
reproduction on resistant varieties after repeated exposures to resistant cotton was also evidenced (Ogallo
et al. 1997), indicating the need to increase the number of the sources of resistance to achieve effective
durable resistance. Indeed, cotton breeding for RKN resistance presently relies on a small number of if
not a unique source of resistance, which make such genotypes vulnerable to resistance breakdown.
Alternating sources of resistance, or pyramiding resistance factors constitutes a way to mitigate this
problem.
Breeding for nematode resistance in cotton has been an arduous task. The difficulty in the phenotypic
screening for resistance on a scale compatible with that of breeding programs, and the lack of a clear
understanding of the genetic basis of resistance has made progress difficult. Genetic analyses involving
different sources of resistance point out to the presence of multiple genes. Depending on the source of
resistance and on the crosses used, genes with dominant and others with recessive effects have been
detected; additive effects as well as transgressive segregation have also been shown to occur (Shepherd
1974b; Bezawada et al. 2003; McPherson et al. 2004; Zhang et al. 2007; Wang et al 2008; Ulloa et al
2010).
The efficient transfer of RKN resistance to improved commercial cultivars will largely depend on a
clear knowledge of the genetics of the trait and on the availability of tools to facilitate such a transfer.
Furthermore, the knowledge of resistance gene/locus diversity and of the allelic relations between these
genes/loci is important to achieve a durable high level of resistance.
Molecular genetics tools, and in particular the genetic mapping of resistance genes/loci and the
identification of molecular markers tightly associated with these resistance genes or loci have been useful
in better understanding the genetics of RKN resistance in cotton, in studying the genetic relation between
genes/loci, and represent powerful tools to assist the transfer of resistance for cotton crop improvement.
In recent years, a number of genetic mapping studies have been undertaken aiming at the mapping
resistance of loci and at identifying molecular markers associated with RKN resistance (Bezawada et al
2003; Shen et al 2006; Wang and Roberts 2006; Wang et al 2006; Ynturi et al 2006; Niu et al 2007;
Wang et al 2008; Gutirrez et al 2010; Shen et al 2010). These studies have allowed to clarify the status
of nematode resistance loci in cotton, and to identify molecular markers tightly associated to major
resistance genes that are useful in breeding.
These and other studies (Roberts and Ulloa 2010) point out to chromosome 11 as bearing major
genes in at least two sources of resistance, and have resulted the identification of markers closely
associated to resistance. In Acala NemX, the microsatellite marker CIR316 is closely associated with a
major recessive gene (rkn1) (Wang et al 2006). In the Auburn 623 RNR-derived sources of resistance
(M240 and M120), the same markers is associated with a major dominant gene, Mi-C11(Gutirrez et al
2010, Shen et al 2010). It is not clear whether these loci are allelic or not. Interestingly, Clevewilt 6, one
of the parental lines used to develop the highly resistant Auburn 623 RNR accession, also carries a
resistance QTL associated with marker CIR316 (Bezawada et al 2003, Gutirrez et al 2010). Similarly, a
major QTL for resistance (galling index) was mapped on the same chromosome in M-495, a wild cotton
germplasm line (He et al 2010). In G. barbadense, in addition to these major genes, chromosome 11 has
also been shown to carry a transgressive segregation factor (RKN2) associated with the recessive rkn1
gene. On its own, RKN2 does not impart resistance, but when present with the rkn1 allele, RKN2
increases resistance (Wang et al 2008). The major resistance allele identified by marker CIR316 is not
present in Wild Mexican Jack Jones (WMJJ), the second parental line of Auburn 623 RNR. Instead,
WMJJ carries another locus on chromosome 14, linked to markers BNL 3545 and BNL 3661 that is also
present in Auburn 623 RNR and its derivatives, but not in Clevewilt 6 (Gutirrez et al 2010).
Consistent with previous studies, this latter molecular mapping study also showed that each one of
the major genes/loci is responsible for different resistance mechanisms, that, when present together, lead
to the highest level of resistance. The gene/locus on chromosome 11 primarily impacts root galling, while
the proper allelic combination at locus on chromosome 14 induces a reduced egg production. The
favorable allelic composition at all three markers lead to the highest level of resistance (Gutirrez et al
2010).
IDENTIFICATION OF A NOVEL SOURCE OF RESISTANCE TO RKN IN COTTON
Accessions of Gossypium species, which included modern or obsolete cultivars, breeding lines, and wild
accessions of G. hirustum, G. barbadense, and G. arboreum with known or suspected resistance to RKN
were evaluated for their resistance to a Brazilian isolate of M. incognita race 3 under controlled
conditions in a greenhouse. Resistance was evaluated based on three criteria: galling index (GI), egg
mass index (EMI) and reproduction factor (RF).
Among the accessions tested, reactions to RKN inoculation varied from highly susceptible to
resistant (data not shown). In agreement with previous studies (Shepherd 1983; Robinson and Percival
1997; Robinson et al. 2004), no general trend between species and reaction to inoculation was observed.
Similarly, no relation with geographical origin was evidenced. Most of the accessions that had been
tested in other studies showed responses in agreement with published results. Among the accessions
tested, the G. barbadense accession from Peru CIR1348 showed highly reduced nematode reproduction
(Table 1), and was classified as highly resistant. This accession was as efficient as M-315RNR the
resistant control in reducing nematode reproduction (RF = 0.01 vs. RF = 0.03 for M315RNR). In
addition, this accessions displayed very low galling index (GI = 0) and egg mass index (EMI = 0).
TABLE 1: GALLING INDEX (GI), EGG MASS INDEX (EMI) AND REPRODUCTION FACTOR (RF) PRESENTED BY DIFFERENT GOSSYPIUM SPP. 120 DAYS AFTER INOCULATION WITH 5,000 M.
INCOGNITA EGGS PER PLANT
Accession GI
1
EMI
1
RF
2
FM966 susceptible control G. hirsutum 5 5 14a
M-315RNR resistant control G. hirsutum 0.8 0 0.03b
CIR1348 G. barbadense 0 0 0.01b
1
Mean value (8 repetitions) of GI or EMI. 0: no galls or egg masses, 1: 1-2 galls or egg masses, 2: 3-10 galls or egg masses,
3: 11-30 galls or egg masses, 4: 31-100galls or egg masses, and 5 >100 galls or egg masses per root system.
2
RF = FP/IP, were FP = final nematode population and IP = initial nematode population (IP = 5,000). Mean values
(8 repetitions) were transformed in log (x+1). Means followed by different letters are significantly (P< 0.05) according to
Scott-Knots test.
Accession CIR1348 thus appears to be as resistant to RKN inoculation as the accessions that display
the highest level of resistance known to date (Auburn 623RNR and derivatives). Interestingly, CIR1348
is wild accessions of G. barbadense from Peru. South America, and in particular Peru, is considered to be
the center of origin and diversity of G. barbadense (Giband et al 2010). It is thus expected that a rich
genetic variability is encountered in wild accessions from this region (Westengen et al. 2005), including
for resistance to RKN and other disease or pests. This situation is similar to that of wild accessions and
landraces of G. hirsutum from Mexico, the center of origin of the species, which include the accessions
Wild Mexican Jack Jones and TX-25 in which notable levels of resistance were identified (Shepherd
1983; Robinson and Percival 1997; Robinson et al. 2004).
HISTOLOGICAL CHARACTERIZATION OF THE RESISTANCE REACTION IN ACCESSION CIR1348
The mechanism of the resistant displayed by the highly resistant accession CIR1348 was studied through
the observation of histological sections of root samples using bright-field and UV microscopy.
Stage 2 juvenile (J2) penetration was not affected in accession CIR1348, since similar numbers of J2s
could be observed in the susceptible and resistant accessions. Similar observations were made in the
moderately resistant accession Clevewilt-6 (McClure et al. 1974), in the highly resistant accession M-315
RNR (Jenkins et al. 1995), and in number of other resistant accessions (Faske and Starr 2009). Pre-
existing mechanisms which could impede nematode penetration seems to be apparently absent in cotton.
Rather, in accession CIR1348, as in other RKN-resistant accessions of cotton, it appears that resistance
may result from post-penetration events associated with the blocking or delay of nematode development
and reproduction.
Root sections harvested at 7-21 days after inoculation (DAI) showed major alteration in the cells in
contact with the nematodes. Hypersensitive- response (HR)-like lesions were found around all nematodes
after they penetrated the epidermis and migrated through the cortex, or when they reached the vascular
cylinder. Sections also showed almost entire bodies of nematodes completely surrounded by
autofluorescence or toluidine dark-stained components.
At 21-29 DAI, only a few giant cells were observed, some of them showing multiple nuclei and
reduced thickening of walls. At 21 DAI, strongly deformed J3/J4 juveniles were detected in the vicinity
of the altered giant cells. At 29 DAI, most giant cells had degenerated, and presented a retracted
cytoplasm containing numerous small vacuoles. No adult female with eggs were seen in any of the 34-45
DAI sections that were analyzed.
It thus appears that in CIR1348 at least two different mechanisms could be involved in the expression
of resistance. One mechanism, which occurs at about 7 DAI, blocks or delays the development of J2 that
have penetrated the roots. The second, that involves a mechanism impeding the formation of functional
feeding sites, occurs at about 21 DAI and further impedes the formation of adult females.
Genetic analyses (McPherson et al. 2004; Zhang et al. 2007) point out to a 2-gene model for the
inheritance of resistance to RKN in cotton. In their study, Jenkins et al. (1995) proposed that one gene
acting at an earlier stage is responsible for the mechanism seen at 8 DAI, while the second explains the
later (24 DAI) phenomenon. Molecular mapping data (Ynturi et al. 2006; Gutirrez et al. 2010) support
these hypotheses, and revealed the occurrence of QTLs on chromosomes 11 and 14 to explain the
resistance in cotton accessions (Auburn 634 RNR and M-240 RNR, respectively) which share the same
source of resistance as M-315 RNR. The QTL on chromosome 11 is associated with reduced root galling
index, while that on chromosome 14 is associated with reduced egg production (Gutirrez et al. 2010).
Whether this situation holds true for accession CIR1348 remains to be clarified.
GENETIC ANALYSIS OF THE RESISTANCE IN ACCESSION CIR1348
The highly resistant accession CIR1348 was crossed with a susceptible one (the susceptible control
FM966) to generate F
1
and F
2
plants for further genetics studies. As above, progenies were assessed for
GI, RMI, and RF after RKN inoculation under controlled conditions. The analysis of the response of the
F
1
plants clearly showed that resistance in accession CIR1348 is recessive, the F
1
showing a GI = 5, EMI
= 4.8, and RF = 25, values similar to that of the susceptible parent FM966 (GI = 5, EMI = 5 and
RF = 29).
The results of the F
2
plants were more complex to analyze. Analyses conducted on a reduced number
of plants (n = 18) showed that at least one major recessive gene is involved in the determination of the
phenotype. Nevertheless, these analyses cannot rule out the hypothesis that a second recessive gene, with
a more moderate effect on the phenotype, is also involved in determining the high level of resistance
observed in accession CIR1348. The analysis of a larger number of F
2
plants is underway to clarify this
point.
If the second hypothesis holds true, the situation in accession CIR1348 would be similar to that in
other sources of resistance, with the main difference being that the resistance in the former is recessive,
while it is usually considered dominant (or partially dominant) in the latter.
A number of studies (Shen et al 2006; Wang et al 2006; Gutirrez et al 2010; He et al 2010; Shen et
al 2010) have shown that the SSR marker CIR316, mapped on chromosome 11, is associated to RKN
resistance in a number of accessions. To verify if the same locus is also involved in the resistance in
accession CIR1348, we applied marker CIR316 and analyzed the resulting amplification profile.
While the susceptible and resistant controls (FM966 and M-315RNR, respectively) exhibited the
banding pattern expected for marker CIR316, accession CIR1348 showed an amplification pattern
different from that of both controls. It thus appears that resistance in accession CIR1348 is determined by
allele(s) different from that (those) previously described for known sources of resistance.
The present study on the characterization of the genetics and of the defense mechanisms associated
with this novel source of resistance to RKN in cotton constituted a first step towards its use in crop
improvement.
REFERENCES
[1] Anonymous (2005). Plant variety protection certification no. 200500113 for cotton Acala
[2] NemX HY. Plant Variety Protection Office, Agricultural Marketing Service, US Department of Agriculture, Washington,
DC.
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resistance gene(s) in cotton. The Journal of Cotton Science, 7, 179-184.
[4] Davis, R. F. and Kemerait, R.C. (2009). The multi-year effects of repeatedly growing cotton with moderate resistance to
Meloidogyne incognita. Journal of Nematology, 41, 140-145.
[5] Davis, R.F., Chee, P., W., Lubbers, E. L., and May, O. L. (2011). Registration of GA120R1B3 germplasm line of cotton.
Journal of Plant Registrations, 5, 384-387.
[6] Faske, T. R. and Starr, J. L. (2009). Mechanism of resistance to Meloidogyne incognita in resistant cotton genotypes.
Nematropica, 39, 281-288.
[7] Giband, M., Dessauw, D., and Barroso P. A. V. (2010). Cotton: Taxonomy, Origin, and Domestication. In Wakelyn, P.J. &
Chaudhry M.R (Eds) Cotton: Technology for the 21
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Century (pp 5-17). International Cotton Advisory Committee,
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[8] Gutirrez, O.A., Jenkins, J.J., McCarty, J.C., Wubben, M.J., Hayes, R.W. and Callahan, F.E. (2010). SSR markers closely
associated with genes for resistance to root-knot nematode on chromosomes 11 and 14 of Upland cotton. Theoretical and
Applied Genetics, 121, 1323-1337.
[9] He, Y., Iqbal, N., Shen, X., Davis, R.F., and Chee, P. (2010). QTL mapping of resistance to root-knot nematode in the wild
cotton germplasm line M-495RNR. P. 763. In: Proceedings of the Beltwide Cotton Conference, New Orleans, LA, Jan 47,
2010. National Cotton Council of America, Memphis, TN, USA.
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nematode Fusarium wilt disease complex in cotton. Crop Science, 19, 898-901.
[11] Jeffers, D. P., and Roberts, P. A. (1993). Effect of plant date and host genotype on the root-knot Fusarium wilt disease
complex of cotton. Phytopathology, 83, 645-654.
[12] Jenkins, J. N., Parrott, W. L., Kappelman, A. J., & Shepherd, R. (1979). Registration of JPM 781-783 cotton germplasm.
Crop Science, 19, 932.
[13] Jenkins, J. N., Creech, R. G., Tang, B., Lawrence, G. W., & McCarty, J. C. (1995). Cotton resistance to root-knot nematode
: II. Post-penetration development. Crop Science, 35, 369-373.
[14] Jones, J. E., Beasley, J. P., Dickson, J. I., & Caldwell, W. D.(1988). Registration of four cotton germplasm lines with
resistance to reniform and root-knot nematodes. Crop Science, 28, 199-200.
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LA 887 cotton. Crop Science, 31, 1701.
[16] McClure, M. A., Ellis, K. C., & Nigh, E. L. (1974). Post-infection development and histopathology of Meloidogyne
incognita in resistant cotton. Journal of Nematology, 1, 21-26.
[17] McPherson, G.R., Jenkins, J.N., Watson, C.E, and McCarty, J.C. (2004). Inheritance of root-knot nematode resistance in M-
315 RNR and M78-RNR cotton. Journal of Cotton Science, 8, 154-161.
[18] Niu, C., D.J. Hinchliffe, R.G. Cantrell, C. Wang, P.A. Roberts, and J. Zhang. (2007). Identification of molecular markers
associated with root-knot nematode resistance in upland cotton. Crop Science, 47, 951-960.
[19] Oakley, S. R. (1995). CPCSSD Acala C-225: A new nematode resistant Acala variety for Californias San Joaquin Valley.
In: Proceedings of 1995 Beltwide Cotton Production Research Conference (p. 39). Memphis, TN: National Cotton Council
of America.
[20] Ogallo, J.L., Goodell, P.B., Eckert, J., & Roberts, P.A. (1997). Evaluation of NemX, a new cultivar of cotton with high
resistance to Meloidogyne incognita. Journal of Nematology, 29, 531-537.
[21] Ogallo, J.L., Goodell, P.B., Eckert, J., and Roberts, P.A. (1999) Management of root-knot nematodes with resistant cotton
cv. NemX. Crop Science, 39, 418-421.
[22] Roberts, P.A., and Ulloa, M. (2010). Introgression of root-knot nematode resistance into tetraploid cottons. Crop Science,
50, 940-951.
[23] Robinson, A. F. & Percival, A. E. (1997). Resistance to Meloidogyne incognita raa 3 and Rotylenchulus reniformis in wild
accessions of Gossypium hirsutum and G. barbadense from Mexico. Journal of Nematology, 29, 746-755.
[24] Robinson, A. F., Bridges, A. C., & Percival, E. (2004). New source of resistance to the reniform (Rotylenchus reniformis
Linford and Oliveira) and root-knot (Meloidogyne incognita Kofoid &White, Chitwood) nematode in upland (Gossypium
hirsutum L.) and Sea Island (G. barbadense L.) cotton. The Journal of Cotton Science, 8, 191-197.
[25] Shen, W., Van Becelaere, G., Kumar, P., Davis, R.F., May, O.L., and Chee, P. (2006). QTL mapping for resistance to root-
knot nematodes in M-120 RNR Upland cotton line (Gosspypium hirsutum L.) of the Auburn 623 RNR source. Theoretical
and Applied Genetics, 113, 1539-1549.
[26] Shen,X., He, Y., Lubbers, E.L., Davis, R.F., Nichols, R.L., and Chee, P.W. (2010). Fine mapping QMi-C11 a major QTL
controlling root-knot nematode resistance in Upland cotton. Theoretical and Applied Genetics, 121, 1623-1631.
[27] Shepherd, R. L. (1974a). Registration of Auburn 623 RNR cotton germplasm. Crop Science, 35, 373-375.
[28] Shepherd, R. L. (1974b). Transgressive segregation for root-knot nematode resistance in cotton. Crop Science, 14, 872875.
[29] Shepherd, R. L. (1982). Genetic resistance and its residual effects for control of the root-knot nematode Fusarium wilt
complex in cotton. Crop Science, 22, 1151-1155.
[30] Shepherd, R. L. (1983). New sources of resistance to root-knot nematodes among primitive cottons. Crop Science, 23, 999-
1002.
[31] Shepherd, R. L., McCarty, J. C., Jenkins, J. N., & Parrott, W. L. (1996). Registration of nine cotton gernplasm lines
resistant to root-knot nematode. Crop Science , 36, 820.
[32] Starr, J. L., Koenning, S. R., Kirkpatrick, T. L., Robinson, A. F., Roberts, P. A., and Nichols, R. L. (2007). The future of
nematode management in cotton. Journal of Nematology, 39, 283-294.
[33] Starr, J.L., Smith, C.W., Ripple, K., Zhou, E., Nichols, R.L., and Faske, T.R. (2011). Registration of TAM RKRNR-9 and
TAM RKRNR-12 germplasm lines of upland cotton resistant to reniform and root-knot nematodes. Journal of Plant
Registrations, 5, 393-396.
[34] Ulloa, M., Wang, C., and Roberts, P. A. (2010). Gene action analysis by inheritance and quantitative trait loci mapping of
resistance to root-knot nematodes in cotton. Plant Breeding, 129, 541-550.
[35] Wang, C. and Roberts, P.A. (2006). Development of AFLP and derived CAPS markers for root-knot nematode resistance in
cotton. Euphytica, 152, 185-196.
[36] Wang, C., Ulloa, M., and Roberts, P.A. (2006). Identification and mapping of microsatellite markers linked to a root-knot
nematode resistance gene (rkn1) in Acala NemX cotton. Theoretical and Applied Genetics, 112, 770-777.
[37] Wang C. Ulloa, M., and Roberts, P.A. (2008). A transgressive segregation factor (RKN2) in Gossypium barbadense for
nematode resistance clusters with gene rkn1 in G. hirsutum. Molecular Genetics and Genomics, 279, 41-52.
[38] Williamsom, V. M., and Hussey, R. S. (1996). Nematode pathogenesis and resistance in plants. The Plant Cell, 8, 1735-
1745.
[39] Westengen, O. T, Huamn, Z., and Heun, M. (2005). Genetic diversity and geographic pattern in early South American
cotton domestication. Theoretical and Applied Genetics, 110, 392-402.
[40] Ynturi, P., Jenkins, J.J. MacCarty, J.C., Guttierrez, O.A., and Saha, S. (2006). Association of root-knot nematode resistance
genes with simple sequence repeat markers on two cromosomes in cotton. Crop Breeding and Genetics, 46, 2670-674.
[41] Zhang, J. F., Waddell, C., Sengupta-Gopalan, C., Potenza, C., and Cantrell, R.G. (2007). Diallel analysis of root-knot
nematode resistance based on galling index in upland cotton. Plant Breeding, 126, 164-168.
[42] Zhou, E., Wheeller, T. A., & Starr, J. L. (2000). Root galling and reproduction of Meloidogyne incognita isolates from
Texas on resistant cotton genotypes. Journal of Nematology, 32, 513-518.
Predominance of Resistance Breaking Cotton Leaf

Curl Burewala Virus (ClCuBuv)
in Northwestern India
Prem A. Rajagopalan
1
, Amruta Naik
1
, Prashanth Katturi
1
, Meera Kurulekar
1

Ravi S. Kankanallu
2
and Radhamani Anandalakshmi
1

1
Plant-Virus Interactions Lab, Mahyco Research Center, Maharashtra Hybrid Seeds Company
Limited, Dawalwadi, Post Box no-76, Jalna, Maharashtra431 203, India

2
Vegetable Research Center, Maharashtra Hybrid Seeds Company Limited, Bettanagera Village,
Huskur Post, Bangalore562 123, India
Cotton leaf curl disease (CLCuD) is the most devastating among viral diseases of cotton Gossypium
hirsutum (L.) in northwestern India (Varma and Malathi, 2003). Plants susceptible to the virus show up-
ward and down-ward leaf curling,thickening of veins, enations on the leaf abaxial surface, overall
stunting, and flower and fruit abortion leading to low productivity. CLCuD is caused by a group of
geminivirus species from the genus Begomovirus, in the family Geminiviridae. Cotton infecting
begomoviruses (CBVs) like the majority of Old World begomoviruses, are monopartite having genomes
that consist of only one circular single-stranded DNA (ssDNA) molecule and are associated with
betasatellite (Briddon et al. 2003) and frequently a third component known as alphasatellite (Briddon et
al., 2004). The CBVs complex is transmitted by the whitefly Bemisia tabaci (Genn.)
Though CBVs was first reported from IARI, New Delhi, India in 1989, it was identified as the causal
agent of severe epidemic outbreaks of the CLCuD in Punjab and adjacent SriGanganagar in 1994. Since
then, CBVs has spread to all of the cotton growing regions of north westerns India, where it has become
the limiting factor for cotton production in every season, causing up to 100% yield loss. The major
begomoviruses associated with CLCuD are Cotton leaf curl Rajasthan virus (CLCuRV), Cotton leaf curl
Multan virus (CLCuMuV) and Cotton leaf curl Kokhran virus (CLCuKoV).
During 2002, we commenced a comprehensive study to understand the distribution, diversity and
biological characterization of CBVs in northwestern India. In surveys conducted during 2002-2005, we
noticed predominance of CLCuRV in fields when compared to either CLCuMuV or CLCuKoV. The
cotton hybrids and varieties which were developed and marketed by the different seed companies and
public institutions were showing varying degrees of tolerance to different CBVs.
During our reconnaissance studies in the 2005 cropping season, some of the plants in farmers fields
in SriGanganagar showed the severe symptoms in cotton cultivars which were earlier resistant to
Rajasthan strain (CLCuRV), Later we confirmed that the plants were infected by the e resistance
breaking virus Burewala strain and now recognized as a distinct begomovirus species- Cotton leaf curl
Burewala virus(CLCuBuV).
However, during 2009 -2010, severe and wide spread CLCuD was observed on cotton in the fields of
Bathinda, Abohar, Fazilka, SriGanganagar, and the surrounding Punjab and Rajasthan states and causing
yield loss even up to 100%. Most cotton cultivars previously resistant to CBVs were found to be severely
affected
To identify the specific viral genotype(s) involved in the recent outbreak, begomovirus field isolates
were collected from cotton fields and subjected to DNA sequencing. Partial sequences of 258, as well as
full-length sequences of 30 complete virus genome sequences were determined and sequences were
compared to those isolates from 2003-2008. Based on partial and full length genome sequences, it can be
concluded that the new emergent, resistance-breaking strain, CLCuBuV has become established in
northwestern India.
52
Predominance of Resistance Breaking Cotton Leaf Curl Burewala Virus 305
Nearly 93% (238 out of 258) of the samples were infected with CLCuBuV and full length
characterization studies showed that this virus isolates are prevalent in three putative mutant forms. We
have noticed two major variations when compared to place of origin of the mutant virus. Amrao et al.
(2010) reported the prevalence of three C2 mutants in Vehari, Pakistan, but we were able to collect only
one type of mutant in our surveys. Further Amrao et al. (2010) speculated that, C2 mutation is an escape
mechanism in CLCuD resistant G.hirsutm lines. Interestingly, we have isolated CLCuBuV carrying
intact C2, also from these tolerant lines. How these different mutant viruses are vectored by whiteflies
and how the virus is thriving on different genotypes of cotton from one season to another need to be
looked into for devising effective control measures to combat the spread of CLCuD
REFERENCES
[1] Amrao L, Amin I, Shahid MS, Briddon RW, Mansoor S (2010a) Cotton leaf curl disease in resistant cotton is associated
with a single begomovirus that lacks an intact transcriptional activator protein. Virus Res 152:153163
[2] Briddon RW (2003) Cotton leaf curl disease, a multi component begomovirus complex. Mol Plant Pathology 4:427434
[3] Briddon RW, Bull SE, Amin I, Mansoor S, Bedford ID, Rishi N, Siwatch S, Zafar MY, Abdel-Salam AM, Markham PG,
(2004) Diversity of DNA1; a satellite-like molecule associated with monopartite begomovirusDNA complexes. Virology
324:462474
[4] Varma A, Malathi VG (2003) Emerging geminivirus problems: A serious threat to crop production. Annals of Applied
Biology 142:145164

Cotton Production, Physiology and Economics

Cotton Genotypes Performance
under Rainfed and Irrigated Conditions
in two Regions of Northern Argentina
Marcelo Paytas
1
and Jose Tarrago
2

1
INTA EEA Reconquista, Santa Fe, (3560), Argentina
2
INTA EEA Las Brenas, Chaco, (3722), Argentina
E-mail: mpaytas@correo.inta.gov.ar

AbstractNarrow-row cotton production systems have became popular in Argentina in the last few years. It is mainly
cultivated under rainfed conditions as a low input crop which is challenging and risky. Irrigation can improve the
performance of current genotypes and may reduce the variability in yield produced under rainfed conditions across different
environments. This research was aimed to understand the differences in growth, development and yield of two cotton
cultivars in a narrow row system under rainfed and irrigated conditions.
Experiments were conducted during 2010-11 under rainfed and irrigated conditions at the Research Station of INTA
Reconquista, Santa Fe (2911S, 5942W) and INTA Las Brenas, Chaco (2705S, 6106W). The annual rainfall and its
distribution, temperatures, evaporative demand and soil types differ between both cotton regions. The experimental design in
each location was a split plot design with four replications: two genotypes (NuOpal and DP402) with two moisture levels
(rainfed and irrigated). The results indicated differences between genotypes in terms of days to crop maturity. Earliness was
found for DP402 for both locations compared with NuOpal. However, no significant differences in terms of phenology were
found between rainfed and irrigated conditions due the amount of soil water content available from rainfall for the plant in
both systems. Dry matter production and partitioning to reproductive organs was affected by genotypes and moisture levels.
DP402 with shorter vegetative and reproductive stages produced significant differences in dry matter between moisture levels
than NuOpal with later maturity. Percentage of fruit retention increased by maturity in DP402 compared with NuOpal
under both rainfed and irrigated conditions, although NuOpal produced higher number of nodes and fruiting sites but higher
fruit abortion in the lower part of the plant.
INTRODUCTION
Narrow-row cotton has become popular in Argentina in the last few years reaching about 90% of the
national sowing area. By reducing distance between rows and increasing plant population, plants became
smaller and harvested with stripper machines reducing harvesting costs compared with previous
traditional cotton systems. Cotton is mainly cultivated under dryland conditions as a low input crop
which is challenging and risky. However, irrigation practices can improve the current genotypes
performances and may reduce the variability in yield.
Changing row spacing and plant population has been used to increase yield in many other crops. By
changing the spacing between plants, competition for light, water and nutrients is altered, which can
change fruit number and retention per plant and the size of the plant (Bednarz, 2000). Due to the
influence of environmental conditions on plant growth and development, specific row spacing and
population recommendations for crops may vary. The optimum plant population for any crop is the
population that maximizes yield while optimising resource use (Willey and Heath, 1969). Nowadays,
Argentinean cotton farmers are mainly using 52 cm as row spacing and 220,000 plants per hectare.
Whether this population is optimal or not to produce high yielding cotton with current Bt varieties is
focus of numerous studies.
Boll retention and distribution within a plant play an important role in determining final yield, and
are linked to the allocation of assimilate produced during vegetative growth by the plant. If the
availability of assimilate is adequate to support the developing bolls, then the bolls will be retained
(Constable, 1991; Jenkins et al., 1990a). However, if the demand from growing bolls exceeds the
assimilate supply, the retention of bolls will decline as a result of an increase in the boll shedding (Guinn,
1998; Mason, 1922).
53
Most of the time, research has been done comparing different crop configuration, while in this work
the aim was to maintain the same configuration and vary the inputs of water to increase source
availability for maximizing cotton yield, using two different genotypes and two growing environmental
conditions. Increased resource availability by irrigation may reduce the variability in yield produced
under rainfed conditions across these different environments. This research was aimed to understand the
differences in growth, development and yield of two cotton cultivars in narrow row systems under
rainfed and irrigated conditions.
Experiments were conducted during 2010-11 under rainfed and irrigated conditions at the Research
Station of INTA Reconquista, Santa Fe (2911S, 5942W) and INTA Las Brenas, Chaco (2705S,
6106W), Argentina. The experimental design in each location was a split plot design with four
replications: two cultivars (NuOpal and DP402) were sown at a spacing of 0.52 m between rows having
11 plants per meter. Two soil moisture treatments (irrigated and rainfed) were compared. In both the
locations, irrigated plots received three irrigations at the time of flowering (about 90 mm in Las Brenas
and 60 mm in Reconquista) besudes water from rainfall. Neutron moisture meter measurements were
used to monitor soil moisture content (0-150 cm depth). Harvests for total biomass, biomass partitioning,
radiation interception and yield, as well as mapping, were done at various developmental stages
throughout the season. Meteorological conditions were recorded during the season.
The annual rainfall and its distribution, temperatures, evaporative demand and soil types differ between
both cotton regions. Wetter conditions were found at Reconquista, Santa Fe compared with Las Brenas,
Chaco. Differences between cultivars were observed in terms of days to crop maturity. Earliness was
found for DP402 for both locations compared with NuOpal. However, no significant differences in terms
of phenology were found between rainfed and irrigated conditions due the amount of soil water content
available from rainfall for the plant in both systems.
Dry matter production and partitioning to reproductive organs was affected by genotypes and
moisture levels. DP402 with shorter vegetative and reproductive stages produced significant differences
in dry matter production between moisture levels than NuOpal with later maturity. In both locations,
similar responses were found in terms of dry matter production and partitioning to reproductive organs.
Lower solar radiation interception was found in the lower part of the canopy in NuOpal. Possibly, the
greater vegetative growth in NuOpal may have contributed to reduced boll growth and shedding of
flowers and young bolls lower in the canopy due to poor light infiltration. Percentage of fruit retention in
first fruit positions on the main stem increased by maturity in DP402 compared with NuOpal under both
rainfed and irrigated conditions. NuOpal produced higher number fruits abortions in the lower part of the
plant. It is likely that solar radiation and photosynthesis in low position fruiting sites become a limitation,
with a bigger plant and complete canopy closure resulting in fruit abortions and decrease in the yield
potential in conventional cropping systems (Constable and Rawson, 1980b; Wullschleger and Oosterhuis,
1990a; 1990b).
The longer period to maturity in NuOpal may compensate after reproductive organs in the first few
positions were aborted, with higher number of nodes and fruiting sites on lateral and upper part of the
canopy, increasing final cotton yield in a wet season for both locations. However, DP402 with shorter
vegetative and reproductive period produced higher seed cotton yields than NuOpal (Table 1) in both
locations, with a better crop performance under narrow row systems in a subtropical environment with
humid crop season.
Cotton Genotypes Performance under Rainfed and Irrigated Conditions in two Regions of Northern Argentina 311
TABLE 1: SEED COTTON YIELD (KG*HA-1) FOR TWO GENOTYPES UNDER RAINFED AND IRRIGATED CONDITIONS FOR TWO LOCATIONS IN NORTH ARGENTINA
Location: Reconquista,
Santa Fe
Seed Cotton yield (kg*ha-1) Location: Las Brenas, Chaco Seed Cotton yield (kg*ha-1)
NuOpal-I 3.298 NuOpal-I 2.160
NuOpal-RF 3.045 NuOpal-RF 2.156
DP402-I 3.788 DP402-I 3.231
DP402-RF 3.540 DP402-RF 3.340
Significance * *
I: Irrigated treatment
RF: Rainfed treatment
*Significance (P 0.05)
REFERENCES
[1] Bednarz, C.W., Bridges, D.C. and Brown, S.M. (2000) - Analysis of cotton yield stability across population densities.
Agronomy Journal 92, 128-135.
[2] Constable, G.A. and Rawson, H.M. (1980b) - Carbon production and utilization in cotton - inferences from a carbon
budget. Australian Journal of Plant Physiology 7: 539-553.
[3] Constable, G.A. (1991) - Mapping the Production and Survival of Fruit on Field-Grown Cotton. Agronomy Journal 83:
374-378.
[4] Guinn, G. (1998) - Causes of square and boll shedding. Beltwide Cotton Conferences, pp. 13551364.
[5] Jenkins, J.N., McCarty, J.C. and Parrott, W.L. (1990) - Effectiveness of fruiting sites in cotton - yield. Crop Science 30:
365-369.
[6] Mason, T.G. (1922) - Growth and abscission in Sea Island cotton. Annals of Botany 36: 457-484.
[7] Willey, R. and Heath, S. (1969) - The quantitative relationship between plant population and crop yield. Advances in
Agronomy 21: 281-321.
[8] Wullschleger, S.D. and Oosterhuis, D.M. (1990a) - Photosynthetic and respiratory activity of fruiting forms within the
cotton canopy. Plant Physiology 94: 463-469.
[9] Wullschleger, S.D. and Oosterhuis, D.M. (1990b) - Photosynthetic carbon production and use by developing cotton leaves
and bolls. Crop Science 30: 1259-1264.
The Adaptation of Irrigated Cotton

to the Tropical Dry Season
S.J. Yeates
1,2

1
Principal Research Scientist, CSIROPlant Industry, Ayr, Qld, Australia
2
The Australian Cotton Cooperative Research Centre

AbstractThe reintroduction of cotton to most of the Australian tropics was prevented by insect pests that are
dominant during the wet (summer) season and a perception that the crop could only be grown in the wet season.
Growing cotton during the dry (winter) season has avoided these pests. However the photothermal pattern of the dry
season is the reverse of the wet season and that of spring sown cotton in temperate latitudes. Average night
temperatures are cool mid season (12 to 14
o
C) with extremes < 6
o
C and high temperatures are likely early and late in
the season. Solar radiation is 20% less than at temperate latitudes mid season and could also limit crop growth. It was
not known what yield or fibre quality was possible. Over three seasons two upland Bt-transgenic cultivars and one
Gossypium barbadense cultivar were sown from March to June in field experiments at the Ord River (15.5
o
S). A pot
experiment conducted at Katherine, (14.5
o
S) over two seasons where average ambient minimum temperatures were
4
o
C lower than the field experiments during flowering were compared with temperatures 6
o
C higher by moving plants
into a glasshouse at night. Despite the photothemal constraints, lint yields were at the high end of Australian and
international benchmarks when sown in March and April. The lower temperature and radiation during flowering and
early boll growth for the March and April sowings combined to reduce the crop growth rate during this phase
compared with cotton grown at temperate latitudes. However, assimilate supply was adequate because boll demand
was also lower at this time due to early flowers having slower development, lower retention and smaller bolls.
Increasing late season temperature and radiation permitted yield compensation via an extended flowering period and a
greater contribution to yield from later pollinated flowers on the top and outside of the plant. The Katherine
experiment found boll retention and size was correlated (p < 0.01) with minimum temperature during flowering. Full
yield recovery occurred because cold minimums were episodic. RUE was negatively correlated with average
temperature up to first flower a response not reported previously in cotton and explained some of variation in RUE
measured here and elsewhere. Cool temperatures during fibre development reduced fibre length and strength at March
and April sowings. Further screening may identify cultivars with suitable fibre length and strength in these
conditions.
Introduction
There have been many attempts to grow cotton in the Australian semi-arid tropics (SAT). The region is
vast, approximately 30% of the Australian continent, and largely unutilised for cropping of any species.
The region contains about 66 drainage basins or river catchments; these account for around 60% of
Australias surface water runoff, with significant ground water and arable soils (NLWRA 2001). The
only significant commercial production of cotton in the region occurred at the Ord River between 1963
and 1974. Cotton was grown during the wet season (November to April) with irrigation supplementing
rainfall to finish the crop early in the dry season (April to June) (Hearn, 1975). Despite yields similar to
south-eastern Australia during the same period, cotton production became uneconomic due to poor fibre
quality and resistance of Helicoverpa armigera to insecticides, which resulted in excessive pesticide
usage (Hearn, 1975).
The reintroduction of cotton to the Australian SAT is being assessed via a multidisciplinary study
that evaluates a novel production system designed to avoid the pest management problems of the
previous cotton industry. The new system involves dry season cropping to avoid peak numbers of the
key pests Helicoverpa armigera, Helicoverpa punctigera, Spodoptera litura, Pectinophora gossypiella
and Anomis spp., which characterise the wet season, and incorporates Integrated Pest Management and Bt
transgenic genotypes (Strickland et al.,, 1998). A comparison of the proposed system with the previous
wet season system is shown in Table 1. Sowing of cotton crops from March 1st is desirable in the Ord
River and much of the Australian SAT as they flower during the cooler months of May to August
avoiding key insect pests (Strickland et al.,, 1998 & 2003). Once the first field is sown in a valley all
54
cotton must be sown within five weeks of that date to minimise the number of generations of
Helicoverpa armigeria exposed to Bt proteins (Monsanto and Cotton Australia, 2010). Pest management
research to date demonstrates effective insect management was achieved by adopting this system,
requiring only 3.5 insecticide sprays per crop (Strickland et al.,, 1998; Annells and Strickland, 2003)
compared with 40 for the 1970s industry (Hearn, 1975).
TABLE 1: KEY ELEMENTS OF A NOVEL COTTON PRODUCTION SYSTEM FOR THE ORIA CONTRASTED WITH THE PREVIOUSLY UNSUCCESSFUL SYSTEM OF
THE 1970S (ADAPTED FROM STRICKLAND ET AL., 1998)
1970s Industry New Industry
Wet season planting window that was long November to
February.
Dry season (winter) cropping, with a narrow planting window
(5 weeks) in March April.
Flowering from wet season (February) to early dry season
(May).
Flowering in low pest months of May to August.
Conventional cultivars Bt transgenic cultivars
Broad spectrum insecticides IPM systems
No pesticide resistance management Pre-emptive Bt resistance management
The key agronomic change in this proposed production system is the requirement for a five week
planting window that can commence on March 1
st
. Hence it is pertinent to ask why sowing after February
was not practiced previously in the Ord? Firstly, there was a perception that cotton growth and
development during the coldest months of May-August would be poor, delaying boll set until
temperatures increased and pushing harvest into the wet season. Results and recommendations on sowing
date were contradictory (Toms, 1963; Stern, 1965; Thomson, 1965; Hearn, 1975) prompting the
conclusion the possibility of March sowings warrants further investigation (Thomson, 1965). Secondly,
larger modern pickers combined with all weather storage of seed cotton are now available and reduce the
possibility of a long harvest and ginning season and UV light damage to fibre that occurred in the 1970s
(Hearn, 1975). Thirdly, prior to 1972, water storage capacity was insufficient to irrigate large areas of a
fully irrigated dry season crop but the irrigation system capacity is now expanded.
Growing cotton in the dry season creates new challenges for crop growth and the timing of farming
operations. A possible growing season of sowing in April, when trafficability is least affected by wet
season rain, then picking in October is the reverse of temperate Australia in terms of temperature and
daylength where cotton is usually sown in October and picked in April. Figure 1 compares the dry season
in the ORIA (Kununurra, 15
o
S) with temperate summer cotton at Narrabri, NSW (30
o
S), for monthly
rainfall, maximum and minimum temperature and solar radiation. Growing season rainfall is much less at
Kununurra (Fig. 1A), although rainfall prior to sowing is higher and may cause difficulties with land
preparation and sowing operations. It will be important to pick promptly at Kununurra as rainfall
increases significantly each month after October.
Monthly temperatures (Fig. 1B) are higher early and late in the season, while mid season minimum
temperatures are cooler averaging 14
o
C with extremes below 10
o
C (Cook and Russell, 1983) which could
be problematic for fibre quality and boll growth (Gipson and Ray 1970; Hearn 1994) and would delay
crop development (Constable and Shaw, 1988). High temperatures during September and October could
also be detrimental to boll growth (Hearn, 1994), but should enhance boll desiccation and improve
defoliant efficacy.
Potential daily photosynthesis is lower during flowering and boll growth at Kununurra because daily
radiation is about 80% of Narrabri during this phase (Fig. 1C) (Hearn, 1994). However, it is not known
whether reduced daily radiation will translate into lower yields as cooler temperatures may compensate
via slower development rate and less night respiration

Fig. 1: Climatic Comparison the Proposed Tropical Dry or Winter Growing Season in the Ord River (April to October) and the Temperate Summer
Growing Season at Narrabri 30oS (October to April) A) Mean Monthly Rainfall; B) Average Monthly Temperatures,
with Possible Development States Shown for the Ord River Based on Degree day Sums (Constable and Shaw 1988); C)
mean daily Radiation for each Month. Where Narrabri, --- Ord River.
There is very little literature reporting cotton grown during the dry season in the SAT worldwide.
Cotton is known to be grown during the dry season in several tropical regions, such as eastern Asia,
Central America, Colombia, Sudan, and Malawi. In most cases production is near the coast or large lakes
where temperature extremes are minimised and these countries have lower economic yield expectations
than Australia (Hearn, 1995). Hence, there is a need to develop and evaluate the dry season production
system outlined in Table 1, as a prerequisite to assessing the feasibility of reintroducing cotton into the
Australian SAT. The Ord River is suitable for this evaluation as it is one of the few valleys north of 21
o
S
developed for irrigation and expansion of the cropping area was planned for the near future (Yeates,
2001; Yeates et al., 2002a).
While insect management and crop husbandry research was being conducted separately to this
research (Strickland et al., 1998; Annells and Strickland 2003; Yeates et al., 2002b), the research
reported here addresses the following important crop adaptation issues relevant to dry season cotton
production:
0
20
40
60
80
100
120
140
160
180
200
J
a
n
/
J
u
l
F
e
b
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A
u
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a
r
/
S
e
p
A
p
r
/
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c
t
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a
y
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o
v
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u
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l/
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a
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c
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a
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u
n
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e
a
n

m
o
n
t
h
l
y

r
a
i
n
f
a
l
l

(
m
m
)
Pick
Plant
A
0
5
10
15
20
25
30
35
40
Apr/Oct May/Nov Jun/Dec Jul/Jan Aug/Feb Sep/Mar Oct/Apr
M
e
a
n

M
o
n
t
h
l
y

T
e
m
p
e
r
a
t
u
r
e

(
o
C
)
1st quare 1st Flower Cut Out
1st Open Boll Maturity
B
10
12
14
16
18
20
22
24
26
Apr/Oct May/Nov Jun/Dec Jul/Jan Aug/Feb Sep/Mar Oct/Apr
M
e
a
n

M
o
n
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y

S
o
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a
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R
a
d
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a
t
i
o
n
(
M
J
/
m
2
/
d
a
y
)
C
Does the photothermal regime of the tropical dry season affect crop development or limit the
conversion of radiation to dry matter and its partitioning.
What yield and quality is possible using modern genotypes and management given the potential
limitations of temperature and radiation in the dry season?
What is the optimum sowing window for yield and quality given sowing must commence after
March 1 to avoid insect pests and there must be sufficient time to pick by before the start of the
wet season?
The research described here integrates four papers (Yeates et al., 2010a,b,c and Yeates et al., 2011) and
some previously unpublished research into one document.
Field Experiments
Sowing date by cultivar experiments conducted over three seasons at the Frank Wise Institute, 13 km
NW of Kununurra WA, Australia (15
o
39S, 128
o
43E) in the Ord River Irrigation Area were used to
collect relevant data. These experiments are described in detail in Yeates et al., (2010a). To summarise,
the Gossypium barbadense cultivar Pima S7 was compared with two Bt transgenic Gossypium hirsutum
(upland) cultivars: Siokra L23i and Sicot 50i (producing the Monsanto Cry1Ac protein). In the first
season, the non Bt transgenic equivalent of the upland cultivars (Siokra L23 and CS50) were sown.
Where data are combined for the three seasons these cultivars are referred to as L23 and S50. In each of
the 3 seasons these cultivars were sown on 4 occasions (main plots): 27 to 29 March, 21 to 29 April, 15
to 23 May and 9 to 14 June; there were 4 replications. The experiments were furrow-irrigated. The crop
was sowing a at 90 cm row spacing on wide beds accommodating two rows per bed. Plots were 6 rows
wide and 20m in length. Rows were in an east west direction.
Boll period was measured by tagging 30 recently pollinated (white or pink) first position flowers in
each plot on 3 occasions with the date and node number recorded, which represented the flowers on
lower, middle and upper part of the main-stem. The date of tagging was identified by different coloured
tags Bolls were hand picked on alternate days, the number of bolls and date picked was recorded and the
boll period calculated as the time from tagging to the median open day. Seed cotton was machine
harvested from 13m of a centre row of each plot. Above ground biomass from 1m
2
from each plot was
partitioned into stems, leaves, squares, flowers and bolls prior to drying at 80
o
C for 3 4 days in a fan
forced oven. Biomass was partitioned at early squaring, at first flower, at approximately 30 and 60 days
after first flower and when approximately 60% of the bolls were open. The final biomass sampling was
made prior to chemical defoliation. The measurement of RUE was described in detail in Yeates et al.,
(2010b). To determine the effect of temperature on RUE, the average RUE calculated for the different
growth phases for each sowing month of each variety was plotted against the average minimum,
maximum and mean temperature for the duration of the growth phase. Biomass was converted to a
glucose equivalent using the method of Wall et al. (1994).
Pot Experiment
The experiments were located at the Katherine Research Station, 4 km east of Katherine (14
o
28' S,
132
o
18' E), Northern Territory, Australia (see Yeates et al., 2011, for more details). Due to greater
distance from the ocean Katherine has a greater probability of cooler dry season minimums than the Ord
River, with similar maximum temperatures, photoperiod and monthly radiation (Cook and Russell,
1983). Minimum temperature was manipulated by protecting plants in a glasshouse at night during
flowering (warm plants) and comparing these with plants grown at ambient temperatures at night
(cool plants). Glasshouse temperatures were maintained approximately 5
o
C above ambient to ensure
similar daily variation in minimum temperature to plants grown at ambient temperatures. The only
exceptions were when ambient temperatures were < 6
o
C, on these nights glasshouse temperatures were
not permitted to fall below 10.3
o
C and when ambient temperatures were warm, glasshouse minimum
temperatures did not exceed 24
o
C. The experiment was run over two seasons with sowing occurring in
late April. To ensure plants were exposed to the same minimum temperatures prior to flowering all plants
were grown outside until 6 and 7 days prior to first flower in 2003 and 2004 respectively. When the
temperature treatments commenced the warm night plants were moved inside at night for the next 60 and
53 days in 2003 and 2004 respectively; that is at least 15 days after flowering was completed.
Results and Discussion
Lint Yield
For the field experiments Lint yields exceeded 2000 kg/ha when sowing occurred during March and
April for the upland cultivars (Fig. 2). For Pima S7 sowing in March produced the highest lint yields
which were approximately 1800 kg/ha. Yields declined rapidly when sown after mid May. Despite the
photothermal limitations described above these lint yields were at worst in line with recent Australian
commercial irrigated yields (ABS 2006) and commonly reported research yields for irrigated cotton in
temperate Australia and the USA, where lint yield was inflated by laboratory ginning, (e.g. Fritschi et al.,
2003; Hutmacher et al., 2004; Bange and Milroy, 2004) and at the top of international averages
elsewhere (ICAC 2002). The lower yield of the Gossypium barbadense cultivar Pima S7 compared to the
Gossypium hirsutum cultivars was consistent with research and commercial experience with this species
from temperate regions (Fritschi et al., 2003). Hence sowing in March or April of either species should
meet current commercial expectations if repeated reliably on a larger scale, provided fibre quality
standards are achieved. Moreover sowing before May would ensure picking could occur before the onset
of the wet season (Yeates et al., 2010a).

Fig 2: The Effect of Sowing Date on Average Lint Yield for Three Seasons for Upland Varieties and Pima S7. Bars show Range of
Yields. Adapted from Yeates et al. 2010a. How were Yields Achieved?
Final crop biomass did not limit yield. For the highest yielding March and April sowings final crop
biomass was mostly > 1000 g /m
2
(Fig 3). This was similar to the maximum values reported for irrigated
cotton in temperate Australia (Sadras 1996; Bange and Milroy 2004) and the USA (Fritschi et al., 2003).
These biomasses were also 25 to 60 % higher than for dry season cotton but similar to wet season cotton
grown in the 1960s at this location (Stern, 1965; Thomson, 1965).
0
500
1000
1500
2000
2500
27 to 29 March 21 to 29 Apri l 15 to 23 May 9 to 14 June
sowing Date
L
i
n
t

Y
i
e
l
d

(
k
g
/
h
a
)
Upland
Pima S7

Fig. 3: Linear Biomass Accumulation Showing Growth Rate as Slope of Fitted lines for the Highest Yielding March and
April Sowings. Presented is One Upland Cultivar and Pima S7. where = March sowing, = April sowing and
DAS = Days after Sowing. Adapted from Yeates et al. (2010b)
An important finding was that biomass was accumulated differently in the tropical dry season than in
temperate (~ 30
o
lat.) and tropical wet season grown cotton. Fig. 5 shows for the highest yielding March
and April sowings, final biomass was accumulated by sustaining a modest growth rate of 6.9 to 12.3
g/m
2
/d for a long period (78 to 134 days) between first squaring and boll opening. This contrasts with
similar yielding crops in temperate climates where growth was characterised by shorter period (25 to 60
days) with a greater maximum growth rate of 15 to 20 g/m
2
/d (Kennedy and Hutchison, 2001; Bange and
Milroy, 2004) and lower yielding wet season cotton, with a similar vegetative biomass to this study,
where a maximum growth rate of 7 to 16 g/m
2
/d was maintained for 50 to 60 days (Stern 1965; Thomson
1965).
There was positive correlation between yield and greater horizontal fruiting, that is more 2
nd
position
(P2) and 3
rd
position (P3+) bolls and the lack of correlation between the number of first position (P1)
bolls and their retention and yield (Table 2). This is a departure from spring sown upland cotton in
temperate climates where P1 bolls and their retention are regularly monitored due to their positive
association with yield (Constable, 1991; Kerby and Hake, 1996). The relationship between yield and P1
retention found for temperate regions is related to the need for earlier maturity due to a growing season
length that is defined by cool temperature (Kerby and Hake, 1996). In temperate climates the growth of
L23 - 1997
March
slope = 9.21g/m
2
/d
R
2
= 0.994
April
slope= 7.90 g/m
2
/d
R
2
= 0.998
0
200
400
600
800
1000
1200
0 20 40 60 80 100 120 140 160 180
D
r
y

w
t

(
g
/
m
2
)
Pima S7-1997
March
slope = 7.42 g/m
2
/d
R
2
= 0.988
April
slope = 7.63 g/m
2
/d
R
2
= 0.952
0
200
400
600
800
1000
1200
0 20 40 60 80 100 120 140 160 180
L23 - 1996
March
slope = 8.44 g/m
2
/d
R
2
= 0.987
April
slope = 11.26g/m
2
/d
R
2
= 0.998
0
200
400
600
800
1000
1200
0 20 40 60 80 100 120 140 160 180
D
r
y

w
t

(
g
/
m
2
)
Pima S7- 1996
April
slope = 9.82 g/m
2
/d
R
2
= 0.97
March
y = 12.3 g/m
2
/d
R
2
= 0.992
0
200
400
600
800
1000
1200
0 20 40 60 80 100 120 140 160 180
L23 - 1995
March
slope = 6.19 g/m
2
/d
R
2
= 0.976
April
slope = 7.47 g/m
2
/d
R
2
= 0.999
0
200
400
600
800
1000
1200
0 20 40 60 80 100 120 140 160 180
DAS
D
r
y

w
t

(
g
/
m
2
)
Pima S7- 1995
April
yslope= 6.07 g/m
2
/d
R
2
= 0.98
March
slope = 7.59 g/m
2
/d
R
2
= 0.98
0
200
400
600
800
1000
1200
0 20 40 60 80 100 120 140 160 180
DAS
bottom P1 bolls often coincides with favourable climatic conditions, which are the longest photoperiods
and highest solar radiation for the growing season. Hence, P1 bolls were larger at each fruiting branch
and had a greater chance of survival (Constable, 1991). There is also greater likelihood of sub-optimal
temperatures for later developing bolls making compensation from the loss of lower fruit risky. The
tropical dry season is the reverse with the pollination of early flowers coinciding with less favourable
climatic conditions, that is, cool to cold night temperatures and relatively low radiation, while later
developing bolls were exposed to rising temperatures and radiation (Fig. 1).
TABLE 2: LINEAR REGRESSION COEFFICIENTS FOR THE RELATIONSHIP BETWEEN YIELD COMPONENTS OR PLANT MAPPING VARIABLES AND LINT YIELD. DATA
FOR THE THREE SEASONS COMBINED. WHERE, ** P<0.01, * P<0.05 AND NS = NOT SIGNIFICANT,
POSITION 3+ = (POSITION 3 +POSITION 4 + ADVENTITIOUS BOLLS). ADAPTED FROM YEATES ET AL. 2010A
Yield Component / Plant Mapping Variable Lint Yield (n=48 for each cultivar)
L23 S50 Pima S7
Position 1 bolls (m
-2
) ns ns ns
Position 2 bolls (m
-2
) 0.41** 0.36** 0.38**
Position 3+ bolls (m
-2
) 0.34** 0.36** 0.45**
Boll development was slower for bolls from early P1 flowers when sown in March and April
compared with May or June sowings (Fig. 4). The boll development period was at least 20 days less for
later flowers for the March and April sowings. This pattern of boll period with plant position and sowing
date reflected the temperatures during boll development. That is boll growth from the earliest flowers on
March and April sown plants coincided with the coolest night temperatures (Fig. 1).

Fig. 4: The Impact of Sowing Month on Boll Periods for: Early Flowering P1 Bolls (1
st
Three Fruiting Branches),
P1 Bolls Mid Canopy P1 Bolls From late Flowers or Top of the Canopy
A reduced contribution to yield from P1 flowers combined with a longer boll period may have had a
positive impact on yield for the March and April sowings by reducing boll demand for assimilate early in
flowering, a time when assimilate supply was limited by low solar radiation and cool night temperatures
(Fig 1). Compensation for the loss of early flowers occurred due to a greater production of horizontal
fruiting sites that flowered later when temperatures were warmer and radiation higher. The positive
relation between yield and time-to-maturity supports this hypothesis (Yeates et al., 2010a).
0
10
20
30
40
50
60
70
80
90
March April May June
Month Sown
B
o
l
l

P
e
r
i
o
d

(
d
)
0
10
20
30
40
50
60
70
80
90
March April May June
Month Sown
B
o
l
l

P
e
r
i
o
d

(
d
)
Because the field experiments were grown using Integrated Pest Management where pest number
thresholds are used as a trigger for control measures it was difficult to determine whether cool minimum
temperatures were the only factor in the reduced contribution of early bolls to yield. Other possible
causes such as water logging, nutrient stress can also occur. Moreover in the field there was no warm
night control to compare with.
The pot experiment found that yield was not reduced by ambient minimum temperatures observed
during flowering that averaged 10.2
o
C and 12.6
o
C compared with minimums that were 5
o
C higher. This
experiment showed the reduced contribution to yield from early first position flowers in the tropical dry
season was due to low minimum temperatures near anthesis and not biotic causes such as pests. Fig. 5
shows that boll weight and retention of P1 bolls was reduced in proportion to minimum temperature
during flowering. Yield compensation from cold minimums during flowering occurred via two
processes. Firstly, for all fruiting positions boll survival was significantly correlated with minimum
temperature near anthesis. Because minimum temperatures in the field were variable, greater boll
retention and weight followed periods of warmer minimums (Yeates et al., 2011). Secondly, the greater
contribution to yield from later pollinated flowers reflected the general increase in temperature and
radiation later in flowering that typifies the tropical dry season (Fig. 1).

Fig. 5: The Impact of Average Minimum Temperature During the Flowering Period on Seed Cotton per Boll and
Boll Number on First Position Bolls. Data from Experiment at Katherine
For all varieties the glucose equivalent RUE changed significantly (p<0.05) with crop ontogeny ;
being highest in the squaring to flowering period (1.88 to 2.34 g/MJ) then declining to a minimum late in
boll growth of 1.2 to 1.4 g/MJ (Yeates et al., 2010b). From first square to first flower the linear
correlation between RUE and average temperature was highly significant (p<0.01) (Fig. 6). The lines
fitted for the upland cultivars Siokra L23i and Sicot 50i were not significantly different hence their
responses were combined (Fig. 6A). The correlation between temperature and RUE after early flowering
Seed Cotton = 0.24T + 2.0661
R
2
= 0.98
0
1
2
3
4
5
6
7
9 10 11 12 13 14 15 16 17 18
Average Minimum Temperature
o
C
S
e
e
d

C
o
t
t
o
n

(
p
e
r

b
o
l
l
)
Boll Number = 0.0887T + 1.1783
R
2
= 0.65
0
0.5
1
1.5
2
2.5
3
9 10 11 12 13 14 15 16 17 18
Average Minimum Temperature
o
C
B
o
l
l

N
u
m
b
e
r

(
p
e
r

p
l
a
n
t
)
was not significant, although the temperature range was smaller (data not presented). This reduction of
RUE with average temperature up to first flower has not been reported previously in cotton and explains
some of variation in RUE observed here (Yeates et al., 2010b) and elsewhere (Sadras, 1996; Bange and
Milroy, 2004). The development of this relationship was possible because the temperature range
observed for these sowing months was greater than could be generated reliably over the first square to
first flower phase in temperate latitudes (~30
o
) or in the tropical wet season. Low temperatures have
reduced RUE in peanut (Bell et al., 1992), sorghum (Hammer and Vanderlip, 1989) and maize (Andrade
et al., 1993). The effect of lower temperatures on RUE would be expected to occur in cotton via their
effect on photosynthesis (Peng and Krieg, 1991).

Fig. 6: The Effect of Average Temperature on RUE for A) Upland Cultivars and B) Pima S7 between First Square and First Flower. RUE Was Calculated for
Each Sowing Date from the Linear Regression between Accumulated Biomass and Intercepted PAR. Fitted Lines, their Equations and Regression
Coefficients are Shown. Where ** = P < 0.01, Tav = Average Temperature between First Square and First Flower. From Yeates et al. (2010b)
Low temperatures are more likely to reduce RUE when cotton is sown in May or June because the
greatest proportion of growth up to early flowering at these sowing dates coincides with the cooler
months of June and July (Yeates et al., 2010a). This observation appears to be supported by the generally
lower biomass (flowering and maturity) of the May and June sowings (Yeates et al., 2010b). For these
experiments, monthly temperatures were near ( 2
o
C) the long-term average (Yeates et al., 2010a), hence
the RUE for cotton sown in April could also be reduced in seasons with lower than average minima
during flowering.
For the upland cultivars the range in RUE measured here was similar to elsewhere using the glucose
equivalent (GE) and dry weight only methods of calculating biomass (Sadras, 1996; Bange and Milroy,
2004; Rosenthal and Gerik, 1991) but lower than the peak values of 3 g/MJ/m
2
reported by Milroy and
Bange (2003), where leaf nitrogen concentration was high. The RUE for Pima S7 (Gossypium
barbadense) of 1.2 to 2.3 g (GE) / MJ (Fig. 4.2) is the first for this species.

0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
20 21 22 23 24 25 26 27
R
U
E

(
g

M
J

-
1
)
A
Upland RUE=0.223*Tav-3.43
R
2
=0. 53**
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
20 21 22 23 24 25 26 27
Average Temperature (C
o
)
R
U
E

(
g

M
J

-
1
)
B
Pima S7 RUE
=0.265*Tav-4.01
R
2
=0.86**
Yeates et al. (2010c) found fibre length and strength at the highest yielding March and April sowings
were low to marginal compared with market preference values. This was due to cool temperatures during
fibre development. The cultivar differences observed here suggest wider screening may identify
Gossypium hirsutum cultivars with suitable fibre length and strength in these conditions. The commercial
prospects for Gossypium barbadense are doubtful unless longer and stronger fibre types are identified.
CROP MANAGEMENT IMPLICATIONS
The emphasis in crop monitoring will need to shift from measuring and ensuring high P1 boll
retention and then protecting these bolls because of their high contribution to yield in temperate climates,
to monitoring all fruiting positions to ensure the production of new fruiting sites is sufficient to permit
yield compensation when needed.
Achieving a balance between yield compensation via fruiting sites toward the top and outside of the
plant and appropriate vegetative growth will require careful management. Over use of growth regulators
or insufficient irrigation or nutrient deficiency will inhibit compensatory growth and reduce yield. On the
other hand a luxurious supply of water and or nutrients combined with an insufficient amount of growth
regulator could lead to excessive or rank vegetative growth. Concurrent growth regulator research has
shown that avoidance of high doses during fruiting site production is essential to permit compensation
via the production of additional fruiting sites (Yeates et al., 2002b). Concurrent studies (S.J. Yeates; J.
Moulden, AGWA unpublished data) found lowering of plant density to assist the growth of these fruiting
sites is unlikely to be an option because it only increased the ratio of out side bolls to top bolls.
ACKNOWLEDGEMENT
The financial support from the Australian Cotton Cooperative Research Centre made this research
possible. I would also like to thank Dr Greg Constable CSIRO for his cotton physiology expertise,
Stewart Addison, Geoff Strickland and their staff from Agriculture WA for entomological assistance,
Warren Muller of CSIRO-MIS for statistical assistance and Kellie Cooper of CSIRO for HVI analysis.
REFERENCES
[1] ABS. (2006) The Australian Bureau of Statistics, Agricultural Commodities Australia 7121.0 -2004-05, p19.
www.abs.gov.au
[2] Andrade, F.H., Uhart, S.A. and Cirilo, A., (1993) Temperature affects Radiation Use Efficiency in Maize - Field Crops
Res., 32: 17-25.
[3] Annells, A.J. and Strickland, G.A., (2003) Assessing the Feasibility for Cotton Production in Tropical Australia: Systems
for Helicoverpa spp. Management - In: Swanepoel A., (Ed.), Proceedings of the World Cotton Research Conference-3:
Cotton Production for the new millennium. 9-13 March 2003, Cape Town, South Africa, pp. 905-912.
[4] Bange, M.P. and Milroy, S.P., (2004) Growth and partitioning of diverse cotton cultivars - Field Crops Res., 87: 73-87.
[5] Bell, M.J., Wright, G.C. and Hammer, G.L., (1992) Night Temperature Affects Radiation-Use Efficiency in Peanut - Crop
Sci., 32: 1329-1335.
[6] Constable, G.A. and Shaw, A.J. (1988) Temperature Requirements for Cotton Agfact ,P5.3.5, Department of
Agriculture NSW, Australia.
[7] Constable, G.A. (1991) Mapping the Production and Survival of Fruit on Field Grown Cotton - Agron. J., 83: 374-378
[8] Cook, L.J. and Russell, J.S. (1983) The Climate of seven CSIRO Field Stations in Northern Australia - Division of
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Which Carbon Footprint Tool

for the Cotton Supply Chain
F. Visser, P. Dargush and C. Smith
School of Agriculture and Food Sciences, University of Queensland, Brisbane, Australia
E-mail: f.visser@uqconnect.edu.au

AbstractAppropriate and user-friendly tools are required by members of the textile supply chain, including
farmers, to measure the carbon footprint levels of raw materials and products. The purpose of the study is therefore to
identify the most appropriate carbon footprint calculator to be used by cotton suppliers to meet the emerging
sustainability measurement requirements from brand owners and retailers in the textile supply chain.
The textile sector has adopted a Life Cycle Analysis (LCA) approach to evaluate the sustainability levels of raw
materials. Currently there is interest in the development of standards and methods that are suitable for the calculation
of the carbon footprint levels of products in particular.
The first step in terms of the methodology was to identify a range of applicable carbon footprint calculators that
may be used by industry for a cotton crop. Nine calculators were identified and they were analysed following an
outcome-based approach.The next phase comprised of an actual case study on an irrigated cotton farm in the
Goondiwindi district in Australia where the field data was gathered to populate the nine calculators. The reporting
framework was developed that incorporates both on-farm and off-farm emission sources and that is also aligned to a
Life Cycle Analysis (LCA) approach. The results show that total nett carbon footprint for the same cotton from the
same farm varies from 828 kg CO
2
e/ha to 4703 kg CO
2
e/ha according to the different calculators. It appears that that
there are big differences in the methodologies that are being applied, especially for soil carbon emissions, and results
even vary for common standardised emission indicators like fuel use. We conclude there is a compelling need for an
internationally standardised format and methodology for crop-level carbon footprint calculations for the textile supply
chain. Although these are all reputable CFP tools, applied to the same farm data, they generate vastly different
results. These variations in outcomes are mainly due to differences in structure and methodologies applied. Further
research needs to be undertaken to apply process-based models to validate the accuracy of soil emission results from
CFP calculators.
INTRODUCTION
Appropriate and user-friendly tools are required by members of the textile supply chain, including
farmers, to measure the carbon footprint levels of raw materials and products. The purpose of the study is
therefore to identify the most appropriate tool to meet the emerging sustainability measurement
requirements from brand owners and retailers in the textile supply chain, in particular for GHG
emissions.
For the purposes of this study a products carbon footprint or GHG inventory is defined as the total
greenhouse gas emissions caused directly or indirectly by a product, in this case raw cotton up to the farm
gate (Russell, 2011)
Considerable interest currently exists in the determination of the carbon footprint levels of products,
as borne out by:
The new GHG Product Accounting and Reporting Standard by The Greenhouse Gas Protocol
Initiative published in September 2011(World Resources Institute & World Business Council for
Sustainable Development 2011),
The new ISO 14067 Standard, Carbon Footprints of Products, to be published in 2012 (PCF
World Forum 2011),
The current review of the PAS 2050:2008 Standard (British Standards Institution 2011), which
regulates the assessment of GHG emissions of goods and services,
55
And the working paper issued by the World Resources Institute in January 2011 regarding
proposed accounting and reporting steps for GHG inventories for agricultural products (Russell
2011).
WHY THE INTEREST IN PRODUCT CARBON FOOTPRINT (CFP) ANALYSES?
There appears to be four main reasons:
Global Warming/Climate Change
One can only manage that what you can measure, and a number of standards and protocols have been
developed internationally to guide the quantification of GHGs in order to mitigate its effect on global
warming and climate change. The initial focus was at national level and organisational or industry level
(The Greenhouse Protocol Initiative, 2010) but has now developed to products and the quantification of
their contribution to GHG emissions.
Economic and Legal Considerations
Various programmes have been launched at regional, national or global levels to either penalise or
encourage GHG emitters to reduce their levels, or to support projects or programmes that have the ability
to sequester carbon from the atmosphere. A case in point in Australia is the proposed Clean Energy Bill
(Department of Climate Change and Energy Efficiency, 2011b) that will levy a carbon price / tax of $23
ton CO2e against certain categories of emitters. On the supporting side is the Carbon Farming Initiative
or Carbon Credits Bill 2011 (Department of Climate Change and Energy Efficiency, 2011a) that will
enable farmers to sell offsets or abatements generated under certain conditions or practices. There may
also be legal requirements whereby products have to declare their carbon footprints as is now being
introduced in France (The Economist, 2011).
Product Differentiation
The quantification and labelling of a products carbon footprint has become an established from of
differentiation in the market place. As of April 2010 there were 5500 carbon footprint labels in the
United Kingdom (Planet Ark, 2010). Among retailers Tesco took the lead in food carbon footprint labels,
and the first labels were introduced in their stores in 2007. The first product was a box of Simple instant
Quaker Oats, which was soon followed by a packet of Walkers Crisps and a range of drinks from
Innocent Smoothies (Miller, 2008; Environmental Leader, 2010). Authorities have stepped in to regulate
and standardise the introduction of such schemes, in particular in France and Japan, South Korea and
Thailand (The Economist, 2011). The measurement of carbon footprints may also be a requirement to
meet certain product certification standards.
Part of Sustainability Measurement
Various initiatives and programs have been launched in the recent past to develop sets of appropriate
indicators the measure the sustainability levels of products in accordance with the three social, economic
and environmental pillars. It is desirable that a scientifically grounded and outcomes based methodology
be applied, as opposed to a criteria or practice based methodology (Von Wirn-Lehr, 2001). Of specific
relevance to the former is the ongoing Stewardship Index for Specialty Crops (SISC) in the United States
where a second round of pilot studies are being planned to test the crop-level metrics that have been
developed (Stewardship Index, 2011).
An indication of the different sustainability indicators that have been developed by the main
certification programs or standards in the textile supply chain is given in Table 1.
Which Carbon Footprint Tool for the Cotton Supply Chain 325
TABLE 1: SUSTAINABILITY INDICATORS AND STANDARDS IN THE TEXTILE SUPPLY CHAIN

As can be seen above, greenhouse gas emission levels / pollution is one of the key environmental
indicators to be measured in an outcomes based product sustainability calculation. GHG calculations are
closely aligned with energy use as the majority of emissions sources are energy use derived, like fuel and
electricity consumption for example.
At this point no calculators or mechanisms are available whereby the actual complete sustainability
level or rating of a product can be determined by industry on an outcomes or scientific basis. The only
outcomes-based sustainability calculator for crops that has been released is the ongoing Fieldprint
Calculator from the Field to Market initiative in the US. The five indicators that have been incorporated
at this stage are land use, soil loss, irrigation water use, energy use and climate impact (GHG). Further
indicators in the pipeline include soil quality, water quality, biodiversity and the required social and
economic indicators (The Keystone Alliance for Sustainable Agriculture, 2010). In terms of the
Stewardship Index mentioned above, the first calculators that will be trialed are for water use efficiency,
soil health, nutrient use and energy use (Stewardship Index, 2011).
Eco
I n d ex
Su s t a in a ble
Ap pa r el
Coa lit ion
Pa t a gon ia
Ch r on .
ULE
8 8 0
BCI Cr a d le
t o
Cr a d le
Blu e
Sign
Oeko-
Tex
10 0 0
GOTS
Land Use Intensity *
Water Use & Quality * * * * *
Waste * * * *
Biodiversity * *
Chem. Tox. People * * *
Chem. Tox. Environm. * * *
Energy Use * * * * *
GHG * * *
Social / Labour * * * *
Non Renewables
Pollution
Sust. Governance *
Env. Management * * *
Work Force *
Customers/ Suppliers *
Community *
Pesticides *
Soil Health *
Product Quality * *
Product Safety * *
Resource Productivity *
Consumer safety *
Air Pollution * *
Water Pollution * * *
Worker H & S * *
Noise Pollution *
Child Labour *
Chemical Inputs *
Storage, Pack. &
Transport
*
Chemical Residues *
Tech. Specifications *

Such practical tools or mechanisms are required by industry to calculate and integrate the outcomes
from the various indicators and metrics due to the complexity involved and to provide scientific integrity
and rigor to the results. Supply agreements could potentially be negotiated on the basis of such outcomes,
and consequently be contested as well.
Greenhouse Gasses (GHG), and closely aligned energy use, are the only indicators for which a
number of such tools or calculators have been developed. The purpose of the study is to identify the most
appropriate carbon footprint calculator to be used by cotton suppliers to meet the emerging sustainability
measurement requirements from brand owners and retailers in the textile supply chain.
CARBON FOOTPRINT CALCULATORS
A number of carbon footprint calculators have been developed for agriculture. The majority of these are
designed to generate farm-level results, but generally these can be adapted to provide crop-level outputs
by only providing the actual inputs of the crop in question.
Classification of Calculators
There are a number of ways in which CFP calculators can be classified for comparison purposes. The
IPCC Good Practice Guidelines distinguishes between three levels of complexity, and corresponding
accuracy, for such methodologies:
Tier 1: Simple, emission factor-based approach, where emissions are calculated by multiplying
activity data by an appropriate emission factor. Tier 1 emission factors are international or regional
defaults.
Tier 2: More specific emission factors or more refined empirical estimation methodologies.
Tier 3: Dynamic bio-geophysical simulation models using multi-year time series and context-specific
parameterization (IPCC National Greenhouse Gas Inventories, 2006).
This is not deemed to be an appropriate classification as it appears that the calculators apply a
combination of tier level approaches depending on the different emission sources and data availability.
Calculators can also be classified in terms of transparency, methodology, effectiveness of
communication, source data, and structure (Kim and Neff, 2009). For the purpose of this study an
outcomes-based approach was adopted, in line with recent global trends in product sustainability
measurement. Accordingly the outcomes from a range of CFP calculators are compared by populating
them with the same, site specific set of data inputs. The aforementioned potential differences are de facto
dealt with in analyzing of the variances in the results.
Structure Requirements for CFP Calculators
As mentioned above, in calculating a products carbon footprint one needs to include emissions caused
directly as well as indirectly by that product. The Greenhouse Gas Protocol defines direct and indirect
emissions as follows:
Direct GHG emissions are emissions from sources that are owned or controlled by the reporting
entity (farm).
Indirect GHG emissions are emissions that are a consequence of the activities of the reporting
entity, but occur at sources owned or controlled by another entity (The Greenhouse Gas Protocol
Initiative, 2001).
The GHG Protocol further categorizes these direct and indirect emissions into three broad scopes:
Scope 1: All direct GHG emissions (On farm)
Scope 2: Indirect GHG emissions from consumption of purchased electricity, heat or steam.
Scope 3: Other indirect emissions, such as the extraction and production of purchased materials)
(The Greenhouse Gas Protocol Initiative, 2001)
The following calculator format (Table-2) comparison framework was developed based on the above
guidelines and the type of results generated by the calculators in question.
As will be shown, the less advanced calculators do not address all the emission sources indicated
above. It is not mandatory to include Scope 3 emissions under the Corporate GHG Standard, but
encouraged where these are deemed to be significant. The calculators generally do include bio-chemical
production as the level of use of fertilisers and chemicals are directly under the control of farm
management and an integral part of the carbon footprint of a crop, and certainly significant in terms of
emission levels, particularly in the case of fertilizers (Russell, 2011). The emissions associated with the
manufacture of capital equipment (tractors, etc.) is also a Scope 3 emission and could also be included in
principle, but none of the calculators make provision for this.
TABLE 2: THE CALCULATOR FORMAT BASED ON GHG PROTOCOL INITIATIVE
Type OF Emissions Scope Comments/ Examples
Direct On-Farm Emissions Scope 1
Energy Usage Mainly fuel use like diesel, petrol, gas
Soil / Crop Residue Emissions
Soil N
2
O Related to fertilizer application
Nett Soil CO
2
Related to whole carbon-cycle in soil
Crop Residue / Stubble Related to tillage practices
Nett Soil / Residue Emissions
Nett On-Farm Emissions
Indirect Off-Farm Emissions
Purchased Electricity Scope 2
Bio-Chemical Production Scope 3
Fertilizer Production Embedded energy in fertilizer production
Chemicals Production Incl. herbicide, fungicide and pesticide
Total Off-Farm Emissions
Total (Nett) Emissions
The one area where there is still a considerable amount of uncertainty and variation in the types of
methodologies applied is the level of carbon soil emissions, resulting predominantly from the different
types of cultivation practices adopted by farmers. The main gap in the methodology is that no standard
has been developed for this type of emission as opposed to most of the other emission categories, and
appears to be a main contributor to variations in footprint results. But the emissions from non-
mechanical sources, such as soils and livestock, are more challenging. Consensus best practices for
dealing with these challenges do not yet exist. The GHG Protocol intends to develop a consensus-based
GHG accounting and reporting protocol for the agriculture sector. (World Resources Institute, 2011) A
working paper has been issued to this effect (Russell 2011). Types of methodologies being applied to
calculate carbon soil emissions is presented in Table 3.
Most of the calculators appear to be based on empirical models (Table 4) although it is not always
possible to ascertain which methodology the results are based on. The above mentioned framework is
also aligned with Life Cycle Analysis (LCA) methodology. The life cycle under investigation is the
production process on the farm and the product boundary is raw cotton at the farm gate. The upstream /
indirect emission sources taken into consideration are the generation of electricity and the emissions
involved in the manufacture of the fertilisers and chemicals. The environmental impact category is
restricted to GHG emissions.

TABLE 3: METHODOLOGIES APPLIED TO SOIL CARBON EMISSION
Methodology Description Comments
Field Measurements Including lab measurements of soil density Time-consuming Expensive
Emission Factors-based Models e.g., tonnes CO2 emitted per ha wheat farming Simple and inexpensive Low accuracy
Empirical Models
From statistical links between GHG data and
management factors
Medium accuracy
Inexpensive, most common
Processed-based Models
Mathematical simulation models of CO
2

emission / fixation process
Expensive, time consuming
Too complex for industry tool
Source: (Russell 2011)
TABLE 4: CFP CALCULATORS THEIR DESCRIPTION AND SOURCE
Calculator Description Source
Lincoln Uni Carbon Calc (NZ) Mainly mixed farming tool http://www2.lincoln.ac.nz/carboncalculator/
Cotton GHG Calculator (Aus) For Cotton Ind., mainly farm-level tool http://www.isr.qut.edu.au/tools/index.jsp
FarmGas GHG Calculator (Aus) Broad acre GHG tool, no energy calc. http://farmgas.farminstitute.org.au
Fieldprint Calculator (US) For commodity crops sust. calculator http://www.fieldtomarket.org/fieldprint-
calculator/
Cool Farm Tool (UK) Funded by Unilever, global application http://www.unilever.com/aboutus/supplier/su
stainablesourcing/tools/
US Cropland Calculator (US) GIS-based empirical method http://surf.kbs.msu.edu/ghgcalculator/
Veggie Carbon Calculator (Aus) For hort. but can apply to broad acre http://www.vegiecarbontool.com/
NCEA USQ (Aus) USQ case study on cotton energy use http://energy.ncea.biz/
ACSC USQ (Aus) USQ case study on cotton GHG levels http://eprints.usq.edu.au/7101/
In the context of this study it is important for any proposed CFP methodology to also adhere to the
principles of LCA methodology. The reason being that LCA has been identified as the mechanism
(specifically PAS 2050) that will be used to measure product sustainability levels in the textile supply
chain by major initiatives like the Eco Index, who has the support of the major brand owners and retailers
like Wal-Mart, Marks & Spencer, Levis, Adidas, Nike, etc (Eco Index, 2011). Since 2008, PAS 2050
has been the only LCA-based product carbon footprint standard. As far as agriculture is concerned it
specifically states that:
Changes in the carbon content of soils, either emissions or sequestration, shall be excluded from
the assessment of GHG emissions under this PAS
Agricultural emissions include emissions from fertilizers (e.g. N
2
O emissions arising from the
application of nitrogen fertilizer and emissions arising from the production of the fertilizer)
The GHG emissions arising from the production of capital goods used in the life cycle of a
product shall be excluded from the assessment of GHG emissions. (Carbon Trust, 2008)
Therefore, in terms of a comparison between the above framework and LCA principles it appears that
it is correct to include both soil N
2
O emissions and the production emissions of fertilizers, and that it is
order not to incorporate emissions from capitals goods manufacturing. Although PAS 2050 does not
require the inclusion of soil carbon emissions, it is very significant that the Greenhouse Gas Protocols
Product Accounting and Reporting Standard released recently in September 2011 actually includes
carbon stock and defines it as CO
2
emissions and removals resulting from a carbon stock change
occurring within and between land use categories (World Resources Institute & World Business Council
for Sustainable Development 2011). It will therefore be important to note what the new ISO 14067
Standard on the Carbon Footprints of Products will stipulate when it is released in 2012 (PCF World
Forum, 2011).
LCA calculations rely predominantly on various databases for emission data, but these do not
typically provide reliable data on soil carbon emissions, as can be explained by the above. As indicated
this may need to be improved in future and carbon footprint calculators can play a valuable role in this
regard. Soil carbon emissions can represent a significant part of a carbon footprint of a crop, be it as a
debit or a credit. It is furthermore important to be able to quantify potential carbon sequestration levels in
the interests of promoting conservation agriculture and for farmers to be able to take up potential offset
opportunities with programs like the Carbon Framing Initiative (Department of Climate Change and
Energy Efficiency, 2011a).
Selection of CFP Calculators
The following criteria were applied in selecting suitable CFP calculators from those applicable to
agriculture:
Recognised and currently in use in agriculture in Australia
Internationally applied in regions where major textile brand owners and retailers are based
Will generate results in a format that will be applicable to LCA evaluation mechanisms in the
textile supply chain, in particular the Eco Index,
Applicable as an industrybased tool
Based on a level of complexity that takes into account most of the emissions sources indicated
above
DATA COLLECTION
The focus of the study is to identify the most appropriate CFP calculator to determine the carbon
footprint levels of Australian cotton, mainly to meet sustainability requirements in the textile supply
chain. The aim is therefore not to provide a detailed analysis of the actual emission levels cotton
production but to identify the most appropriate methodology to achieve this. Nevertheless the cotton farm
used for the study was selected on the basis of being fairly representative of Australian cotton farms with
regards to soil types, climatic conditions and farming practices.
The procedure was to populate all 9 calculators with the same base data from the farm for the
2010/11 growing season. The calculators differ markedly in the type of information required and a
questionnaire was developed to cover the complete range of data required, which were grouped in to the
following categories: energy use, fertiliser use, crop chemicals, cultivation practice, residue management,
crop rotation, land use change, soil characteristics, and water use / irrigation. Some of the common base
data applied are listed in Table 5.
TABLE 5: COMMON BASE DATA
Indicator Value
Area cotton production 300 ha
Diesel use / crop 40 000 l
Tillage practice Reduced till
N application rate 220 N / ha
Chemicals applications 3 per crop
Rainfall 650 mm / a
Soil bulk density 1.7g / cm3
Soil carbon content 0.5 %
Soil clay fraction 55 %
Dry land / Irrigated Irrigated
RESULTS
The methodology to calculate an organization or products carbon footprint has been standardized by the
initial Greenhouse Gas Protocol, the IPCC guidelines and subsequently the ISO 14067-1 standard, upon
which the Carbon Trust Standard and PAS 2050 are also based. Therefore in particular for standard
items like fuel use and fertilizer applications one would expect that the same base data would render
similar results across the calculators. This is not the case. As mentioned earlier the one emission source in
agriculture that there has not been a standard methodology for is soil emissions, and these results vary
considerably as can be expected.
Table 6 below shows the carbon footprint results by emission source and calculator from the selected
cotton farm, with the corresponding chart below:

TABLE 6: COMPARATIVE ANALYSIS (KG CO
2
/ HA OF VARIOUS GHG CALCULATORS
Lincoln Uni. Cotton GHG Farm Gas Field Print Cool Farm US Cropland Veggie Carb NCEA - USQ ACSC - USQ
Source of Emissions Carbon Calc Calc - QUT GHG Calc Calculator Tool Calc - MSU Calculator Cotton Cotton
Direct On-Farm Emissions
Energy / Fuel CO
2
821 360 141 357 90 360 460 322
Soil / Crop Residue Emissions
Soil N
2
O Emissions 1241 77 828 1665 950 3650 1336 300 2244
Crop Residue / Stubble 66
Nett Soil CO
2
Emissions 158 -1141 60
Nett Soil / Residue Emissions 1241 77 828 1823 -125 3710 1336 300 2244
Nett On-Farm Emissions 2062 437 828 1964 232 3800 1696 760 2566

Indirect Off-Farm Emissions
Purchased Electricity 1109 1137 555 1329
Bio-Chemical Production
Fertiliser Production 844 480 766 572 80 804 803
Chemicals Production 58 62 925 615
Total Off-Farm Emissions 844 480 0 824 1743 80 1137 2284 2747
TOTAL (Nett) EMISSIONS 2906 917 828 2788 1975 3880 2833 3044 5313
Model: 300 ha cotton farm in Goondiwindi - Reduced till production, 220 Kg N per hectare, 3 pesticide applications per
crop, 40 000 L diesel per crop, no petrol.
It is immediately apparent and alarming that the calculators generate such a considerable difference
in results from the same cropping data. This is mainly due to two reasons: a) Inconsistencies is structure,
not all the emissions sources are taken into consideration although these are fairly well defined in the
standards, apart from soil carbon emissions; b) Differences in methodologies, again especially in the case
of soil emissions, although there are considerable variances even for standardized outcomes like fuel
use. It must be borne in mind that the calculators in question have all been developed by reputable
institutions and their outcomes could all therefore be regarded as credible and viable alternatives for
farmers to use. Therefore if a cotton ginner includes carbon footprint levels as a buying criteria for a
premium grade/ price, farmer A could qualify with a level of 917 kg CO
2
e/kg by using the Cotton GHG
calculator, whereas farmer B may be excluded at a level of 3880 kg CO
2
e/ha by using the US Cropland
calculator although they may be following exactly the same farming practices under the same conditions.
The specific emission components are dealt with in more detail below.
Energy/ Fuel Use

Fig. 1: Energy used in the Different Calculators
CO
2
Fuel use is one of the standardised emission indicators. In this case the farm used 40 000L of diesel
to grow the 300 hectares of cotton during the 2010 / 2011 planting season (Fig.1). NCEA(USQ) and
ACSC(USQ) are actually two cotton case studies that determined the GHG emission levels of two cotton
operations. As such these studies can be used as benchmarks for the current operation, and when
converted to 300 hectares the NCEA operation would have used 47 700L and the ACSC operation 31
620L which also explains these differences in emission levels. As a standardised item one can actually
determine what the emission levels should be for this amount of diesel use by applying the IPCC energy
content and emission factors for mobile combustion diesel usage. This equates to 360 kg CO
2
e/ha as
indicated by the line on the above graph (2006 IPCC National Greenhouse Gas Inventories 2006). It is
therefore unclear what the results for the Lincoln University, Fieldprint and US Cropland calculators
could be based on.
Soil N
2
O Emissions

Fig. 2: Nitrous Oxide Emissions
Soil nitrous oxide emissions mainly result from fertiliser applications (Fig. 2). As most of the
calculators do not do a separate calculation for soil carbon emissions, some of those emissions may be
included under this section, as well as emissions from stubble decay, although these are less significant.
If one applies the IPCC default of 1.25% direct N
2
O emissions resulting from N fertiliser applications, it
equates to 341 kg CO
2
e/ha. As mentioned although some of calculators may add in some emissions from
crop residues and soil carbon, it still remains uncertain what the amount of 3 650 kg CO
2
e/ha in the case
of the US Cropland calculator could be based on. In the case of the two USQ case studies, the emissions
were not measured but derived from industry estimates and reports.
Crop Residue / Stubble
The Cool Farm Tool is the only calculator that specifies the emissions resulting from different crop
residue management practices. In the case of this study the reduced till option was selected where more
than 30% of the crop residue is left on the surface with the next planting. The corresponding emissions
are 66 kg CO
2
e/ha. The Grains Greenhouse Accounting calculator from the University of Melbourne
estimates this to be 48 kg CO
2
e/ha (Echard, 2009).
332
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No-till practices will more than compensate for fuel use according to the Cool Farm Tool results. Again
the variance in the results raises concern for any reliable industry or product level comparisons where the
methodologies are not stipulated or standardised.
Purchased Electricity
There is relatively little variance in the emission results from purchased electricity for the four calculators
that quantify this. Purchased electricity is a mandatory Scope 2 emission and should be part of any
product CFP calculation. In the case of agriculture it is mainly used for irrigation and can make up a
considerable part of a farms total carbon footprint, as is the case with these results where it is
comparable to fuel use.
Although one may expect that is another fairly standardised indicator, emission factors vary by
country and according to the look-up table in the Cool farm Tool it varies from 0.0 in the case of Norway
to 2.5 kg CO
2
/kWh for India. In the case of Australia there are regional variances from 0.23 for
Tasmania to 1.23 kg CO
2
/kWh for Victoria (NGERS (Measurement) Technical Guidelines, 2010).
Bio-Chemical Production
Fertilisers

Fig. 5: Emissions Associated with Fertilizer Production
Most of the calculators account for the Scope 3 emissions associated with the manufacture of
fertilisers (Fig 5). The FarmGas calculator is mainly focussed on emissions from farming practices and in
the case of the Veggie Carbon calculator it only deals with Scope 1 and 2 emissions, although PAS 2050
requires the inclusion of these emissions.
Again the results are relatively consistent compared to some of the other emission indicators. In the
interest of comparability, Urea was used as the standard fertiliser in the calculators as it is commonly
used in Australia along with anhydrous ammonia. However, these calculations do not entail a simple
application of a standard conversion rate for fertilisers in general, as the different fertilisers attract
different emission factors. Only the Cool Farm Tool and the Fieldprint calculators make a specific
distinction between the different types of fertilisers used. The table below shows the differences in
emission levels between using 220 kg N/ha of Urea and Anhydrous Ammonia for soil N
2
O emissions and
those from the production of the fertilisers:

334
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335
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ions are of
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his regard.

To gauge to total effect of fertiliser applications one should combine the soil N
2
O levels with the
fertiliser production emissions, which makes it the single biggest contributing factor to a crops total
carbon footprint. As irrigated cotton in mainly grown in fairly arid environments, electricity (or diesel)
use for irrigation will always be an important factor, but can also be affected by rainfall as the growing
and rainfall seasons often coincide for cotton.
One of the major contributions that CFP calculators can make is to show the potential benefits that
conservation farming practices can have to buffer or offset other emissions associated with growing a
crop, as clearly shown above. It was mentioned earlier than in the event of a no-till practice; the carbon
credit will have increased to 1750 kgs CO
2
per hectare according to the calculators estimate. One will
expect fuel use to be a significant contributor in capital intensive farming operations, as is the case with
cotton farming in Australia.
CONCLUSION
The following conclusions are drawn from the results:
There is a compelling need for an internationally standardised format and methodology for crop-
level carbon footprint (CFP) calculations for the textile supply chain
CFP tools, applied to the same farm data, generated vastly different results. Variations in
outcomes are mainly due to differences in structure and methodologies applied.
CFP calculators can play a valuable role in quantifying and acknowledging the positive outcomes
that conservation farming practices can have on mitigating the emission levels of farm products
No common methodology exists in particular for the calculation of soil carbon emissions
Further research needs to be undertaken to apply process-based models to validate the accuracy
of soil emission results from CFP calculators.
REFERENCES
[1] IPCC (2006) - IPCC National Greenhouse Gas Inventories 2006, 2006 IPCC Guidelines for National Greenhouse Gas
Inventories, 2006 IPCC National Greenhouse Gas Inventories Programme and Institute for Global Environmental
Strategies, Japan
[2] British Standards Institution. (2011) - PAS 2050 Research Report, British Standards Institution, London.
[3] Carbon Trust. (2008) - PAS 2050: 2008 - Specification for the assessment of the life cycle greenhouse gas emissions of
goods and services, Carbon Trust, London.
[4] DoCCaE Efficiency (2011a) - Carbon Credits Bill. Department of Climate Change and Energy Efficiency, Australian
Government.
[5] DoCCaE Efficiency (2011b) - Clean Energy Bill Department of Climate Change and Energy Efficiency, Australian
Government.
[6] Echard, R. (2009) - Grains Greenhouse Accounting, University of Melbourne, viewed 21 September 2011.
[7] Eco Index. (2011) - Eco Index Apparel Tool, Sustainable Apparel Coalition, viewed 15 September 2011,
<http://www.ecoindexbeta.org/>.
[8] Environmental Leader. (2010) - 72% of UK consumers: Give us carbon footprint labels on food, Environmental Leader,
viewed 12 January 2010.
[9] Johnson, J.M.F., Franzluebbers, A.J., Weyers, S.L. and Reicosky, D.C. (2007) - Agricultural opportunities to mitigate
greenhouse gas emissions, Environmental Poll., 150: 10724.
[10] Kim, B, Neff, R. (2009). Measurement and communication of greenhouse gas emissions from U.S. food consumption via
carbon calculators, Ecol. Econ., 69: 18696.
[11] Miller, V. (2008) - Weighing in, Guardian, 8 February 2008, p. 2.
[12] NGERS (2010) - NGERS (Measurement) Technical Guidelines, Department of Climate Change and Energy Efficiency,
Canberra.
[13] PCF World Forum. (2011) - ISO 14067 - Carbon Footprint of Products, PCF World Forum, viewed 12 September 2011.
[14] Planet Ark. (2010) - Carbon Reduction Label - Frequently asked questions, Planet Ark, viewed 26 July 2010.
[15] Russell, S. (2011) - Corporate Greenhouse Gas Inventories for the Agricultural Sector: Proposed accounting and reporting
steps, World Resources Institute, Washington.
[16] Stewardship Index. (2011) - Stewardship Index for Speciality Crops, Stewardship Index, viewed 27 July 2010.
[17] The Carbon Trust. (2010) - Footprint Expert, Carbon Trust, viewed 24 May 2010.
[18] The Economist. (2011) - Following the footprints, The Economist, vol. Q2 2011.
[19] The Greenhouse Gas Protocol Initiative. (2001) - Corporate Accounting and Reporting Standard, World Resources Institute
& World Business Council for Sustainable Development, Washington.
[20] The Greenhouse Protocol Initiative. (2010) - Measurement of GHG emissions, The Greenhouse Protocol Initiative, viewed
24 August 2010.
[21] The Keystone Alliance for Sustainable Agriculture. (2010) - Field to Market, The Keystone Center, Keystone.
[22] Von Wirn-Lehr, S. (2001). Sustainability in agriculture - An evaluation of principal goal-oriented concepts to close the gap
between theory and practice, Agric. Ecosys. Environ., 84: 115129.
[23] World Resources Institute & World Business Council for Sustainable Development. (2011) - The Greenhouse Gas Protocol
Initiative: Product accounting and reporting standard, World Resources Institute & World Business Council for
Sustainable Development, Washington.
Studies on the Seed Cotton Yield, Growth and Yield

Contributing Characters of New Bt Cotton Hybrids
under Varied Agronomic Manipulations
Kulvir Singh, Harmandeep Singh, R.K. Gumber and Pankaj Rathore
PAU, Regional Research Station, Faridkot151 203, Punjab

AbstractA field experiment was conducted at Punjab Agricultural University, Regional Research Station, Faridkot
during Kharif 2010 in a split plot design. The treatments comprised three Bt cotton hybrids (MRC 7361, Bioseed 6488
and RCH 134) in main, two plant geometries (67.5 x 75 cm & 67.5 x 90 cm) in sub and three nutrient levels {100%
recommended dose (RD) (150kg N, 30kg P/ha); 125% RD & 150% RD} in sub-sub plots replicated thrice. The soil of
the experimental site was loamy textured with slightly alkaline pH (8.7), normal EC (0.40 m mhos/cm), low in OC
(0.48%), low in available P (7.5 Kg/ha) but high in available K (750 Kg/ha). Among Bt hybrids, MRC 7361 had
heavier and more bolls per plant and recorded significantly higher seed cotton yield (2795 kg/ha) than Bioseed 6488
(2217 kg/ha) and RCH 134 (1897 kg/ha). Further leaf area index (5.91) was greater in case of MRC 7361 which
contributed to improved yield contributing parameters and significantly higher seed cotton yield (SCY). Ginning
outturn (%), lint and seed yield was also found to be significantly better for MRC 7361 than Bioseed 6488 and
RCH 134 hybrids. Fertilizer use efficiency (4.97 kg SCY/kg fertilizer applied) and water productivity (532.4 g/m
3
)
were significantly higher in MRC 7361 than the other two hybrids RCH 134 had the least values for LAI (3.80), FUE
(3.24 kg SCY/kg fertilizer applied) and water productivity (361.3 g/m
3
). Highest net returns of Rs. 88971/ha were
observed with MRC 7361 than Bioseed 6488 (Rs. 67301/ha) and RCH 134 (Rs. 55287/ha). Plant geometry had no
significant effect on yield. However, number of bolls/plant was significantly better under wider than closer spacing.
Among nutrient levels, 150% RD fertilizer produced 35.3 % higher seed cotton yield (2591 kg/ha) than 100% RD
fertilizer (1914 kg/ha) though it was at par with 125% RD fertilizer (2404 kg/ha). Significant improvement in number
of monopods and sympods per plant coupled with higher LAI (5.16) with 150% RD was mainly responsible for higher
yield than the 100% RD fertilizer level. Thus, significantly highest net returns (Rs. 80912/ha) were recorded for 150%
than 100% RD (Rs. 56351) and was similar to 125% RD.
Keywords: Bt cotton, hybrids, Seed cotton yield, Leaf Area Index, Plant geometry, Nutrient levels, Fertilizer use
efficiency and Water productivity.
INTRODUCTION
Cotton is an important cash crop in South-western zone of Punjab grown during Kharif season. It plays a
significant role in state economy and occupied an area of 511 thousand ha with a production of 2006
thousand bales and lint yield of 667 kg/ha during 2009-10 (Anonymous, 2011). Owing to fast growth and
better performance in terms of high seed cotton yield, Bt cotton hybrids have become popular among
farmers and is presently covering about 90 per cent of the cotton acreage in the state. Apart from
improvement in yield, Bt cotton hybrids have also lowered the pest incidence and reduced environmental
pollution by limited use of insecticides. Development of much efficient hybrids has a crucial role to play
for increasing the seed cotton yield in the present era of Bt cotton. The productivity of cotton can be
improved by identifying high yielding genotypes coupled with suitable agronomic practices. Among
different agronomic manipulations, selection of potential genotypes having high yield contributing
characters, optimum plant stand and suitable fertilization plays crucial role in increasing the productivity
of cotton. Therefore, the present study was undertaken to identify high yielding Bt cotton hybrids and
identify their agronomic requirements under the site specific agro-climatic conditions of Faridkot.
The field experiment was conducted during the Kharif 2010 at Research farm of Punjab Agricultural
University, Regional Research Station, Faridkot (30
0
40N and 74
0
44E), a typical representative of
South-Western zone (Zone IV) in Punjab. The soil of the experimental field was loamy in texture,
alkaline in soil reaction (pH 8.7), normal EC (0.40 m mhos/cm), low in OC (0.48%), low in available P
(7.5 kg/ha) but high in available K (750 kg/ha). The experiment conducted in a split split plot design
56
Studies on the Seed Cotton Yield, Growth and Yield Contributing Characters of New Bt Cotton Hybrids 339
comprised of three Bt hybrids (MRC 7361, Bioseed 6488 and RCH 134) in main plots, two plant
geometries (67.5 x 75 cm and 67.5 x 90 cm) in sub plots and three nutrient levels { 100% of RD (150kg
N, 30kg P/ha), 125% of RD (187.5kg N, 37.5kg P/ha) and 150% of RD (225 kg N, 45kg P/ha)} in sub-
sub plots replicated thrice. The crop was sown in first fortnight of May by dibbling 3-4 seeds/hill which
were later thinned to one seedling per hill. Full dose of phosphorus was applied before sowing while
nitrogen dose was given in two splits, first half at the time of thinning and the remaining at flowering
stage. Other production and protection measures were applied as per recommendations given by Punjab
Agricultural University. The data were analyzed statistically following procedures outlined by Cheema
and Singh (1991).
Effect of Bt Cotton Hybrids
TABLE 1: GROWTH, YIELD AND YIELD CONTRIBUTING CHARACTERS OF BT COTTON HYBRIDS UNDER DIFFERENT PLANT GEOMETRY AND NUTRIENT LEVELS
Treatments Plant
Height
(cm)
LAI Monopods/Plant Sympods/
Plant
Bolls/
Plant
Boll
Weight
(g)
Seed Cotton
Yield
(Kg/ha)
Lint
Yield
(kg/ha)
Biomass
(q/ha)
GOT
(%)
Bt Cotton Hybrids
Bioseed 6488 121.5 4.97 2.56 21.8 54.6 3.91 2217 748.5 122.4 33.7
RCH 134 107.0 3.80 2.46 22.2 47.8 4.50 1897 649.8 118.2 34.1
MRC 7361 147.2 5.91 3.08 29.5 57.8 5.41 2795 978.4 162.9 35.0
CD (0.05) 8.1 0.42 0.35 0.92 4.3 0.23 257 97.3 12.0 0.67
Plant Geometry
67.5 x 75 cm 127.4
5.05
2.66 23.2 51.3 4.50 2218 761.1 135.1 34.2
67.5 x 90 cm 123.1
4.74
2.74 25.8 55.5 4.71 2387 823.3 133.9 34.4
CD (0.05) 3.7 NS NS 2.39 3.2 NS NS NS NS NS
Nutrient Levels
100 % RDF 121.3 4.42 2.51 22.3 51.7 4.46 1914 660.7 130.5 34.4
125 % RDF 125.1 5.11 2.79 24.8 53.6 4.62 2404 828.7 133.9 34.3
150 % RDF 129.4 5.16 2.80 26.4 55.0 4.75 2591 887.2 139.1 34.2
CD (0.05) 5.0 0.39 0.21 2.41 NS NS 184 61.4 NS NS
The tested Bt cotton hybrids differed significantly for seed cotton yield (SCY) as well as for yield
contributing characters. Data in the Table 1 indicated that MRC 7361 produced significantly higher seed
cotton yield as compared to Bioseed 6488 and RCH 134 due to more number of monopods, sympods and
bolls per plant. Nehra et al. (2006) at Ganganagar, Rajasthan reported significant differences for SCY
among the different Bt cotton hybrids due to more number of bolls per plant. Singh et al. (2007) also
found significant differences for seed cotton yield among the tested Bt cotton hybrids. The present
investigations indicated an increase in yield of MRC 7361 over Bioseed 6488 and RCH 134 to the tune of
26.1 and 47.3 percent, respectively. Among Bt hybrids, MRC 7361 had heavier bolls than the rest.
Besides smaller boll size, RCH 134 Bt had significantly fewer bolls per plant compared to the other
hybrids. The biomass production was significantly higher for MRC 7361 as compared to other hybrids
was probable due to taller plants and vigorous growing habit. As a result of this, MRC 7361 exhibited
significantly higher leaf area index than Bioseed 6488 and RCH 134 (Table-2). All of the above said
factors further culminated to give higher fertilizer use efficiency (FUE) and water productivity (WP)
indices for MRC 7361 than the rest of the hybrids. Ginning out turn (%) for MRC 7361 was found to be
significantly better than Bioseed 6488 and RCH 134. Similarly, lint and seed yield was also
significantly superior in MRC 7361 than Bioseed 6488 and RCH 134. Significantly higher net returns
of Rs. 88971/ha were recorded with MRC 7361 as compared to Bioseed 6488 (Rs. 67301/ha) and RCH
134 (Rs. 55287/ha). Higher B: C ratio also indicated superiority of MRC 7361 (3.88) than rest of the
hybrids.
Effect of Plant Geometries
The data given in Table 1 suggest that plant geometries did not effect SCY significantly. Brar et al.
(2008) also reported non-significant differences for seed cotton yield with respect to plant geometries.
However in present investigations, the number of sympods and boll number per plant was significantly
more under wider geometry i.e 67.5 x 90 cm but converse was true for plant height. Narayana et al
(2007) reported that the number of bolls/plant was significantly better under wider plant geometry of 120
x 60cm than the closer geometry (90 x60 and 90 x 90cm). However, non-significant differences were
observed for LAI, GOT, Lint and seed yield (Table-1) and FUE and WP.
TABLE 2: EFFECT OF DIFFERENT TREATMENTS ON FERTILIZER USE EFFICIENCY (FUE) AND WATER PRODUCTIVITY (WP)
Treatments LAI FUE
(kg Seed Cotton Yield/ kg Fert. Applied)
WP (g/m
3
)
Bt Cotton Hybrids
Bioseed 6488 4.97 3.75 422.3
RCH 134 3.80 3.24 361.3
MRC 7361 5.91 4.90 532.4
CD (0.05) 0.42 0.43 48.9
Nutrient levels
(150kg N, 30kg P/ha) 100 % RD 4.42 4.09 364.7
(187.5kg N, 37.5kg P/ha) 125 % RD 5.11 4.11 457.9
(225 Kg N, 45kg P/ha) 150 % RD 5.16 3.69 493.5
CD (0.05) 0.39 0.32 35.2
Effect of Nutrient Levels
Among nutrient levels, 150% RD produced 35.3 percent higher yield than 100 % RD fertilizer and was
equivalent to 125% RD (Table-1) significantly, increased number of monopods and sympods per plant
were also observed with 150% RD than the recommended fertilizer level. Consequently, better indices
for LAI (5.16) as compared to 100% RD nutrient levels might have reflected its positive effect on SCY.
Singh and Rathore (2007) also reported significant improvement in SCY with increased N level.
However, in the present studies, number of bolls per plant, boll weight and dry matter was not affected by
nutrient levels. Lint and seed yield under high N levels was significantly better than the 100 % RD
level (Table 1). Significantly higher net returns under high nutrition levels (Rs. 74297-80912) than 100%
RD (Rs. 56351/ha) and consequently higher B:C ratio (3.34-3.54) also supported the favour of higher
nutrient applications. These results are in conformity with the findings of Biradar et al. (2010) who
reported higher returns with enhanced level of nutrition (150% RD) than recommended (100% RD) level
of nutrition.
REFERENCES
[1] Anonymous. (2011) - Punjab Agricultural University, Ludhiana. Package of practices for Kharif Crops.
[2] Brar, J.S., B.S. Sidhu, K.S. Sekhon and G.S. Buttar (2008) - Response of Bt cotton (Gossypium hirsutum L.) to plant
geometry and nutrient combinations in sandy loam soil. J. Cotton Res. & Dev., 22(1): 5961.
[3] Biradar, V., Rao, S. and Hosamani, V. (2010) - Economics of late sown Bt cotton (Gossypium hirsutum L.) as influenced by
different plant spacing, fertilizer levels and NAA applications under irrigation. International J. of Agric. Sci.,
6(1): 196198.
[4] Cheema, H.S. and Singh, B. (1991) - Software: Statistical package CPCS-1 Deptt. of Math, Statistics and Physics; Punjab
Agricultural University, Ludhiana.
[5] Narayana, E., Hema, K., Srinivasulu, K., Prasad, N.V.V.S.D. and Rao, N.H.P. (2007) - Agronomic evaluation of Gossypium
hirsutum hybrids for varied spacings and nitrogen levels in vertisols under rainfed conditions. J. Cotton Res. & Dev.
21(2): 197200.
[6] Nehra, P.L., Kumawat, P.D. and Nehra, K.C. (2006) - Response of promising Gossypium hirsutum hybrids to fertilizer
levels in irrigated north western plain zone of Rajasthan. J. Cotton Res. & Dev. 20(1): 8788.
[7] Singh, K. and Rathore, P. (2007) - Effect of different spacings and nitrogen levels on growth and yield attributes of
American cotton (Gossypium hirsutum L.) genotypes. J. Cotton Res. & Dev. 21(2): 178179.
[8] Singh, K., Jindal, V., Singh, V. and Rathore, P. (2007) - Performance of Bt cotton hybrids under different geometrical
arrangements. J. Cotton Res. & Dev. 21(1): 4144.
Evaluation of Cotton Genotypes for High Density

Planting Systems on Rainfed Vertisols
of Central India
M.V. Venugopalan, A.H. Prakash, K.R. Kranthi, Rachana Deshmukh,
M.S. Yadav and N.R. Tandulkar
Central Institute for Cotton Research, P.O. Box. No. 2, Shankarnagar, Nagpur440010, India

AbstractPromising genotypes of both Gossypium hirsutum and Gossypium arboreum were evaluated for their
performance at higher planting densities in a field trial under rainfed conditions at the Central Institute for Cotton
Research, Nagpur, India during 2010-2011. The central region is characterized by a hot, dry sub-humid climate and
the soil was a Typic Haplustert, low in organic C and available P but rich in available K. Five genotypes of
G. hirsutum (Anjali, CCH 724, NISC 50, PKV 081 and CNH 120MB) evaluated at 5 crop densities viz. 60x30,
45x20, 45x13.5, 30x30 and 30x20 cm. Similarly five genotypes of G. arboreum (AKA 07, CINA 404, PA 255, PA 08,
JK 5) were also evaluated at 5 crop densities viz. 60x15, 45x13.5, 45x10, 30x20 and 30x15 cm. Results indicated that
among G. hirsutum, genotype PKV 081 was found most suitable for high density planting at 30x20 cm and 45x13.6
cm (both corresponding to 166000 plants/ha) in terms of yield, morphology, earliness, tolerance to sucking pests and
boll weight. In G. arboreum, CINA 404 gave the highest yield of 2174 kg seed cotton/ha at 45x10 cm (222000
plants/ha). However, other high yielding genotypes viz., JK5 (1842 kg seed cotton/ha) and AKA07 (1815 kg seed
cotton/ha) were dwarf and more compact than CINA 404. Across genotypes, a spacing of 45x13.5 cm for G. hirsutum
and 45x10 cm for G. arboreum was optimum for planting short compact plant types. In both G. hirsutum and G.
arboreum genotypes the boll weight and harvest index decreased with increasing plant density. The latter in turn,
decreased nutrient utilization efficiency. Nevertheless, in all the genotypes of both species, the nutrient uptake
efficiency and partial factor productivity for N increased with increase in planting density. The partial factor
productivity of N ranged from 10.6 to 21.8 kg seed cotton/kg N applied and 18.3 to 22.8 seed cotton/kg N applied in G.
hirsutum and G. arboreum genotypes respectively and PKV 081 and CINA 404 recorded the highest values.
INTRODUCTION
Manipulation of planting density, plant population and spatial arrangement of cotton plants continues to
be topics of cotton research worldwide and India is no exception. It is widely accepted that increasing
planting density is an option to increase yield or profits and also to improve input use efficiency. The
availability and acceptance of effective alternate weed and insect pest management strategies and spindle
type pickers has rekindled interest on high density cotton planting systems (Reddy et al., 2009). With
increase in planting density the yield per unit area generally to an upper limit or optima later plateaus and
ultimately declines. The optimum plant density under this parabolic relationship will depend upon the
genotype characteristics, properties of soil, climatic parameters and management regime (Silvertooth et
al., 1999). The earliness usually associated with high density planting makes this system suitable for
rainfed Vertisols where the cotton crop invariably experiences terminal moisture stress.
Before the advent of cotton hybrid and the adoption of seed dibbling techniques, varieties of
G. hirsutum and G. arboreum were planted at populations ranging from 55000 to 89000 plants/ha
(Bonde, 1992). Cotton breeding efforts in India were not focused on developing plant types specially
suited for high density planting (CICR, 2011). The present study was therefore formulated to evaluate
promising genotypes of both G. hirsutum and G. arboreum on rainfed Vertisols with specific objectives
of identifying genotypes amenable to high density planting and to optimize planting density/crop
geometry for yield maximization under high density planting.
Separate field experiments were conducted under rainfed conditions during the kharif (monsoon) season
of 2010-2011 at the experimental farm of Central Institute for Cotton Research, Nagpur. The site is a
representative of Agro eco sub region 10.2 (hot dry sub-humid climate) dominated by Vertisols and
57
Vertic intergrades. As against a mean seasonal rainfall of 760 mm, the season received 1005 mm rainfall
spread over 53 days. The soil at the experimental site was a Typic Haplustert slightly alkaline in reaction
low in organic C (0.39%) low in Olsens available P (10.3 kg/ha) and high in available K (630 kg/ha).
Two sets of experiments were laid out in split plot design with 3 replications. In the first set the main plot
comprised of 5 crop geometries viz., 60x30 cm (55,555 plants/ha), 30x30 cm, 45x20 cm (both 11,111
plants/ha), 30x20 cm and 45x13.5 cm (both 1,66,666 plants/ha) and 5 genotypes of G. hirsutum viz.,
Anjali, CNH 120MB, PKV 081, NISC 50 and CCH 724 as sub-plots. In the second set, the main plot
comprised of 5 crop geometries viz., 60x15 cm (11,111 plants/ha), 45x13.5 cm, 30x20 cm (both 1,66,666
plants/ha), 45x10 cm and 30x15 cm (both 2,22000 plants/ha and 5 genotypes AKA 7, CINA 404, PA
255, PA 08 and JK 5 formed the sub plots. The crop was sown on and raised using the recommended
package of practices.
At maturity, five plants per plot were harvested, separated into leaves, stem, carpels, seed, lint and
other floral parts and dried to calculate dry matter yield. N content in various plant parts was estimated
following wet and digestion and total N uptake was calculated. Seed cotton yield was recorded for the
entire net plot. Nitrogen Use Efficiency (NUE or N response or Partial factor productivity of N) and
its components, N uptake efficiency (N Upt E) and N utilization efficiency (N Uti E), were calculated on
a kg kg
-1
basis according to Moll et al. (1982), where: NUE = N Upt E x N Uti E and in turn N Upt E =
Nt/ Na and N Uti E= SCY/ Nt, where Nt, Na and SCY denote N uptake, N applied and seed cotton yield,
respectively. Data was subjected to ANOVA (Gomez and Gomez, 1984).
Yield and Yield Attributes
Hirsutum
The effect of spacing, genotype and genotype x spacing interaction on seed cotton yield, number of
bolls/m
2
and boll weight was significant (Table 1). Averaged across genotypes, the seed cotton yield was
significantly higher in the spacings of 45x13.5 cm and 30x20 cm, both accommodating 166000 plants/ha.
Jagannathan and Venkitaswamy (1996) also reported that dwarf compact genotypes responded
favourably to a population of 1,11,000 plants/ha on Vertisols. Since within a given population both the
crop geometries (spacings) produced similar yields, a rectangular spacing (45x13.5 cm or 45x20 cm)
which facilities inter-culture was as good as a more square type geometry (30x20cm or 30x30 cm).
The highest mean yield (1203 kg/ha) was obtained with 45x13.5 cm spacing (166000 plants/ha)
which in turn was 38% higher than that obtained with the recommended (60x30 cm spacing). Among the
5 genotypes PKV 081 was significantly superior to the other genotypes. Genotype x spacing interaction
was also significant indicating that genotypes Anjali, PKV 081 and CCH 724 were more amenable to
closer spacings i.e. higher planting densities (166000 plants/ha). Rossi et al., (2007) also reported
significant genotype x spacing interaction in Spain.
TABLE 1 EFFECT OF PLANTING DENSITY ON YIELD AND YIELD ATTRIBUTES IN G. HIRSUTUM GENOTYPES
Spacing (cm) 60x30 45x20 30x 30 45x 13.5 30x20 Mean
Population/ha 55000 111000 111000 166000 166000
Genotypes Seed cotton yield (kg/ha)
Anjali 502 847 853 966 796 793
CNH120MB 1030 976 1138 1250 1289 1137
PKV 081 1200 1714 1418 1921 1967 1644
NISC 50 1056 890 1103 1016 1052 1023
CCH 724 679 843 681 864 835 781
Mean 893 1054 1039 1203 1188
CD at 5% Spacing 97.636 Genotypes 70.205 Interaction 170.710
Table1(Contd.)
Evaluation of Cotton Genotypes for High Density Planting Systems on Rainfed Vertisols of Central India 343
Table1Contd.
Number of bolls/m
2

Anjali 15.3 24.1 28.4 28.6 25.8 24.4
CNH120MB 34.2 45.0 53.7 82.0 70.1 57.0
PKV 081 34.5 56.1 58.0 98.7 65.5 62.6
NISC 50 28.1 27.1 32.1 42.4 36.0 33.1
CCH 724 15.3 37.0 41.3 31.4 43.3 33.7
Mean 25.7 37.9 42.7 56.6 48.1
Boll Weight (g)
Anjali 3.27 3.52 3.21 3.33 2.74 3.22
CNH120MB 2.31 2.56 2.07 2.33 1.93 2.24
PKV 081 3.49 3.32 2.69 3.25 3.08 3.17
NISC 50 3.53 3.27 3.35 2.52 2.97 3.13
CCH 724 3.33 3.04 2.32 2.78 1.98 2.69
Mean 3.19 3.14 2.84 2.72 2.55
In all the genotypes the boll number per plant decreased with closer spacing due to greater inter-plant
competition. However, the increase in the number of plants per unit area at closer spacing compensated
for this decline and hence the boll number/m
2
were significantly higher at all the closer spacings
compared to the recommended (60x30 cm) spacing. Jost and Cothern (2000) also observed a decrease in
boll number per plant but an increase in boll number per unit area at elevated populations.
The highest yielding cultivar, PKV 081 had the highest boll number/m
2
. The genotype x spacing
interaction was also significant and unlike other genotypes PKV 081 produced almost thrice the number
of bolls/m
2
at 45x13.5 cm over the currently recommended spacing of 60x30 cm. In general the boll
weight declined at closer spacing. There was significant variation among genotypes. Genotypes Anjali,
PKV 081 and NISC 50 produced significantly bigger bolls than CNH 120MB and CCH 724.
Arboreum
All the genotypes of G. arboreum responded significantly to higher planting density (Table 2) and the
mean yield increase the highest planting density (45x10 cm) was 45.4% over the recommended one
(60x15 cm). Among the genotypes CINA 404 yielded significantly higher than all other genotypes but in
terms of microbiological parameters (height, monopodia etc.) AKA 7 and JK 5 were more amenable to
HDPS. Genotypes AKA7 and JK5 were more determinate and the boll bursting was uniform. Earlier
(Belot and Salles 2010) suggested that termination in fruiting after the production of 5-7 bolls/plant is
essential for the success of high density planting. Genotype x plant density interaction was also
significant for yield. The yield increase at higher plant density was primarily due to more number of
bolls/m
2
since the effect of boll weight was not significant. Across genotypes the boll number/m
2
was
significantly higher at 45x10 cm spacing. Among genotypes, CINA 404 produced significantly larger
bolls over the other genotypes. Low boll weight is an inherent limitation in G. arboreum cotton and an
increase in boll weight may directly contribute to an increase in yield.
Nutrient use Efficiency Parameters
Hirsutum
The N response ratio or N use efficiency or partial factor productivity is the simplest measure of
efficiency. It provides a measure of crop yield per unit of N applied. In different genotypes of N ranged
from 10.6 to 21.8 and PKV 081 recorded the highest values (Table 3). In general there was a gradual
increase in N response from 11.9 kg/ha at 55000 plants/ha (60x30 cm) to 13.9-14.1 at 111000 plants/ha
and 20.4-23.9 at 166000 plants/ha. This N response is a product of N uptake efficiency (N uptake/N
applied) and N utilization efficiency (seed cotton yield/N uptake). Among genotypes, the N uptake
efficiency was the highest for PKV 081. The N uptake efficiency as distinctly higher at 111000 plants/ha
(0.91-1.00) and 55000 plants/ha (0.91-1.00) compared to 0.73 at the recommended 55000 plants/ha. N
utilization efficiency declined slightly with increase in planting density.
TABLE 2: EFFECT OF PLANTING DENSITY ON YIELD AND YIELD ATTRIBUTES IN G. ARBOREUM GENOTYPES
Spacing (cm) 60x15 45x13.5 30x 20 45x 10 30x15 Mean
Population/ha 111000 166000 166000 222000 222000
Genotypes Seed cotton yield (kg/ha)
AKA-07 1163 1349 1456 1815 1419 1441
CINA-404 1430 1550 1610 2173 1772 1707
PA-255 1259 1595 1349 1625 1226 1411
PA-08 1090 1318 1455 1509 1479 1370
JK-5 1223 1452 1151 1842 1734 1480
Mean 1233 1453 1404 1793 1526
Number of bolls/m
2

AKA-07 50.0 58.1 67.3 76.5 65.4 63.4
CINA-404 59.8 71.0 75.6 82.6 75.2 72.9
PA-255 54.3 77.5 77.5 77.7 71.5 71.7
PA-08 55.5 62.7 58.1 93.7 66.6 67.3
JK-5 46.9 70.1 63.6 117.2 86.3 76.8
Mean 53.3 67.9 68.4 89.5 73.0
Boll weight (g)
AKA-07 2.29 2.37 2.43 2.24 2.07 2.28
CINA-404 2.47 2.45 2.67 2.22 2.41 2.44
PA-255 1.91 1.98 1.80 2.48 2.31 2.10
PA-08 2.30 2.05 2.12 2.05 1.72 2.05
JK-5 2.41 2.10 1.73 2.11 2.07 2.08
Mean 2.28 2.19 2.15 2.22 2.12
CD at 5% Spacing NS Genotypes 0.173 Interaction NS
N utilization efficiency is a product of HI and N biomass production efficiency (Ortiz Monasterio
et al., 1997). In the present study there is a reduction in HI at higher planting density which would have
decreased the N utilization efficiency. Darawsheh et al., (2009) also observed that the partitioning of
assimilates to reproductive parts was lower in narrow row high plant density systems. For improved N
utilization efficiency genotypes which maintain high HI even at increased planting densities would be
ideal. Genotype with higher harvest index viz., PKV 081 also had a higher N utilization efficiency.
TABLE 3: EFFECT OF PLANTING DENSITY ON N USE EFFICIENCY PARAMETERS IN G. HIRSUTUM GENOTYPES
Spacing (cm) 60x30 45x20 30x 30 45x 13.5 30x20 Mean
Population/ha 55000 111000 111000 166000 166000
Genotypes N use Efficiency (N Response Ratio or Partial Factor Productivity)
Anjali 6.69 11.29 12.88 11.37 10.61 10.57
CNH120MB 13.73 13.01 16.67 15.19 17.19 15.16
PKV 081 16.00 22.85 25.61 18.91 25.61 21.80
NISC 50 14.08 11.87 13.55 14.71 13.55 13.55
CCH 724 9.05 11.24 9.08 9.08 11.52 9.99
Mean 11.91 14.05 15.56 13.85 15.70
N uptake efficiency
Anjali 0.46 0.84 0.78 0.72 0.73 0.70
CNH120MB 0.84 0.88 1.02 1.04 1.14 0.98
PKV 081 0.84 1.19 1.34 1.06 1.37 1.16
NISC 50 0.90 0.91 1.10 1.04 1.04 1.00
CCH 724 0.63 0.73 0.75 0.67 0.73 0.70
Mean 0.73 0.91 1.00 0.91 1.00
N Utilization Efficiency
Anjali 14.655 13.47 16.55 15.815 14.54 15.01
CNH120MB 16.366 14.75 16.27 14.557 15.14 15.42
PKV 081 19.006 19.22 19.08 17.841 18.73 18.78
NISC 50 15.697 13.03 12.36 14.102 12.98 13.63
CCH 724 14.392 15.36 12.11 13.619 15.84 14.26
Mean 16.02 15.17 15.27 15.19 15.45
Evaluation of Cotton Genotypes for High Density Planting Systems on Rainfed Vertisols of Central India 345
Arboreum
The N response ratio or partial factor productivity of N ranged from 18.3 to 22.8 and CINA 404 recorded
the highest values (Table 4). Typical value of Nitrogen use efficiency under Indian condition was 5.7 kg
lint 1kg N corresponding to around 17.1 kg seed cotton/kg N (ICAC 2008). In general there was a
gradual increase in N response from 16.4 kg/ha at 111000 plants/ha (60x15 cm) to 19.4-19.5 at 55000
plants/ha and 20.4-23.9 at 222000 plants/ha. This N response is a product of N uptake efficiency (N
uptake/N applied) and N utilization efficiency (seed cotton yield/N uptake). Where there was little
variation (range 0.95 to 1.18) in N uptake efficiency among genotypes, the uptake efficiency as distinctly
higher at 166000 plants/ha (0.97-1.09) and 222000 plants/ha (1.13-1.17) compared to 0.81 at the
recommended 111000 plants/ha. Genotype with higher harvest index viz., AKA 7 and JK 5 had higher N
utilization efficiency. Increase in population to 222000 plants/ha tended to decrease the N utilization
efficiency.
TABLE 4: EFFECT OF PLANTING DENSITY ON N USE EFFICIENCY PARAMETERS IN G. ARBOREUM GENOTYPES
Spacing (cm) 60x15 45x13.5 30x 20 45x 10 30x15 Mean
Population/ha 111000 166000 166000 222000 222000
Genotypes N use Efficiency (N Response Ratio or Partial Factor Productivity)
AKA-07 15.51 17.99 24.20 19.41 18.92 19.21
CINA-404 19.05 20.67 28.99 21.47 23.63 22.76
PA-255 16.79 21.27 21.67 17.99 16.35 18.81
PA-08 14.53 17.57 20.12 19.40 19.72 18.27
JK-5 16.31 19.36 24.56 19.36 23.12 20.54
Mean 16.44 19.37 23.91 19.53 20.35
N uptake Efficiency
AKA-07 0.75 0.92 1.06 1.01 1.02 0.95
CINA-404 0.99 0.98 1.40 1.22 1.30 1.18
PA-255 0.82 1.01 1.14 1.28 1.07 1.06
PA-08 0.75 0.97 1.05 1.08 1.07 0.98
JK-5 0.73 0.94 1.17 0.85 1.21 0.98
Mean 0.81 0.97 1.17 1.09 1.13
N utilization Efficiency
AKA-07 20.71 19.64 22.86 19.30 18.56 20.22
CINA-404 19.33 21.00 20.73 17.58 18.12 19.35
PA-255 20.60 21.03 18.98 14.01 15.27 17.98
PA-08 19.37 18.03 19.08 18.04 18.43 18.59
JK-5 22.39 20.61 20.93 22.85 19.12 21.18
Mean 20.48 20.06 20.52 18.36 17.90
CONCLUSION
On rainfed Vertisols, the genotype PKV 081 was found most suitable for high density planting system
(HDPS) (166000 plants per ha) based on yield (1921 kg/ha), morphological features, earliness, tolerance
to sucking pests and boll weight. In G. arboreum, on basis of yield, CINA 404 (2174 kg/ha) performed
well under under HDPS (222000 plants/ha). However other high yielding genotypes viz., JK 5 (1842
kg/ha) and AKA 7 (1815 kg/ha) were dwarf and more compact than CINA 404. Across the genotypes the
spacing of 45x13.5 cm (166000 plants/ha) was optimum for short compact types. Similar a spacing of
45x10 cm (222000 plants/ha) was optimum for short compact plant types of G.arboreum.
REFERENCES
[1] Belot, J.L. and Salles, A.O. (2010) - Second-cycle cotton grown with high plant density in Mato Grosso, Brazil. The ICAC
Recorder 28(2): 915.
[2] Bonde, W.C. (1992) - Achievements in cotton production technology. Edited by A.K. Basu and S.S. Narayanan, 1992.
Achievements of AICCIP (196792), 176 pp.
[3] CICR (2011) - CICR vision 2030. Central Institute for Cotton Research, Nagpur.
[4] Darawsheh, M.K., Khah, E.M., Avivalakis, G., Chachalis, D. and Fatbardh Sallaku (2009) - Cotton row spacing and plant
density cropping systems I. Effects on accumulation and partitioning of dry mass and LAI. Journal of food, Agriculture and
Environment, Vol. 7 (3&4), July-October 2009.
[5] Gomez, K.A. and Gomez, A.A. (1984) - Statistical Procedures for Agricultural Research, 2nd edn. John Wiley,
New York, USA.
[6] ICAC (2008) - Nitrogen fertilization in cotton. The ICAC Recorder. 26(1) 813.
[7] Jagannathan, N.T. and Venkitaswamy, R. (1996) - Effect of plant density and nutrient levels of new cotton varieties.
Madras agric. J. 83(3): 159161.
[8] Jost, P.H. and Cothern, J.T. (2000) - Growth and yield comparisons of cotton planted in conventional and ultra-narrow row
spacings. Published in Crop Sci. 40: 430435.
[9] Moll, R.H., Kamprath, E.J., and Jackson, W.A. (1982) - Analysis and interpretation of factors which contribute to
efficiency of N utilization. Agron. J., 74: 562564.
[10] Ortiz-Monasterio, R.J.I., Sayre, K.D., Rajaram, S., McMahon, M. (1997) - Genetic progress in wheat yield and N use
efficiency under four N rates. Crop Sci., 37, 898904.
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irrigated environment: plant population and lint yield. The Journal of Cotton Science 13: 4855.
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World Cotton Conference 4, (Sept. 10-14, 2007). Lubbock, Texas. wcrc.confese.com/wcrc/2007/tech program/p
1772HTM.
[13] Silvertooth, J.C., Edmisten, K.L., and McCarty, W.H. (1999) - Production practices. In C.W. Smith (ed.) Cotton: Origin,
history, technology and production. John Wiley and Sons, New York. pp. 463465.
Pruning and Detopping Studies in Bt-Cotton

S.S. Hallikeri
Associate Professor of Agronomy,
Agricultural Research Station, Dharwad farm, Dharwad580007, Karnataka, India

AbstractExcess growth management is a challenging task in cotton because of its indeterminate growth habit.
Failure of reproductive phase into seed cotton leads to a sudden spurt of vegetative growth. Various management
strategies to minimize cotton growth are detopping, pruning of monopodials at their early period of 40-50 DAS and
planting geometry Field studies were undertaken at ARS, Dharwad from 2007 to 2009 on medium deep black soils in
a split-split-plot design with detopping at 80 DAS and no detopping as main plots, retaining of only one, two, three or
all monopodia as sub-plots and planting geometry of 90x 30 cm, 90 x 60 cm as sub-sub-plots. Averaged over years,
detopping had no significant effect on seed cotton yield (SCY) (2116 kg/ha) when compared with no detopping (2149
kg/ha). However, detopping reduced and number of sympodial branches. Further, pruning of monopodials, too had no
effect on yield. Planting geometry with closer spacing of 90 x 30 cm produced significantly higher seed cotton yield
(2264 kg/ha) than 90 x 60 cm (2001 kg/ha). Productivity per plant, number of bolls per plant and boll weight
significantly increased at wider spacing due to reduced population levels. It is concluded that growth-reducing
function like detopping can be adopted under excess growth situations without affecting productivity.
INTRODUCTION
Cotton, the white gold' or the 'king of the fibre, as it is often referred to, still holds its position high.
World over, cotton is gradually assuming the status of a preferred fibre even for fashion fabrics. Cotton
cultivation needs to be sustainable, offering livelihood security to millions of people in the country.
Though release of Bt-cotton in the country increased the productivity of the crop several other production
factors may help further upgrade the productivity. Some situations such as high soil fertility and
irrigation water management practices lead to more of vegetative growth and resultant competition for
natural resources. Thus adjacent plants are affected and reproductive phase gets delayed. Growth
modification practices therefore become important by converting its phase of vegetative to reproductive
growth. This can be achieved by detoping and pruning. Detopping is removal of top terminal portion
after prominent vegetative growth stag. This may encourage growth of already formed sympodials as
well as formation and development of fruiting bodies. Detopping may also help reduce sucking pests and
bollworm infestation, as it avoids fresh growth out let which is unwanted at particular stage. Hence,
detopping of cotton after required growth may result in booming of cotton production. In view of this
hypothesis, field studies were conducted for 3 years to study the response of Bt-cotton when pruned and
detopped under different planting patterns.
Field experiments were conducted for three years from 2007 to 2009 at Agriculture Research Station,
University of Agricultural Sciences, Dharwad, Karnataka. Soils were medium deep black and neutral in
their pH (7.6) with low available N, medium in available P and high exchangeable K. Experiment was
laid out in split-split plot design with three replications. Main plots comprised detopping and no
detopping with different levels of pruning of monopodials in the subplots. Two planting methods (90 x
30 and 90 x 60 cm) were in the sub-sub plots. Experiment was sown every year in the month of June
July. Recommended doses of 80:40:40 kg NPK was applied with half of the N and entire P & K as basal.
The remaining N was applied 60 DAS. Detopping of top terminal portion was done at 80 DAS in the
detopping treatments. Pruning of monopodials were undertaken at 50 DAS as per the treatments.
Standard recommendations for plant protection, weed control, nutrient management were followed.
Growth and yield parameters were recorded at the time of harvesting. Data on various parameters
recorded from the field experiment were statistically analyzed using MSTAT-C programme and the level
of significance used in F test was at p=0.05.
58
Effect of Detopping
Detopping of cotton at 80 DAS significantly affected the growth parameters viz., plant height and
sympodial branches at various crop growth stages (Table-1). Detopping at 80 DAS significantly
decreased plant height compared to no detopping. Decrease in plant height was due to termination of
apical dominance (Brar et al., 2000 and Vekatakrishnan and Pothiraj, 1994). However, detopping had no
effect on the production of monopodial branches. On the other hand, detopping reduced the number of
sympodial branches at harvest when compared with no detopping. Effect of detopping was seen at both
vertical and horizontal growth of cotton.
TABLE 1: EFFECT OF PRUNING AND DETOPPING ON GROWTH AND YIELD PARAMETERS OF COTTON (MEAN OF 2007 2008 AND 2009)
Treatments Plant ht. Mono
Podia
Sym
Podia
Bolls/ Plant Boll wt. Yield/ Plant Yield kg/ha
Main plot (D)
Detopping at 80 DAS 87.0 1.48 14.1 27.2 3.89 98.4 2116
No Detopping 101.8 1.54 16.9 27.9 3.95 97.5 2149
S.Em. + 0.8 0.03 0.1 0.4 0.02 0.7 42
CD @ 5% 2.9 NS 0.3 NS NS NS NS
Sub plot (P)
Retaining of 1-monopodium 96.0 1.11 15.8 27.3 3.97 96.6 2098
Retaining of 2-monopodia 94.1 1.40 15.7 26.8 3.94 95.4 2129
Retaining of 3-monopodia 93.1 1.55 15.4 27.8 3.94 98.3 2096
Retaining all monopodia (check) 94.5 1.98 15.2 28.3 3.83 101.5 2209
S.Em. + 1.2 0.06 0.2 0.6 0.04 2.1 37
CD @ 5% NS 0.18 NS NS 0.1 NS NS
Sub sub plot (S)
90 x 30 cm 95.8 1.45 15.3 23.0 3.87 80.3 2264
90 x 60 cm 93.0 1.57 15.7 32.1 3.97 115.6 2001
S.Em. + 0.9 0.05 0.1 0.4 0.03 1.5 26
CD @ 5% 2.4 NS 0.41 1.2 0.07 4.1 73

Interaction (DXPXS) NS NS * NS NS NS NS
Canopy modification by detopping of cotton had no influence on seed cotton yield (SCY) as
compared to no detopping. Even though the plant was condensed vertically in the detopped treatment,
yield per hectare was not affected. Similarly, yield parameters of cotton viz., yield per plant, boll weight,
number of bolls per plant, number of green bolls per plant were also not significantly affected due to
detopping. These results are similar to those observed by Brar et al. (2002) for cotton cultivars and Rao
and Lakshminarayan (1985) for hybrids. This indicates that the cotton crop canopy can be modified by
reducing its spread and height without reducing the productivity of Bt-cotton. Excess growth in cotton is
a result of high fertility (N-fertilizers) and availability of soil moisture. The effects of detopping are more
pronounced in longer duration cottons with fruiting period extending beyond 150-180 DAS. Beneficial
effect of detopping may not be possible in cotton with less luxuriant growth (Rao and Lakshminarayan,
1985).
In years of heavy rainfall and under high fertility conditions, hybrid cotton attains unmanageable
growth and vegetative crop canopy which has got adverse effect on reproductive phases of cotton directly
and encourages more and more incidence of bollworms indirectly and it further results in ineffective
pesticide spraying due to higher canopy levels. Application of higher quantity of nitrogenous fertilizers
also can cause more vegetative growth and increases the competition for scarce inputs. Canopy
management practices thus plays an important role and helps to encourage reproductive phase. As cotton
is indeterminate in growth habit detopping of the plant after suitable vegetative stage (80 DAS)
encourages the better growth of already developed sympodial branches (Brar et al., 2000) and
disallowing the pest of cotton for feeding and egg laying (Singh and Sandhu, 1996).
Pruning and Detopping Studies in Bt-Cotton 349
Effect of Pruning of Monopodials
Unlike detopping pruning of monopodials to retain specific numbers did not significantly affect either
plant height or sympodial numbers. Pruning of monopodial branches also had no significant on SCY.
However, there was a yield reduction to an extent of 3.6 to 5.0 per cent when compared with no pruning
(checks). Similar trend was also observed with yield per plant and number of bolls per plant. Objective of
horizontal coverage of monopodials was checked thus allowing the higher populations per unit area.
Hence pruning of monopodial branches did not offer any yield advantage in increase yield.
Planting Geometry Relations
Mean of three years data on productivity indicated that Bt-cotton can be accommodated at higher
population levels (90x30 cm) when comparing with lower populations (90x60 cm) (Table-2). Closer
spaced plants had fever bolls and lower per plant yield than the wide spaced plants. Yield per unit area in
closer spaced treatments was greater compared to wider spaced treatments. Plant height significantly
increased at 90x 30 cm but reduced it s sympodial numbers when compared with 90x60 cm.
TABLE 2: EFFECT OF PRUNING AND DETOPPING ON YIELD (KG/HA) OF COTTON (MEAN OF 2007 2008 AND 2009)
Treatments Main plot (D) Detopping at 80 DAS No detopping Mean of sub plots
90 x 30 cm 90 x 60 cm Mean 90 x 30 cm 90 x 60 cm Mean
Retaining of 1-monopodium 2178 1983 2080 2245 1984 2115 2098
Retaining of 2-monopodia 2273 2093 2183 2241 1908 2074 2129
Retaining of 3-monopodia 2207 1882 2045 2240 2052 2146 2096
Retaining all monopodia (check) 2307 2007 2157 2422 2099 2261 2209
Mean 2241 1991 2116 2287 2011 2149
Comparisons for means of S.Em. + CD @ 5%
Main plot (D) 42 NS
Sub plot (P) 37 NS
Sub sub plot (S) 26 73
M X PX S 75 NS

The interaction effects were not significant. Detopping and no pruning at 90 X 30 cm spacing recorded
the highest seed cotton yield (2422 kg/ha) compared to other treatment interactions. Data of this study
indicates that either detopping or pruning of monopodia had no advantage with any of the planting
geometry. Hence from the results it can be concluded that detopping had no advantage to produce higher
yield. But growth can be checked with detopping without affecting yield. Pruning of monopodia had no
advantage and some time reduced the seed cotton yield. Hence retention of all monopodia would be an
ideal practice under rainfed conditions.
REFERENCES
[1] Brar, A.S., Singh, A. and Singh, T. (2000) - Response of hybrid cotton (Gossypium hirsutum) to nitrogen and canopy
modification practices. Indian J. Agron., 45: 395400.
[2] Brar, A.S., Singh, N. and Deol, J.S. (2002) - Influence of plant spacing and growth modification practices in yield and its
attributing characters of two cotton cultivars (G. hirsutum). J. Research, Punjab Agriculture University, 39 (2): 181183.
[3] Singh, Jai and Sandhu, B.S. (1996) - Effect of detopping in cotton on the efficacy of chemical control of boll worms.
Journal of Indian Society of Cotton Improvement, 21: 5657.
[4] Rao, D.V.M. and Lakshminarayan (1985) - Effect of physical and chemical growth regulation of plant growth on the
development and yield of two cotton hybrids. Cotton Development Journal 15: 5558.
[5] Venkatakrishnan, A.S. and Pothiraj, P. (1994)-Effect of nutrient management, foliar spray and detopping on yield attributes
and seed cotton yield. Madras Agriculture Journal, 81 (9): 519520.
Input use Efficiency, Productivity, Profitability

and Sustainability of Bt Cotton Based Multi Tier
System with Nutrient Levels
K. Sankaranarayanan
1
, P. Nalayini
2
, C.S. Praharaj
3
and N. Gopalakrishnan
4
1,2
Senior Scientists, Agronomy,

Central Institute for Cotton Research,
Regional Station, Coimbatore641003
3
Principal Scientist, Agronomy, Indian Institute of Pulses Research, Kanpur (UP)
4
Assistant Director General (Commercial Crops),
Indian Council of Agricultural Research, New Delhi

AbstractField studies were conducted to assess the performance of Bt cotton based vegetables multi tier systems
with different nutrient levels to intercrops during winter season (Aug-Feb) of 2007-08 and 2008-09 at Coimbatore,
Tamil Nadu. The treatment combinations comprised of three Bt cotton based multi tier systems (Bt cotton + coriander
+ vegetable cowpea + cluster bean) (in addition to recommended level of nutrient to Bt cotton) combined with 50, 75
and 100 % of recommended levels of nutrient to intercrops. Bt cotton + radish + beet root + coriander and.Bt cotton +
radish + cluster bean + beet root) were compared with sole Bt cotton. Pooled analysis of two years data revealed that
crop growth characteristics, yield attributes, yield and quality parameters of Bt cotton were not significantly affected
by multi-tier systems and nutrient levels. Seed cotton yield of ranged from 2565 to 2912 kg/ha, whereas sole cotton
yield was 2593 kg /ha. The significantly highest LAI of 2.21, partial factor productivity (24.4 kg/kg) and economics
nutrient use efficiency (1.80 kg /Rs) were achieved with multi tier system involving Bt cotton with coriander,
vegetable cowpea and cluster bean and application of 50 % of nutrient to intercrops. The results on light interception
(78.6%), weed smothering efficiency (43.8%), water use efficiency ( 90.8 kg/ha-cm), water productivity (Rs 20.9/M
3
),
relative production efficiency (135.5%), gross return (Rs1, 40,270/ha), seed cotton equivalent yield (61.0 q/ha), land
equivalent ratio (2.43), area time equivalent ratio (1.72), diversity index (3.03), uptake of nitrogen (183.4 kg/ha),
phosphorus (36.3 kg/ha) and soil available nitrogen (192.4 kg/ha) were significantly highest with multi tier system
involving Bt cotton with coriander, vegetable cowpea and cluster bean with 100 per cent nutrient to intercrops.
However, the system is labour intensive, resulted on enhanced cost of cultivation; hence multi tier planting of Bt
cotton + radish + beet root + coriander with application of 100 % nutrient to intercrops registered the significantly
highest net profit (Rs 93, 098/ha), benefit cost ratio (3.23), per day profitablity (620.7) and relative economic efficiency
(191%).
INTRODUCTION
In multiple cropping systems, use of resources like sunlight, nutrient and water is more efficient leading
to increased biological diversity and higher production stability. Monocropping is exception while
mixture (of species) is the rule of nature. Reviewing cropping system in South Asia, it was observed that
intercropping is a well established practice covering over 12 m ha (Woodhead et al., 1994). India ranks
first in world acreage (10.2 m ha) with almost 33 % of total cotton area. Since cotton is a crop of
relatively longer duration, its slow initial growth offers a vast scope for cultivation of suitable vegetable
intercrops. Our requirement on vegetables has increased to about 127.2 million tonnes to meet the
nutritional requirement of an estimated 1200 million people expected by 2020-21. Although, the
productivity levels of vegetables increased manifolds, it is not sufficient to feed ever increasing
population as a result of increased demands. This will complicate the issue of price rise further leading to
increased costs of vegetables. Thus, keeping in view of climatic vulnerability, market fluctuation of
vegetable crops and better resources use, Bt cotton is chosen as a candidate crop and Bt cotton based
system with multiple vegetable intercrops is aimed. The multi tier systems includes different proportion
of short duration vegetables, make intensive cropping, hence recommended nutrient levels to sole Bt
cotton may not be sufficient to multi tier cropping. Thus, additional nutrition to intercrops is needed to
make multi tier systems sustainable.
59
Input use Efficiency, Productivity, Profitability and Sustainability of Bt Cotton Based Multi Tier System 351
A field trial was conducted during winter season (August to February) of 2007-08 and 2008-09 at Central
Institute for Cotton Research, Regional Station, Coimbatore. The experimental soil was clay loam in
texture, low in available N (214 kg/ha), medium in available P (16.8 kg P
2
O
5
ha
-1
) and high in
exchangeable K (812 kg K
2
O ha
-1
) with a pH of 8.5. The experiment was laid out in a randomized block
design with three replications. The treatment combinations comprised of three Bt cotton based multi tier
systems (with recommended level of nutrient to Bt cotton combined with 50, 75 and 100 % of
recommended nutrient levels to intercrops. The three multi-tier system were S1.Bt cotton + coriander +
vegetable cowpea + cluster bean (nutrient levels to intercrop, 11.0:21.8:6.4, 16.4:32.7:9.5 and
21.9:43.6:12.6 kg N, P
2
O
5
& K
2
O/ha respectively for 50, 75 and 100%), S
2
. Bt cotton + radish + beet root
+ coriander (nutrient levels to intercrop, 10.7:29.6:17.2, 16.0:44.3:25.8 and 21.3:59.1:34.3kg N, P
2
O
5
&
K
2
O/ha respectively) and S
3.
Bt cotton + radish + cluster bean + beet root (nutrient levels to intercrop,
13.7:35.5:20.2, 20.5:53.3:30.2 and 27.3:71.0:40.3 kg N, P
2
O
5
& K
2
O/ha, respectively) were compared
with sole Bt cotton. Bt cotton (Gossypium hirsutum L.) hybrid RCH20 Bt, coriander (Coriandrum
sativum L.) cultivar SURABHI, radish (Raphenus sativus L.) cultivar PUSA CHETKI, beet root (Beta
vulgaries L.) cultivar DDR, cluster bean (Cyamopsis teragonolaba L.) cultivar PUSA NAVBAHAR,
and vegetable cowpea (Vigna unguiculata L.) Walp) cultivar CO2 were included in the trial. The doses
of fertiliser level to inter crops were arrived by adjusting the recommended level of nutrients to plant
population in a particular intercropping system. N was applied in two equal splits, first at the time of
planting and second 40 DAS (days after sowing), while entire P and K were applied as basal at the time
of planting. Bt cotton equivalent yields were worked out by equating prices of component crops yield as
suggested by De et al. (1976). Benefit cost analysis was also made for different systems to select the
viable and remunerative combination.
The field was ploughed once with tractor drawn mould board plough and then harrowed twice. Bt
Hybrid RCH 20 was planted at 120 x 45 cm, where two ridges at 60 cm apart were formed, and various
intercrops (three crops) were planted on 4 sides of the 2 ridges in sequence. Radish, coriander and other
vegetables (beet root, cluster bean and vegetable cowpea) were planted at intra-row spacing of 10, 15 and
20 cm, respectively. Crop received a total rainfall of 438.2 and 436.8 mm during 2007-08 and 2008-09
where as effective rainfall was only 275.9 and 264.2 mm, respectively (measured by the FAO method
suggested by Brouwer and Heibloem (1986). In addition to effective rainfall, water need of the crop was
supplemented by irrigation. No additional irrigation was provided for the intercropping systems (similar
to sole Bt cotton crop). Bt cotton hybrid seeds treated with imidachloprid were taken up for planting.
Endosulfan (2.5 l/ha) was used to contain the menace of leaf eating caterpillar in radish at 25 and 20
DAS, profenophos (1.25 l/ha) at 62 and 68 DAS in Bt cotton and larvin (1 kg/ha) at 140 and 136 DAS
had been used for Bt cotton for control of sucking pest and boll worms, respectively, in 2007-08 and
2008-09. Growth characters, yield attributes, main crop (seed cotton) and intercrop yields were recorded
during the course of investigation. Area time equivalent ratio (ATER) was calculated from the land
equivalent ratio (LER) using the formula (Heibsch, 1980):
ATER = LER x Dc x Dt
-1
where, Dc is time taken by crop, Dt is time taken by whole system.
Fibre quality parameters (2.5% span length, maturity ratio, uniformity ratio, micronaire, fibre
strength and fibre elongation) were also analyzed. Fibre quality index (FQI= LT/M, where L, 2.5% span
length in mm, T, fibre bundle tenacity at 3.2 mm gauge in g tex
-1
and M, micronaire Value in in
-1
),
count (C = 0.196 x FQI 16) and count strength product (CSP=1.740 x FQI + 1600) were also worked
out. All the quality parameters were analyzed by using the High Volume Instrument (Statex-Fibrotex
model HVI).
Relative production efficiency (RPE) was calculated using the following formula:
RPE = (EYD-EYE)*100/EYE, Where EYD is the equivalent yield under improved/diversified
system, while EYE is the existing system yield. Relative Economic Efficiency (REE) is a comparative
measure of economic gains over the existing system. It is expressed in percentage (REE = (DNR-
ENR)*100 ENR
-1
), where DNR is the net return obtained under improved/diversified system, while ENR
is net return in the existing system. Pooled analysis of the two years data was done to obtain reliable
assessment of multi-tier intercropping. Gross return, net return and benefit: cost analysis was also derived
on the basis of prevailing market price of inputs and outputs.
Bt Cotton
Growth Characters and Yield
TABLE 1: GROWTH CHARACTERS, YIELD ATTRIBUTES AND SEED COTTON YIELD AS INFLUENCED BY MULTI TIER SYSTEMS AND NUTRIENT LEVELS
Treatments Plant
Height (cm)
No of
Squar
e
No.
Sympodia
No.
Monopodia
No. of
Nodes
LAI No of
Bursted
Bolls
Boll
Weight (g)
Seed Bt
Cotton
Yield
(kg/ha)
T
1
.50%RDF(IC).+C+
Co+V.C+C.b
107.3 7.9 20.1 2.4 23.4 2.52 27.7 5.3 2643
T
2
.75%RDF(IC)+C+
Co+V.C+C.b
108.3 3.8 20.8 2.5 24.3 2.35 29.1 5.3 2864
T
3
.100%RDF(IC)+C
+Co+V.C+C.b
117.3 3.5 21.7 2.5 25.4 2.59 31.6 4.9 2912
T
4
.50% RDF(IC).
+C+ R+ B+ Co
99.0 7.3 19.1 2.3 22.7 2.22 28.4 5.4 2781
T
5
.75% RDF(IC)+
C+ R+ B+ Co
104.6 4.6 19.4 2.3 23.2 2.27 29.1 5.2 2844
T
6
.100% RDF(IC)+
C+ R+ B+ Co
112.9 8.3 20.5 2.6 24.8 3.01 30.3 5.3 2896
T
7
.50% RDF(IC)+
C+ R+ C.b+ B
103.5 5.1 19.5 2.1 23.7 2.52 24.1 5.5 2565
T
8
.75% RDF(IC)+C
+R + C.b+ B
113.0 4.9 21.3 2.5 25.5 2.68 26.7 5.5 2735
T
9
.100% RDF(IC)+
C+ R+ C.b +B
113.7 4.0 19.4 2.3 23.2 2.56 30.1 5.3 2901
T
10
.Sole Bt cotton 106.6 9.9 20.5 2.6 23.7 2.85 27.6 5.2 2593
SEd 7.1 4.0 1.2 0.3 1.2 0.44 3.9 0.2 110.7
CD (5%) NS NS NS NS NS NS NS NS NS
Analysis of pooled data on biometric observations recorded at 45, 90 and 120 DAS and at harvest, yield
attributes and seed cotton yield (Table 1) showed that none of the attributes were significantly influenced
by the combination of multi-tier cropping systems and nutrient levels to intercrops. Seed cotton yield for
different systems ranged from 2565 to 2912 kg/ ha and sole cotton yield of 2593 kg/ ha was recorded.
Since the component vegetable inter-crops were harvested early (coriander within 30- 45 DAS, radish at
45 DAS, vegetable cowpea, cluster beans, and beet root within 75 DAS) and nutrient demand met by
fertilizer application to intercrops none of the above crops competed with the main crop of Bt cotton
during the growth and development. As a result, statistically similar growth characters, yield attributes
and seed cotton yield were recorded in Bt cotton with different inter/ sole crop systems. Thus, intensive
cropping systems through crop mixes with application of required nutrient levels to intercrops was
successful as the components in the system have different nutrient and moisture requirement, varied
feeding zones in the soil profile, differential growth duration for enabling the utilization of natural
resources optimally. It was also reported that seed cotton was not adversely influenced by intercropping
with cowpea and onion (Chowdhury and Singh, 1983).

Intercrops
Observation on growth characters of intercrop showed that the tallest plant (59.1 cm at 45 DAS) with
cluster bean and longest extractable tap root (21.2 cm) and highest root volume (385.2 cc) with radish at
45 DAS and greatest leaf area (5042 cm
2
at 45 DAS) with vegetable cowpea. Yield (Kg ha
-1
) of vegetable
intercrops (Table 2) varied for coriander (2465 to 3508), vegetable cowpea (1646 to 2019), cluster bean
(2504 to 4985), beet root (639 to 1471), radish (2846 to 4122). Pure crop yield (t/ha) of vegetables,
coriander, vegetable cowpea, cluster bean, beet root and radish were 12.2, 3.9, 6.5, 15.2 and 19.9
respectively. As a result, net return (x000 Rs/ ha) calculated from intercrops varied in coriander (26.65
to 39.33), vegetable cowpea (4.08 to 5.49), cluster bean (6.18 to 14.90), beet root (0.728 to 4.52), radish
(8.63 to 13.33). Similarly, B:C ratio varied for coriander (8.5 to 11.8), vegetable cowpea (1.6 ), cluster
bean (1.6 to 1.7), beet root (1.22 to 2.1), radish (3.1 to 3.6). Thus, the economics of intercrops alone
revealed that coriander harvested under Bt cotton + beet root + coriander + radish with 100 % RDF (T
6
)
registered the highest gross return (42, 973 Rs/ ha), net return (39, 328 Rs/ ha), benefit cost ratio (11.8)
and seed cotton equivalent yield (18.68 q/ha). While calculating cost of cultivation for intercrops, cost of
seeds, sowing charges, fertiliser and harvest charges of intercrops were taken into count, hence cost of
cultivation is less with intercrops, which favoured high net return, benefit cost ratio and seed cotton
equivalent yield.
TABLE 2: GROWTH CHARACTERS, YIELD AND ECONOMICS OF BT COTTON AND INTERCROPS OF MULTI TIER SYSTEM AT RECOMMENDED LEVEL OF NUTRIENT APPLICATION
Characters Plant
Height
at
45 DAS
(cm)
Root Length
(cm) at 45
DAS
Root Volume
(ml) at 45
DAS
LA at 45
DAS
Yield
(kg/ha)
Gross
Return
(Rs/ha)
CC
(Rs/ha)
Net
Return
(Rs/ha)
B/C
Ratio
SCEY
(kg/ha)
T3.Bt cotton 31.5 19.8 11.6 1375 2919 67143 28979 38164 2.3 2912
Coriander 19.6 7.8 23.2 2816.0 2761 33818 3645 30173 9.3 1470
V.cowpea 28.5 16.7 19.6 5012.0 2019 13930 8444 5486 1.6 606
Cluster bean 47.8 15.4 10.3 2064.0 3678 25380 15100 10279 1.7 1103
T6.Bt cotton 30.2 15.7 11.6 1386 2903 66778 28913 37865 2.3 2896
Beet root 25.1 7.7 75.6 812.0 1471 8825 4302 4523 2.1 384
Coriander 26.4 7.4 19.3 2743.0 3508 42973 3645 39328 11.8 1868
radish 19.8 13.6 310.3 1126.0 3619 16286 4903 11383 3.3 708
T9.Bt cotton 26.6 18.9 14.3 1315 2920 67164 29010 38154 2.3 2901
Beet root 23.1 6.3 74.6 1124.0 887 5323 3951 1372 1.3 231
Cluster bean 46.1 16.8 10.1 1854.0 4894 33770 19873 13897 1.7 1468
Radish 23.6 21.2 364.6 1324.0 4122 18551 5217 13334 3.6 807
T10.Bt
cotton
28.1 18.2 14.6 2816 2590 59580 27588 31992 2.2 2593
Price: cotton (Rs23.0/kg), radish (Rs 4.5/kg), vegetable cowpea (Rs 6.90 /kg), beet root (Rs 6.0 /kg), cluster bean (Rs6.90 /kg), coriander
(Rs 12.25/kg) (prevailing price of experimental periods)
Quality Parameters
The modification/ changes of management practices might have specific impact on Bt cotton quality
parameters. Pooled analysis indicated that none of the Bt cotton quality parameters were influenced by
intercropping system and nutrient levels to intercrops. Mean fibre quality values recorded under the inter-
crops varied for 2.5 % span length (30.9 to 33.3 mm), maturity ratio (0.80 to 0.85), uniformity ratio (47
to 52 %), micronaire (4.5 to 4.9 in
-1
), fibre strength (21.0 to 22.1 g/tex), FQI (299 to 341), count (43 to
51 %) and CSP (2120 to 2193). Fibre quality is genetically inherited and therefore the response was not
distinct (Bhuva et al., 1995). Similarly, Mohammad et al. (2001) noticed that fibre qualities indices such
as ginning percentage (GOT), fibre length, fibre fineness, maturity co-efficient and fibre strength were
not altered by intercropping.

Multi tier (Intercropping) system
The advantages of this are physically reflected from the differential growth characters. For example,
mean plant height, root length and root volume observed at 45 DAS were 21.2, 17.9 cm and 334.9 cc
with radish, 27.5, 15.6 cm and 23.5 cc with vegetable cowpea, 24.1, 6.9 cm and 76.6 cc with beet root,
45.0, 16.2 cm and 11.2 cc with cluster bean, 21.8, 7.2 cm and 21.0 cc with coriander and 31.7, 19.1 cm
and 13.0 cc with base crop Bt cotton. These variations resulted in formation of a multi-tier (Table 2). LAI
of the different system was also worked out at 45 DAS by including leaf area of base crop (Bt cotton) and
multi-tier forming intercrops (vegetables). LAI was significantly influenced by the multi-tier systems as
highest of LAI of 2.2 was with Bt cotton + coriander + v.cowpea + cluster bean with 50 % RDF (T
1
)
which in fact was 4.6 times than that in sole Bt cotton (0.48). Pooled mean data on per cent of light
interception showed that highest value of 78.6 was measured with Bt cotton + coriander + vegetable
cowpea + cluster bean with 100 % RDF.
Production Efficiency
TABLE 3: GROWTH CHARACTERS, WEED SMOTHERING, INPUT USE EFFICIENCY AND PRODUCTION EFFICIENCY OF MULTI TIER SYSTEMS
Treatments Root
Volume
(cc) at
45 DAS
LAI
at 45
DAS
Weed
dry
Weight
(kg/ha)
WSE
(%)
LI
(%)
WUE
(kg/ha-cm)
WPY
(Rs/M3)
RPE
(%)
Total
Nutrients
(kg/ha)
PFP
(kg/kg)
ENUE
(Kg/Rs)
T
1
.50%RDF(IC).+
C+Co+V.C+C.b
65.4 2.21 482 41.6 74.0 77.2 17.8 100.3 219.1 23.7 1.77
T
2
.75%RDF(IC)+C
+Co+V.C+C.b
74.0 2.08 502 39.2 76.8 86.7 19.9 124.8 238.6 24.4 1.80
T
3
.100%RDF(IC)+
C+Co+V.C+C.b
64.7 2.09 464 43.8 78.6 90.8 20.9 135.5 258.1 23.6 1.72
T
4
.50% RDF(IC).
+C+ R+ B+ Co
405.9 1.28 615 25.5 58.6 75.8 17.4 96.6 237.4 21.4 1.60
T
5
.75% RDF(IC)+
C+ R+ B+ Co
395.5 1.20 586 29.1 62.5 82.1 18.9 113.0 266.1 20.7 1.53
T
6
.100% RDF(IC)+
C+ R+ B+ Co
416.8 1.12 562 32.0 63.6 87.3 20.1 126.4 294.7 19.9 1.45
T
7
.50% RDF(IC)+
C+ R+ C.b+ B
491.8 1.17 582 29.5 64.2 69.1 15.9 79.2 249.3 18.6 1.38
T
8
.75%
RDF(IC)+C +R +
C.b+ B
470.0 1.08 526 36.3 66.3 78.7 18.1 104.2 284.0 18.6 1.37
T
9
.100% RDF(IC)+
C+ R+ C.b +B
463.6 1.04 512 38.0 68.1 80.8 18.6 109.5 318.6 17.0 1.24
T
10
.Sole Bt cotton 14.6 0.48 826 0.0 38.7 38.5 8.9 0.0 180.0 14.4 1.12
SEd 16.2 0.10 58.3 7.6 1.5 6.1 1.1 0.2
CD(5%) 32.7 0.20 118 15.4 3.1 12.3 2.3 0.40
Various intercropping with nutrient levels efficiency parameters measured through relative production
(RPE) and economic efficiency (REE), land equivalent ratio (LER), diversity index (DI) and area time
equivalent ratio (ATER) were also significantly improved with multi-tier intercropping system in
comparison to sole Bt cotton. The enhanced efficiency for multi-tier system varied for relative production
efficiency (79.2 to 135.5 percent), relative economic efficiency (69.3 to 191.0), land equivalent ratio
(1.51 to 2.43), diversity index (2.46 to 3.03) and area time equivalent ratio (1.20 to 1.72). Amongst the
intercropping systems evaluated, multi tier system applied 100 % RDF to Bt cotton + coriander +
vegetable cowpea + cluster bean (T
3
) was having the highest RPE, LER, ATER and DI of 135.5%, 2.43,
1.72 and 3.03 respectively (Table 3). Thus, the increased system efficiency was due to additional yield
and return realised from intercrops by application of 100% RDF to intercrops. It was also reported that
effective land use efficiency was reflected by higher land equivalent ratio and was higher with
intercropping systems (Chelliah and Gopalaswamy, 2000). Diversity index (DI) indicated the system was
more diversified and sustainable one. Many times, sustaining yield from a farming system in totality may
be of prime consideration for farmers under resource scarce condition than maximizing yield or income
from a single crop. The most interesting biological and economic aspect of multi-tier intercropping is the
potential for compensation among the components of the system, often referred to as biological or
economic buffering in the system that leads to greater stability in (total) yields of component crops.
ATER provides more realistic comparison of the yield advantage of intercropping over monocropping
since it takes into account both area and time taken by the component crops in an intercropping.
Input Use Efficiency
Water
Being a costly and scarce resource, irrigation water and its availability for agriculture is expected to go
down further due to increased domestic and industrial demands. Water use efficiency (WUE, kg ha
-1
cm
-1
)
and water productivity (WPY, Rs m
-3
) and weed smothering efficiency (WSE %) were influenced
significantly by multi-tier cropping systems. The significantly highest water use efficiency of 90.8 kg ha
-
1
cm
-1
, water productivity of Rs 20.9 m
-3
of water and 43.8 % weed smothering efficiency were calculated
for 100 % RDF with Bt cotton + coriander + vegetable cowpea + cluster bean (T
3
) (Table 4). Sole Bt
cotton system produced only 38.5 kg by using one hectare centimetre of water while gross return of Rs
8.9 per hectare was realized for m
3
of water (1000 litres) used. Thus, the most efficient cropping pattern
is one which is capable of giving maximum return per unit quantity of water on a long term sustainable
basis (Jain, 2008).
TABLE 4: AVAILABLE MAJOR NUTRIENTS STATUS OF POST HARVEST SOIL AND NUTRIENT UPTAKE AS INFLUENCED BY MULTI TIER SYSTEMS AND NUTRIENT LEVELS
Treatments Nutrient Uptake (kg/ha) Available Nutrients (kg/ha)
N P K N P
2
O
5
K
2
O
T
1
.50%RDF(IC).+C+Co+V.C+C.b 156.7 30.7 156.1 187.4 17.1 728.4
T
2
.75%RDF(IC)+C+Co+V.C+C.b 178.0 35.3 181.0 184.8 17.5 725.6
T
3
.100%RDF(IC)+C+Co+V.C+C.b 183.4 36.3 187.2 192.4 16.9 720.8
T
4
.50% RDF(IC). +C+ R+ B+ Co 125.3 23.2 114.8 167.3 19.3 684.3
T
5
.75% RDF(IC)+ C+ R+ B+ Co 131.5 24.2 122.2 162.8 18.6 652.8
T
6
.100% RDF(IC)+ C+ R+ B+ Co 136.5 25.0 127.3 168.5 20.4 675.6
T
7
.50% RDF(IC)+ C+ R+ C.b+ B 141.8 29.0 148.5 176.2 17.4 712.6
T
8
.75% RDF(IC)+C +R + C.b+ B 158.1 32.3 168.7 178.4 17 714.6
T
9
.100% RDF(IC)+ C+ R+ C.b +B 163.0 33.3 172.5 172.9 17.9 736.8
T
10
.Sole Bt cotton 101.1 19.1 83.1 182.3 17.9 706.3
SEd 7.5 4.1 8.1 7.1 1.8 18.2
CD(5%) 15.2 8.2 16.4 14.3 NS 36.8
Nutrients
Nutrient use efficiency of multitier system and nutrient application was assessed by calculating the partial
factor productivity (kg ha
-1
) and economics of nutrient use efficiency (kg Rs
-1
) and found that both the
indices were significantly influenced by multi-tier cropping systems. Partial factor productivity (kg/ha) of
multi- tier cropping systems with nutrient levels were higher and the degree of advantage varied from
118.0 to 169.4%. Multitier intercropping systems with levels of nutrients was increased the economics of
nutrient use efficiency by 110.7 to 160.7% due to effective utilisation of resources by the components in
the mixture which in turn produced higher yield and better economic nutrient use efficiency. Multi-tier
intercropping of Bt cotton + coriander + vegetable cowpea + cluster bean with 75 % RDF (T
2
) also gave
the highest partial factor productivity (PFP) of nutrient (24.4 kg per kg of nutrients) and economics of
nutrient use efficiency of 1.80 kg seed Bt cotton per rupees invested on fertiliser were arrived (Table 4).

Light
Unlike rainfall and nutrients, use of solar energy is limited to be captured and stored for latter use in the
way that other natural resources are managed as light is instantaneously available and needs to be
instantaneously intercepted and used. In the current investigation, the highest percentage of light
interception was observed at 45 DAS by 100 % RDF with multi-tier system of Bt cotton intercropped
with coriander, v.cowpea and cluster bean (T
3
, 78.6 %), and was followed by that 75 % RDF with
coriander, vegetable cowpea and cluster bean (T
2
, 76.8 %) (Table 3). High foliage producing capacity of
coriander, vegetable cowpea and cluster bean resulted in high light interception in the above systems.
Under multiple cropping situations, the component crops were grown in such a way that competition for
light was minimized and total interception increased. Since the least light interception (38.7) was
observed with sole Bt cotton, the efficiency was not much higher in sole Bt cotton mainly because of
slow ground coverage by Bt cotton foliage.
Labour
Labour requirement of the multi-tier cropping with nutrient levels ranged from 329 to 543 man days in
comparison to sole Bt cotton with 268 man days. Labour intensiveness associated with multi-tier systems
was analysed and it was observed that labour use efficiency in term of gross return per labour was higher
with systems. Amongst intercropping systems, the highest labour use efficiency of Rs 387.8 per labour
(of 8 hours) was arrived at with Bt cotton + radish + beet root + coriander with 100 % RDF (T
6
) and sole
Bt cotton system had value of Rs 222.4/ labour.
NUTRIENT AVAILABLE AND UPTAKE
Multi-tier intercropping system is highly intensive in nature and their impact on fertility of the soil needs
to be assessed. In the present study, nutrient available in post harvest soil and uptake of nutrients by
different systems varied significantly except for soil available P (Table 4). Distinctive feature of pulses is
the capability to fix atmospheric N
2
biologically by prime modulators. Significantly higher available N
(192.4 kg /ha) in 100 % RDF with multi-tier system of Bt cotton intercropped with coriander, vegetable
cowpea and cluster bean (Table 4) was recorded. The treatment combination (T
3
) had legumes (vegetable
cowpea and cluster bean) in addition 100% RDF was applied to intercrops that resulted in native soil
enrichment. Thus, growing of legumes, as an intercrop was beneficial to soil health and soil fertility
(Basu, 1992). Significantly least soil available N (162.8 kg/ha) and K (652.8 kg/ha) were estimated with
the combination consisting of multi-tier one viz., Bt cotton + radish + beet root + coriander with 75 %
RDF (T
5
) which might be due to non legume intercrops and more uptake of nutrients, might have reduced
soil available nitrogen considerably.
NPK uptake of multi-tier system with different levels of nutrient application in the present trial
ranged from 125.3 to183.4, 23.2 to 36.3 and 114.8 to 187.2 kg per hectare for N, P and K respectively.
Results revealed that 100 % RDF with multi-tier system of coriander, vegetable cowpea and cluster bean
(T
3
) had significantly higher NPK than sole Bt cotton. The ability of intercropping systems to make more
efficient use than sole crops was evident both for soluble and non-soluble nutrients. Because of different
root growth pattern of component species, intercropping also explore entire soil mass in the rooting zone.
This was also ascribed to the more availability and absorption of nutrient under intercropping situation.
Similarly, maximum N, P and K uptake were under cotton + blackgram than sole cotton (Harisudan,
2004).
Economics
The multi-tier system with application of nutrient to intercrops significantly increased gross return, net
return, BCR, per day profitability and seed cotton equivalent yield. This enhanced the gross return in the
range of 179.1 to 235.4%, net return by 169.3 to 291.0%, BCR by 94.0 to 149.5%, and per day
profitability by 169.0 to 291.0% and seed cotton equivalent yield by 179.1 to 235.4% in comparison to
sole Bt cotton. Highest gross return and seed cotton equivalent yield were obtained with multi-tier system
of coriander, vegetable cowpea and cluster bean and 100% RDF (T
3
, Table 5). However, the system is
labour intensive, resulted in enhanced cost of cultivation; hence multi tier planting of Bt cotton + radish +
beet root + coriander with application of 100 % nutrient to intercrops registered the significantly highest
net profit (Rs 93, 098/ha), benefit cost ratio (3.23), per day profitablity (620.7) and relative economic
efficiency (191%). Intensification of crop on time and space dimension in the system (T
6
) (by selecting
short duration, non competitive crops), application of 100 % of recommended levels of nutrient to
intercrops (in addition to main crop) and method of planting adopted could not suppress growth of the
base crop and produce statistically as much as equal seed cotton yield (2896 kg/ ha) as that of sole crop in
addition to supplementing it by vegetable yield. On the similar lines, cropping system did not influence
seed cotton yield but additional yield of intercrops makes a system more remunerative than sole cotton
(Sepat and Ahlawat, 2010).
TABLE 5: ECONOMICS AND EFFICIENCY OF LAND, LABOUR, TIME AND DIVERSITY AS INFLUENCED BY MULTI TIER SYSTEMS AND NUTRIENT LEVELS
Treatments Gross
Return
(Rs/ha)
Net
Return
(Rs/ha)
B/Cratio SCEY
(q/ha)
Labour
(Man
Days/ha)
Per Day
Profitablity
REE
(%)
LER ATER DI
T
1
.50%RDF(IC).+C+
Co+V.C+C.b
119334 69680 2.40 51.9 445 464.5 117.8 2.03 1.47 2.85
T
2
.75%RDF(IC)+C+
Co+V.C+C.b
133931 79280 2.45 58.2 506 528.5 147.8 2.34 1.67 2.98
T
3
.100%RDF(IC)+C+
Co+V.C+C.b
140270 84102 2.50 61.0 520 560.7 162.9 2.43 1.72 3.03
T
4
.50% RDF(IC).
+C+ R+ B+ Co
117115 77723 2.97 50.9 329 518.2 142.9 1.51 1.20 2.48
T
5
.75% RDF(IC)+ C+
R+ B+ Co
126863 86154 3.12 55.2 341 574.4 169.3 1.61 1.25 2.65
T
6
.100% RDF(IC)+
C+ R+ B+ Co
134861 93098 3.23 58.6 348 620.7 191.0 1.68 1.28 2.74
T
7
.50% RDF(IC)+ C+
R+ C.b+ B
106725 54153 2.03 46.4 485 361.0 69.3 1.85 1.39 2.46
T
8
.75% RDF(IC)+C
+R + C.b+ B
121661 65136 2.15 52.9 530 434.2 103.6 2.09 1.53 2.67
T
9
.100% RDF(IC)+
C+ R+ C.b +B
124808 66757 2.15 54.3 543 445.0 108.7 2.14 1.59 2.59
T
10
.Sole Bt cotton 59580 31992 2.16 25.9 268 213.3 0.0 1.00 1.00 1.00
SEd 8258 6729 0.10 3.31 46.4 14.10 0.19 0.16 0.25
CD(5%) 16718 13622 0.20 6.7 94.0 28.6 0.38 0.32 0.50
SCEY-Seed cotton equivalent yield, LER- land equivalent ratio, ATER- Area time equivalent ratio, DI- Diversity index
Multitier cropping systems are dynamic interactive practices aimed at better use of the production
components such as soil, water, air space, solar radiation and all other inputs on sustainable basis. Bt
cotton + radish + beet root + coriander with application of 100 % nutrient to intercrops registered the
significantly highest net profit, benefit cost ratio, per day profitability and relative economic efficiency.
Thus, higher yield and profit could be realized with the introduction of multi-tier cropping in a unique
tier-arrangement in Bt cotton hybrids with application of recommended level of nutrient to intercrops in
addition to cotton nutrient level under irrigated condition, the study suggested.
REFERENCES
[1] Basu, A.K. (1992)-Integrated nutrient supply system in cotton based cropping system. Fert. News 37, 4754.
[2] Bhuva, K.S., Sukhadia N.M., Malavia D.P. (1995) - Intercropping in upland cotton (Gossypium hirsutum) with pulse and
oil seed crops under rainfed conditions. Indian Journal of Agronomy 40, 9597.
[3] Brouwer, C., Heibloem M. (1986) - Irrigation water needs, Irrigation water management training manual No.3, FAO,
Rome, Italy.
[4] Chelliah, N., Gopalaswamy N. (2000) - Effect of intercropping and foliar nutrition on the productivity of summer irrigated
cotton. Madras Agriculture Journal 87, 267270.
[5] Chowdhury, R.K., Singh BP.(1983) - Intercropping of pulses A good bet. Field Crop Abstracts 30, 900.
[6] Harisudan, C. (2004) - Nutrient management in cotton + Black gram intercropping system. M. Sc thesis, Tamil Nadu
Agricultural University, Coimbatore, India.
[7] Heibsch, CK. (1980) - Effect of N fertilization and crop duration on equivalency ratios in intercrops versus monoculture
comparisons. Ph.D Thesis, North Carolina State University, Raleigh, N.C., USA.
[8] Jain, T C. (2008) - New Paradigm in Agronomic Research and Development. Indian Journal of Agronomy 53: 241244.
[9] Mohammad Bismillah Khan, Mahboob Akhtar, Abdul Khaliq. (2001) - Effect of planting patterns and different
intercropping systems on the productivity of cotton (Gossypium hirsutum L.) under irrigated conditions of Faisalabad.
International Journal of Agriculture & Biology 3: 432435.
[10] Seema Sepat, Ahlawat I.P.S. (2010) - Effect of cropping system, phosphorus sources and level on phosphorus uptake and
yield of cotton (Gossypium hirsutum). Extended summary In: XIX national symposium on Resource Management
approaches Towards Livelihood Security24 Dec. 2010 Bengaluru, Karnataka, organised by Indian Society of Agronomy,
Indian Council of Agricultural Research and University of Agricultural Sciences.
[11] Woodhead, T., Huke, R., Huke E. (1994) - Areas, location and on-going collaborative research for the rice-wheat system in
Asia, Bangkok, Thailand. FAO Bullet. 6897.
Effects of Prolonged and Integrated Use

of Organics and Inorganics
on the Performance of Cotton
S.N. Upperi
1
and V.B. Kuligoud
2

1
Extension Leader, College of Agriculture, Bheemaraynagudi, UAS, Raichur;
2
Associate Professor, College of Agriculture, Bheemaraynagudi, UAS, Dharwad
AbstractExperiment was conducted to evaluate the prolonged and integrated use of organic and inorganic nutrient
sources on the performance of hybrid cotton and leaf reddening during growing seasons of 1999, 2000 and 2001 at
ARS, Bheemarayanagudi, Karnataka. The experiment was laid out in Split plot design with four organics viz.,
incorporation of cotton stalk @ 5 t /ha, vermicompost @ 2.5 t/ ha and FYM @ 10 t /ha, along with graded levels of in
organics application viz. 50 percent RDF ( 75:37.5:37.5 kg NPK/ha), 50 percent RDF + MgSO4 @ 20 kg/ ha and 100
percent RDF + MgSO4 @ 20 kg/ ha. The anthocyanin content ( mg/ g of FW) of the leaf samples were decreased from
0.27 with 50 per cent RDF to 0.06 with 100 per cent RDF + MgSO4 @ 20 kg/ ha) thereby revealed decrease in leaf
reddening with increased nutrition and supply of Mg. This improvement in leaf greenness enhanced cotton yield
significantly from supply of 50 % RDF (25.66 q /ha ) to the application of 50 percent RDF + MgSO4 @ 20 kg/ha (
28.41 q/ ha) and 100 per cent RDF + MgSO4 @ 20 kg/ha ( 32.17 q/ ha). Results also revealed significant increase in
organic carbon contents (from 0.42 to 0.63), available P
2
O
5
( 24.39 and 28.50 kg/ ha) and available potassium (from
522.22 to 561.889 kg /ha) with the application of vermicompost @ 2.5 t/ ha.
Keywords: Cotton, Anthocyanin, organics, kapas yield and available nutrients.
INTRODUCTION
Cotton an economic fiber crop of India has low average yields (526 kg/ha) compared to the world
average (761 kg/ha) and that of Australia (1991 kg/ha). Lower yields of India, are attributed to cultivation
of cotton under vagaries of monsoon, soil fertility stress and management factors. Supplying optimal
quantities of mineral nutrients of macro and micronutrients to growing crops is one way to improve crop
yields (Zubillage et al., 2002). The concentration of mineral nutrients in the soil solution varies over a
wide range, depending on many factors such as pH, soil organic matter and fertilizer application
(Marschner, 1986). Depleted soil nutrient status under intensive cultivation, will not sustain the
productivity in the long run, unless they are replenished timely with both organics and inorganics. India
presently uses 15 million tons of nutrients in the form of chemical fertilizers. Supplying the same through
organic sources alone is impossible. The chemical fertilizers should be used judiciously and synchronized
the demand of the crop uptake by supplying right doses of inorganics with organics (Bekunda et al.,
1997). Green manures and FYM applied along with a reduced rate of NPK were able to reduce the
mineral fertilizers used by 50%. When cotton stalks are chopped and incorporated into the soil, 48% of
the N, 41% of the phosphorus and 74% of the potassium taken from the soil by the cotton plant was
returned to the soil. Vermicomposts are finely divided peat-like materials with high porosity, aeration,
drainage, water-holding capacity (Edwards and Burrows, 1988) and contained more mineral elements than
commercial plant growth media, and many of these elements were changed to forms that could be readily
taken up by the plants, such as nitrates, available phosphorus, and soluble potassium, calcium, and
magnesium.
With these views, an experiment was conducted to study the impact of different organic sources and
graded levels of fertilisers on the productivity of irrigated cotton.
60
A field experiment was conducted at ARS Bheemarayanagudi, (Karnataka) during 1999, 2000 and 2001
in a split plot design replicated thrice. The main plots comprised cotton stalk (5 t/ha), vermicompost (10
t/ha) and FYM (10 t/ha) along with a control. Sub-plots were 50%, 50% RDF + MgSO
4.

Chopped small pieces of cotton stalks, vermicompost and FYM were weighed and incorporated into
the soil, 15 days before sowing. Imidacloprid treated seeds were dibbled on the ridges of the furrow at a
spacing of 30 x 20 cm. Irrigation were given as and when crop demanded usually once in 25 days. The
recommended dose of fertilizers were applied around the plants and irrigated. Hand weeding and cultural
operations like spraying were carried out as and when needed. The leaf samples were collected at 60 days
for chlorophyll and at 90 days for anthocyanin estimation. Half of the N and full dose of P fertilizers
were applied at the time of sowing and the rest N and K at 30 and 60 days after sowing. Seed cotton was
picked treatment wise and yield was recorded.
Soil Analysis
Soil samples collected before and after the experiment were processed and analysed for pH, EC, organic
carbon, N, P, K, S and micronutrients status. The initial soil samples and the FYM nutrient content is
presented in Table 1. Available N was determined by alkaline potassium permanganate distillation
method as described by Subbaiah and Asija (1959). Soil pH, electrical conductivity, available phosphorus
content of the soil was determined by Olsens method, available potassium was determined by Flame
photometer, after extracting the soil with neutral normal ammonium acetate, Calcium and Magnesium by
EDTA method, available sulphur was determined by turbidometric method using 0.15% CaCl
2
as an
extractant, as outlined by Jackson (1973). Available micronutrients were determined by atomic
absorption spectrophotometer after DTPA (diethylene triamine penta acetic acid) extraction as proposed
by Lindsay and Norvell (1978).
Chlorophyll and Anthocyanin OD values were determined by the procedure outlined by
spectrophotometer at 663 and 525 nm respectively.
TABLE 1: ANALYTICAL RESULTS OF SOIL AND FYM
Parameters Soil FYM
1. pH 8.54 7.10
2.Electrical Conductivity 0.19 dSm
-1
0.58
3. Exchangeable Ca+Mg 56 (C mol (p
+
)/kg) -
4. Available K
2
O 540 kg/ha 0.64 %
5. Available P
2
O
5
36.2 kg/ha 0.40%
6. Available N 240 kg/ha 0.90 %
7. Available sulfur 48 kg/ha 0.32 %
DTPA Extractable Micronutrients (ppm)
Zn 0.6 101
Cu 0.36 82
Mn 2.98 152
Fe 5.9 266
The pooled data in Table 2 indicated that the application of organics and graded level of inorganic
fertilizers increased the yields significantly over the years. The highest significant mean yields of 3114
and 3043 kg/ha, recorded with the application of FYM and vermicompost respectively over control
(2497) and cotton stalk (2714 kg/ha), attributed to the combined effect of both organics and inorganics in
supplement of nutrients, plant growth regulators and increased microbial activities associated. Similar
results were reported by Nenthran et al. (1999) and he was of the opinion that the hormones and growth
regulators of the vermicompost higher the yield. Where has the application of 100% RDF + 20 kg
MgSO
4
recorded significantly increased yields of cotton, over only 50% RDF and 50% RDF + 20 kg
Effects of Prolonged and Integrated Use of Organics and Inorganics 361
MgSO
4.
This may be due to the favorable effect of MgSO
4
for chlorophyll synthesis and ATP formation.
The increased yield of 50% RDF + 20 kg MgSO
4
over only 50% RDF was attributed to the nutrients
released from the cotton stalks. Similar result were reported earlier (Blaise and Ravindran, 2003) wherein
they observed retention of cotton crop residues increased soil organic matter and also possible nutrient
recycling. The interaction of organics and in-organics was not found significant.
TABLE 2: YIELD OF COTTON (Q/HA) AS INFLUENCED BY THE APPLICATION OF THE ORGANICS AND INORGANICS
T
r
e
a
t
m
e
n
t
s

Ist Year IInd Year IIIrd Year Pooled analysis
5
0
%

R
D
F

5
0
%

R
D
F
+

2
0
k
g

M
g
S
O
4

1
0
0

%

R
D
F

+

2
0
k
g

M
g
S
O
4

M
e
a
n

5
0
%

R
D
F

5
0
%

R
D
F
+

2
0
k
g

M
g
S
O
4

1
0
0

%

R
D
F

+

2
0
k
g

M
g
S
O
4

M
e
a
n

5
0
%

R
D
F

5
0
%

R
D
F
+

2
0
k
g

M
g
S
O
4

1
0
0

%

R
D
F

+

2
0
k
g

M
g
S
O
4

M
e
a
n

5
0
%

R
D
F

5
0
%

R
D
F
+

2
0
k
g

M
g
S
O
4

1
0
0

%

R
D
F

+

2
0
k
g

M
g
S
O
4

M
e
a
n

I
n
o
r
g
a
n
i
c

O
r
g
a
n
i
c
s

Control 27.18 30.61 29.22 29.01 18.09 20.64 22.16 20.29 23.13 26.26 32.15 28.28 22.8 25.92 27.85 25.52
Cotton Stalks @
5t/ha
32.84 32.13 33.81 32.93 19.43 20.94 24.26 21.55 25.7 29.54 37.19 30.81 26 23.55 31.77 28.44
Vermi compost
@2.5t/ha
29.42 31.05 34.14 31.53 20.91 23.8 27.62 24.11 30.18 33.61 39.91 34.57 26.76 29.55 33.71 30.31
FYM @ 10t/ha 30.75 32.61 35.99 33.12 21.28 23.76 25.48 28.49 29.17 35.31 44.54 36.34 27.06 30.65 35.34 31.02
Mean 30.5 31.6 33.29 19.91 22.48 24.88 27.05 31.18 38.2 25.65 28.41 32.17
Comparison SEM CD (0.05) SEM CD (0.05) SEM CD (0.05) SEM CD (0.05)
Organics 1.33 4.219 0.417 1.022 1.68 4.11 0.73 2.52
In-organics 1.15 1.5 0.361 0.882 1.45 3.56 1.38 3.91
Interaction 2.31 3.02 0.72 NS 2.91 NS 1.29 NS
The results in the Table 3 indicated that the application of graded doses of RDF with MgSO
4

recorded lower significant anthocyanin (0.1 mg/g) content compared to 50% RDF. This may be due to
higher contents of N, P, and Mg. Similar results were reported by Chakravorty (1980). However,
significant differences were not observed with organics. This finding is in accordance with the work of
Dhopte and Lall (1987) and they indicated that healthy leaves of cotton contained 2% N, 1.7 P (ppm),
155 K (ppm), 30.4 Ca (me/l.), 18.3 Mg (me/l.) and higher micronutrients compared to partly red.
TABLE 3: EFFECT OF APPLICATION OF ORGANICS AND INORGANICS ON THE ANTHOCYANIN AND CHLOROPHYLL CONTENT OF THE COTTON LEAVES, AVERAGED OVER YEARS
Anthocyanin (mg/g FW) Chlorophyll (mg/g FW)
50%
RDF
50% RDF+ 20kg
MgSO
4

100 % RDF +
20kg MgSO
4

Mean 50% RDF 50% RDF+
20kg MgSO
4

100 % RDF +
20kg MgSO
4

Mean
0.24 0.17 0.11 0.18 3.49 3.55 4.01 3.70
0.27 0.15 0.09 0.17 3.48 3.50 3.94 3.64
0.26 0.17 0.10 0.18 3.27 3.46 4.36 3.70
0.22 0.12 0.06 0.13 4.29 3.85 5.16 4.44
0.25 0.17 0.10 0.16 3.68 3.60 4.37 3.87
CD 5 %
Organics 0.107 0.612
Inorganics 0.089 0.327
Interaction 0.179 0.653

Application of 100% RDF+20 kg MgSO
4
increased the chlorophyll content (4.37) significantly over
50% RDF (Table 3), which is due to increased chlorophyll content, where N and Mg being the
components of chlorophyll with higher contents of N, P, K and Mg as revealed by Dhopte and Lall
(1987). However the application of organics had no significant effect on chlorophyll content, which may
be due to availability of both macro and micro nutrients in the plant systems.
The plant nutrient contents mentioned in Table 4 indicated significant differences in N content of the
leaves with the treatments, which varied from 2.17 to 2.83%. This is due to synchronized nutrient uptake
due to solubalising effect of organics. Similar results were reported by Mahavishnan et al. (2005).
However the treatment differences were not observed in case of P

and K

content inspite of their higher
contents with graded doses of nutrients. The soil sample collected after harvest of the crop was analysed.
Organic carbon content varied from 0.48 to 0.63% and did not differ significantly among treatments.
However the available P
2
O
5
and K
2
O differed significantly due to organic sources. Application of FYM
recorded (28.5 kg/ha) and vermicompost (24.4 kg/ha) recorded significantly higher available P compared
to the cotton stalk (20.93 kg/ha) and control (15.45 kg/ha). Similarly singnificant available K contents
were recorded with vermicompost (561.9 kg/ha) followed by FYM (551.6 kg/ha) than control (522.2
kg/ha) and cotton stalk (543.9 kg/ha). Application of organics sustained the availability of P
2
O
5
and K
2
O
compared to the soil values before the experiment.
TABLE 4: EFFECT OF APPLICATION OF ORGANICS AND INORGANICS ON THE NUTRIENT STATUS OF COTTON LEAVES AT 90 DAS
Treatments Per cent Nitrogen Per cent Phosphorous Per cent Potassium
50%
RDF
50%
RDF+
20kg
MgSO
4

100 %
RDF +
20kg
MgSO
4

Mean 50%
RDF
50%
RDF+
20kg
Mgso
4

100 %
RDF +
20kg
MgSO
4

Mean 50%
RDF
50%
RDF+
20kg
MgSO
4

100 %
RDF +
20kg
MgSO
4

Mean
Control 2.17 2.32 2.49 2.33 0.22 0.32 0.19 0.25 1.32 1.16 1.67 1.37
Cotton stalk
@ 5t/ha
2.60 2.62 2.65 2.62 0.25 0.24 0.22 0.24 1.47 1.47 1.43 1.458
Vermi
compost
@2.5t/ha
2.74 2.73 2.71 2.76 0.19 0.26 0.17 0.21 1.24 1.39 1.23 1.26
FYM @
10t/ha
2.66 2.72 2.82 2.74 0.22 0.22 0.19 0.21 1.51 1.45 0.96 1.31
Mean 2.55 2.59 2.69 2.61 0.22 0.26 0.19 1.39 1.35 1.313
Comparison SEM CD 5 % CD 5 % CD 5 %
Organics 0.010 0.034 NS NS
In organics 0.010 0.030 NS NS
Interaction 0.019 0.057 NS NS
CONCLUSION
Application of organics not only reduced the cost of fertilizer (50% RDF) but also increased the nutrients
availability and sustained higher production levels.
REFERENCES
[1] Bekunda, M.A., Bationo A. and SSali H. (1997). Soil fertility management in Africa. Wisconsin, USA.
[2] Blaise, D. and Ravindran, C.D. (2003) Influence of tillage and residue management on growth and yield of cotton grown
on a Vertisol over 5 years in a semi-arid region of India. Soil Tillage Res., 70: 163173.
[3] Chakravorty, S.C. (1980). Chemical factors responsible for red leaf disease in G. hirutum cotton. Proc. 76
th
Indian, Sci.
cong. Part III PP 120 Ab no. 100 Agri.Sci. section.
[4] Dhopte, A.M. and. Lall, S.B. (1987). Relative efficacy of antitranspirant, growth regulators and mineral nutrients in control
of leaf reddening in hirsutum cotton under dryland conditions. Ann. Plant Physiology. 1(1): 5671.
[5] Edwards, C.A. and Barrows, I. (1988). The potential of earth worm compost and plant growth media. In earth worm in
Environmental and waste management Ed. C.A, SPB Academic public. b.v. The Netherland 211220.
[6] Jackson M.L. (1973). Soil chemical analysis. Prentice Hall of India, Pvt. Ltd., New Delhi, India.
[7] Lindsay, W.L. and Norvell, W.A. (1978). Development of a DTPA soil test for zinc, iron, manganese and copper. Soil. Sci.
Soc. America J., 42: 421428
Effects of Prolonged and Integrated Use of Organics and Inorganics 363
[8] Marschner H. (1986). Functions of mineral nutrients: macronutrients. In: Haynes RJ, editor. Mineral Nutrition of Higher
Plants. Academic Press, Orlando, FL., 195267.
[9] Mahavishnan, K., Mangal Prasad and Bhanu Rekha, K. (2005). Integrated nutrient management in cotton-sunflower
cropping system in the sandy loam soils of north India. Journal of Tropical Agriculture 43(12): 2932.
[10] Nenthran, N.N., Jayaprasad, K.V. and Kale R.D. (1999). China aster cultivation using vermicompost as organic
amendment. Crop Research Hissar. 17: 209215.
[11] Subbaiah B.V. And G.L. Asija. (1959). A rapid procedure for the estimation of the available nitrogen in soils. Curr. Sci.
25: 259260.
[12] Zubillaga MM, Aristi J.P, and Lavado R.S. (2002). Effect of phosphorus and nitrogen fertilization on sunflower (Helianthus
annus L) nitrogen uptake and yield. J Agron Crop Sci. 188: 26774.
Response of Cotton to Bio Boron

and its Use Efficiency in Vertic Ustropept Soil
of Tamil Nadu, India
P. Janaki
1
and S.

Meena
1
1
Department of Soil Science & Agricultural Chemistry, Tamil Nadu Agricultural University,
Coimbatore641003, Tamil Nadu, India,
E-mail: janakibalamurugan@rediffmail.com
AbstractA field investigation was carried out to study the effect of different boron (B) sources on yield and its use
efficiency by hybrid cotton on Vertic Ustropept of Tamil Nadu, India. The experiment with hybrid cotton (RCHB
708Bt XL) was conducted during the winter season (August 2007 to January 2008) Treatments included supply of B
through different organic manures, borax and control besides including the foliar spray of B. Results showed that the
supply of B through organic and inorganic sources produced significantly higher seed cotton yield (SCY) than B alone.
Manure application had a positive influence on through slow and continued supply of B along with other nutrients to cotton.
The agronomic efficiency was high for the organic B source treatments. This could be attributed to the higher absorption
of N from soil and translocation to the fruiting organs from vegetative part by the applied B.
Keywords: Bio Boron, cotton, B use Efficiency, seed cotton yield
INTRODUCTION
In India, 33 % soils are deficient in B (Singh, 2006) Cotton requires B to support the processes of growth
and development of cotton fibre in the boll (Stewart, 1986). Organic matter plays an important role in
controlling B concentration in the soil solution and that it has a prominent effect on reducing B uptake by
plants (Yermiyagu et al., 2001). Foliar applications sometimes needed due to heavy rain or the absence of
irrigation facilities besides the toxicity of B application caused by the narrow range of B requirement
between deficiency and sufficiency of B by the crops. Janaki et al. (2005) found that the application of
recommended dose of NPK plus Zn @ 25 kg ha
-1
and B foliar spray twice @ 0.3 kg ha
-1
during early and
peak boll formation stages is beneficial in increasing the seed cotton yield (SCY). Hence the supply of B
through organic sources is beneficial for agricultural crops besides improving the efficiency. Therefore, a
study was conducted to study the effect of different boron sources on yield and its use efficiency by
hybrid cotton in Vertic Ustropept soils of Tamil Nadu, India.
METHODOLOGY
The experiment with hybrid cotton (RCHB 708Bt XL) was conducted during the winter season (August
2007 to January 2008) in Salem district of Tamil Nadu, India. The soil of the experimental fields belongs
to Periyanaickenpalayam series with the available NPK status of 165:46:497 kg ha
-1
. The experimental
field was clay loam in texture with hot water soluble B content of 0.78 mg kg
-1
soil. The irrigation water
used for the experiment had EC of 3.09 dS m
-1
and B content 0.29 meq/lit.
The treatments compromised T
1
: Absolute control; T
2
:FYM 5 t ha
-1
; T
3
: Poultry manure 5 t ha
-1
; T
4
:
vermicompost 5 t ha
-1
; T
5
: cotton waste compost 5 t ha
-1
; T
6
: Ground nut husk compost 5 t ha
-1
; T
7
: T
2
+
borax @ 5 kg/ha as basal; T
8
: Borax @ 5 kg/ha as basal; T
9
: T
8
+ Foliar spray of B @ 0.3 % (2 times at
45 DAS and 75 DAS) and T
10
: Foliar spray of B @ 0.3 % (2 times at 45 DAS and 75 DAS) for cotton
with three replications in a randomized block design. The cotton hybrid was sown at a spacing of 0.90 m
between rows and 0.60 m between plants and the plot size is 20 m
2
. Nitrogen was applied in two splits,
half at 25 days after sowing (DAS) along with entire amounts of P and K. Nutrient fertilizers were spot
applied. The remaining N was applied 60 DAS. The growth and yield parameters were determined at boll
bursting stage and combined yield of all six pickings were recorded from each treatment plots.
61
Response of Cotton to Bio Boron and its Use Efficiency in Vertic Ustropept Soil of Tamil Nadu, India 365
RESULTS
Seed Cotton Yield
Boron application significantly enhanced the SCY than the control treatment (Table 1). The per cent
increase in yield due to the application of organic and inorganic B sources ranged from 5.8 to 16.5 and
21.9 to 27.8 respectively. Among organic sources, ground nut husk performed better than the other
sources. Application of 5 kg borax as basal plus two foliar sprays of B produced higher SCY among
inorganic sources. This showed a positive effect of B on influencing the SCY. Howard et al. (1998)
reported that the efficiency of foliar applied B is higher than soil application when deficient condition of
B is suspected. B applied through foliar spray enhanced the seed cotton yield by efficiently translocating
the sugars from leaves to fruiting organs and by utilizing applied fertilizer N. Similar findings were
reported by Anderson and Bosewell (1968). Supply of B through organic and inorganic sources produced
significantly higher SCY than B alone. This showed the positive influence of manure application on
increasing SCY probably due to slow and continued supply of B along with other nutrients to cotton.
Similar findings were reported by Padole et al. (1998).
TABLE 1: INFLUENCE OF B SOURCES ON YIELD, YIELD ATTRIBUTES AND QUALITY PARAMETERS OF HYBRID COTTON
Treatments Seed Cotton
Yield
(kg ha
-1
)
No. of
Bolls/
Plant
Plant Height
(cm)
Ginning
Percentage
(kg ha
-1
)
Lint
Index
Uniformity
Ratio (%)
T
1
2861 78 156 32.3 0.35 44
T
2
3027 79 160 38.3 0.37 46
T
3
3093 78 164 38.6 0.37 45
T
4
3032 86 164 36.2 0.36 46
T
5
3233 80 164 39.4 0.36 46
T
6
3333 84 160 38.9 0.37 45
T
7
3509 95 162 39.9 0.38 47
T
8
3488 91 165 40.4 0.38 47
T
9
3655 98 163 38.7 0.39 47
T
10
3613 96 160 39.3 0.38 47
Mean 3284 87 161.8 38.2 0.37 46
CD (5%) 385** 6.2* NS 2.9* 0.02* NS
In cotton production, number of bolls per plant and its weight assume practical significance as they
are directly related to productivity. In this study, boll weight was increased with the application of B.
Basal borax plus foliar spray produced higher boll biomass per plant and SCY and was similar with the
organic B sources treatments.
Seed Lint Characters
Ginning percentage is a measure of lint yield from seed cotton. All the treatments except control
performed well in increasing the ginning percentage. Roberts et al. (2000) reported that the application of
B (0.22 kg ha
-1
) as foliar spray increased the lint yield by 8 %. Lint index is the weight of lint obtained
from 100 g of seed cotton and similar results as that of ginning percentage was observed.
B concentration of Index Leaf
The B content was high in the soil application of borax plus B foliar spray at squaring and boll formation
stages. This is in accordance with the earlier findings of Heitholt (1994) who reported the beneficial
effect of B spray on leaf blade B concentration. B concentration in index leaf increased from squaring to
boll formation stage suggesting high requirement at boll development phase to produce quality fibre.
Significant relationship of B content with N content was observed in this study (Fig. 1). The
relationship was positive between B and N at squaring and boll formation stages whereas the relationship
of B with P and B with K was significant only at squaring and boll formation stages, respectively. The
366
increase i
translocat
B Uptake
Treat
T
T
T
T
T
T
T
T
T
T
Me
CD
Accumula
was signif
which rec
in B concen
tion and utiliz
Fig. 1: R
tments
T
1

T
2

T
3

T
4

T
5

T
6

T
7

T
8

T
9

T
10

ean
(5%)
ation of B by
ficantly influ
ceived soil ap
World Cot
ntration of i
zation of N t
Relationship betwe
TABLE 2: INFLUENC
Leaf Stem
23.2 78.6
30.4 82.1
27.4 82.2
29.0 79.5
31.8 81.8
33.7 83.0
34.7 93.8
35.7 97.5
36.3 94.4
34.1 93.3
31.6 86.6
NS NS
y different pa
uenced by the
pplication of
tton Research C
index leaf d
towards SCY
en the B Content a
CE OF B SOURCES ON B
m Root
6 4.5
1 4.7
2 4.4
5 4.8
8 5.0
0 4.8
8 5.8
5 5.3
4 5.5
3 4.6
6 4.9
S NS
arts of the co
e treatments.
f Borax plus
Conference on T
decreased th
Y, B (Miley e
and Macro Nutrien
B UPTAKE BY DIFFEREN
B Uptake
Seed
95.8
95.2
98.9
99.0
101.6
105.6
135.5
131.1
155.3
131.0
114.9
50.6*
otton is prese
. Uptake of B
foliar spray
Technologies fo
he N concen
et al. 1969).
nts Content in Inde
NT PARTS OF HYBRID C
at Harvest (g
Lint bu
68.5 9.
92.5 12
98.4 13
95.0 16
110.4 16
110.9 15
125.5 21
124.3 23
127.2 24
125.0 22
107.8 17
30.5* 6.9
ented in Tab
B in the seed
of B twice (
or Prosperity
ntration and
ex Leaf of Bt Hybri
COTTON AT HARVEST
ha
-1
)
ur Seed
.4 1
2.2 1
3.0 1
6.0 1
6.2 2
5.8 2
1.4 2
3.1 2
4.3 2
2.1 2
7.3 2
9* 50
le 2. B uptak
d cotton was
(Table 2). In
is due to t

d Cotton
d cotton
64.4
87.7
97.3
94.0
11.9
16.6
61.0
55.4
82.5
55.9
22.7
0.6**
ke in seed, li
higher in the
crease in B a
the higher
Total
280.0
317.1
324.3
323.3
346.8
353.9
416.7
417.0
443.0
410.1
363.2
70.4**
int and bur
e treatment
addition to

the soil in
plus folia
inorganic
Among th
accumulat
BORON USE
Agronomi
per unit o
efficiency
the total b
total biom
The agron
the higher
applied B
(1969). T
recovery a
amount of
Agronomi
indicates t
requiring
Soil Availab
The soil a
(Table 3)
stage. Thi
the retent
Positive e
balance sh
applicatio
improving
inorganic
Response of
ncreased B u
ar spray of
B sources a
he different p
ted low B. B
E EFFICIENCY
ic efficiency
f applied nu
y of nutrient
biomass prod
mass at harve
nomic efficie
r absorption
B. The influe
The fraction
and is calcul
f nutrient. Sim
ic efficiency
that the inclu
crops, not on
ble Boron Sta
available B
. The mean
is showed th
tion of added
effect of org
heet at harve
on to high B
g fibre qualit
B sources.
f Cotton to Bio B
uptake and 5
B twice trea
and 20.4 per
parts, the see
B accumulatio
AND RECOVER
y is usually r
trient. The u
applied is m
duced is a so
st. This para
ency was high
of N from s
ence of B o
of applied n
lated as the r
milar to AE,
Fig. 2: Effect of
y and appare
uding B rich
nly increased
atus
content at d
B content in
he release of
d B in soils
ganic B sour
est (Table 3
requiring cro
ty. This is a b
Boron and its U
58 % increas
atment than
r cent for or
ed and lint a
on was in the
RY
reported on t
ultimate aim
measured in t
ource for sin
ameter is deri
h for the orga
soil and trans
n nitrogen t
nutrient that
ratio of actu
the apparent
f B Sources on the
ent B recov
organic sour
d the SCY bu
different grow
n soil decrea
f B from ma
and may he
rces on main
). Hence, in
ops may elim
boon for orga
se Efficiency in
sed B uptake
to control.
rganic B sou
accumulated
e order of see
the basis of
of growing c
terms of SCY
nk (SCY), th
ived for boro
anic B source
slocation to
translocation
t is absorbed
al uptake mi
B recovery w
e B use Efficiency a
ery was hig
rces in the m
ut also impro
wth stages w
ased from fl
anure by its d
elp in reduc
ntaining the
cluding B ri
minate B defi
anic farming
Vertic Ustropep
e was record
An average
urces than c
higher amou
ed > lint > st
additional y
cotton crop i
Y produced
he nutrient u
on.
e treatments (
the fruiting
n and utilizat
d by a crop
inus uptake i
was high in th
and its Recovery i
gh for the or
manure prepa
oved the B ut
was influenc
owering to b
decompositio
ing the leach
B availabil
ich organics
ficiency there
g practices as
pt Soil of Tamil
ded in the so
e increased
ontrol was o
unts of B wh
tem >leaf > b
yield (over n
is to get high
per unit of a
se efficiency
(Fig. 2). This
organs from
tion was rep
is expressed
in unfertilize
he organic B s
in Hybrid Cotton
rganic B sou
aration and it
tilization effi
ced significa
boll formatio
on. Applicat
hing losses
lity in soil i
in the manu
e by reducing
s it warrants
Nadu, India
il application
B uptake o
obtained in
hile the stem
bur > root.
no nutrient ap
her SCY and
applied nutri
y is also calc
s could be at
m vegetative p
ported by M
d as apparen
ed plots, to t
source treatm
urce treatme
ts application
iciency.
antly by the
on and then
tion of FYM
(Sharma et
is confirmed
ure preparati
g the yield lo
frequent app
367
n of borax
of 50.6 for
this study.
m and seed
pplication)
d hence the
ient. Since
culated for
ttributed to
part by the
Miley et al.
nt nutrient
the applied
ments.

ents which
n to high B
treatments
at harvest
M increased
al., 2006).
d by the B
ion and its
oss besides
plication of
TABLE 3: INFLUENCE OF B SOURCES ON B AVAILABILITY AND B BALANCE IN SOIL AT DIFFERENT GROWTH STAGES
Treatments B availability (mg kg
-1
) B balance in soil
(g ha
-1
) at harvest Squaring Boll formation Harvest
T
1
0.88 0.69 0.51 0.280
T
2
1.15 0.70 0.62 0.241
T
3
1.04 0.74 0.62 0.275
T
4
1.10 0.74 0.64 0.261
T
5
1.16 0.78 0.61 0.248
T
6
0.46 0.78 0.64 0.196
T
7
1.45 0.96 0.76 -0.148
T
8
1.55 0.93 0.80 -0.148
T
9
1.53 0.93 0.81 -0.122
T
10
1.05 0.74 0.64 0.410
Mean 1.14 0.80 0.67 -
CD (5%) 0.35* 0.17** 0.13** -
CONCLUSION
It is concluded from the study that the soil application of borax (5 kg ha
-1
) alongwith 0.3 kg B ha
-1
as
foliar spray twice at 45 and 75 DAS recorded high dry matter production and SCY. All other B applied
treatments produced similar biomass and SCY. This showed that the supply of B to cotton can be met by the
organic B sources by integrating B rich organic wastes in the compost production. This also paves a way for
the organic cotton production where the frequent use of inorganic B sources is warranted. However, a
detailed research is required in this division in future.
REFERENCES
[1] Anderson, O.E. and Bosewell, F.E. (1968) - Boron and manganese effects on cotton yield, lint quality, and earliness of
harvest. Agron. J., 60: 488493.
[2] Heitholt, J.J. (1994) - Supplemental boron, boll retention percentage, ovary carbohydrates, and lint yield in modern cotton
genotypes. Agron. J., 86: 492497.
[3] Howard, D.C., Gwathmey, C.O., Roberts, R.K. and Lessman, G.M. (1998) - Potassium fertilization of cotton produced on a
low K soil with contrasting tillage system. J. Prod. Agric., 11: 7479.
[4] Janaki, P., Arivalagan, T., Malarkodi., M. and Meena, S. (2005) - Integrated nutrient management for cotton Brinjal
cropping sequence. J. Agrl. Res. Mgt., 4(Suppl.): 8789.
[5] Miley, W.N., Hardy, G,W., Sturgis, M.B. and Sedberry, J.E. (1969) - Influence of boron, nitrogen and potassium on yield,
Nutrient uptake, and abnormalities of cotton. Agron. J., 61: 913.
[6] Padole, V.R., Deshmukh, P.W., Nikesar, R.J. and Bansode, N.V. (1998) - Effect of organics and in organics on yield and
quality of cotton grown on vertisol. Pkv. Res. J, 22: 68
[7] Roberts, R.K., Ufersman, J.M. and Howard, D.D. (2000) - Soil and foliar applied boron in cotton production: An Economic
Analysis. The J. Cotton Sci. 4: 171177.
[8] Sharma K.R., Srivastava, P.C., Srivastava, P.

and Singh, V.P.
(
2006) -
.
Effect of farmyard manure application on boron
adsorption-desorption characteristics of some soils. Chemosphere. 65(5): 769777
[9] Singh, M.V. (2006) - Emerging Boron Deficiency in Soils and Crops in India and Its Management. In: Proceedings of
18
th
World Congress of Soil Science held at Philadelphia, Pennsylvania, USA during July 915, 2006, pp. 104.
[10] Stewart, J. McD. (1986) - Integrated events in the flower and fruit. In: J.R. Mauney and J.McD. Stewart (ed). Cotton
Physiology, Cotton foundation, Cotton Council of America, Memphis, pp.261297.
[11] Yermiyahu, U., Keren, R. and Chen, Y. (2001) - Boron uptake by plants as affected by organic matter. In: Plant Nutrition -
Food security and sustainability of agro-ecosystems, Edited: W.J. Horst et al., 92: 852853
A Thermal Optimum Approach to Irrigation

Scheduling in Australian Drip Irrigated Cotton
W.C. Conaty
1,4,5
, J.E. Neilsen
1,2
, J.R. Mahan
3
, B.G. Sutton
4
and D.K.Y. Tan
4

1
CSIRO Plant Industry and Cotton Catchment Communities CRC,
Locked Bag 59 Narrabri NSW 2390 Australia;
2
Monsanto Australia, Level 12, 600 St Kilda Rd, Melbourne VIC 3004
3
USDA/ARS Plant Stress and Water Conservation Laboratory,
3810 4th St, Lubbock, TX 79415, USA;

4
Faculty of Food, Agriculture and Natural Resources,
The University of Sydney, Sydney NSW 2006
e-mail: warren.conaty@csiro.au
AbstractTo optimise irrigation water application there is a need for efficient and targeted irrigation scheduling
systems. A thermal optimum approach to irrigation scheduling uses the principle that plant performance is optimised
when a plant is maintained within its optimal temperature range through timely irrigation. The advantage of such an
irrigation scheduling system is that it is based directly on plant stress physiology, and not on indirect measurements
of plant stress such as soil water and/or environmental load. Researchers at the USDA/ARS have previously used the
relationship between canopy temperature (T
c
) and plant water status to schedule irrigations based on a temperature-
time threshold system. These systems command irrigations when the crops T
c
exceeds its optimal temperature
threshold (TT) for a pre-determined period of stress time (ST), i.e. time the T
c
> TT. The aim of this research was to
assess the utility of this thermal optimum approach to irrigation scheduling in Australian drip irrigation systems. At
the Australian Cotton Research Institute at Narrabri, Australia, field based T
c
was continuously monitored with
infra-red thermometers (Zytemp TN901, Hsinchu, Taiwan) in drip irrigated cotton with varying soil water deficits.
The optimum temperature of the cotton cultivar Sicot 70BRF was assessed by both crop (lint yield) and leaf level (gas
exchange) responses to T
c
. Data showed a single relationship where yield reductions occurred beyond T
c
of ~ 28 C,
validating previous work on the species-specific optimal temperature of cotton. In drip systems, which have the
capacity to rapidly supply water at precise volumes, the ST threshold for peak lint yield was observed at 5.0 h daily ST.
Future work should include further field validation, investigations in furrow irrigated systems and an integrated
approach to stress detection, which should consider both the degree and duration of time T
c
> 28 C. The thermal
optimum approach to irrigation scheduling offers potential for plant based precision irrigation scheduling. This study
provides the TT and ST thresholds, as well as a research overview in irrigation scheduling using a thermal optimum
approach in a high yield context.
INTRODUCTION
Irrigation scheduling serves to ensure that water is available in sufficient quantity at the time when it is
needed by the plant. A thermal optimum approach to irrigation scheduling uses the principle that plant
performance is optimised when a plant is maintained within its normative temperature range through
irrigation water application (Mahan et al., 2005). This approach is based on the theory that as soil water
availability declines, transpiration is reduced, and hence, canopy temperatures rise. The advantage of
such an irrigation scheduling system is that it is based directly on plant stress physiology, and not on
indirect measurements of plant stress: soil moisture conditions or environmental load. The use of canopy
temperatures as an indicator of plant stress is well established (Idso, 1982; Jackson et al., 1981).
Irrigation scheduling based on the measurement of plant temperatures is the focus of this paper. One such
irrigation scheduling system is BIOTIC (Biologically Identified Optimum Temperature Interactive
Console), described by Mahan et al. (2005).
In an effort to develop early indicators of plant water stress, the optimum plant temperature,
estimated from the thermal dependence of the Michaelis constant of an enzyme (K
M
), and the range of
temperatures conducive to optimal metabolic activity, the thermal kinetic window (TKW), of cotton was
defined (Burke et al., 1988; Upchurch and Mahan, 1988). This was supported by the finding that, through
evaporative cooling from transpiration, well watered cotton plants preferentially maintained their leaf
62
temperature (T
l
) at about 28C (Hatfield et al., 1987). Plants exhibit homoeothermic behaviour, where
they will preferentially maintain their in vivo temperature at a specific temperature, and growth occurs at
these plant temperatures (Burke and Upchurch, 1989; Mahan and Upchurch, 1988). This finding led
USDA researchers to develop BIOTIC, an irrigation scheduling tool that manages the timing of
irrigations using T
c
measurements and a specific time threshold (Upchurch et al., 1996).
The BIOTIC continually measures the T
c
of the target crop with an infra-red thermometer (IRT).
After each measurement, the T
c
is compared with a pre-determined threshold of water stress T
c
, where if
the T
c
is greater than this threshold, the crop is considered thermally stressed. The crop can then be
irrigated if relative humidity is not limiting transpirational cooling. Although this approach provided
precise and rapid alleviation of water stress, the approach needed to be modified for use in systems with
longer irrigation intervals (3-7 d) (Mahan et al., 2005). Under such circumstances, a time threshold is
required in the decision making process. The time threshold defines the average amount of time that the
canopy temperature of a well watered crop can exceed the temperature threshold, even in the absence of a
water deficit. Irrigation is appropriate when the temperature and time thresholds have been reached, and
relative humidity is not limiting transpirational cooling (Mahan et al., 2005).
The BIOTIC approach to water stress detection is limited by the availability of energy to raise the
plant temperature above the optimum, water availability for transpirational cooling, and potential
incorrect water stress detection during periods of high vapour pressure deficit that may not allow for
transpirational cooling to the temperature optimum.
This paper aims to describe the implementation of BIOTIC irrigation scheduling in cotton
(Gossypium hirsutum) in the semi-arid cotton growing region of Narrabri in New South Wales, Australia.
The temperature threshold was assessed through a dual-level approach, where the optimum temperature
of the Australian cotton cultivar Sicot 70BRF was considered in terms of leaf-level gas exchange and
crop level yield- T
c
response. The time threshold will also be determined through yield- T
c
associations.
This is the first study adapting the BIOTIC protocol to Australian conditions, and will provide a
foundation for the development of the BIOTIC theory to irrigation scheduling in the Australian cotton
industry.
Site Description
A field experiment was conducted at the Australian Cotton Research Institute (ACRI) in Narrabri (30 S,
150E) NSW Australia. The climate of this region is characterised by mild winters, hot summers and
summer-dominant rainfall patterns, with an annual average of 642mm (Bureau of Meteorology, 2009).
The soil of the site was a uniform grey cracking clay (Australian soil taxonomy: Grey Vertosol; USDA
soil taxonomy: Typic Haplustert). Weather data is shown in Table 1.
TABLE 1: WEATHER CONDITIONS OBSERVED FROM EMERGENCE TO MATURITY, MEASURED 0.4 KM FROM THE EXPERIMENTAL FIELD SITE
Weather variables
Average T
max
(C) 30.4
Average T
min
(C) 15.6
T
max
>36 C (d) 13
Accumulated day degrees 2115
Solar radiation (MJ m
-2
d
-1
) 23.6
Precipitation (mm) 354
Average daily RH
max
(%) 75.9
Experimental Design and Plot Management
The cotton cultivar Sicot 70BRF was planted on 5 October 2007 in a randomised complete block design
with five replicates of surface drip irrigation treatments based on daily evapotranspiration (ET
o
) rates.
Plots were six rows of surface drip-irrigated cotton with an additional 2 rows buffer around each plot.
Plots were irrigated daily to every three days, depending on daily ET
o
and in-season rainfall. Five
A Thermal Optimum Approach to Irrigation Scheduling in Australian Drip Irrigated Cotton 371
irrigation treatments included a control or theoretical optimal (nominally 100% daily water requirement
of control applied - Treatment 4), an excessive (nominally 125% of control daily water requirement of
control applied - Treatment 5) and three deficit (nominally 65%, 75% and 90% of control daily water
requirement applied - Treatments 3, 2 and 1) irrigation regimes. Daily irrigation rates were calculated
according to Allen et al. (1998) where the daily water requirement (crop evapotranspiration) is equal to
the product of a ET
O
and a locally calibrated crop factor (K
C
). ET
O
was calculated using on site weather
station data and the Penman-Monteith equation (Allen et al., 1998). The K
C
used was based on recent
experimental data (not published) conducted by the Commonwealth Scientific and Industrial Research
Organisation (CSIRO) agronomic management program over a number of growing seasons. Management
for all field experiments followed current high-input commercial practices outlined by Hearn and Fitt
(1992). Each experiment was managed according to its individual requirements (e.g. with respect to pest
control), with all plots receiving the same management regime.
Gas Exchange-leaf Temperature Relationships
Leaf carbon assimilation (A) and stomatal conductance (g
s
) were measured using an infra-red gas
analyzer (IRGA), Portable Photosynthesis System; Li-COR
model 6400-XT (Li-COR Biosciences,

Lincoln, Nebraska, USA). Measurements were taken on the youngest fully expanded leaf in all plots of
the theoretical optimal (control) (Treatment 4), excessive (Treatment 5) and the largest soil moisture
deficit (Treatment 1) irrigation treatments. A range of irrigation treatments was used, ensuring an array of
studied T
l
and corresponding gas exchange rates. The T
l
were measured with a chromel-constantan
thermocouple junction located within the sensor head of the Li-6400 (Li-COR, 2004a).
As gas exchange is affected by light intensity, humidity, temperature, carbon dioxide and time of
day, the Li-6400 was matched to ambient conditions and held constant for the time period of
measurements. This resulted in cuvette relative humidity controlled at 50 - 70%, carbon dioxide
maintained at 380 mol (CO
2
) mol
-1
air, photosynthetic active radiation (PAR) set to 2000 mol m
-2
s
-1

and air temperatures ranging from 23 to 42 C (depending on daily ambient conditions). Equations for
calculating A (in mol (CO
2
) m
-2
s
-1
) and g
s
(in mol (H
2
O) m
-2
s
-1
) are given in the Li-COR Biosciences
manual (Li-COR, 2004b).
Yield- canopy Temperature and Yield- stress Time Relationships
Mechanically-picked seed cotton weight data was recorded from one centre row of each plot. The gin
turn-out (per cent lint of seed cotton) and subsequent lint yield was then calculated from a sub sample of
the seed cotton.
Wireless, battery-operated Smart Crop
TM
infrared thermometers (Smartfield Inc., Lubbock, TX,
U.S.A.) were placed in four replicates of the experiments. The SmartCrop system uses a Zytemp model
TN901 infrared thermometer (IRT) (Zytemp, Hsinchu, Taiwan). The remote IRTs measure average
output temperature within the field of view at a one minute intervals, and transmit a 15 min average
temperature to a base station via a low power radio link. Data were collected throughout the season from
flowering through to crop maturity (80 DAS to 178 DAS). Sensors were positioned and maintained
periodically at 15 cm above the canopy pointing south (to reduce the effects of specular reflectance) at an
angle of 70 for the duration of the measurement period.
Stress time (ST) is defined as the daily period of time canopy temperatures exceed 28 C. This was
calculated daily from T
c
data, and is represented as a daily seasonal average.
Statistical Analysis
All data were analysed in GenStat 11
th
edition. Differences in treatments were assessed at the 95%
confidence level (P=0.05) on normally distributed data.

RESULTS
Temperature Threshold
A split-line regression was fitted to the lint yield- T
c
(R
2
=0.86) (Fig. 1), where yield reductions were
observed when seasonal average day-time T
c
> 27.7 0.9 C. Although a split line regression was fitted
to the data, it is not expected that lint yield will remain constant at canopy temperatures below about 25
C. Although the data set was range limited, crops with average T
c
below 25 C may not accumulate the
required day-degrees for crop maturity, and would be outside the range of seasonal canopy temperatures
experienced in a commercial Australian production setting. Despite this, because of the range of data
available, this regression provided the best fit to the data.

Fig. 1: Split-line Regression of Lint Yield- canopy Temperature (T
c
) Response Showing Lint Yield Reductions
when T
c
> 27.7 C (R
2
= 0.86; when x 27.67, y = 3320.5; when x > 27.67, y = -413.4x +14757.21).

Fig. 2: (a) Polynomial Regression (P<0.001) of Leaf Net Carbon Assimilation (A) and Leaf Temperature (Tl) Peaking at Tl = 29.3C (R2=0.41; y= -0.52x2
+30.50x -407.83); and (b) Polynomial Regression (P<0.001) of Stomatal Conductance (gs) and Tl Peaking at Tl = 29.1C (R2=0.48; y= -0.019x2 +1.09x -
15.07). Vertical Bars Represent two Standard Errors of the Mean.
The measured gas exchange parameters exhibited a second order polynomial response to
temperature. The model-calculated peak A occurred at 29.3 C, with an observed standard error of 3.61
mol (CO
2
) m
2
s
-1
(R
2
= 0.41; P<0.001) (Fig. 2a). The model calculated peak g
s
at 29.1 C, with an
T
c
(C)
24 26 28 30 32
L
i
n
t

y
i
e
l
d

(
k
g

h
a
-
1
)
1000
1500
2000
2500
3000
3500
4000
(a)
A

(
m
o
l

(
C
O
2
)

m
2

s
-
1
)
15
20
25
30
35
40
45
50
(b)
T
l
(C)
24 26 28 30 32 34 36
g
s

(
m
o
l

(
H
2
O
)

m
2

s
-
1
)
0.0
0.2
0.4
0.6
0.8
1.0
1.2
observed standard error of 0.124 mol (H
2
O) m
2
s
-1
(R
2
= 0.50; P<0.001) (Fig. 2b). Although the fit of
these regressions was not particularly strong, obvious trends in gas exchange were observed with peak A
and g
s
occurring at ~ 29C. These regressions are weakened by the limitation in the range of data as field-
based data were collected over a relatively narrow (but representative of commercial irrigated Australian
cotton systems) range of T
l
(26 34 C). Hence, there are no data providing leverage to the regression
with low A and g
s
due to low or high T
l
. The range of T
l
that represent A equal to that of the calculated
peak assimilation (29.3C) occurred between 27.5 and 31.2C, whilst the range for peak stomatal
conductance (29.1C) occurred between 26.8 and 30.5C.
Time Threshold
Average T
c
and ST displayed a positive linear relationship (R
2
= 0.97; P<0.001), where average ST
increased by approximately 1 h for every 1 C increase in average T
c
(Fig. 3). Using this relationship, a
temperature threshold of 28 C is expected to produce an average daily ST of 5.3 h. Because average
daily ST is calculated from T
c
, and associations are auto correlated, this value can only be considered a
rough estimate of the ST threshold for irrigation, required to produce peak yields. The association
between average daily ST and lint yield showed similar results to the approach used in Fig. 3. A
quadratic relationship was fitted to average daily ST and lint yield (R
2
= 0.89; P<0.001; y = -81.49x
2
+
618.98x + 2293.06), where peak lint yields were observed between 2.6 and 5.0 ST h, with an average of
3.8 ST h. This suggests that in practice, lint yield reductions occurred when average daily ST > 5.0 h.

Fig. 3: Association between Canopy Temperature (Tc) and Average Daily Stress Time (ST) (P<0.001) (R2 = 0.97;
Y = 1.05x -24.12). when Tc = 28 C, Average Daily ST = 5.3 h.
DISCUSSION
In arid environments, plant temperatures can range from significantly less than air temperature under
optimal water status to significantly higher than air temperature when plant water status is less than
optimal (Mahan et al., 2010). Therefore, T
c
can potentially be used to infer transpiration rates, and
provide a basis for determining plant water stress. Average T
c
in this study reflected this trend, where T
c

increased with decreasing water supply. Using a split-line regression (Fig. 1), lint yield reductions
occurred when T
c
> 28 C. On the semi-arid Texas High Plains, Wanjura et al. (1990; 1992; 1988) took a
different approach to determining the temperature threshold. They hypothesised that maximising the
amount of time T
c
were at the optimum or within the TKW, lint yields should also be maximised.
Wanjura et al. (1990; 1992; 1988) showed that when cotton irrigations were scheduled, if the average T
c

during a 15 min period exceeded a pre-determined temperature threshold of 26, 28, 30 or 32 C, lint
yields were consistently highest in the 28 C temperature threshold.
5.3 h ST
T
c
(C)
24 26 28 30 32
S
t
r
e
s
s

t
i
m
e

(
h
)
0
2
4
6
8
10
Gas exchange provide a measure of the degree of drought stress imposed on a crop and the response
of leaf gas exchange measurements have been used to detect and quantify water stress (Baker et al.,
2007). Therefore, leaf A and g
s
were used as surrogates for plant performance at a given T
l
. The peak in
gas exchange parameters, both A and g
s
, occurred at T
l
of ~ 29 C (Fig. 2). This initially suggested that,
when measured in the same cultivar, the optimum for gas exchange in field grown Australian cotton may
be slightly higher than the historical temperature optimum (Burke, 1990), and the threshold for yield
reductions due to excessively high T
c
(Fig. 1). However, the range of T
l
that produced optimum gas
exchange rates equal to that of the peak at 29 C occurred between 26.8 C and 31.2 C. This range in
optimum temperatures was similar to the TKW for cotton (23.5 C to 32 C) and encompassed the
optimum temperature for cotton metabolism (28 C) as outlined by Burke et al. (1988). Therefore, we
conclude that the thermal optimum for the Australian cotton cultivar Sicot 70BRF is about 28 C, which
is consistent with the optimum temperature of cotton growth and metabolism outlined in a recent review
by Burke and Wanjura (2010).
The ST threshold was also evaluated in this study. The ST threshold represents the site-specific
average daily period of time that T
c
can be expected to exceed the thermal optimum, regardless of soil
water availability (Mahan et al., 2005). Using a temperature threshold of 28 C, T
c
-ST associations
showed a corresponding ST of 5.3 h (Fig. 3), while yield-ST associations showed a reduction in yield
when ST > 5.0 h. This suggests that in practice, peak yields can be achieved in drip irrigated cotton
grown at Narrabri with a ST threshold of up to 5.0 h. On the semi-arid Texas High Plains, Wanjura et al.
(2006) observed a negative linear response of lint yield with ST where an increase in 1 h ST resulted in a
decrease of 343 kg lint ha
-1
(19% reduction ST h
-1
). In this environment, the highest lint yields were
observed at about 5.5 h ST, and a curvilinear response was not observed. However, this response was
observed over a smaller range of ST (6 9 h ST compared with 2 9 h ST in our study), and when our
data are restricted to this range, a linear response is observed (R
2
= 0.79) where an increase in 1 h ST
resulted in a 513 kg lint ha
-1
yield reduction (15% reduction ST h
-1
).
CONCLUSION
This paper represents the first step in developing the temperature and time thresholds required for
efficient plant based irrigation scheduling in the semi-arid cotton growing region of Narrabri, Australia. It
is recommended that a temperature threshold of 28 C and a time threshold of 5 h are employed in the
system, to produce peak lint yields with the most efficient use of water. This approach is limited to
irrigation systems that have a high capacity to supply water at small time intervals (drip/sprinkler
irrigation), and is site specific. Future work should further investigate the utility of the system in
Australian production systems, such as furrow and large deficit irrigation systems, and may need to
assess the environmental conditions where canopy temperatures are inefficient in detecting plant water
stress, such as high vapour pressure deficit environments.
ACKNOWLEDGEMENT
This study was funded by the Cotton Catchment Communities Cooperative Research Centre and the
Cooperative Research Centre for Irrigation Futures, with further support from the University of Sydney.
Thanks to Merry Errington, Nicola Cottee, Jo Price, Jono Cuell, Mitch Cuell, Martyn Tann and Meghan
Smith in Narrabri for technical assistance. We are grateful to Steven Yeates for agronomic advice and
providing the crop factor. Further thanks to Michael Bange and Greg Constable, for advice and reviewing
the manuscript.
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Kinetic Window" Criteria -Beltwide Cotton Conference. pp. 183185.

Efficient Water Management Technology

for Sustainable Cotton Production in Central India
V. Kumar
1
, R.G. Patil
2
and J.G. Patel
3

1
Research Scientist (Cotton) & Head, NAU, Surat
2
Research Scientist (SWM), NAU, Navsari
3
Associate Research Scientist, NAU, Bharuch
AbstractThree research achievements are considered as milestones of Indian agriculture. The first was introduction
of Norin 10- Brevor 14 dwarfing gene in wheat which lead to green revolution in mid sixties. The second was
development of worlds first successful hybrid of cotton Hybrid-4 which lead to white revolution in early seventies and
third is introduction of cry gene (Bt) in cotton which is considered the beginning of gene revolution in early years of
current century.
To attain self sufficiency in agriculture, government paid utmost attention to the development of irrigation
projects, fertilizer factories and pesticide industry and framed policies conducive to the overall development of
agriculture in India. The outcome is that today, the country is self sufficient in most agricultural commodities. All this
has been possible because of high yielding varieties/hybrids and input intensive production technologies. To harvest
more and more, injudicious use of inputs especially water, fertilizer and pesticides became a fancy. Consequently,
water resources have either dried up or become unusable due to salt ingress, soils have become sick/deficient in organic
matter, and multiple nutrients and lower microbial activity, resulting in a declining/steady crop productivity.
Declining water availability for agriculture, frequent droughts and unprecedented floods warrant long term and
strategic research programmes. Therefore efficient use of water becomes of paramount importance.
Central zone of India, comprising the states of Madhya Pradesh, Gujarat and Maharashtra has cotton area which
is mainly rain dependent (70-75%). The productivity is low in this zone vis-a-vis Northern zone which is 100 per cent
irrigated and Southern zone which is about 40-45 per cent irrigated. The technology has over the years, been devised to
(i) conserve rain water in rainfed tract through use of tillage, land configuration and mulch (ii) water management in
irrigated tract through drainage and scheduling of irrigation based on soil moisture depletion, critical growth stages
and climatological changes and (iii) other approaches for efficient water use through planting methods, tolerant
variety, planting time, intercropping and use of (agro) chemicals. Efficient use of available water and fertilizers for
better seed cotton yields and saving in irrigation water are discussed.
INTRODUCTION
Cotton has been a crop of importance from the days of yore. Excavations of Indus valley civilization are
testimony of cotton use more than 5000 years ago. Much has changed over centuries, over decades and
much over recent years. The first big change was in 1971 when the first hybrid cotton Hybrid 4 (H4) was
released for commercial cultivation. This transformed the entire cotton scenario in the country whether it
was research, cultivation practices, seed and pesticide industry or it was production and productivity. The
second big change is more recent when cotton hybrids were released for cultivation in 2002.
Today, India produces about 30.0 m bales of cotton from nearly 10.0 m ha with an average
productivity of around 500 kg lint/ha. Yet it is much below the world average of >700 kg (Anonymous,
2010). Even with best productivity of 560 kg/ha, it ranks 24
th
amongst cotton producing countries
(Venugopalan et al., 2009). This calls for immediate attention why Indian cotton yields, despite Bts and
hybrids, have not even touched world average, leave aside China and Israel. Is it production technology
or is it water or both? Data presented in Table 1 would make it amply clear that water plays a
predominant role in cotton productivity. Gujarat contributed 81 per cent production from 60 per cent
irrigated area during 2006-09 against 19 per cent from 40 per cent unirrigated area during the same
period. The average irrigated productivity was 743 kg against 262 kg of unirrigated being +34.8 per cent
and -52.5 per cent, respectively, over average productivity (551 kg) of the state.
63
Efficient Water Management Technology for Sustainable Cotton Production in Central India 377
That does not undermine the importance of production technology. In fact, there are farmers
harvesting as much as 2000 kg lint/ha in irrigated areas of Kutch, Rajkot and Narmada districts of
Gujarat against 743 kg of state irrigated average. Hence, this gap is to be narrowed but the real challenge
is for rainfed areas.
TABLE 1: AREA, PRODUCTION AND PRODUCTIVITY OF COTTON IN GUJARAT (AV. 2006-09)
Particulars Irrigated Rainfed Total
Area (M ha) 1.431 60) 0.952 (40) 2.383
Production (M Bales) 6.257 81) 1.465 (19) 7.722
Productivity(kg/ha) 743 (+34.8) 262(-52.2) 551
Figures in parenthesis are per cent over state total. Source: Statistical Unit, Dept. of Agriculture, Gandhinagar
Since independence in 1947, the irrigated area under cotton in India has grown by little over five
fold, so is the lint yield (Sivanappan, 2004). Currently, the country grows cotton on 11.0 m ha having
wide range of soils and agroclimatic conditions. Nearly 65 per cent of cotton area is unirrigated, majority
of it is in central zone (75%) followed by South zone (60%). The central zone comprises Maharashtra,
Gujarat and Madhya Pradesh occupies 66 per cent of the cotton area in the country contributing 58.6 per
cent production to the national kitty (Table 2).
TABLE 2: STATE WISE AREA, PRODUCTION AND PRODUCTIVITY OF COTTON (2008-09)
State Area (M ha) Production (M Bales) Productivity (Lint kg/ha)
North Zone [100] 1.215 (13.0) 3.900 (13.4) 546 (+ 3.8)
Central Zone [24] 6.205 (66.2) 1.700 (58.6) 466 (-11.4)
South Zone [40] 1.855 (19.8) 6.700 (23.1) 614 (+ 16.7)
Others 0.098 1.400
India [35] 9.373 29.00 526
Figures in parenthesis are percentage of the country. [ ]% irrigated area
The productivity in Maharashtra and Karnataka irrigated area which are predominantly unirrigated is
abysmally low vis--vis Gujarat, Andhra Pradesh and Tamil Nadu. Central India is especially prone to the
vagaries of weather where insufficient rainfall, uneven distribution and even failures are not uncommon.
Therefore, the strategies have to be multipronged.
MOISTURE CONSERVATION IN RAINFED TRACT
Tillage
TABLE 3: EFFECT OF DEPTH OF TILLAGE ON YIELD OF COTTON G.COT.HY.12 UNDER SOUTH GUJARAT CONDITIONS
Treatment Seed Cotton (kg/ha)
Surat (Irri.) Bharuch (Rainfed)
Depth of Tillage
10 cm 2115 1514
20 cm 2417 1785
30 cm 2509 1648
S. Em. 51 54
C.D. at 5% 152 170
Anon., 2009
Tillage for moisture conservation is an age old practice traditionally done by bullock drawn plough.
Development of mold board and disc plough lead to intensive use in cotton, as well (Kairon et al., 2002).
In central and southern India, deep ploughing once in 2-3 years is common practice (Blaise and
Ravindran, 2003). Preparatory criss-cross harrowing and planking, repeated interculturing (3-5) by
bullock drawn hoe or mechanized cultivator is practiced by cotton farmers. However, due to variations in
soil, climate, rainfall, weediness and type of cottons grown, tillage practices also vary. Reduced tillage
was found to be better or equal to conventional tillage (Blaise et al., 2005) in rainfed central India. In
sandy loam soils of North India, deep ploughing followed by conventional tillage was much better than
no tillage or conventional tillage (Anon., 2003). Bt hybrids performed better with reduced tillage than
conventional tillage (Blaise, 2011 and Jalota et al., 2008). Tillage at 20 cm depth was found optimum in
rainfed as well as irrigated Vertisols of South Gujarat (Table 3).
Land Configuration
Effective management of soil moisture is crucial for realizing potential productivity of hybrid cotton.
Making beds (7.5 m) and furrows (20 cm) had been found effective in conserving moisture and
enhancing yield in Gujarat (Table 4).
TABLE 4: EFFECT OF LAND CONFIGURATION ON RAINFED COTTON EQUIVALENT YIELD (POOLED OVER 3 YEARS)
Treatment Bed width Mean
Depth of furrow 5.0 m 7.5 m 10.0 m
20 cm 1153 1284 1061 1166
30 cm 1169 1285 1095 1183
Mean 1161 1284 1077
CD at 5 %( Bed) 92
Achhalia (Anon., 2005).
Forming beds (120 to 180 cm wide) and furrows on a grade for in situ water harvesting was found to
be efficient in deep black soils with 700-850 mm rainfall (Venkateshwarlu, 1980). Opening of furrows
after every row of cotton between 30-45 days after sowing and mulching with crop residue were found
promising in Maharashtra (Giri et al., 2008). Opening of alternate furrows in sole crop after last
interculture enhanced yield of seed cotton, intercropping black gram further improved WUE and returns
(Patel et al., 2008). Shinde et al., (2009) however, reported opening of furrows in every row after last
interculture to be the most efficient.
Mulch
Practice of mulching is in vogue for a long time. It is an important technique known to reduce
evaporation, increase average soil moisture and yield. Besides biomulches, plastic mulches have become
handy due to inherent advantage in weed suppression, maintaining soil temperature, faster mineralization
in addition to efficient moisture conservation.
Use of black plastic mulch was found advantageous in increasing seed cotton yield to an extent of
22% in rainfed Vertisols and 92% in coastal salt affected soils (Table-5)
TABLE 5: ADVANTAGES OF MULCHING IN COTTON
Location Variety Soil Type Mulch Material Increase in Yield (%)
Bharuch G.Cot.-10 Clay (Rainfed) Grass
Black plastic Soil
8
22
12
Danti G.Cot.Hy-8 Clay (Coastal salt affected) (Rainfed) Grass
Black plastic
62
92
Source: Bharuch (Raman et al (1995). Danti Patel et al (1996).
Straw mulching and biomulching with sunnhemp were superior to opening furrows in alternate rows
in improving WUE at Akola and Nagpur (TMC, 2008).
WATER MANAGEMENT IN IRRIGATED TRACT
Drainage
Cotton requires equal attention for drainage, as for irrigation. Because it is sensitive to root aeration, no
water stagnation should be allowed to occur. In cotton growing areas of South Gujarat, water stagnation
occurs during the monsoons. Opening 22.5 cm deep furrows after every 2 or 4 rows of cotton (120 cm
apart) increased seed cotton yield by 53 or 42 per cent respectively, at Tanchha (Fig-1). In coastal salt
affected rainfed soils at Danti providing drain of 0.6 m depth every 2.4 m significantly enhanced seed
cotton yield. At Parbhani, providing drain at 75 m distance enhanced cotton equivalent yield by 70 per
cent over normal planting without drain (Anon. 2007).

Source: (Anon., 2005)
Fig. 1: Effect of Drainage on Seed Cotton Yield at Tanchha
ASSESSING WATER REQUIREMENT
Cotton is mostly grown as kharif crop and needs 2 to 10 irrigations after cessation of rainfall under
different soils and climate of western India. Thus, consumptive use varies around 650 to 1000 mm at
different places (Bonde and Shanmungam, 1990). Mohan and Arumagan (1994) worked out crop
coefficient (Kc) of cotton to be 0.46, 0.70, 1.01 and 0.39 at four different growth stages. The breakup of
water use at different growth stages of crop under normal condition is 3.8 mm/day upto 1
st
flowering, 8.5
mm/day upto peak bloom and 5.1 mm/day thereafter, till harvest (Kairon et al., 2002).
Scheduling of Irrigation
Ever increasing demand of irrigation water coupled with depleting ground water sources, expanding
domestic and industrial demand calls for efficient use of water. Normally cotton is grown by flood
irrigation technique wherein efficiency of water is low, as also of other associated inputs. Different
methods for scheduling of irrigation so as to achieve efficient use of water have been recommended.
Soil Moisture Depletion Approach
Under surface method, the crop is irrigated mainly through furrows, occasionally in border strip. The
scheduling of irrigation at 75 per cent available moisture depletion was found to be optimum throughout
the country (Table-6) except in sandy loam soil of Hisar (25% ASM) and loamy soils of Madurai (50%
ASM) (Anon,. 1971-72 to 1977-79).

TABLE 6: SEED COTTON YIELD AS AFFECTED BY DIFFERENT SOIL MOISTURE REGIMES
Location Soil Type Seed Cotton (kg/ha)
Available Soil-moisture Depletion (%)
25 50 75 100
Coimbatore Sandy-clay loam 2340
(13)
2560
(9)
2930
(5)
2870
(4)
Siruguppa Medium black clay 2970
(14)
3270
(11)
3210
(8)
-
Hisar Sandy-loam 1180
(4)
1040
(2)
950
(1)
-
Rahuri Black clay 1910
(4)
1950
(3)
1890
(2)
-
Madurai Loam - 2000
(14)
1590
(9)
1140
(7)
Bhavanisagar Clay loam 975
(18)
965
(13)
975
(9)
900
(5)
Dharwad Medium black clay 2470
(12)
2585
(9)
2565
(7)
-
( ) = Numbers of irrigation Anon,. (1971-72 to 1977-79)
Critical Growth Stages Approach
Cultivation of cotton with protective irrigation is one of the mechanisms to enhance productivity and this
should be based on critical growth stages (Grimes et al., 1968 and Buttar et al., 2005). In loamy-silty/clay
loam soil of Sriganganagar, three irrigations at 45 DAS, flowering and boll development stage were
found advantageous (Anon., 2005-06). Shinde et al. (2009) also reported similar findings from Parbhani.
However, in high rainfall area of South Gujarat, two irrigation at flowering and boll development were
sufficient with equal yield and WUE of surface irrigation at 0.8 IW/CPE (Table 7).
TABLE 7: YIELD AND IRRIGATION USE EFFICIENCY OF COTTON AS AFFECTED BY IRRIGATION AT CRITICAL GROWTH STAGES
Treatment Seed cotton
(kg/ha)
Irrigation Use Efficiency
(kg/ha/mm)
Pooled Avg.
Rainfed control 1222 -
Irrigation at flowering 1390 17.3
Irrigation at boll development stage 1487 18.6
Irrigation at 25 days after boll development 1354 16.9
Irrigation at flowering and boll development stage 1692 10.6
Irrigation at boll development and 25 days after boll development
stage
1560 9.8
Irrigation at flowering, boll development and 25 days after boll
development stage
1485 6.2
Irrigation at 0.8 IW/CPE 1660 10.3
S.Em 50 -
CD at 5% 142 -
(Patel et al., 2008)
Climatological Approach
The temperature requirement of cotton varies from 15C (Doorenbos and Kassam, 1979) to 43C
(Kulandaivelu, 1980 and Thind, 2007) depending upto stage of the crop. Therefore, there cannot be a
better approach than climatological data for surface, drip and sprinkler irrigation.
Surface Irrigation
Surface irrigation based on IW/CPE ratio 0.6 was found to be better for South Gujarat (Patel et al., 1995)
and 0.75 0.80 for Rahuri and Sriganganagar (Anon., 1977-78 and 1981-83) (Table 8).

Sprinkler
Sprinklers have not got much acceptance amongst cotton farmers because of high sensitivity of pollen to
water. Burke (2003) observed that single spray reduced seed set by 55 per cent. Additional sprays
resulted in further losses and flower shedding. The findings of Sriganganagar also substantiate this
(Anon., 2002-03 and 2003-04).
TABLE 8: EFFECT OF IW/ CPE RATIOS ON SEED COTTON YIELD
Location Soil type Mean Seed Cotton Yield (kg/ha) at IW/CPE ratio
0.3 0.4/ 0.45 0.6 0.75/ 0.8 0.9 C.D. @5%
Rahuri Black clay loam -
-
2003
(48)
2475
(58)
2640
(64)
2518
(70)
-
-
Sriganga nagar Silt
laom
1800
(35)
2000
(49)
-
-
2180
(63)
-
-
121
-
Surat Black clayey -
-
2267
(3)
2299
(5)
2021
(6)
-
-
NS
-
Achhalia Clay 752
(48)
1009
(56)
1112
(64)
1243
(72)
-
-
-
-
( ) = Depth of irrigation in cm Source: Anon. (1977-78, 1981-83) Patel et al. (1995), Anon.(2006-07)
Drip
TABLE 9: ADVANTAGES OF DRIP TECHNOLOGIES OVER CONVENTIONAL METHOD OF IRRIGATION IN COTTON
Location Variety/
Hybrid
Soil type Technolog
y
Water
saving (%)
Fertilizer
saving (%)
Yield
increase
(%)
Source
Gujarat
Khandha

G Cot Hy.8 Clay Drip 40 - 33 Anon. (1995)
Surat

G Cot Hy.6 Clay Drip 40 - - Anon. (1999)
Junagadh

G Cot Hy. 10 Clay Drip,
fertigation
20 25 30 Malavia et al.,
(1999)
Danti

Hy G Cot.8 Clay (salt
affected)
Drip +
Mulch
(Paired
row)
- - 54 Anon. (2002)
Maharashtra
Rahuri NHH-44 Clay Drip,
fertigation
(liquid
fertilizer)
- 25 12 Mane et al.,
(1996)
Parbhani NHH-44 Clay Drip,
normal
plain
10 - 22 Tekale et al.,
(1999)
Parbhani

NHH-44 Clay Drip,
paired row
32 - 12 Bharambe
et al., (2002)
Akola

AHM-68 Clay Drip,
planting
densities
(0.8 ETC)
- - Higher yield
with less
density
Dahatonde and
Deshmukh
(2003)
Rajasthan
Sriganganagar

LH-144 Loam to silty
clay loam
Drip,
normal
planting
10 - 4 Anon.
(2002-03)
Raskar (2009) found distinct benefit of drip over furrow (paired row) irrigation in terms of yield
(11.4%), WUE (61.6%) and saving in water (71.6%) at Rahuri.
Padmakumari and Sivanappan (1979) observed distinct advantage of drip over surface method in silty
clay loam of Coimbatore as early as 1979. Extensive studies carried out in Gujarat found that water
saving due to drip ranged from 20 per cent in Saurashtra to as high as 76 per cent in middle Gujarat.
Along with saving, drip irrigation also increased cotton yield by 30 per cent under normal soil condition
of South Gujarat. However, this increase was 54 per cent in coastal salt affected soils (Anon., 1995;
Anon., 1999; Malavia et al., 1999 and Anon., 2002). In Maharashtra, savings in irrigation water (Banke,
1996; Bharambe et al., 2002) upto 50 per cent and increase in seed cotton yield from 10 to 32 per cent
Takale, et al., 1999; Mane et al., 1996) and savings in fertilizer (Banke, 1996; Mane et al., 1996) have
been widely reported. Dahatonde and Deshmukh (2003) observed similar advantage of drip with less
planting densities. (Table 9).
OTHER CROP/WATER MANAGEMENT PRACTICES
In addition to moisture conservation and irrigation water management practices there are ways and
means through which water requirement of cotton could be better managed for yield advantage.
Right Variety
TABLE 10: PERFORMANCE OF TOP TEN BT COTTON ENTRIES IN IRRIGATED AND UNIRRIGATED SITUATION IN GUJARAT
Irrigated (Surat & Junagadh) Rainfed (Bharuch, Dhanduka & Amreli) Average Across Locations
Hybrids Seed Cotton
(kg/ha)
Hybrids Seed Cotton (kg/ha) Hybrids Seed Cotton
(kg/ha)
RCH-138 3023 KDCHH-9632 BGII 2429 RCH-377 2572
PRCH-31 2966 KCH-14 BGII 2425 RCH-138 2561
ABCH-1065 BGII 2925 GK-205 2378 KCH-14 BGII 2532
RCH-377 2894 RCH-377 2357 PRCH-31 2529
TULASI-4 BGII 2849 KDCHH-9821 2274 KDCHH-9821 2484
AKKA 2826 RCH-138 2253 GK-205 2463
AKKA BGII 2802 MRC7247 BGII 2244 AKKA 2462
KDCHH-9821 2800 PRCH-31 2237 AKKA BGII 2397
TULASI-9 2741 AKKA 2220 MRC-7351 2374
IT-923 2720 RCH-515 BGII 2185 MRC-7305 BGII 2371
Source: Patel et al. (2010)
Since about 90 per cent cotton area in the country is under Bt hybrids, it is indeed a difficult task for a
farmer to make a choice out of 1340 Bt varieties. Water requirement of different varieties vary according
to their stature, yielding ability, duration besides soils and climatic conditions. An attempt made under
TMC MM I 1.4 across five locations in Gujarat indicated that out of ninety hybrids tested, five of the
top ten Bt hybrids were common in both rainfed and irrigated conditions indicating their wider
adaptability (Patel et al., 2010). The difference in yield in the two situation was 4-7 q/ha (Table 10). The
third best Bt hybrid in irrigated did not find a place in top ten in rainfed. Similarly, the best performing
hybrid in rainfed did not come up in top ten in irrigated. Thus, selection of variety could prove to be very
important for profitable yield in a given situation.
Date of Sowing
TABLE 11: SEED COTTON YIELD AS INFLUENCED BY TIME OF SOWING
Sowing Time No. of Bolls/ plant Seed Cotton (kg/ha)
16
th
Met. Week (16-22 April) 73.92 2200
18
th
Met. Week (30-03 May) 75.79 2248
20
th
Met. Week(14-20 May) 84.23 2409
22
th
Met. Week (28-03 June) 76.54 2292
24
th
Met. Week(11-17 June) 70.00 1974
26
th
Met. Week (25-01 July) 66.61 2004
CD @ 5 % 1.75 47.0
Pachora, Maharashtra Source: Lalage et al.,(2004)
Date of sowing is crucial in managing water relation of cotton plant. The normal sowing time is after
onset of monsoon in central zone. Withdrawal of monsoon in mid September causes serious problem at
peak flowering and boll development stages due to limited non availability of water. This results in either
shedding and/or poor boll development. Such a situation can be avoided by planting at the right time.
Planting cotton in third week of May was most advantageous for cotton in Vidarbha, Khandesh and
Marathwada pockets of Maharashtra (Table-11).
Similarly in Gujarat, the best planting time for cotton was mid May, if irrigation is available to avoid
terminal stress at flowering (75 DAS) and boll development stage (90 DAS) (Anon., 2002a).
Intercropping
Cotton is a long duration crop of 160-180 days and has slow initial growth. Given the spacing adopted,
there remains scope for intercropping in irrigated as also in rainfed cotton. This ensures better returns and
assurance and higher input use efficiency (Willey, 1979). Cotton + green gram, cotton + pigeonpea,
cotton + black gram, cotton + soybean, cotton + maize, cotton + groundnut, cotton + sesamum are
recommended and being practiced in central zone (Table 12).
TABLE 12: INTERCROPPING IN COTTON
Treatments Nanded Rahuri
Seed Cotton
(kg/ha) 2007-09
Cotton Equivalent
(kg/ha)
Seed Cotton
(kg/ha) 2007-09
Cotton Equivalent
(kg/ha)
T1-Sole Bt cotton 2552 2552 1832 1832
T2- Cotton + Pigeon pea (4 :2) 1849 2893 1768 2381
T3- Cotton + Soya bean (1:1) 2156 2949 2202 2163
T4- Cotton + Green Gram (1:1) 2291 2720 2276 2412
T5- Paired row planting cotton
(60:120 cm)
2493 2493 1952 1952
T6-Paired row cotton +Green Gram 2249 2909 1978 2117
T7- Cotton + Cluster Bean (1:1) 2215 2717 1715 2342
T8- Cotton + Black Gram(1:1) 2146 2593 2043 2066
T9- Cotton + Sesamum (1:1) -- -- -- --
T10- Cotton + Green Gram (1:3) in
150 cm rows spacing
-- -- -- --
SEm 57.12 67.00 26.8 26.8
CD at 5% 158.1 185.4 46.5 46.5
Source Anon., 2010
Use of Antitranspirants/ Osmoprotectants
TABLE 13: EFFECT OF ANTITRANSPIRANTS ON YIELD OF BUDDED COTTON G. COT. 101
Year Kaoline 6% Kaoline 4% PMA 5
mg/l
PMA 2.5
mg/l
Film
Forming
Water
spray
Unsprayed
control
CD @
5%
1999-00 1242 1294 1399 1714 1600 1198 1106 146
2000-01 967 1008 1253 1284 1228 847 743 241
TABLE 14: STUDIES ON AMELIORATION OF WATER STRESS THROUGH USE OF OSMOPROTECTANTS
Treatments Seed Cotton Yield (kg/ha) Average Percent over Control
Glycine betaine 0.3% 1619 12.5
Calcium Chloride 0.25% 1549 7.6
Potassium nitrate 0.5% 1511 5.0
Potassium nitrate 1.0% 1525 6.0
Calcium Chloride 0.25+ 0.5%KNO3 1522 5.8
Thioglycolic acid 100 mg/l 1528 6.2
Mercaptaethyl amine 100mg/l 1694 17.7
Thiourea 500mg/l 1575 9.5
Water 1506 4.6
Control 1439
Surat, Anon., 2008
Antitranspirants havent found wide spread use in agricultural crops. Yet, they continue to be
researchable lot. Experiments carried out in rainfed area of middle Gujarat clearly showed benefit of
using phenyl mercuric acetate (2.5 mg/l) and kaoline 4 per cent on perennial budded cotton (Anon., 2002)
(Table 13). Use of osmoprotectants to ameliorate stress did not indicate consistent effect, nevertheless
mercapataethyl amine (100 mg/l) brought about 17.7 per cent increase in yield over rainfed control in a
three year trial (Table 14).
SUMMARY
India grows cotton on 10.0 m ha having wider range of soil and agroclimatic condition. Wide variability
does exist in productivity in irrigated and unirrigated cotton. Whereas this gap is to be narrowed, the
productivity of irrigated has to be increased to attain the targets of XII FYP (39.0 m bales?). Water
management could play a major role in this. Making beds and furrows and opening of furrows was found
effective in conserving moisture. Deep tillage at least once in 2-3 years followed by conventional tillage
has been found better than conventional tillage.
Drainage in water logging prone areas is advised. The size of drain depends upon the soils and area.
Mulching by crop residue and/or black plastic (25 micron) has proved very effective in conserving
moisture, checking evaporation, suppressing weeds, therefore efficient utilization of resources. Irrigation
based on ASM depletion IW/CPE ratio /critical growth stages saved water and enhanced water use
efficiency. Drip irrigation with (out) fertigation extensively established their usefulness in saving 20-76
per cent water, increase 10-32 per cent yield with better WUE depending upon soils and climatic
conditions. Other approaches for efficient water management are usually advanced date of sowing,
adoption of intercrops and selection of right variety.
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[44] Venugopalan, M.V., Sankaranarayan, K., Blaise, D., Nalayani, P., Prahraj, C.S. and Gangaiah, B. (2009) - Bt cotton
(Gossypium sp.) in India and its agronomic requirement.-A review. Indian J. Agron., 54(4): 343360.
[45] Willey, R.W. (1979) - Intercropping: its Importance and research needs part I. Competition and yield advantages. Field
Crop Abstract 32: 110.
Biodegradable Polyethylene MulchingA New

Approach for Moisture Conservation, Weed Control
and Enhanced Productivity of Winter Irrigated Cotton-
Maize System
P. Nalayini
1
, K. Sankaranarayanan
1
, K. Velmourougane
2
and M. Suveetha
1
1
Central Institute for Cotton Research, Regional Station, Coimbatore641003, India
2
Central Institute for Cotton Research, Nagpur, Maharashtra440010, India
AbstractField experiment was conducted during 2008-09 and 2009-10 cropping season under winter irrigated
(August February) cotton followed by maize (March-May) at Coimbatore, Tamilnadu. Eight mulch treatments
including polyethylene mulching, biodegradable polyethylene, sugarcane trash, maize stover, gunny sheet, coir waste
(subsoil), surface coir waste application were compared with no mulch control under three moisture regimes, 0.4 ETc
through drip, 0.8 ETc through drip and conventional irrigation. The results indicated that the cotton (RCHB 708 Bt)
responded significantly to moisture regimes and mulches. The crop responded up to 0.8 ETc under no mulch
condition. However, under mulched situation, the seed cotton yield started decreasing beyond 0.4 ETc. Among the
mulches, polyethylene and biodegradable polyethylene mulching were similar and better than the other mulches.
Among the combinations, polyethylene mulching with drip irrigation at 0.4 ETc recorded 5641 kg /ha and was on par
with biodegradable polyethylene (5241 kg/ha) at the same moisture level of 0.4 ETc. The water requirement for ELS
cotton RCHB 708 Bt was 464, 630 and 800 mm at 0.4 ETc, 0.8 ETc and conventional irrigation, respectively. The
zero tilled rotation maize grown after cotton was also significantly influenced due to moisture regime and mulch
treatments.
INTRODUCTION
Use of polyethylene mulching for moisture conservation, weed control and enhancing the productivity of
agricultural crops has been documented and gaining global significance now than ever before as it
reduces the water loss, improves efficiency of water and thus helps in saving the precious resource. Drip
irrigation used in combination with plastic mulch needs water to meet the crop water requirement as the
other losses of water are kept under minimum thereby increasing the use efficiency of water. Yield of
cotton increased by 1.9 and 2.1fold following zero tilled rotation maize due to poly ethylene mulching as
compared to conventional method without mulching (Nalayini et al., 2009). However, polyethylene film
cannot biodegrade naturally and it can be made biodegradable by adding a small quantity (4%) of pro-
degradant additive (a patented product from UK) at the time of manufacturing the poly film. This
technology produces plastic which degrades by a process of oxo- degradation. The very small quantity of
added additive changes the behavior of the plastic products. The plastic after the desired usage will not be
just a fragment, but will be consumed by bacteria and fungi after the additive has reduced the molecular
structure to sub 40,000 daltons which permits living micro-organisms access to the carbon and hydrogen
and the transformed plastic behaves in the same way as natures wastes. It is assimilated by the same
bacteria and fungi and they convert the degraded plastic to cell biomass just like lignocelluloses such as
straw, leaves and twigs (Michael Stephenson, 2009). Oxo-biodegradable plastics sheets can be
programmed at manufacturing stage to degrade soon after the harvest or until the mission is
accomplished. Field trial in Australia using biodegradable mulch film on Capsicum spp. has shown that
biodegradable plastic performs as well as polyethylene film and offers advantage of being able to be
ploughed into the ground after harvest which can reduce the disposal costs while enriching the soil with
carbon on degradation (Anon, 2004). The use of biodegradable mulch films seems to be a promising
alternative because the films can degrade right in the field and the amount of waste ending up in landfills
can be avoided (Kijchavengkul et al., 2008).
64
Biodegradable Polyethylene MulchingA New Approach for Moisture Conservation, Weed Control 387
Polyethylene mulching standardized for cotton (30 or 50 ) poses environmental threats as it is non-
degradable. Hence, if a substitute with biodegradable nature and with advantages of poly ethylene, it is
the need of the hour and so this study was attempted.
Field experiments were conducted consecutively during 2008-09 and 2009-10 with cotton in winter
(August-February) cotton followed by maize in the summer (February-April) season. Raised bed 1.5 m
wide and 9 m long, were made using tractor drawn broad bed furrow (BBF) maker with 30 cm furrow for
irrigation. The polyethylene sheet of 30 thickness with silver colour top layer and black colour bottom
layer was used. The sheet was spread uniformly over the raised bed and the edges were sealed with soil,
leaving 30 cm from both the edges. Two sowing lines were fixed using sowing rope having markings at
60 cm apart. The holes were made at 60 cm apart using a 5 cm diameter GI pipe (by gently pressing the
GI pipe, holes were made easily as the thickness of the film was very less). Sowing was done carefully
inside the sowing holes and the sowing irrigation was given immediately. For biodegradable
polyethylene mulching, the same procedure was followed. In the polyethylene mulch sheet at the time of
manufacturing, an additive (d2w, a patented product supplied by Symphony Environmental Technologies
PIC. a global supplier of pro-degradant additive.) was added at the time of manufacturing at the rate of
4% concentration to make the biodegradable polyethylene used in the experiment. For sub soil coir
treatment, the coir waste was applied before sowing using the tractor drawn subsoil coir pith applicator
which could open the furrow at the centre of the raised bed and placed the coir wastes and covered it with
soil. For sugarcane trash, maize stover and surface coir mulching, 50 % of the mulch materials were
applied 15 DAS and the remainder applied 30 DAS above the entire raised bed. Sugarcane trash and
maize stover were cut into 10-15 cm using chop cutter before application. The gunny sheet was spread in
between the two sowing rows. Sowing (RCHB 708 Bt) was done on 29 August 2008 and 5 September
2009. The design used was split plot with eight mulch treatments, control (no mulch), sub soil coir pith
(20 t/ha), maize stover (50 t/ha), sugar cane trash (50 t/ha), surface coir (20 t/ha), gunny sheet,
biodegradable poly mulching and poly mulching in the main plots with three moisture regimes viz.,
conventional irrigation, drip irrigation at 0.4 ETc and drip irrigation at 0.8 ETc in the sub plot. The soil of
the experimental sites was low in available nitrogen (168 to174 kg/ha), high in available P (27.6 to 28.6
kg/ha) and high in available K (625 to 654 kg/ha). The infiltration rate of the experimental soil was
1.88cm/hr and moisture content (%) at field capacity and permanent wilting point was 23.4 and 17.4
respectively. Fertilizers (120: 26.4: 50 kgs NPK/ha) were applied uniformly to all the treatments. Full
dose of phosphatic fertilizer and of nitrogenous and potassic fertilizers were given as basal at sowing
and the remaining dose of N and K were given in three equal splits at monthly interval. Drip lines were
fitted with one lateral for two rows and one dripper for two crop hills.
Calculation of Water to be Applied through Drip System
The volume of water to be given on alternate days through drip was calculated using the following
formula.
( ) A K K E V
c p p
= 7 . 0

(1)
Where, V is Volume of water to be given (liters) / dripper Ep is Pan evaporation (mm)
K
p
is Pan Co-efficient (0.7) and K
c
is Crop Co-efficient (0.45, 0.75, 1.15 and 0.75 for initial (0-25
DAS), development stage (26-70 DAS), boll development (71-120 DAS) and maturity stage (121-
harvest), respectively).
Based on the above formula, a ready reackoner was prepared and the irrigation was scheduled on
alternate days based on the open pan evaporimeter reading and the exact quantity of water given was
measured using water meter. For control plot, the water to be irrigated was measured through a V
notch. After the harvest of cotton, stalks were cut below the cotyledon leaves to avoid re growth of
cotton. Holes were made 5 cm away from the cotton rows at 20 cm (plant to plant) and the maize hybrid,
CORH M4 was sown.
Growth Attributes
Growth attributes like, plant height, leaves/plant and dry matter accumulation were influenced
significantly due to mulches and moisture regimes (Table 1). All these growth attributes were higher
under drip at 0.8 ETc and was similar to drip at 0.4 ETc and significantly higher than conventional
irrigation. Except surface coir pith application and no mulch control all other mulches recorded
significantly taller plants, more number of leaves and higher dry matter accumulation and were on par
with each other.
TABLE 1: GROWTH AND YIELD ATTRIBUTES OF EXTRA LONG STAPLE BT COTTON CV. RCHB 708 BT AS INFLUENCED BY MOISTURE REGIMES AND MULCHES
Treatments Plant ht (cm) Leaves/
Plant
Sympodia/
Plant
Bolls/Plant Boll wt.
(g/boll)
Mulches
No mulch control 118.7 229.5 22.7 44.7 5.90
Sub soil coir pith (2 kg/ m
2
) 145.7 261.5 24.4 49.1 5.90
Maize stover (5 kg / m
2
) 146.2 263.0 24.6 51.7 6.13
Sugarcane trash (5 kg / m
2
) 147.2 264.0 24.2 50.7 6.00
Surface coir pith (2 kg/ m
2
) 120.5 245.0 23.1 45.2 5.83
Gunny sheet mulching 145.0 260.0 24.5 49.8 6.20
Biodegradable poly mulching 152.5 288.2 25.5 72.3 6.10
Poly ethylene mulching 154.2 295.4 26.2 79.4 6.30
CD(P = 0.05) 16.8 41.5 1.35 7.92 NS
Moisture regimes
0.4 ETc (Drip) 152.0 285.2 24.9 59.0 6.25
0.8 ETc (Drip) 156.2 297.0 25.3 56.6 5.93
Conventional irrigation 115.5 207.8 23.0 50.5 5.98
CD(P = 0.05) 7.34 18.12 0.59 3.45 NS
CD (p=0.05) for interaction NS NS NS 13.30 NS
Yield Attributes
TABLE 2: BOLL NUMBERS IN EXTRA LONG STAPLE BT COTTON CV. RCHB 708 BT AS INFLUENCED BY INTERACTION BETWEEN MOISTURE REGIMES AND MULCHES
Mulches Moisture Regimes
Drip
(0.4 ETc)
Drip
(0.8 ETc)
Conventional
Irrigation

Mean
No mulch control 40.3 54.3 39.5 44.7
2
) 53.9 48.1 45.2 49.1
2
) 54.7 52.9 47.5 51.7
2
) 52.2 49.0 50.8 50.7
2
) 49.1 46.0 40.5 45.2
Gunny sheet mulching 54.4 50.6 44.4 49.8
Biodegradable poly mulching 78.2 72.9 65.9 72.3
Poly ethylene mulching 89.3 78.7 70.0 79.4
Mean 59.0 56.6 50.5
CD (p=0.05) for mulches 7.92
CD (p=0.05) for moisture regimes 3.45
CD (P=0.05) for interaction 13.30
The ELS cotton RCHB 708 Bt responded significantly to the type of mulches, moisture regimes and their
interaction (Table 2). The results indicated that under no mulch condition, the ELS Bt cotton responded
up to 0.8 ETc through drip which recorded 54.3 bolls/plant as against 39.5 bolls/plant at conventional
irrigation. In general wherever mulches were applied, the response was obtained up to 0.4 ETc and the
bolls number started decreasing beyond 0.4 ETc. Polyethylene mulching and biodegradable polyethylene
mulching recorded higher number of bolls/plant and was on par and significantly superior to the rest of
the treatments. Boll weight was not significantly altered either due to mulches or due to moisture
regimes.
Seed Cotton Yield (SCY)
SCY followed trend similar to that of boll numbers. The interaction effect between mulches and moisture
regimes was significant (Table 3). Under no mulch condition, RCHB 708 Bt responded up to 0.8 ETc
through drip and SCY was 44.5 % greater over conventional irrigation. However, under mulched
condition, the response was seen only up to 0.4 ETc and the SCY started decreasing at 0.8 ETc. Among
the mulches, polyethylene mulching and biodegradable poly ethylene mulching recorded significantly
higher SCY than the other mulches. Among the combinations, irrigation through drip at 0.4 ETc with
polyethylene mulching recorded the highest seed cotton yield of 5641kg/ha and was on par with
biodegradable polyethylene mulching (5234 kg/ha) at the same moisture level and polyethylene mulching
under conventional irrigation and biodegradable polyethylene mulching under conventional irrigation.
Thus, we infer that when polyethylene mulching and biodegradable polyethylene mulching are used even
the conventional irrigation is sufficient to get the desired yield. In terms of conserving moisture further
we can combine the polyethylene or biodegradable polyethylene with drip irrigation. Higher yield and
better growth under mulched condition at lower moisture regime were due to better moisture
conservation and favourable condition for plant growth. Ramakrishna et al. (2006) reported 94.5% higher
yield in groundnut due to polyethylene mulching. Elias and Goldhamer (1991) have reported 39%
enhanced yield due to mulching in cotton crop in California, USA.
TABLE 3: SEED COTTON YIELD IN EXTRA LONG STAPLE BT COTTON CV. RCHB 708 BT AS INFLUENCED BY INTERACTION BETWEEN MOISTURE REGIMES AND MULCHES
Mulches Moisture regimes
Drip
(0.4 ETc)
Drip
(0.8 ETc)
Conventional
Irrigation

Mean
No mulch control 3175 4070 3421 3555
2
) 4314 4067 4092 4158
2
) 4254 4215 3979 4149
2
) 4208 4077 4002 4096
2
) 4111 3809 3442 3787
Gunny sheet mulching 4354 4250 3994 4199
Biodegradable poly mulching 5234 4647 4679 4853
Poly ethylene mulching 5641 5418 5357 5472
Mean 4411 4319 4121 4284
CD (p=0.05) for interaction 928.5
Water Use Efficiency
TABLE 4: DRY MATTER PRODUCTION, WATER USE EFFICIENCY AND NUTRIENTS UP TAKE OF BT COTTON CV. RCHB 708 BT AS INFLUENCED BY MOISTURE REGIMES AND MULCHES
Treatments DMP
(t/ha)
WUE* Nutrients Uptake (kg/ha)
N P
2
O
5
K
2
O
Mulches
No mulch control 4.80 58.4 120.0 25.4 134.4
2
) 6.30 70.2 158.1 34.0 217.9
2
) 6.39 68.9 160.4 35.5 215.5
2
) 6.45 68.7 162.5 35.8 216.4
2
) 5.15 63.3 127.2 26.3 164.8
Gunny sheet mulching 6.42 69.2 161.8 34.0 215.4
Biodegradable poly mulching 6.61 81.8 172.4 35.6 218.1
Poly ethylene mulching 6.98 91.6 177.3 35.4 230.3
CD(P = 0.05) 1.68 19.35 1.41 15.38
Moisture regimes
0.4 ETc (Drip) 6.59 95.2 165.4 35.9 210.3
0.8 ETc (Drip) 6.75 68.6 170.8 36.4 216.0
Conventional irrigation 5.05 50.7 128.7 25.9 178.5
CD(P = 0.05) 0.73 8.44 0.62 6.72
CD (p=0.05) for interaction NS NS NS 24.10
*WUE = water use efficiency (Kg of seed cotton/ha cm of water used)
The water use efficiency as measured by the ratio of seed cotton produced to the water consumed by the
cotton crop ranged from 58.4 to 91.6 kg/ha cm, with the highest values recorded for the poly ethylene
mulching followed by biodegradable poly mulching (Table 4) and the least with no mulch. Among the
moisture regimes, drip at 0.4 ETc recorded the highest WUE which was probably due to better moisture
conservation due to mulches at lower moisture regime of 0.4 ETc.
Uptake of Nutrients
The uptake of nutrients by ELS cotton, RCHB 708 was influenced significantly by mulches and moisture
regimes (Table 4). Higher temperature and faster mineralization under mulching might have caused
increased uptake of nutrients and the favourable rhizospheric and phyllospheric microbes such as
Diazotrophs, Facultative methylotrophs, Azospirillum, Phosphorus Solubilizing Bacteria (PSB) and
Vesicular Arbuscular Mycorrhizae (VAM) were higher by 2 to 3 fold due to mulching (Nalayini et al.,
2009). The enhanced temperature under mulching coupled with higher available soil moisture regime
was highly favourable for the rhizosphere and phyllosphere microorganism. The temperature increase
due to mulching has been reported earlier (Chen and Yin, 1989 and Chakraborty and Sadhu, 1994). All
other mulches except no mulching and surface coir recorded significantly higher uptake of N, P
2
O
5
and

K
2
O. The significant reduction in uptake of these nutrients in surface coir application might be due to
higher C/N ratio of the coir material when it is applied on the surface of the soil which caused stunted
growth and lesser dry matter accumulation by the cotton crop. However, when the coir was placed deep
using coir pith applicator, it was as good as application of other plant wastes like sugarcane trash or
maize stover mulching. Among the moisture regimes, drip at 0.4 ETc and 0.8 ETc were similar and
significantly superior to conventional irrigation.
Fibre Quality
Fibre quality parameters (Span length, strength, micronaire, uniformity and maturity ratio) were not
influenced significantly due to the mulches or moisture regimes.
Zero Tilled Rotation Maize
TABLE 5: GRAIN YIELD OF ZERO TILLED ROTATION MAIZE AFTER THE HARVEST OF COTTON AS INFLUENCED BY MOISTURE REGIMES AND MULCHES

Mulches
Moisture regimes
Drip
(0.4 Etc)
Drip
(0.8 Etc)
Conventional
Irrigation
Mean
No mulch control 4.00 4.80 4.13 4.31
Sub soil coir pith 6.27 6.23 6.20 6.23
Maize stover 6.23 6.40 6.23 6.29
Sugarcane trash 6.30 6.40 6.27 6.32
Surface coir pith 2.70 2.63 2.70 2.68
Gunny sheet mulching 6.37 6.40 6.13 6.30
Biodegradable poly mulching 6.23 6.77 6.27 6.42
Poly ethylene mulching 6.57 6.40 6.50 6.49
Mean 5.58 5.75 5.55
CD (p=0.05) for interaction 0.32
The treatment surface coir application resulted in the lowest maize grain yield (Table 5). This may be due
to wider C/N ratio of the coir waste (how much) when applied on the surface. All the mulches except
surface coir mulching recorded higher maize grain yield over no mulch control might be due to moisture
conservation and also due to supply of nutrients due to degradation of crop waste applied as mulches over
period of six to eight months. Among the moisture regimes, drip at 0.8 ETc recorded the highest maize
grain yield followed by drip at 0.4 Etc and conventional method. The efficient moisture conservation
achieved at lower moisture regime of 0.4 ETc for cotton crop could not be achieved for maize crop as the
applied mulches started degradation over a period of time.
CONCLUSION
Our studies suggest biodegradable polyethylene was as good as polyethylene mulch for Bt cotton. Crop
based mulches like sugarcane trash mulching, maize stover mulching and sub-soil application of coir
waste could be considered as next only to polyethylene and biodegradable polyethylene mulching.
Surface coir application was not useful. Under no mulch condition ELS cotton responded up to 0.8 ETc.
while under mulched condition, the crop responded only up to 0.4 ETc and the yield level started
decreasing at 0.8 ETc. Hence, whenever mulching is used we can reduce irrigation water requirement by
half. Polyethylene mulching and biodegradable polyethylene mulching with irrigation at 0.4 ETc through
drip system was similar to conventional irrigation with mulch. Hence, whenever polyethylene mulching
and biodegradable polyethylene mulching were used, even the conventional irrigation is sufficient to get
the desired yield.
REFERENCES
[1] Anon, 2004. Degradable Plastics-Breaking news Down Under. Australian Government Department of the Environment and
Heritage, News letter, Issue 2, 15.
[2] Chakraborty, R.C. and Sadhu, M.K.1994. Effect of mulch type and colour on growth and yield of tomato (Lycopersicum
esculentum) Indian Journal of Agricultural Sciences 64: 608612.
[3] Chen, Q.E. and Yin, J.S.1989. Effects of plastic mulch on soil properties and cotton growth in saline alkali soils. Journal
of Soil Science China 20: 13.
[4] Elias, Fereres and Goldhamer, David A. 1991. Plastic mulch increases cotton yield, reduces need for pre-season irrigation.
California Agriculture 45(1): 2528
[5] Kijchavengkul,T.,Auras,R.,Rubino,M.,Ngouajic,M.,Fernendez,R.T.2008.Assessment of aliphatic aromatic copolyester
biodegradable mulch films.Part I: field study. Chemosphere 71(5), 942953.
[6] Michael Stephenson.2009. Biodegradable plastics. In: Proceedings of the Plas Tec2008- International Conference on
Plastics and Environment held during 22
nd
-23
rd February
, 2008 at Chennai pp 127144. Organized by Chennai Plastics
Manufacturers and merchants Association.
[7] Nalayini, P., Raja, R and Anderson A. Kumar.2006.Evapotranspiration based scheduling of irrigation through drip for
cotton (Gossypium hirsutum). Indian Journal of Agronomy 51(3): 232235.
[8] Nalayini, P., Anandham, R., Sankaranarayanan. K and Rajendran, T.P. (2009) -Polyethylene mulching for enhancing crop
productivity and water use efficiency in cotton (Gossypium hirsutum) and maize (zea mays) cropping system. Indian
Journal of Agronomy 54(4): 409419.
[9] Ramakrishna, A., Hoang Minh Tam., Suhas P. Wani, Tranh dinh Long. 2006. Effect of mulch on soil temperature,
moisture, weeds infestation and yield of groundnut in northern Vietnam. Field Crops Research 95: 115125.
Comparative Study of Different Weeding Methods

on Cotton Crop under Drip Irrigation System
Dil Baugh Muhammad
1
, M.N. Afzal
2
, I. Raza
2
and P.L. Dupont
3

1
Senior Scientific Officer/Head, Agronomy Section, Central Cotton Research Institute, Multan
2
Scientific Officer, Agronomy Section, Central Cotton Research Institute, Multan
3
Director, M/S. Jaffer Agro Services (Pvt.) Ltd.
AbstractWeeds are the most efficient users of resources due to their different kinds, intensity, fast growth and soil
coverage habits. It is important to keep weeds below the critical threshold level where the use of tractor drawn
implements is not possible. Field experiment was conducted in two consecutive years (2009 & 2010) to evaluate the
effect of different weed control measures on weed intensity, seed cotton yield, its components and fibre characteristics
under drip irrigation. Treatments comprised pre-emergence application of Stomp 330E (Pendimethalin 33%) @ 2.5
litre/ha, Dual Gold 960EC (s-metolachlor) @ 2.0 litre/ha, and post emergence alone with Mera -71SG (Ammonium
salt of Glyphosate) @ 2.0 kg /ha, or following the pre-emergence treatment, respectively. (Stomp 330E + Mera -71SG,
Dual Gold 960EC + Mera-71S), plastic mulching, manual weeding and untreated check. Treatments were arranged in
a randomized complete block design with four replications. Cotton cultivar CIM-573 was dibbled manually on top of
the beds at the Experimental Research Area of Central Cotton Research Institute, Multan on silty loam soil. Results
indicated that among chemical weeding treatments, the greatest yield gains were obtained by pre emergences followed
by post emergence than pre emergence alone and post emergence alone. Furthermore, chemical weeding methods
significantly broad and narrow leaved weeds than untreated. Dual Gold 960EC + Mera -71SG resulted in 98.5 and
98.6 % broad and narrow leaved weeds control, respectively at 60 days after sowing over untreated. Among
treatments, manual weeding (3 times) produced the maximum seed cotton yield among the treatments which was
167.8% more than untreated. Whereas, plastic mulching resulted in the lowest (49.5%) increase in seed cotton yield
over untreated. Fiber characteristics were not affected by any of the treatments.
Keywords: Cotton (Gossypium hirsutum L.), Pre- and post-emergence weedicides, Weed intensity, Seed cotton
yield. Drip irrigation
INTRODUCTION
TABLE 1: THE MAJOR WEED FLORA OF COTTON IN SOUTHERN PUNJAB, PAKISTAN
Botanical Name Common name Local Name
Euphorbia prostrata Petty spurge Dhodak
Convolvulus arvensis Field bindweed Lehli
Cynodon dactylon pers Bermuda grass Khabbal ghass
Cyperus rotundus Purple nutsedge Deela
Portulaca oleracea Common purslane Kulfa
Sorghum halepense Johnsongrass Baru
Trianthema monogyna Horse purslane It-Sit
Amaranthus viridis Green amaranth Chulai
Echinochloa colonum Jungle rice Swanki ghass
Setaria viridis Green foxtail Loomar ghass
Euphorbia helioscopia Sun spurge Dhodak
Corchorus tridens Wild jute Jangli Patsun
Digeria arvensis Kundra Tandla
Tribulus terrestris Pucturevine Bhakhra
Cotton (Gossypium hirsutum L.) is the leading fibre and cash crop of Pakistan and major source of
foreign exchange earning. The crop always remains at risks and suffers production constraints due to
many factors including weeds, insect pests and diseases. Weeds are responsible for losses in cotton yield
to an extent of 34-61.4% (Ahmad, 1995) Therefore, to enhance productivity weeds should be controlled
either by cultural, mechanical, biological and chemical methods. The major weed flora observed in cotton
fields of southern Punjab of Pakistan are presented in Table-1.
65
Comparative Study of Different Weeding Methods 393
In developing countries, farmers are reluctant to adopt chemical weed control methods due to lack of
comprehensive knowledge about its use. Weed management becomes complicated when mechanical
weeding is not possible and manual weeding becomes very expensive. Furthermore, manual labourers are
not available when needed. The highest weed control was achieved by the application of pre emergence
weedicide (Pendimethalin) followed by post emergence weedicide (Glyphosate) application with two
hand weedings and two hoeings at 20 and 40 days (Deshpande et al., 2006). Similarly, application of S-
metolachlor and Glyphosate-TM AEPOST increased cotton lint yield (Clewis et al., 2008). Where as,
others use plastic mulching to alter soil temperature, crop water use, reducing weed competition for other
resources, improving crop quality and development and enhancing yields (Lamont, 2005). The present
study was conducted to evaluate the effect of different weed control measures on yield, yield components
and fibre characteristics of cotton crop under drip irrigation.
Field experiments were conducted during 2009 and 2010 at the Agronomic Research Area of Central
Cotton Research Institute, Multan. The experiment were laid out in a randomized complete block design
with four replications on well prepared soil and bed furrows were made 75cm apart by tractor driven
implements. Bed shaping was done having ditches 75cm apart from each other and from top 45cm on
raised sides to support irrigation water by drip. Cotton cultivar CIM-573 was dibbled manually on inner
side of raised edges at the top of the beds on 17 May 2009 and 12 May 2010 on silt loam soil. Thinning
was done 22 days after sowing (DAS) by making single plant per hill. Stomp 330E @ 2.5 litre/ha
-1

(Pendimethalin 0.825 kg a.i ha
-1
) and Dual Gold 960EC @ 2.0 litre/ha
-1
(S-metolachlor 1.92 kg a.i/ ha
-1
)
were sprayed soon after planting on moist soil with knapsack hand sprayer. The post emergence
herbicide Mera -71SG (Ammonium salt of Glyphosate) @ 2.0 kg /ha was sprayed 35 DAS in specific
treatments as protective spray by using shield with spray nozzle. Manual weeding was done at 24, 40 and
52 DAS during 2009 and 25, 41 and 55 DAS during 2010. Removal of weeds by manual weeding from
all treatments including control plots was done 70 DAS. All kinds of uprooted weeds were removed from
the field. The data were statistically analyzed by using Fishers analysis of variance techniques and least
significance difference test at 5% probability level applied to compare the significance of the treatment
means (Steel and Torrie, 1997).
Data presented in Table 2 showed that all the pre-emergence and mechanical (mulching & manual) weed
control methods gave significant weed control over untreated. The dry weight of weeds 30 DAS with
Stomp 330E and dual Gold 960EC was reduced by 75 to 86 than the untreated. Among pre-emergence
weedicides, Dual Gold 960EC produced better weed control as compared to Stomp 330E. Mulching
method resulted in 100.00 and 95.8% control of broad and narrow leaved weeds, respectively over
untreated. Manual weeding resulted in 89.7 and 91.0% broad and narrow leave weeds control over
untreated, respectively. In case of mechanical weed control methods; mulching gave 10.3% broad leave
weeds and 4.8% narrow leave weeds control higher than manual weeding. Similar results were also found
by earlier researchers (Ali et al., 2005).
TABLE 2: DRY WEIGHT OF WEEDS AND WEED CONTROL %AGE 30 DAYS AFTER SOWING
Dry weight (g m
-2
) %age Weed Control
Broad
Leaves
Narrow
Leaves
Broad
Leaves
Narrow
Leaves
Stomp 330E @ 2.5 L ha
-1
at sowing 15.6 24.4 81.6 75.1
Dual Gold 960EC @ 2.0 L ha
-1
at sowing 11.6 14.4 86.3 85.3
Mulching (Plastic) at sowing 0.00 4.1 100.00 95.8
Manual weeding 8.7 8.8 89.7 91.0
Untreated check 84.6 97.9 - -
C.D 5% 2.270 2.433 - -
Post-emergence weedicide application

gave near total control of both broad and narrow leaved weeds
over untreated, respectively at 60 DAS (Table-3). Interactive effect of Stomp 330E as pre-emergence and
Mera 71SG @ 2.0 kg ha
-1
as post emergence also resulted in 98.0 and 98.3% broad and narrow leave
weeds control over untreated, respectively. Similarly, Dual Gold 960EC pre-emergence along with Mera
71SG post-emergence gave effective weed control over untreated, respectively. Mulching and manual
method of weeding reduced weed biomass significantly compared to untreated. Sheikh et al. (2006)
reported similar results.
TABLE 3: DRY WEIGHT OF WEEDS AND WEED CONTROL %AGE 60 DAYS AFTER SOWING
Treatments Dry Weight (g m
-2
) % Age Weed Control
Broad
Leaves
Narrow
Leaves
Broad
Leaves
Narrow
Leaves
-1
at sowing 122.2 78.5 40.3 63.5
-1
at sowing 116.6 72.7 43.1 66.2
-1
37 days

after sowing
3.2 3.2 98.4 98.5
-1
+
-1
4.1 3.6 98.0 98.3
-1
+
-1
3.0 3.1 98.5 98.6
Mulching (Plastic) at sowing 1.7 21.4 99.2 90.1
Manual weeding 4.7 4.8 97.7 97.8
Untreated check 204.8 215.1 - -
C.D 5% 2.588 1.792 - -
Plant growth and seed cotton yield were significantly better with pre-emergence, post-emergence,
mulching and manual weed control methods alone or in combination than the untreated (Table-4). Stomp
330E, Dual Gold 960EC and Mera 71SG alone produced 91.7, 101.4 and 63.1% higher seed cotton yield
over untreated, respectively. Among weedicides, Mera 71SG (post emergence) applied at day 35 after
sowing gave less increase in seed cotton yield as compared to pre-emergence weedicides due to initial
weed-crop competition. Pre-emergence weedicides (Stomp 330E and Dual Gold 960EC) alongwith post-
emergence weedicide increased seed cotton yield by 116.2 and 141.5% over untreated, respectively.
Among treatments, manual weeding showed maximum increase in seed cotton yield (167.8%) over
untreated. Mulching (Plastic film) gave smallest (49.5%) increase in seed cotton yield over untreated.
This was probably due to low evaporation and less aeration which may have affected root expansion and
limiting plant growth. These results are in line with the findings of other scientists including Cheema et
al. 2005 and Dilbaugh et al. 2009).
TABLE 4: SEED COTTON YIELD AND ITS COMPONENTS INFLUENCED BY DIFFERENT WEED CONTROL METHODS ON BED-FURROW PLANTING UNDER DRIP IRRIGATION
Treatments Plant height cm) Number of bolls m
-2
Boll weight (g) Seed cotton yield (kg ha
-1
)
-1
83.8 68 2.81 1641
-1
87.5 73 2.80 1724
-1
68.2 56 2.76 1396
-1
+
-1
86.6 79 2.78 1851
-1
+
-1
88.8 88 2.80 2067
Mulching (Plastic) 67.1 54 2.78 1280
Manual weeding 91.5 96 2.85 2292
Untreated check 61.3 35 2.68 856
C.D 5% 7.530 3.647 NS 256.418
The economics of all the weeding methods tested in this study are presented in Table 5 showed that
under mechanical methods manual weeding (3 times) gave maximum net profit Rs.107549 ha
-1
followed
by chemical weeding control method (Dual Gold 960EC + Mera 71) produced maximum net profit of
Rs.92777 ha
-1
. It is clear from the results that use of pre-emergence with post-emergence weedicide is
necessary for profitable cotton production on bed-furrow under drip irrigation system. Ali et al (2005),
Cheema et al. (2005) and Dilbaugh et al. (2009) reported similar results on cotton planted under bed-
furrow planting technique.
Comparative Study of Different Weeding Methods 395
TABLE 5: ECONOMICS ANALYSIS OF DIFFERENT WEEDING METHODS
Weeding Method Input Cost of Variable
Factor (Rs ha
-1
)
Value of Increased Seed Cotton Yield
over no Weeding (Rs. ha
-1
)
Net
Profit
Stomp 330E@ 2.5 L ha
-1
1795 62564.5 60769.5
Dual Gold 960EC@ 2.0 L ha
-1
1540 69179.6 67639.5
Mera 71SG@ 2.0 kg ha
-1
2200 43038.0 40838.0
-1
+ Mera
71SG @ 2.0 kg ha
-1

3995 79301.5 75306.5
-1
+
-1

3740 9516.7 92776.7
Mulching (plastic) 3848 33792.8 29944.8
Manual weeding 6900 114449.2 107549.2
Untreated check - - -
Stomp 330E = Rs.718/lit.
Dual Gold 960EC = Rs.770/lit.
Mera 71SG = Rs.1100/kg
Manual weeding = Rs.115/day
Price of Seed cotton = Rs.3188/40 kg
With regard to fiber characteristics, staple length (25.9 -27.1 mm) micronaire (5.2 to 5.4) and fiber
strength (27.1 to 28.2 g/tex) of cotton lint was not influenced by different weeding management methods
either by chemical, manual or mulching.
CONCLUSION
Weed management under drip irrigation requires special attention. Weeds can be effectively controlled
by application of pre and post emergence weedicides. Mera 71SG (post-emergence) weedicide was
effective in control of perennial weeds. Though plastic mulching controlled weeds effectively, it did not
result in yield gains obtained with manual and chemical treatments. Net profit was the most with manual
weeding followed by chemical weed control.

REFERENCES
[1] Ahmad, Z. (1995) - Twenty-five years research activities at Central Cotton Research Institute, Multan from 197075,
pp. 6-7.
[2] Ali, M.A., Sabir, S., YAr, K., Ali, M. and Saeed, M. (2005) - Cultural and chemical weed management in cotton for higher
profitability in Pakistan. J. Agric. Res. 43(1): 51-59.
[3] Cheema, M.S., Akhtar, M. and Iqbal, M.S. (2005) - Evaluation of mechanical, chemical and manual weed control methods
in cotton. Pak. J. Weed Sci. Res. 11(3-4): 137-140.
[4] Clewis, S.B., Millere, D.K., Koger, C.H., Baughman, T.A., Price, A.J., Portefield, D. and Wilcut, J.W. (2008) - Weed
management and crop response with glyphosate, S- metolachlor, Trifloxysulfuron, Prometryn and Msma in glyphosate
resistant cotton. Weed Tech. 22(1): 160-167.
[5] Deshpande, R.M., Pawar, W.S., Mankar, P.S., Bobde, P.N. and Chimote, A.N. (2006) - Integrated weed management in
rainfed cotton (Gossypium hirsutum L.). Ind. J. Agron. 51(1): 68-69.
[6] Dilbaugh, M., Afzal, M.N., Raza, I. and Mian, M.A. (2009) - Effect of mechanical and chemical weed control on the
productivity of cotton. Pak. J. Weed Sci. Res. 15(2-3): 117-122.
[7] Lamont, W.J. (2005) - Plastics. Modifying microclimate for the production of vegetable crops. Hort Tech. 15: 477-481.
[8] Sheikh, M.A., Saleem, A. and Malik, N.A. (2006) - Integrated weed management and its effects on seed cotton yield in
cotton (Gossypium hirsutum L.) crop. Pak. J. Weed Sci. Res. 12(1/2): 111-117.
[9] Steel, R.G.D. and Torrie, J.H. (1997) - Principles and procedures of Statistics. A biometric approach, 2
nd
edition, McGraw
Hill Book Co. Inc. Tokyo.
Comparative Efficiency and Economic Viability

of Herbicides for Controlling Weeds in Bt Cotton
(Gossypium hirsutum L.)
J.G. Patel, V.C. Raj, V.P. Usadadiya, R.R. Parmar, C.M. Sutaria,
R.L. Leva and V. Kumar
Main Cotton Research Station, Navsari Agricultural University, Surat395007 India
INTRODUCTION
Cotton is the most important commercial crop of India, with an important role in the Indian economy.
Being a long duration and widely spaced crop, loss of seed cotton yield in India due to weeds ranged
from 50 to 85 per cent (Singh and Madhukar, 1981). Besides initial slow growth, continuous rains make
the soil wet and sticky and trafficability is poor. As a result, weeds grow profusely. Thus, weed
management plays a crucial role. Weeds being naturally hardy compete well with crop for moisture,
nutrients, light and space resulting in poor yield of crop. Looking to the variety of weed flora, feasibility
and conveniences, it is difficult to adopt any single method of weed management, thereby emphasizing
integrated approach in this regard. Keeping this in view, studies were conducted to compare efficiency
and feasibility of herbicides for weed control in Bt cotton.
A field experiment was conducted at Main Cotton Research Station, Navsari Agricultural University,
Surat under irrigated condition during 2007-08 to 2009-10. The soil of experimental site is deep black,
low in organic carbon (0.36), medium in available P (32.7 kg/ha) and high in exchangeable K (536
kg/ha). The experiment comprised ten treatments of weed management viz., weedy check (control),
farmers practices (Hand weeding at 20, 40 and 60 DAS and interculture at 45 and 90 DAS) and different
rates of herbicides (pendimethalin and fluchloralin as pre-emergence and Quizalofop ethyl as post
emergence). These treatments were replicated thrice in a randomized block design. Cotton (RCH-2 Bt)
was sown in the last week of June with a spacing of 120 cm between rows and 45 cm between plants.
Nitrogen was applied through urea in four equal split doses (60 kg/ha) at an interval of 30 days. The crop
was irrigated twice at 20 days interval after cessation of monsoon. Weed population was counted from 1
m
2
area at 30 and 60 DAS. Weed dry weight from the same fixed area in each plot was also recorded at
30 and 60 DAS and at harvest. Observations related to cotton growth and yields were also recorded.
Weed Density, Dry Weed Weight and Weed Control Efficiency
The maximum dry weight of weed was recorded at harvest in the weedy check (Table 1). All the
herbicides and cultural operation treatments decreased significantly weed density than weedy check. Low
weed density as well as dry weed weight was recorded in the pendimethalin (1.0 kg a.i./ha pre
emergence) along with hand weeding at 30 and 60 DAS treatment followed by fluchloralin (0.75 and 1.0
kg a.i./ha pre-emergence) with hand weeding 30 & 60 DAS and farmers practices. Weed control
efficiency was the highest under treatment of pendimethalin (1.0 kg/ha) + hand weeding at 30 and 60
DAS followed by fluchloralin (0.75 or 1.0 kg/ha) + hand weeding at 30 and 60 DAS and local practices.
The higher weed control efficiency with the herbicide treatments was attributed due to lower weed dry
bio-mass when compared to the non-herbicide treatments.
66
Comparative Efficiency and Economic Viability of Herbicides for Controlling Weeds 397
TABLE 1: WEED DENSITY, WEED DRY WEIGHT AND WEED CONTROL EFFICIENCY AS AFFECTED BY DIFFERENT WEED MANAGEMENT TREATMENTS
Treatments
30 DAS 60 DAS
Dry weed
Weight
(kg/ha)
Weed Control
Efficiency (%)
Weed
Density
(no/m
2
)
Dry Weed
Weight
(g/m
2
)
Weed
Density
(no/m
2
)
Dry Weed
Weight
(g/m
2
)
T
1
:Unweeded control 48.3 27.6 75.7 133.8 1478 --
T
2
:Farmers practices (HW at 20,40 &
60 DAS + IC at 45 & 90 DAS)
27.3 16.9 20.5 45.0 449 69.62
T
3
:Pendimethalin @ 0.75 kg a.i. /ha
pre-em.+ HW at 30 & 60 DAS
23.8 16.7 21.8 46.6 470 68.20
T
4
:Fluchloralin @ 0.75 kg a.i./ha pre-
em. + HW at 30 & 60 DAS
24.2 14.7 22.1 43.0 431 70.84
T
5
:Pendimethalin @ 1.00 kg a.i./ha pre-
em. + HW at 30 & 60 DAS
20.5 14.1 21.0 40.2 401 72.87
T
6
em. + HW at 30 & 60 DAS
21.3 15.8 20.5 44.1 442 70.09
T
7
:Pendimethalin @ 0.75 kg a.i./ha pre-
em.+ HW at 30 & 60 DAS
25.0 18.5 26.3 54.0 545 63.13
T
8
: Fluchloralin @ 0.75 kg a.i./ha pre-
em. + Quizalofop-ethyl @ 0.04 kg
a.i./ha at 30 & 60 DAS
22.5 15.6 28.0 64.2 641 56.63
T
9
:Pendimethalin @ 1.00 kg a.i./ha
pre-em. + Quizalofop-ethyl @ 0.05 kg
a.i./ha at 30 & 60 DAS
20.8 13.2 25.7 65.3 656 55.62
T
10
em. Quizalofop-ethyl @ 0.05 kg a.i./ha
at 30 & 60 DAS
17.8 14.5 23.0 52.3 525 64.48
S. Em. + 2.42 1.77 2.90 5.77 62.6 --
C D (P=0.05) 7.77 5.67 9.27 18.46 186.1 --
Growth, Yield and Yield Attributes
TABLE 2: GROWTH, YIELD AND ECONOMICS OF BT COTTON (RCH-2) AS INFLUENCED BY DIFFERENT WEED MANAGEMENT PRACTICES
Treatments
Plant Height
at Harvest
(cm)
No. of
Sympodial
Branches/ Plant
No. of
Bolls/
Plant
Boll
Weight
(g)
Seed
Cotton
Yield
(kg/ha)
Net
Return
(Rs/ha)
BCR
T
1
:Unweeded control 85.6 13.6 25.2 3.7 1780 29091 2.60
T
2
:Farmers practices (HW at 20,40
& 60 DAS+ IC at 45 & 90 DAS)
96.3 16.1 31.9 3.9 2353 41262 3.00
T
3
:Pendimethalin @ 0.75 kg a.i. /ha
94.4 15.7 35.1 4.0 2575 47623 3.30
T
4
:Fluchloralin @ 0.75 kg a.i./ha
pre-em. + HW at 30 & 60 DAS
92.4 15.6 32.0 3.9 2315 40738 3.00
T
5
97.7 17.6 38.9 4.0 2753 51667 3.50
T
6
:Fluchloralin @ 1.00 kg a.i./ha
96.4 16.1 35.8 3.8 2545 46184 3.30
T
7
93.5 14.7 31.0 3.9 2152 36768 2.80
T
8
: Fluchloralin @ 0.75 kg a.i./ha
pre-em. + Quizalofop-ethyl @ 0.04
kg a.i./ha at 30 & 60 DAS
90.3 15.1 30.1 3.9 2128 36212 2.80
T
9
: Pendimethalin @ 1.00 kg a.i./ha
92.2 15.1 34.3 3.9 2504 45745 3.20
T
10
: Fluchloralin @ 1.00 kg a.i./ha
95.1 16.5 36.3 4.0 2629 48788 3.40
S. Em. + 2.9 0.63 1.55 0.11 79.4 - -
C D (P=0.05) NS 1.79 4.62 NS 224 - -
Most of the growth and yield attributes were affected remarkably due to different weed management
treatments (Table 2). All herbicidal treatments as well as hand weeding at 20, 40 and 60 DAS and inter-
culturing at 45 and 90 DAS resulted in more number of sympodial branches, bolls per plant and larger
bolls than the weedy check. The highest seed cotton yield was recorded under the treatment of
pendimethalin (1.0 kg/ha) pre emergence + hand weeding at 30 and 60 DAS and the lowest in the weedy
check treatment. The higher yield under the integrated weed management practices may be attributed to
better weed control and reduced crop weed competition in early growth stage, ultimately increase the
cotton yield.
Economics
In terms of economics, the maximum net monetary returns (Rs.51667/ha) and BCR (3.50) was accrued
under the treatment of pendimethalin @ 1.0 kg/ha as pre-emergence + HW at 30 and 60 DAS.
CONCLUSION
It is concluded that among various weed management practices the treatment with application of
pendimethalin (1.0 kg/ha) pre emergence alongwith two hand weeding dose at 30 and 60 DAS was the
best with respect to growth and yield of cotton crop. The treatment also had low weed infestation. Thus,
an integrated weed management practice integrating chemical and manual control is effective, efficient
and economical to control weeds in Bt cotton.
REFERENCES
[1] Singh, C. and Madhukar, M.J. (1981) - Review on loss of cotton due to weeds. In: Proceeding of Weed Science
Conference, Bangalore, 1981.
Agronomic Management and Benefits

of Glyphosate Tolerant Transgenic
Cotton Hybrids
C. Chinnusamy, C. Nithya, P. Muthukrishnan and S. Jeyaraman
DWSRC, Department of Agronomy, Tamil Nadu Agricultural University,
Coimbatore641003, India
E-mail: chinnusamyc@gmail.com
AbstractCrops made resistant to herbicides by biotechnology are being widely adopted in North America and
entering other parts of the world. Those containing transgenes that impart resistance to post-emergence, non-selective
herbicides such as glyphosate and glufosinate will have a major impact. These products allow the farmer to more
effectively use reduced or no-tillage, eliminate use of some environmentally suspect herbicides and use fewer herbicides
to manage the entire spectrum of weed species. Introduction of herbicide-resistant crops has dramatically changed weed
management in crop production systems. Particularly, glyphosate-resistant cotton technology has been readily adopted by
producers. Further, wide scale introduction of glyphosate-resistant (Round up Ready) cotton cultivars in addition to the
herbicides flumioxazin, pyrithiobac and trifloxysulfuron-sodium has increased options for weed management in cotton. Farmers
have a powerful new tool that, if used wisely, can be incorporated into an integrated pest management strategy to
economically and effectively manage weeds.
Keywords: Agronomic management, herbicide tolerant cotton, weed density, weed dry weight, weed control
efficiency, seed cotton yield
INTRODUCTION
Cotton hybrids are cultivated under wider plant spacing and heavy fertilizer nutrients, which inturn invite
multiple weed species infestation. Due to increased scarcity of labourers, manual weeding is seldom
economical and the currently available pre - emergence herbicides have lesser weed control efficiency in
controlling major problematic weeds like Cyperus rotundus and Cynodon dactylon for a longer initial
growth phase of cotton hybrids. Many pre-emergence herbicides presently used in cotton for weed
control take care of weeds for a limited period. Hence, late emerging weeds escape killing. The available post-
emergence herbicides are mostly non-selective and even directed spray of some herbicides cause
considerable crop damage. So, there is an ample scope for controlling weeds by the application of early
post-emergence herbicide.
Glyphosate resistant cotton was introduced in 1997 and revolutionized weed control in cotton. Round
up Ready cotton provided producers with greater flexibility in the timing of herbicide application and
also offered a broad spectrum of weed control than other systems. Glyphosate and glufosinate have
almost no soil residual activity because they are tightly bound to the organic particles in the soil. This
trait facilitates crop rotation by providing flexibility in selection of potential rotation crops. Glyphosate
tolerant crops will not cause any residual effect on succeeding crops.
Enhanced glyphosate resistant (Round up Ready Flex) cotton was introduced in 2006. RR Flex
cotton exhibits both vegetative and reproductive tolerance to glyphosate to be applied POST over the top
at any growth stage, without risk of boll abortion. The technology has allowed growers to reduce or
eliminate soil-applied herbicides and to abandon cultivation. It also has allowed a shift to conservation
tillage. Glyphosate-resistant technology has gained tremendous acceptance in cotton producing states since
commercialization, with the majority of acreage planted to cultivars containing this herbicide-resistant trait.
Increased dosages and an extended application time are beneficial since glyphosate provides broad-spectrum
control of many annual and perennial grasses, sedges, and broadleaf weeds. RRF cotton has potential benefits,
including an expanded window for POE glyphosate applications, enhanced application flexibility and
67
convenience, increased production efficiencies, less dependence on selective spray equipment and the ability
to tailor herbicide applications to the intensity of weed infestation instead of cotton growth stage. Therefore,
studies were conducted to test selective early post emergence herbicides for weed control in cotton.
Field experiments were conducted at Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu
during winter seasons of 2009-10 and 2010-11 with the objectives, to evaluate the weed control
efficiency of glyphosate on herbicide tolerant transgenic stacked cotton and to study the agronomic
management and benefits of herbicide tolerant transgenic cotton. The experiments were laid out in
randomized block design with treatments replicated thrice. In which, the treatments consisted of
glyphosate at 900 (T
1
), 1350 (T
2
), 1800 (T
3
), 2700 (T
4
) and 3600 (T
5
) and 5400 (T
6
) g a.e ha
-1
and hand
weeding twice at 15 and 30 days after sowing DAS (T
7
) along with unweeded check (T
8
) were imposed.
Glyphosate spraying was done twice (25 and 65 DAS) in case of herbicide tolerant transgenic cotton.
Agronomic Benefits of Herbicide Tolerant Cotton
Broader Spectrum of Weed Control
TABLE 1: EFFECT OF GLYPHOSATE APPLICATION ON TOTAL WEED DENSITY (NO. M
-2
), DRY WEIGHT (G M
-2
) AND WEED CONTROL EFFICIENCY (WCE %)
IN HERBICIDE TOLERANT TRANSGENIC COTTON
Treatments Winter, 2009-10 Winter, 2010-11
25 DAFHS 25 DASHS 25 DAFHS 25 DASHS
Density Dry
Weight
WCE Density Dry
Weight
WCE Density Dry
Weight
WCE Density Dry
Weight
WCE
Gly. 900 g a.e ha
-1
3.31
(9.00)
2.57
(4.58)
93.36 3.26
(8.67)
3.05
(7.33)
92.79 3.99
(14.0)
2.80
(5.28)
91.27 4.13
(13.0)
2.91
(6.47)
92.33
Gly. 1350 g a.e
ha
1

2.94
(6.67)
2.33
(3.43)
95.02 2.81
(6.00)
2.45
(4.13)
95.95 3.74
(12.0)
2.65
(5.08)
92.44 3.77
(10.0)
2.35
(3.57)
95.96
Gly. 1800 g a.e
ha
-1

2.43
(4.00)
1.84
(1.26)
97.97 2.58
(4.67)
1.97
(1.90)
98.16 2.94
(6.67)
2.29
(2.92)
95.12 3.22
(8.33)
1.98
(1.93)
98.00
Gly. 2700 g a.e
ha
-1

2.00
(2.00)
1.77
(1.03)
98.37 2.23
(3.00)
1.87
(1.50)
98.55 2.23
(3.00)
2.11
(2.21)
96.29 2.66
(7.00)
1.90
(1.60)
98.42
Gly. 3600 g a.e
ha
-1

1.80
(1.33)
1.76
(1.00)
98.41 1.80
(1.33)
1.82
(1.02)
98.75 1.80
(1.33)
1.92
(1.58)
97.34 2.55
(7.00)
1.81
(1.27)
98.83
Gly. 5400 g a.e
ha
-1

1.41
(0.00)
1.41
(0.00)
100.0 1.61
(0.00)
1.41
(0.00)
99.70 1.41
(0.00)
1.85
(0.22)
97.81 2.08
(2.67)
1.64
(0.52)
99.54
HW on 15 & 30
DAS
5.46
(28.00)
2.26
(3.12)
95.48 7.22
(50.3)
5.56
(29.2)
71.24 6.53
(40.7)
4.34
(17.0)
74.91 7. 36
(50.0)
6.08
(35.0)
56.67
Unweeded check 10.60
(110.7)
8.42
(69.0)
- 9.98
(97.7)
10.16
(101.3)
- 11.01
(119.3)
8.36
(67.9)
- 11.04
(109.7)
9.07
(80.3)
-
SEd 0.32 0.25 - 0.29 0.29 - 0.30 0.21 - 0.35 0.16 -
CD (P=0.05) 0.68 0.45 - 0.63 0.65 - 0.62 0.44 - 0.75 0.34 -
Figures in parenthesis are original values; DAFHS - Days after first herbicide spray; DASHS - Days after second herbicide
spray
Non-selective herbicides such as glyphosate and glufosinate aid in broadening the spectrum of weeds
controlled, which is particularly important in no-till (NT) systems, and those weedy fields. Weed flora
of the experimental field predominantly consisted of twelve species of broad-leaved weeds, five species of
grasses and a sedge weed. Dominant among grassy weeds were Dactyloctenium aegyptium Beauv. and
Cynodon dactylon (L.) Pers. Trianthema portulacastrum (L.), Cleome gynandra (L.), Digera arvensis (Forsk)
and Parthenium hysterophorus (L.) were the dominant ones among the broad-leaved weeds. Cyperus
rotundus (L.) was the only sedge present in the experimental fields. Experimental results revealed that
application of glyphosate at 2700 g a.e ha
-1
recorded lower weed density, dry weight and higher weed control
efficiency when compared to other doses of glyphosate and hand weeding method (Table 1). According to
Agronomic Management and Benefits of Glyphosate Tolerant Transgenic Cotton Hybrids 401
France et al., (1997) in addition, the systemic activity of glyphosate also helped with the control of
perennial weeds and their perennial vegetative structures such as stolons and rhizomes. It was especially
true for control of perennial grassy species such as quackgrass (Elytrigia repens (L.) Beauv.), foxtail
barley (Hordeum jubatum) and johnsongrass (Sorghum halepense (L.) Pers.). Koger and Reddy (2005)
also found that glyphosate provided marginal or no control of weeds such as Bermuda grass (Cynodon
dactylon (L.) Pers.), hemp dogbane (Apocynum cannabinum L.), hemp sesbania, Ipomoea species, horse
nettle (Solanum carolinense L.) and tropical spiderwort (Commelina benghalensis L.).
Less Carry-over Effect of Herbicides
Glyphosate and glufosinate have almost no soil residual activity because they are tightly bound to the
organic particles in the soil. Hence, there are few restrictions for planting or replanting intervals or
injuries to the subsequent crops. This trait facilitates crop rotation by providing flexibility in selection of
potential rotation crops. Herbicide tolerant crops (HTC) will not cause any residual effect on succeeding
crops (Chinnusamy, 2010).
TABLE 2: RESIDUAL EFFECT OF HERBICIDES ON PLANT HEIGHT (CM) AND DRY MATTER PRODUCTION (DMP) (KG HA
-1
) OF SUCCEEDING CROPS AT 60 DAS
Sunflower Soybean Pearl millet Sunflower Soybean Pearl Millet
Plant
Height
DMP Plant
Height
DMP Plant
Height
DMP Plant
Height
DMP Plant
Height
DMP Plant
Height
DMP
Gly. 900 g a.e ha
-1
154.8 4382 75.8 2080 190.4 3178 138.7 4281 75.4 2287 200.7 2888
Gly. 1350 g a.e ha
1
153.4 4536 76.6 2210 195.5 3385 142.2 4684 78.8 2504 220.0 3317
Gly. 1800 g a.e ha
-1
157.6 4459 77.5 2566 192.3 3283 148.3 4563 76.4 2412 212.8 3460
Gly. 2700 g a.e ha
-1
151.6 4596 79.8 2185 185.0 3290 144.9 4652 75.8 2582 208.0 3458
Gly. 3600 g a.e ha
-1
150.6 4410 78.0 2347 189.7 3369 140.3 4368 77.8 2664 218.9 3124
Gly. 5400 g a.e ha
-1
149.8 4659 76.3 2750 188.1 3477 143.2 4487 77.0 2475 202.6 2915
HW on 15 & 30 DAS 148.9 4350 75.8 2166 179.5 2992 136.8 4276 76.3 2364 198.4 2892
Unweeded check 148.0 4253 74.8 2078 177.4 2871 135.8 4186 75.2 2154 194.7 2774
SEd 12.5 178 4.6 135 15.8 142 11.6 184 4.2 136 16.8 139
CD (P=0.05) NS NS NS NS NS NS NS NS NS NS NS NS
Succeeding crops like sunflower, soybean and pearl millet were sown after cotton crop in the
treatment blocks to assess the carry over effect of Potassium salt of Glyphosate. Observations were
recorded on germination percentage and vigour for all the treatments. Treatments difference found to be
non-significant for both the parameters hence there was normal growth and development of succeeding
crops (Table 2). The results are in line with the findings of Nadanassababady et al. (2000) who had reported
that bioassay of herbicide residues indicated that none of the herbicides evaluated for the chemical
control of weeds in cotton persisted in the soil to the level of affecting the germination and growth of
succeeding crops like finger millet and cucumber.
Reduced Crop Injury
TABLE 3: PER CENT RATING OF PHYTOTOXIC EFFECTS IN HERBICIDE TOLERANT TRANSGENIC COTTON
Treatments Winter 2009-10 Winter 2010-11
7 DAHS 14 DAHS 21 DAHS 7 DAHS 14 DAHS 21 DAHS
Glyphosate 900 g a.e ha
-1
0.0 0.0 0.0 0.0 0.0 0.0
-1
0.0 0.0 0.0 0.0 0.0 0.0
-1
0.0 0.0 0.0 0.0 0.0 0.0
-1
0.0 0.0 0.0 0.0 0.0 0.0
-1
3.0 3.0 2.0 3.0 3.0 2.0
-1
4.0 4.0 3.0 4.0 4.0 3.0
HW on 15 and 30 DAS 0.0 0.0 0.0 0.0 0.0 0.0
Unweeded check 0.0 0.0 0.0 0.0 0.0 0.0
Data not statistically analysed
Various post-emergence type herbicides used for weed control in soybean, canola, or corn can cause crop
injury and ultimately yield loss. Crop injury is more severe when the crop is under stress or unfavorable
environmental conditions occur. In contrast, crop injury is reduced with the use of herbicide tolerant
crops. The phytotoxicity symptoms were not observed in cotton with glyphosate at lower doses viz., 900,
1350, 1800 and 2700 g a.e ha
-1
. Higher doses viz. 3600 and 5400 g a.e ha
-1
were noticed with
phytotoxicity symptoms at early stages of herbicide application (Table 3). Glyphosate cause almost no
crop injury, compared to some traditional herbicides (e.g., lactofen, chlorimuron), especially when
applied to cotton. The greatest benet to growers is the broad-spectrum weed control with post
emergence application of glyphosate to cotton without crop injury as earlier reported by Wilcut et al.
(1996).
Use of Environmentally Safe Herbicides
TABLE 4: EFFECT ON WEED MANAGEMENT METHODS ON SOIL BACTERIA (X 10
-6
CFU G
-1
), FUNGI (X 10
-4
CFU G
-1
) AND ACTINOMYCETES (X 10
-2
CFU G
-1
)
IN HERBICIDE TOLERANT TRANSGENIC COTTON
Bacteria Fungi Actinomycetes Bacteria Fungi Actinomycetes
1
DASS
45
DASS
1
DASS
45
DASS
1
DASS
45
DASS
1
DASS
45
DASS
1
DASS
45
DASS
1
DASS
45
DASS
Gly. 900 g a.e ha
-1
21.60 24.45 21.62 22.42 10.54 12.32 23.61 25.24 19.74 22.34 11.05 12.02
Gly. 1350 g a.e ha
1
22.10 24.21 20.52 22.24 10.74 12.14 23.24 25.14 21.25 23.40 10.42 11.72
Gly. 1800 g a.e ha
-1
23.00 23.83 21.71 21.64 10.82 12.32 23.64 24.86 20.54 24.60 10.78 11.56
Gly. 2700 g a.e ha
-1
22.62 24.34 22.54 21.20 10.95 11.53 24.29 25.64 20.62 23.38 10.57 11.62
Gly. 3600 g a.e ha
-1
21.12 20.47 20.68 20.04 10.47 10.58 22.44 21.63 19.42 19.93 10.70 10.47
Gly. 5400 g a.e ha
-1
20.70 19.64 20.84 19.45 10.34 9.24 20.71 20.27 18.54 18.70 10.26 10.06
HW on 15 & 30 DAS 24.61 25.55 23.74 25.24 10.65 12.05 25.56 25.92 21.24 22.64 11.24 12.05
Unweeded check 25.22 26.82 25.24 25.35 11.05 12.75 24.39 26.24 22.05 23.35 11.33 12.57
SEd 2.06 2.09 1.92 2.00 0.82 1.07 1.92 2.21 2.15 2.08 1.20 1.04
CD (P=0.05) 4.10 4.15 3.87 3.92 1.58 2.05 3.78 4.34 4.20 4.15 2.14 2.00
DASSDays after second spray
In general, glyphosate and glufosinate have lower toxicity to humans and animals compared to some
other herbicides. Since they are absorbed by the organic particles in the soil and decompose rapidly, they
pose little danger for leaching and contamination of ground water or toxicity to wildlife (Knezevic and
Cassman, 2003). Glyphosate applied at doses like 900, 1350, 1800 and 2700 g a.e ha
-1
recorded more
number of bacteria, fungi and actinomycetes (Table 4). This might be due to glyphosate applied directly
on the weeds that added organic material to the soil. Microbial population increases during
decomposition of organic material. Haney et al. (2000) reported that glyphosate was available to soil and
rhizosphere microbial communities as a substrate for direct metabolism leading to increased microbial
biomass activity. Higher doses of glyphosate with 3600 and 5400 g a.e ha
-1
led to slight reduction in
microbial population as observed at initial stages and recovered within 45 days. These results corroborate
the observations of Weaver et al. (2007). Glyphosate had only small and transient effects on the soil
microbial community, even when applied at greater than field rates.
Mode of Action for Resistance Management
Since the discovery and report of triazine resistance almost 40 years ago, weed resistance to herbicides
has been well documented. For example, there are 40 dicot and 15 monocot species known to have
biotypes resistant to triazine herbicides. Also, at least 44 weed species have been reported to have
biotypes resistant to one or more of 15 other herbicides or herbicide families (Heap, 2001). The list of
herbicide-resistant weeds will continue to grow, especially with repeated use of herbicides with the same
mode of action. Many of the selective herbicides in corn and soybean have similar or identical
mechanisms of action such as the inhibition of enzyme acetolactate synthase (ALS) or the inhibition of
acetyl-co-enzyme-A-carboxylase (ACCase). Therefore, HTC particularly cotton (e.g., glyphosate and
glufosinate) can provide a new mode of action when used in an IWM program as an aid in resistance
management.

Crop Management Flexibility
The HT technology is simple to use. It requires neither special skills nor training. The technology does
not have major restrictions and is flexible, which is probably one of the reasons for such wide adoption
by producers. In particular, crops that are tolerant to broad-spectrum herbicides such as glyphosate
extend the period of herbicide application for effective weed control, which is helpful in dealing with
rainy and windy days during the optimal periods for weed control measures. In contrast, poor weather
during the critical period for weed control can greatly limit the effectiveness of more selective herbicides
(Peterson et al., 2002). According to Chinnusamy (2010), total weed density was significantly lowered
with post-emergence application of Glyphosate at 2700, 3600 and 5400 g a.e/ha in transgenic cotton
hybrids when compared to hand weeding (15 and 25 DAS). Keeling et al. (1998) also observed that,
weed control is often excellent (95 %) with the application glyphosate as post emergence in cotton.
Increased Yield and Income
TABLE 5: EFFECT OF GLYPHOSATE APPLICATION ON YIELD ATTRIBUTES AND SEED COTTON YIELD OF HERBICIDE TOLERANT TRANSGENIC COTTON
Sympodial
Branches Plant
-1

No. of
Bolls
Plant
-1

Boll
Weight
(g Boll
-1
)
Seed
cotton
Yield
(kg ha
-1
)
Sympodial
Branches Plant
-1
No. of
Bolls
Plant
-1

Boll
Weight
(g boll
-1
)
Seed
Cotton
Yield
(kg ha
-1
)
Gly. 900 g a.e ha
-1
18.05 57.3 5.10 2607 17.54 50.6 4.92 2470
Gly. 1350 g a.e ha
1
18.64 58.3 5.23 2841 17.82 52.5 5.02 2575
Gly. 1800 g a.e ha
-1
19.84 59.7 5.72 2984 20.12 55.6 5.31 2846
Gly. 2700 g a.e ha
-1
22.51 63.3 5.86 3195 21.46 60.3 5.74 3092
Gly. 3600 g a.e ha
-1
21.37 58.3 5.70 3114 19.64 57.1 5.34 3023
Gly. 5400 g a.e ha
-1
20.58 59.0 5.32 2849 18.28 54.9 4.98 2753
HW on 15 & 30 DAS 17.88 53.7 4.85 2504 15.84 49.2 4.82 2323
Unweeded check 10.35 35.7 4.06 839 9.47 30.5 4.27 713
SEd 1.36 2.9 0.24 157 1.19 3.3 0.25 144
CD (P=0.05) 2.68 5.7 0.42 322 2.34 6.6 0.48 286
Cotton crop being slow in its initial growth and is grown with wider spacing, is always encountered with
severe weed competition during early stage, which results in low yield. Broad spectrums of weeds with
wider adaptability to extremities of climatic, edaphic and biotic stresses are observed in the cotton fields.
High persistence nature of weeds is attributed to their ability of high seed production and seed viability.
Hand weeding or hoeing twice is the most commonly adopted method of weed control in cotton.
However, complete weed control could not be achieved by using any single method alone. Herbicidal
weed control seems to be a competitive and promising way to control weeds at initial stages of crop
growth.
Higher yield of HT transgenic cotton recorded with glyphosate at 2700 g a.e ha
-1
over hand weeding
twice during both the seasons during winter 2009-10 and winter 2010-11 (Table 5). It could be attributed
to efficient control of weeds during the cropping period. The findings are in accordance with observation of
Main et al. (2007) who had earlier reported that Roundup Ready Flex cotton could provide producers with
acceptable weed control without compromising cotton yield. Glyphosate at 2700 g a.e ha
-1
applied twice (25
and 65 DAS) with lower weed density, weed dry weight and higher weed control efficiency, might be the
reason for higher productivity. Cotton productivity is mainly decided by the weed control efficiency of weed
management methods as earlier observed by Grichar et al. (2004) who have noted that, trends in cotton yield
reected through weed control, which was further proved through glyphosate application system provided 96
per cent control of weeds, producing greater than 950 kg ha
-1
of seed cotton.

Glyphosate at 2700 g a.e ha
-1
recorded higher gross and net returns and B:C ratio in HT transgenic
cotton. In transgenic cotton, higher gross return was obtained with hand weeding twice, whereas higher
net return and return per rupee invested were recorded with pendimethalin at 1.0 kg ha
-1
+ hand weeding
(Fig.1). PoE spray of Glyphosate (2700 g a.e ha
-1
) twice (25 and 45 DAS) resulted in complete control of
broad spectrum weeds with higher seed cotton yield and net income in HT transgenic cotton during
winter season. Treatments, hand weeding twice (25 and 45 DAS) or pre-emergence pendimethalin ( 1.0 kg a.i.
ha
-1
) 3 DAS alongwith one hand weeding ( 45 DAS) have higher weed control efficiency and seed cotton
yield of transgenic cotton with better economic returns. Power weeder weeding on 25 DAS + one hand
weeding 45 DAS is a promising alternative weed management method for winter irrigated transgenic
cotton with higher seed cotton yield and better economic returns.

Fig. 1: Weed Management Methods on Economics in Cotton
CONCLUSION
HTC are strongly impacting weed management choices. In many crops their use will decrease the cost of
effective weed management in the short to medium term. Their use will speed adoption of conservation
tillage, greatly reduce the environmental damage of farming by reducing soil erosion by both wind and
water and reduce herbicide use. Herbicide resistance and new weed species problems that arise as a result
of this technology will be dealt with by traditional methods, such as rotating herbicides, mixing
herbicides, and rotating crops. However, they offer the farmer a powerful new tool that, if used wisely,
can be incorporated into an integrated pest management strategy that can be used for many years to more
economically and effectively manage weeds.
REFERENCES
[1] Chinnusamy, C. (2010) - Evaluation of bio-efficacy, residue, phytotoxicity and carryover of potassium salt of glyphosate
formulation MON 76366 on transgenic stacked cotton hybrids (Mon 15985 x Mon 88913). Interim report, DWSRC,
Department of Agronomy, Tamil Nadu Agricultural University, Coimbatore.
[2] Franz, J.E., Mao, M.K. and Sikorski, J.A. (1997) - Glyphosate: A Unique Global Hebricide. Monograph 189. Washington,
D.C. American Chemical Society.
[3] Grichar, W.J, Besler, B.A., Brewer, K.D. and Minton, B.W. (2004) - Using soil-applied herbicides in combination with
glyphosate in a glyphosate-resistant cotton herbicide program. Crop Prot., 23: 10071010.
[4] Haney, R.L., Senseman, S.A., Hons, F.M. and Zuberer, D.A. (2000) - Effect of glyphosate on soil microbial activity and
biomass. Weed Sci., 48: 8993.
[5] Heap, I. (2001) - International survey of herbicide resistant weeds. Online. HRAC, NAHRAC and WSSA.
[6] Keeling, J.W., Dotray, P.A., Osborne, T.S. and Asher, B.S. (1998) - Annual and perennial weed management strategies in
Roundup Ready cotton with Roundup Ultra. Proceedings of the Southern Weed Science Society, Champaign, IL, Vol. 51,
pp.49.
[7] Knezevic, S.Z. and Cassman, K.G. (2003) - Use of herbicide-tolerant crops as a component of an integrated weed
management program. Online. Crop Management doi: 10.1094/CM-2003-031701MG.
[8] Koger, C.H. and R.N. Reddy. (2005) - Glyphosate efficacy, absorption and translocation in pitted moringglory (Ipomoea
lacunose). Weed Sci. 53: 277283.
[9] Main, C.L., A.J. Michael and E.C. Murdock. (2007) - Weed response and tolerance of enhanced glyphosate-resistant cotton
to glyphosate. J. Cotton Sci., 11: 104109.
[10] Nadanassababady, T. Kandasamy, O.S. and Ramesh, G. (2000) -Integration of pre and non-selective post-emergence
herbicides and cultural method for weed control in cotton and its effect on succeeding crops. Trop. Agric. Res., 12: 217
225.
[11] Peterson, J.M, K.G., Cassman and Cantrell, R. (2002). Changes in cultural practices of farmers in southeast Nebraska as a
result of their adoption of transgenic crops. J. Ext. 40: 1.
[12] Weaver, M.A, Krutz, L.J., Zablotowicz, R.M. and Reddy, K.N. (2007) - Effects of glyphosate on soil microbial
communities and its mineralization in a Mississippi soil. Pest Manag Sci. 63(4): 38893.
[13] Wilcut, J.W., Coble, H.D., York, A.C. and Monks, D.W. (1996) - The niche for herbicide-resistant crops in U.S.
agriculture. In: Herbicide-Resistant Crops: Agricultural, Environmental, Economic, Regulatory, and Technical Aspects.
S.O. Duke (ed.) CRC Press, Boca Raton, FL. pp. 213230.
Evaluation of Pyrithiobac Alone and

in Combination with Grassy Herbicides
on Weed Control in Cotton
A.S. Rao
Integrated Weed Management Unit, Regional Agricultural Research Station,
Acharya N.G. Ranga Agricultural University, Lam, Guntur522034, A.P.

AbstractA field experiment was conducted during kharif 2009-10 and 2010-11 at Regional Agricultural Research
Station, Lam, Guntur (A.P.) to evaluate pyrithiobac alone and in combination with grassy herbicides on weed control
in cotton and also to characterize their nature of interaction. The experiment was conducted in a randomized block
design with three replications. Results indicated that all the weed control treatments significantly reduced weed dry
weight over unweeded check. Tank mixing of pyrithiobac with grassy herbicides was superior to application of post
emergence herbicides alone. Among the treatments, post emergence application of pyrithiobac 32 g + quizalofop ethyl
25g/ha recorded the lowest weed dry weight and highest seed cotton yield (2270 kg/ha) and net monetary returns
(Rs.44,245/-) and benefit cost ratio (0.95)and was on par with other herbicide mixtures. All the herbicide combinations
were found to be synergistic.
INTRODUCTION
Cotton being a wide row spaced and relatively slow growing crop in initial stages is subjected to severe
weed competition and reduces yield to an extent of 60% (Sadangi and Barik, 2007). Present
recommendation of pre emergence herbicide (pendimethalin) application followed by two or three inter-
cultivations is a common practice (Prabhau et al., 2010). However, most often due to incessant rains
during kharif season and inter-cultivation is not possible due to excessive moisture in black cotton soils.
Though, paraquat is used as directed spray (Srinivasulu and Rao, 2000; Srinivasulu et al., 2004), it causes
plant injury. Therefore, farmers seek for selective post emergence broad spectrum herbicide/herbicide
mixtures in order to control all weeds in a single spray. Pyrithiobac and grassy herbicides (clodinafop
propargyl, fenoxaprop ethyl, quizalofop ethyl) are systemic herbicides with a specific target site of
action. Pyrithiobac an ALS herbicide inhibits Acetolactate synthase, a key enzyme in the biosynthesis of
branched chain amino acids. Whereas the grassy herbicides, clodinafop propargyl, fenoxaprop ethyl,
quizalofop ethyl are Acc ase inhibitors, which inhibit the enzyme Acetyl CoA Carboxylase (Accase), a
key enzyme in the biosynthesis of fatty acid synthesis. Thus, tank mixing of herbicides with two
distinctly different target sites of action has a potential for effective weed control. Since the type of
interaction between pyrithiobac and grassy herbicides vary considerably depending on herbicide
chemistry and weed species, it was attempted to characterize the nature of interaction between
pyrithiobac and grassy herbicides. Further, information pertaining to tank mixing of selective post
emergence herbicides on cotton is scanty (Snipes and Allen, 1996). With this in view, the present
experiment was conducted to evaluate the post emergence herbicide pyrithiobac alone and in
combination with grassy herbicides.
A field experiment was conducted at Acharya N. G. Ranga Agricultural University, Regional
Agricultural Research Station, Lam, Guntur (A.P.) during kharif 2009-10 and 2010-11. The soil of the
experimental field was clay loam in texture, medium in available N and P but high in available K with a
pH 7.7. The experiment consisting of seven treatments arranged in a randomized block design with three
replications. Cotton (RCH-2 Bt) was sown with a spacing of 90x60 cm. The post emergence herbicides,
20 DAS (days after sowing), were sprayed with a knap sack sprayer fitted with flat fan nozzle using a
spray volume of 500 l/ha. All the recommended package of practices other than weed control was
68
Evaluation of Pyrithiobac Alone and in Combination with Grassy Herbicides on Weed Control in Cotton 407
adopted to raise the experimental crop. The data on density was recorded at 30, 60 DAS and final picking
and dry weight of weeds was recorded at 60 DAS and data were subjected to square root ( )
transformation before statistical analysis to normalise their distribution (Panse and Sukhatme, 1978). The
method described by Colby (1967) was used to calculate the expected response of herbicide
combinations. Expected and observed values were compared using the CD values calculated for the
observed data (Rao and Reddy, 1999). If the observed response of an herbicide combination was either
significantly lower or greater than the expected value, then combination was declared antagonistic or
synergistic, respectively. Combinations were considered to be additive (no interaction) when observed
and expected responses were not significant.
RESULTS AND DISCUSSIONS
Effect on Weeds
The dominant weed flora of the experimental plots consisted of grasses (Echinochloa colona,
Dactyloctenium aegyptium, Panicum repense, Dinebra retroflexa) Sedges (Cyperus rotundus), and broad
leaf weeds (Trianthema portulacastrum, Cleome viscosa, Phyllanthus niruri, Digera arvensis, Physalis
minima, Abutilon indicum)
All the weed control treatments significantly reduced the density and dry weight of weeds as
compared to weedy check at all stages of observations (Table 1). Among the treatments, pyrithiobac
alone or in combination with grassy herbicides was found to be superior to application of grassy
herbicides alone at 60 DAS in reducing the weed dry weight. Among the herbicide combinations, lowest
weed growth was observed with treatment pyrithiobac (32 g) + quizalofop ethyl (25g/ha) and was on par
with other combinations. The reduced weed growth in these treatments might be due to broad spectrum
control of weeds due to combined effect of pyrithiobac and grassy herbicides.
TABLE 1: EFFECT OF DIFFERENT TREATMENTS ON DENSITY AND DRY WEIGHT OF WEEDS IN COTTON (POOLED DATA OF TWO YEARS)
Treatments Dose
(g/ha)
Time of
Application
(DAS)
Total Weed Density
(no./m
2
) at
Total Weed
Dry Weight
(g/m
2
)
at 60 DAS
Weed Dry Weight
Percent Reduction
30 DAS 60 DAS Final
Picking
Observed
(%)
Expected
(%)
T1 - Unweeded
check
- - 16.7
(298.7)
14.2
(215.7)
9.3 (86.5) 20.2 (423.3) 0 -
T2 - Pyrithiobac 63 20 10.3
(108.7)
10.1
(115.8)
7.6 (58.5) 14.5 (228.3) 28.2 -
T3 - Clodinafop
propargyl
60 20 12.2
(158.2)
11.2
(131.2)
8.1 (67.0) 16.7 (295.0) 17.3 -
T4 - Fenoxaprop
ethyl
65 20 12.8
(167.8)
10.5
(112.3)
8.2 (69.5) 16.9 (294.2) 16.3 -
T5 - Quizalofop
ethyl
50 20 12.1
(151.7)
10.1
(105.7)
7.5 (56.5) 17.8 (325.8) 11.9 -
T6 - Pyrithiobac +
Clodinafop propargyl
32+30 20 9.3
(91.7)
08.5
(76.7)
5.9 (36.5) 15.6 (260.0) 22.6 40.6
T7 - Pyrithiobac +
Fenoxaprop ethyl
32+28 20 9.2
(90.2)
08.7
(78.7)
05.6
(32.5)
14.5 (223.3) 28.2 39.9
T8 - Pyrithiobac +
quizalofop ethyl
32+25 20 8.5
(76.3)
08.1
(68.2)
04.9
(25.7)
14.5 (220.8) 28.2 39.9
SED 0.68 0.57 0.45 0.75 0.75
CD (0.05%) 1.95 1.65 1.36 2.14 2.14
Note: DAS: Days after sowing. Data transformed to transformation. Figures in parentheses are original values. Interactions
were considered significant if the difference between the observed and expected values exceeded the appropriate CD value

Herbicide interactions were evaluated by comparing the observed weed dry weight recorded at 60
DAS with that of Colbys expected values. The herbicide combinations consisting of pyrithiobac with all
the grassy herbicides (clodinafop propargyl, fenoxaprop ethyl, quizalofop ethyl) exhibited synergistic
response (Table 1). The observed synergism could be due to differential mechanism of action of these
herbicides resulting in increased phytotoxicity on weeds when applied as tank mixtures.
Effect on Crop
All the herbicides either applied alone or tank mixtures did not injure cotton crop. All the weed control
treatments significantly influenced crop dry weight, boll number, boll weight and seed cotton yield
(Table 2). Among the treatments, the highest seed cotton yield was obtained with post emergence tank
mix application of pyrithiobac and quizalofop ethyl over alone herbicide application but was on par with
other tank mixtures of pyrithiobac with fenoxaprop ethyl and clodinafop propargyl. This treatment
recorded 98per cent higher seed cotton yield than weedy check. The increase in seed cotton yield was due
to effective broad spectrum control of weeds by tank mixing of grassy and broad leaf herbicide, which
resulted in higher crop dry matter, yield components and yield. The results are similar to those reported
by Grichar et al. (2003) on interaction of graminicides applied in combination with pyrithiobac.
TABLE 2: EFFECT OF DIFFERENT TREATMENTS ON YIELD AND YIELD COMPONENTS IN COTTON (POOLED DATA OF TWO YEARS)
Treatments Boll Number/
Plant
Boll Weight (g) Seed Cotton
Yield
(kg/ha)
Net Returns
(Rs/ha)
BCR
(Rs/R)
T1 - Unweeded check 19.2 03.9 1148 920 0.02
T2 - Pyrithiobac 24.3 04.1 1723 22360 0.48
T3 - Clodinafop propargyl 22.9 04.1 1627 19030 0.41
T4 - Fenoxaprop ethyl 24.9 04.3 1807 26230 0.57
T5 - Quizalofop ethyl 25.3 04.3 1876 28490 0.61
T6 - Pyrithiobac +
Clodinafop propargyl
26.5 04.4 2057 36055 0.78
T7 - Pyrithiobac +
Fenoxaprop ethyl
27.7 04.5 2209 42054 0.71
T8 - Pyrithiobac +
Quizalofop ethyl
28.5 04.6 2270 44245 0.95
SED 0.8 0.1 103.3
CD (0.05%) 2.4 0.4 296.6
ECONOMICS
The highest net monetary returns (Rs 44,245/-) and benefit cost ratio (0.95) was obtained with the
treatment pyrithiobac and quizalofop ethyl applied. This was closely followed by pyrithiobac and fenoxa
prop ethyl with net monetary return of Rs. 42,054/- and benefit cost ratio of 0.91.
From this study, it can be concluded that in cotton crop, post emergence tank mix application of
pyrithiobac and quizalofop ethyl applied at 20 DAS was effective and economical without any
phytotoxicity to cotton crop, whenever inter-cultivation is not possible due to incessant rains during
kharif season. The next best treatment is pyrithiobac and fenoxa prop ethyl tank mixture.
REFERENCES
[1] Colby, S.R. (1967) Calculating synergistic and antagonistic response of herbicide combinations. Weeds 15: 2022.
[2] Grichar W. James, Brew, A. Besler, Kevish, D. Brewer and Reobert, G. Lemon (2003) - Interaction of pyrithiobac and
graminicide for weed control in cotton. Weed Technology 17(3): 461466.
[3] Panse, V.G. and Sukatme P.V. (1978) Stastical Methods for Agricultural Workers, ICAR, New Delhi, pp: 152.
[4] Prabhu, G., Halepyati, A.S. and Pujari, B.T. (2010) Weed Management studies in Bt cotton (Gossypium hirsutum) under
irrigated conditions. Proc. of XIX National Symposium on Resourse Management Approach towards livelihood security,
24 December, 2010, UAS, Bangalore, pp: 263.
[5] Rao, S.A. and Reddy, K.N. (1999) Purple nutsedge (Cyperus rotundus) and sickle pod (Senna obtusifolia) response to
glyphosate mixtures with ALS-inhibiting herbicides. Weed Technol., 13(2): 361366.
Evaluation of Pyrithiobac Alone and in Combination with Grassy Herbicides on Weed Control in Cotton 409
[6] Sadangi, P.K. and Barik, K.C. (2007) Effect of weed management practices on nutrient depletion by weeds, yield and
economics winter irrigated cotton (Gossypium hirsutum). Indian J. Agron., 52(2): 172175.
[7] Snipes Charles, E. and Allen Ralph, L. (1996) - Interaction of graminicides applied in combination with pyrithiobac. Weed
Technol., 10(4): 889892.
[8] Srinivasulu, G. and Rao, A.S. (2000) Effect of sequential application of herbicides on weed management in cotton. Proc.
of symposium on Challenges in Agronomic crop management in early 21
st
century held by Society of Agronomists,
ANGRAU, Rnagar, Hyderabad, 2425. May, 2000, pp: 3335
[9] Srinivasulu, K., Hema, K., Rao, A.S. and Rao, K.V. (2004) Evaluation of pre and post emergence herbicides in rain fed
cotton. Intl symposium on Strategies for sustainable cotton production a global vision 2-crop production, 2325. November,
2004, University of Agricultural Sciences, Dharwad, pp: 244245.
Defining Optimal Application Rate and Timing

of Mepiquat Chloride for Cotton Grown
in Conditions that Promote Excessive
Vegetative Growth
G.D. Collins
1
, R. Wells
2
, R. Riar
2
and K.L. Edmisten
2
1
University of Georgia,
2
North Carolina State University, USA
AbstractMepiquat chloride (MC) is commonly used to control vegetative growth and promote early maturity of
cotton. Responses to MC are dependent upon the prevailing environmental conditions, with favorable responses often
observed in conditions that promote vegetative growth or delayed maturity. There have been recent claims of growers
in the Southeastern U.S. using extremely high MC rates on cotton grown in fertile soils and high moisture
environments while failing to adequately control growth. These results raise the question as to whether these high rates
are actually necessary, or should growers be utilizing more aggressive application strategies in order to achieve
optimal growth suppression. Experiments were conducted during 2007 and 2008 to investigate the effects of various
MC application strategies and rates on cotton grown in high-moisture, high-fertility conditions. Treatments consisted
of three MC application strategies (low-rate-multiple, modified early bloom, and early bloom) using normal (1x),
moderate (1.5x), and high (2x) rates, which were compared to a non-treated control (NTC). Results suggested that
normal rates (1x) are equally effective in controlling plant growth or modifying maturity characteristics as the high
rates (1.5x and 2x), therefore the high rates may not be necessary. The less aggressive early bloom strategy controlled
growth similarly to the more aggressive application strategies. However, the modified early bloom strategy may
provide growers with some flexibility in adequately controlling growth in situations where timely MC application to
all cotton hectares at early bloom is challenging. This strategy also requires less equipment trips through the field
compared to the low rate multiple strategy, and it is equally effective in controlling plant growth.
INTRODUCTION
Plant growth regulators (PGRs) are commonly used in Southeastern U.S. cotton production to manage
cotton growth for optimal productivity. The PGR products utilized most in this region contain either MC
(N,N-dimethyl piperidinium chloride) or mepiquat pentaborate (N,N-dimethyl piperidinium pentaborate),
which, if used properly, can positively influence cotton growth and maturity traits in some environments.
Mepiquat-containing PGRs influence cotton growth by obstructing cell elongation (Cordell et al., 2005)
and expansion resulting from inhibiting the biosynthesis of gibberellins (Hake et al., 1991). Plant growth
regulators can have several impacts on the structure of cotton plants. Researchers have found that PGRs
reduced internode length (Reddy et al., 1992), plant height, the number of nodes (Jost and Dollar, 2004;
Nichols et al., 2003; Reddy et al., 1992; Siebert and Stewart, 2006; Wilson et al., 2007), height-to-node
ratio (Johnson and Pettigrew, 2006; Nichols et al., 2003; Oberry et al., 2009), and nodes above white
flower (NAWF) (Johnson and Pettigrew, 2006; Oberry et al., 2009; Pettigrew and Johnson, 2005).
Others have noticed increased boll retention on lower branches (Prince et al., 2000) which can promote
early maturity of the crop (Hake et al., 1991). In addition to early maturity, the effects of PGRs on cotton
growth could potentially improve harvest efficiency (Jost and Dollar, 2004) and insect management
(Edmisten, 2008), while potentially decreasing boll rot (Edmisten, 2008; Jost and Dollar, 2004). The
effect of PGRs on lint yield is inconsistent and varies depending upon other management practices and
the prevailing environmental conditions. Researchers have reported no yield response to mepiquat-
containing PGRs (Edmisten, 1994; Pettigrew and Johnson, 2005), yield losses (Oberry et al., 2009) or
yield increases (Coccaro et al., 2004; Elbehar et al., 1996; Elbehar and Welch, 1996; Johnson and
Pettigrew, 2006; Nichols et al., 2003; Siebert and Stewart, 2006; Wilson et al.,, 2007) in some situations.
Determining the optimal MC application rate or strategy, in terms of achieving optimal growth
suppression or yield, can be difficult as the best rate and application strategy often varies depending upon
the potential for excessive vegetative growth. Conditions that could delay maturity or promote vigorous
69
Defining Optimal Application Rate and Timing of Mepiquat Chloride for Cotton Grown 411
and excessive growth, such as late-planted cotton, high plant populations, and excessive nitrogen and soil
moisture, would likely result in a positive response to MC treatment (Edmisten, 2008; Oberry et al.,
2009). In North Carolina, the currently recommended MC application strategies include the low-rate-
multiple strategy (LRM), the modified early bloom strategy (MEB), and the early bloom strategy (EB)
(Edmisten, 2008). The LRM strategy is the most aggressive strategy, and consists of three to four
applications of low rates of MC beginning at the match-head square (MHS, 7- to 8-leaf cotton with
squares ranging from 3.2 to 6.4 mm in size) growth stage and continuing every seven to 14 days until
early bloom (EB, five to six white blooms per 7.6 m of row) or EB plus seven days. The MEB strategy
consists of at least two, and potentially three, applications of moderate rates of MC at 10 to 14 days prior
to EB, again at EB, and seldomly 10 to 14 days after EB. The EB strategy is the least aggressive strategy,
consisting of at least one, and potentially two, applications of MC at higher rates beginning at EB. In a
study involving narrow-row cotton in North Carolina, Wilson et al., (2007) observed that the LRM and
MEB strategies reduced plant height more than did the EB strategy, and they suggested that these
strategies may be more appropriate for narrow-row systems. Edmisten (1994) reported that a plant
monitoring point system used in conjunction with a LRM strategy required lower MC rates than a
standard LRM strategy. Similar application strategies were implemented across the cotton belt. In
Louisiana, Siebert and Stewart (2006) reported that their MEB strategy increased yield when compared to
cotton receiving no mepiquat during one year. In Tennessee, Craig and Gwathmey (2005) reported that
multiple-application strategies, of both low and high rates, had more impact on plant height than did the
near-bloom applications, although all strategies reduced height when compared to non-treated cotton.
Jost and Dollar (2004) in Georgia, compared multiple-application strategies using different mepiquat-
containing products and rates and found no difference in terms of plant height reduction among
treatments. In a study conducted in Mississippi, Coccaro et al., (2004) found that mepiquat-containing
PGRs applied beginning at the 7- to 8- nodes stage did not control plant height as well as when
applications were initiated at 14 nodes. Oberry et al., (2009) suggested that the yield losses they noticed
in Virginia and South Carolina, may be a result of early applications and/or high seasonal rates, which
may be too aggressive.
Recently, in the Southeastern U.S., there have been some claims or reports of growers utilizing
excessively high rates (0.22 to 0.31 kg a.i. ha
-1
) while failing to adequately control growth. This raises the
question whether these rates were actually necessary, or should growers be utilizing more aggressive
application strategies (multiple applications and/or initiating applications sooner) in order to achieve
optimal growth control. The objective of this experiment was to define the optimal MC application rates
and strategies for cotton grown in conditions that promote extremely excessive vegetative growth, such
as soils with high fertility and moisture capacity or in coastal areas that typically experience frequent
rainfall.
Experiments were conducted during 2007 and 2008 on a Norfolk fine sandy loam soil (fine-loamy,
kaolinitic, thermic, Typic Kandiudult) at an on-farm site in Duplin County near Beulaville, NC. Late-
maturing cultivars, ST 6611B2RF
and DP 164 B2RF
, were planted at a rate of 13.1 seeds row meter

-1

on 2 May 2007 and 6 May 2008, respectively, using a two-row vacuum planter. Plots contained four 12.2
m long rows spaced 0.97 m apart. Treatments consisted of three MC application strategies {LRM, MEB,
and EB} along with a non-treated control (NTC), and three MC rate schematics {0.05 (1x), 0.07 (1.5x)
and 0.1 (2x) kg a.i. ha
-1
} to determine the optimal application strategy and rate for cotton grown in
conditions conducive to excessive vegetative growth. Treatments were replicated four times in a
randomized complete block design. Dry poultry litter was applied at a rate of 13.5 tons ha
-1
prior to
planting to ensure high nitrogen fertility. All other production and pest management practices were
conducted according to the North Carolina Cooperative Extension recommendations for that region
(Bacheler, 2008; Crozier, 2008; Edmisten, 2008; Koenning, 2008; York and Culpepper, 2008).
The LRM treatments received MC {N,N-dimethyl piperidinium chloride (Mepex
, Nufarm Americas
Inc., Burr Ridge IL.)} at the respective total rates divided into four applications beginning at the MHS
growth stage and continuing every seven to 14 days until EB or EB plus seven days. The MEB
treatments received MC at the respective total rates divided into two applications beginning at the MHS
growth stage or MHS plus seven days and again at the EB growth stage. The EB treatments received MC
at the respective total rates in a single application at the EB growth stage. The NTC received no MC at
any time throughout the season.
Treatments were applied using a CO
2
-pressurized backpack sprayer calibrated to deliver 140 L ha
-1

using TeeJet
XR110-02 flat-fan nozzles (TeeJet Technologies, Wheaton, IL). Plant heights, nodes, and
uppermost fully expanded internode length (distance between fourth and fifth true leaf from the top of the
plant) were recorded for six plants in the center two rows of each plot at each MC application, and plant
height, nodes, and NAWF were recorded for six plants in the center two rows of each plot during early
August of each year. Prior to defoliation, the percentage of open bolls was recorded in a randomly chosen
1-m section of row within each plot. Nodes above cracked boll (nodes between highest first position
cracked boll and highest harvestable boll) and angle measurements of the main stalk relative to the
ground were recorded for six plants from the center two rows of each plot. Prior to harvest, plant
mapping data, including boll distribution and growth characteristics, were collected for six plants from
the center two rows of plot.
The center two rows of each experimental unit were mechanically harvested using a two-row spindle
picker on 12 October 2007 and 17 October 2008. Seedcotton weights for each plot were recorded and
sub-samples were collected for high volume instrumentation analysis and lint percentage. Harvest data
included lint yield, micronaire, fiber length, length uniformity, and fiber strength.
Data for growth characteristics, maturity parameters, lint yield, and fiber quality parameters were
subjected to analysis of variance using the general linear model in SAS version 9.1.3 (SAS Institute, Cary
NC). Means of significant main effects and interactions were separated using Fishers Protected LSD at
p<0.05 or 0.1.
TABLE 1: EFFECTS OF MEPIQUAT CHLORIDE (MC) APPLICATION STRATEGIES AND RATES ON PLANT HEIGHT, TOTAL NODES,
NODES ABOVE WHITE FLOWER, AND TOTAL BOLLS PER PLANT IN 2007 AND 2008
MC Application
Strategy
MC Rate at Each
Application
Plant
Height
Total Nodes Per
Plant
Nodes Above White
Flower
Total Bolls Per
Plant

kg ha
-1

____
cm
____

________________________
No.
_________________________

Low Rate Multiple
y
0.01 113.23 bc 18.46 4.17 bc 9.58
0.02 108.99 bc 19.93 4.57 ab 10.1
0.025 108 bc 17.5 3.65 c 11.44
Modified Early
Bloom
x

0.025 113.85 bc 18.58 4.58 ab 10.69
0.04 108.07 bc 20.21 4.04 bc 10.67
0.05 101.51 c 19.53 4.03 bc 11.31
Early Bloom
w
0.05 121.85 b 18.89 4.21 bc 11.9
0.07 112.79 bc 19.72 4.32 bc 11.19
0.1 117.85 b 19.38 4.28 bc 12.52
Non-treated Control
_____
139.5 a 21 5.31 a 10.38
P-value
_____
0.0030 0.3391 0.0402 0.3008
LSD
_____
14.505 NS 0.8118 NS
z
Data are pooled over years. Means within a column followed by dissimilar letters are significantly different based on Fishers
Protected LSD test at = 0.1 or = 0.05.
y
The Low Rate Multiple Strategy consisted of four applications beginning at match-head square, and every 7 to 14 days until
early bloom or early bloom plus 7 days.
x
The Modified Early Bloom Strategy consisted of two applications beginning at match-head square or match-head square plus
7 days, and again at early bloom.
w
The Early Bloom Strategy consisted of a single application at early bloom
There were no interactions between MC treatments and years, therefore all data are reported combined
over years. All MC application strategies and rates within each treatment significantly reduced plant
height compared to the NTC (Table 1) similar to the findings of Jost and Dollar (2004), Nichols et al.,
(2003), Reddy et al., (1992), Siebert and Stewart (2006), and Wilson et al., (2007). Within each
application strategy, the higher MC rates (1.5x and 2x) did not significantly reduce plant height more
than the normal-use (1x) rates. The 2x rate applied according to the MEB strategy significantly reduced
plant height more than both the 1x and 2x rates applied according to the EB strategy. In contrast to the
findings of Nichols et al., (2003), height-to-node ratio (data not shown), and the number of mainstem
nodes were not affected by MC treatment (Table 1). Neither MC application strategy nor rate affected the
total number of bolls per plant, and there was also no effect on the number of sympodial nodes, the node
of first sympodia, the number of vegetative or sympodial bolls, or sympodial boll retention (data not
shown). Similar to the findings of Johnson and Pettigrew (2006), Oberry et al., (2009), and Pettigrew
and Johnson (2005), most MC treatments significantly reduced NAWF when compared to the NTC
indicating that MC may promote earlier maturity. Higher rates applied according to both the MEB and
EB strategies did not reduce NAWF any more than the lower rates.
Regarding boll distribution, the 1.5x and 2x rates applied according to the LRM strategy, and the 1x
and 1.5x rates applied according to the MEB strategy increased the number of bolls on nodes four
through seven compared to the NTC (Table 2). Mepiquat chloride applied according to the EB strategy,
regardless of rate, had no effect on the number of bolls in this node zone, as well as the LRM using the
1x rate and the MEB using the 2x rate. This indicates that the more aggressive application strategies may
enhance maturity more than the less aggressive EB strategy. All MC application strategies and rates
increased the number of bolls on nodes eight through 10. These results are similar to those of Prince et
al., (2000). The number of bolls in this node zone increased as MC rate increased when applied
according to the LRM strategy. Neither MC application strategy nor rate had any effect on the number of
bolls on nodes 11 through 13 and nodes 14 through 16.
TABLE 2: EFFECTS OF MEPIQUAT CHLORIDE (MC) APPLICATION STRATEGIES AND RATES ON BOLL DISTRIBUTION IN 2007 AND 2008.
MC Application
Strategy
MC Rate
at Each Application
Number of Bolls
on Nodes 47
Number of Bolls
on Nodes 810
Number of Bolls
on Nodes 1113
Number of Bolls
on Nodes 1416

kg ha
-1

___________________________________
No.
___________________________________

Low Rate Multiple
y
0.01 2.08 a-d 2.75 d 1.83 1.17
0.02 2.42 ab 2.77 cd 2.13 1.56
0.025 2.29 abc 3.13 ab 2.48 2
Modified Early
Bloom
x

0.025 2.4 ab 3.06 abc 2.23 1.54
0.04 2.48 a 2.98 bcd 2.08 1.31
0.05 2 bcd 3.1 ab 2.67 1.73
Early Bloom
w
0.05 1.88 cd 3.31 a 2.71 1.96
0.07 2.1 a-d 2.92 bcd 2.67 1.92
0.1 2.17 a-d 3.19 ab 2.92 1.85
Non-treated Control
_____
1.7 d 2.35 e 2.42 2
P-value
_____
0.0698 0.0016 0.1165 0.1562
LSD
_____
0.4759 0.2938 NS NS
z
y
x
w
The Early Bloom Strategy consisted of a single application at early bloom.
Most MC application strategies and rates increased the angle of the main stalk relative to the soil
surface compared to the NTC (Table 3), indicating that MC may prevent or reduce the potential for
lodging by reducing plant height and increasing the proportion of bolls retained on lower nodes, resulting
in plants that are less top-heavy. For the LRM and the MEB strategies, the higher MC rates had no
additional effect on stalk angles. Mepiquat chloride at all rates according to the MEB and EB strategies
increased percent open bolls, suggesting that MC may promote an earlier maturity thus earlier harvest.
Within all application strategies, the higher rates did not improve boll opening. Neither MC application
strategy nor rate had any effect on lint yield, lint percentage, or any fiber quality parameter (data not
shown).
TABLE 3: EFFECTS OF MEPIQUAT CHLORIDE (MC) APPLICATION STRATEGIES AND RATES ON STALK ANGLE AND PERCENT OPEN BOLLS IN 2007 AND 2008
MC Application
Strategy
MC Rateat Each
Application
(kg ha
-1)

Angle of Main Stalk Relative
to Soil Surface
(
______
degrees
______
)
Percent Open
Bolls
(
______
%
______
)
Low Rate Multiple
y
0.01 74.38 ab 48.9 ab
0.02 74.88 ab 52.87 a
0.025 74.25 ab 45.97 ab
Modified Early Bloom
x
0.025 72.83 ab 52.85 a
0.04 78.5 a 55.66 a
0.05 80.25 a 54.91 a
Early Bloom
w
0.05 72.63 ab 50.8 a
0.07 65.13 bc 52.78 a
0.1 78 a 50.16 a
Non-treated Control
_____
61.13 c 39.24 b
P-value
_____
0.0586 0.0964
LSD
_____
10.969 9.87
z
y
x
w
The Early Bloom Strategy consisted of a single application at early bloom.
These results are consistent with previous research in that MC can reduce plant height and promote
earlier maturity as seen with the effects on boll distribution in lower node zones, NAWF, and percent
open bolls. In most instances, the normal-use rates, or the 1x rates, had the same effects on plant growth
and maturity as did the higher rates. The higher rates, regardless of the application strategy used, seldom
resulted in any plant modifications that would be advantageous to growers, therefore the normal-use (1x)
rates should suffice in environmental conditions that promote excessive vegetative growth, similar to the
conditions in which this experiment was conducted as supported by the plant height of the NTC.
Although these results suggested that the EB strategy was effective in controlling growth, in these
environmental conditions, the MEB strategy may be more appropriate than the EB strategy because it
could allow growers to control growth with more flexibility due to time constraints of applying MC to all
cotton hectares at the EB stage. This strategy may be especially effective in years when excessive rainfall
limits the ability of equipment to travel through the field. The MEB strategy also requires fewer
equipment passes through the field compared to the LRM strategy, allowing for optimal growth control
while avoiding unnecessary equipment operation costs.
These results are inconsistent with the claims from growers that excessive MC rates controlled plant
height poorly in similar environmental conditions, likely because the growers making these claims
applied MC later than recommended in terms of growth stage. Siebert and Stewart (2006) reported
similar findings, suggesting that excessive rates may not be necessary to adequately control plant growth.
Delaying MC application beyond the appropriate growth stage could potentially result in poor plant
height suppression. Another potential reason for this discrepancy could be that growers likely used
glyphosate-resistant (Round-up Ready
) varieties which have the potential for poor pollination and fruit
abortion when glyphosate is used (Pline et al., 2002). Fruit abortion, especially when it occurs on the
lower nodes (occasionally noted with Round-up Ready
cotton), can cause terminal growth to be slightly

more aggressive, possibly making plant height more difficult to control. This experiment was conducted
using Roundup Ready Flex
varieties, which are tolerant to glyphosate at any growth stage, potentially

allowing more fruit to be retained on lower nodes, which could restrain terminal growth. This
characteristic can often result in less aggressive terminal growth potential, therefore allowing lower MC
rates to suffice in terms of controlling plant growth.

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Effect of Cool Conditions on Cotton Seedlings

D.K.Y. Tan
1
, S. Ormiston
1
, M.P. Bange
2
and J.S. Amthor
1
1
Faculty of Agriculture, Food and Natural Resources, University of Sydney
2
CSIRO Division of Plant Industry and Cotton, Australia
AbstractSuboptimal temperatures reduce cotton germination, seedling emergence, and early vigour, all of which
can lead to poor or delayed stand establishment. Strategies that provide early season low-temperature tolerance might
improve cotton growth when cooler than expected temperatures are experienced early. It could also provide growers
with increased flexibility for planting times especially in cooler regions. Ensuring better establishment under cool
conditions should reduce costs associated with replanting and contribute to greater stand uniformity which will assist
in attaining optimum yield and fibre quality. Plant growth regulators (PGRs) are a potential option of increasing
cotton seedling hardiness against cool temperatures, and we tested whether the PGRs paclobutrazol and trinexapac-
ethyl can improve cold tolerance in cotton seedlings. In this preliminary investigation, we found no evidence that
these PGRs improved cotton germination under cool conditions in the laboratory, whereas they reduced seedling
length under warm conditions.
Keywords: Chilling injury, cotton, germination, establishment, screening techniques, plant growth regulators,
paclobutrazol, trinexapac-ethyl
INTRODUCTION
Cotton (Gossypium hirsutum) is a cold sensitive plant. Chilling injury occurs in cotton seedlings
whenever the temperature drops below 15 C for a few hours during the first several days after
germination (Lauterbach et al., 1999) and temperatures below 20 C slow the germination of cotton seed
and of seedling emergence (Cole and Wheeler, 1974). This emphasises the opportunity to develop
cultivars or management practices that would allow better crop establishment, reducing the costly need to
replant following post-emergence cold shocks, aiding uniformity of crop establishment. Improved
establishment with a reduction in gappy stands leads to improvement in yield and fibre quality. It will
also provide growers with increased flexibility regarding early sowing date options (Constable et al.,
1998).
This work extends previous assessments of genotypic variation of cotton germination and
establishment in response to cold conditions (Duesterhaus, 2000; Duesterhaus et al., 2000; Tuck et al.
2010). Their research suggested that there was genotypic variability in field-grown seedling
establishment in response to cold temperatures, and they developed methods to distinguish differences in
cold tolerance of cultivars in the laboratory using germination tests in both warm and cool conditions and
validated under field conditions. Tuck et al. (2010) successfully used seedling length as an indicator of
cultivar variation however; the approach needs to be validated with a wider range of genotypes and
conditions.
Plant growth regulators (PGRs) and retardants have been suggested for improving plant stress
tolerance by Rademacher (2000). Seed coatings containing PGRs may have the potential to improve cold
tolerance by improving seedling vigour in stressed conditions (Rademacher, 2000). It is hypothesised that
two novel plant growth regulators; paclobutrazol [PBZ (chemical name (2RS, 3RS)-1-(4-chlorophenyl)-
4,4-dimethyl-2-(1H-1,2,3-triazol-1-yl)pentan-3-ol)] and trinexepac-ethyl [TXP (chemical name 4-
(cyclopropyl-alpha-hydroxy-methylene)-3,5-dioxocyclohexanecarboxylic acid ethyl ester)] may be used
as a seed coating to assist in early crop vigour in cold conditions at establishment (Rademacher, 2000).
PBZ is a potent inhibitor of gibberellins biosynthesis and triazole fungicide, and has been shown to
reduce desiccation in seedlings by reducing shoot growth (Gavidia et al., 1997). TXP is an
acylcyclohexanedione that influences the electron transfer chain in plant mitochondria (Heckman et al.,
2002), and interferes with the later stages of gibberellins synthesis reducing plant shoot growth
(Rademacher, 2000). The use of TXP in perennial ryegrass in New Zealand increased harvest index
70
Effect of Cool Conditions on Cotton Seedlings 417
(increased partitioning to the harvestable plant parts), increased seed head density, increased seeds per
spike, improved seed retention and reduced lodging (Chynoweth et al., 2010). No research has been
conducted, however, on the effect of PBZ and TXP on the cold tolerance of cotton seedlings.
This paper reports preliminary investigations to further develop cold tolerance screening methods and
assesses the use of the plant growth regulators PBZ and TXP on cotton germination in warm and cool
laboratory conditions.
Experiments were conducted using seed germination facilities at the Australian Cotton Research Institute
(ACRI) in Narrabri, NSW and the Biomedical Building, University of Sydney, NSW.
To assess the impacts of PBZ and TXP on cotton germination, untreated seeds of the cultivar Sicot
71BRF (CSIRO, Australia) were treated with various rates of the PGRs and then germinated in warm and
cold conditions. Seeds were treated with 0, 4, 20, 40, 80 and 160 g/100 kg of TXP and 0, 4, 8, 16, 32 and
64 ml/100 kg of PBZ.
Four replications of 50 seeds were subjected to both warm and cool germination tests for each seed
treatment. Seedlings with a radicle length of 3 mm or longer were counted as germinated (Wanjura and
Buxton, 1972). Ten seedlings were selected randomly and the length from the hook of the hypocotyl to
the tip of the radicle was measured (hereafter referred to as seedling length). A combination of the warm
and cool germination tests was reported to be a reliable indicator of field performance (Bird and Reyes,
1967). For the warm germination tests seeds are germinated at 30
0
and seed germination percentage and
seedling length recorded after 4 d. For the cold germination tests seeds are germinated at 14
o
C and
seedling lengths are measured after 7 d. A cool-warm vigour index was calculated as the average of the
warm (30
0
C) seedling length on day 4 divided by the cool (14
0
C) seedling length on day 7 (Tuck et al.,
2010). The cool-warm vigour index provided the best prediction of field emergence under cool
conditions in Tuck et al. (2010).
The Sicot 71BRF seed supplied had low germination percentages below 50 % (Tables 1 and 2). PBZ did
not affect the germination percentage at 4 d, but seedling length in the warm germination test at 4 d was
significantly different (Table 1). Seedling length was reduced by up to 20 % in the PBZ treatments
compared with the untreated control and there was a second order polynomial response of seedling length
with PBZ concentration (Figure 1). There were no significant differences in seedling length in the cold
germination treatment on 7 d at 14 C. While there were significant differences in the cool-warm vigour
index, there was no improvement in the index with the application of PBZ over the control.
TABLE 1: EFFECT OF PACLOBUTRAZOLE (PBZ) CONCENTRATION (G/100 KG) ON GERMINATION AND SEEDLING LENGTH AT 30 C ON DAY 4, AT 14 C ON DAY 7 AND COOL-WARM SEEDLING
LENGTH (AVERAGE OF WARM SEEDLING LENGTH (MM) AT 30 C ON DAY 4 AND 14C ON DAY7
PBZ treatment
(mL/100 kg)
Germination Probability
on at 30 C on Day 4
Seedling length (mm)
at 30 C on Day 4
Seedling length (mm)
at 14 C on Day 7
Cool-Warm Seedling
Length (mm)
0 (Control) 0.48 100 13 57
4 0.47 81 11 46
8 0.40 83 11 47
16 0.49 85 11 48
32 0.48 83 13 48
64 0.55 76 11 43
P value 0.071 <0.001 0.5 <0.001
LSD (5%) - 7.8 3.3 4.5
418
Fig. 1: Eff
TXP d
germinatio
length at 7
other treat
TABLE 2: EFFE
TXP Trea
(g/100
0 (Cont
4
20
40
80
160
P val
LSD (5
Neith
evidence t
more tha
concentra
Rademach
seedlings,
fungicidal
ability of
similar 14
fect of Paclobutro
did not signi
on test, how
7 d (Table 2
tments were
CT OF TRINEXAPAC-ET
atment
kg)
Germ
on
trol)
0
lue
5%)
er PBZ nor T
that PBZ or
an TXP com
ations of PB
her, 2000; Ch
, more root h
l effect has b
PBZ to inhi
4x-demethyla
World Cot
zol (PBZ) and Trin
ificantly affe
wever signifi
2). Seedling l
less, but the
HYL (TXP) CONCENTR
SEEDLING LENGTH
mination Proba
at 30 C on D
0.48
0.50
0.50
0.41
0.47
0.53
0.139
-
TPX was ph
TXP enhanc
mpared with
Z and TXP
hynoweth et
hairs and ha
been reported
ibit sterol bio
ation step in
tton Research C
nezapac ethyl (TXP
ect the germi
icant differen
length was s
se difference
RATION (G/100 KG) ON
H (AVERAGE OF WARM
ability
ay 4
Seedl
at
hytotoxic to c
ced cotton ge
h the contr
P (Figure 1),
t al., 2010). S
ad reduced fu
d for PBZ bu
osynthesis is
fungi (Haug
Conference on T
P) Concentration (
ination perce
nces were m
imilar in the
es did not ch
N GERMINATION AND SE
M SEEDLING LENGTH AT
ling Length (m
30 C on Day
100
104
98
91
102
93
0.3
-
cotton seeds
ermination in
rol. Seedling
, as observe
Seeds treated
ungal infecti
ut not TXP (
s related to it
ghan et al., 19
Technologies fo
mL/100 kg) on Co
entage or the
measured in
e control, 4 a
ange the coo
EEDLING LENGTH (MM)
30 C ON DAY 4 AND
mm)
4
Seedling
at 14
(at the conc
n cold condit
g length (g
ed in earlier
d with PBZ a
ons at Day
Haughan et
ts antifungal
988).
or Prosperity

tton Seedling Leng
seedling len
the cold ger
and 20 g/100
ol warm vigo
AT 30 C ON DAY 4,
D 14C ON DAY 7)
g Length (mm)
C on Day 7
13
13
12
11
11
10
0.032
1.9
centrations u
tions. PBZ re
growth) was
studies (Da
and TXP pro
10 at 30 C
al., 1988; Ci
l abilities, th
gth (mm) at 30 C
ngth at 4 d in
rmination fo
0 kg treatmen
our index.
AT 14 C ON DAY 7 A
) Cool-Warm
Length
57
58
55
51
56
51
0.1
-
used), and the
educed seedl
inversely
avis and Cu
oduced stoute
(data not sh
imen et al., 2
hrough inhibi
C on Day 4
n the warm
or seedling
nts, and all
AND COOL-WARM
m Seedling
h (mm)
7
8
5
1
6
1
71

ere was no
ling length
related to
urry, 1991;
er (thicker)
hown). The
2003). The
ition at the
Effect of Cool Conditions on Cotton Seedlings 419
CONCLUSION
The PGRs PBZ and TXP did not improve cold tolerance in cotton seeds and seedlings, but they reduced
cotton seedling lengths in warm conditions. Thus, while the treatments did not improve performance
under cool conditions, they had the potential to reduce performance under warm conditions.
ACKNOWLEDGEMENT
We thank the BION fund (Cotton Seed Distributors and Syngenta) for financial support and Ken Mckee
and Robert Battaglia for seed coating with the plant growth regulators.
REFERENCES
[1] Bange MP, Milroy SP. (2004). Impact of short-term exposure to cold night temperatures on early development of cotton
(Gossypium hirsutum L.). Australian J. Agril. Res. 55: 655664.
[2] Bird LS, Reyes AA. (1967). Effects of cottonseed quality on seed and seedling characteristics. Proceedings of the Beltwide
Cotton Production Research Conference, pp. 199206.
[3] Chynoweth R, Rolston M, BL M. (2010). Plant growth regulators: a success story in perennial ryegrass seed crops. In 'Seed
symposium: Seeds for the Future. Proceedings of a Joint Symposium between the Agronomy Society of New Zealand and
the New Zealand Grassland Association, pp. 4757. (Agronomy Society of New Zealand: Massey University, Palmerston
North, New Zealand).
[4] Cimen I, Basbag S, Temiz M, Sagir A. (2003). Effect of paclobutrazol on earliness of cotton (Gossypium hirsutum). Indian
J. of Agril. Sci. 73: 298300.
[5] Cole DF, Wheeler JE. (1974). Effect of pregermination treatments on germination and growth of cottonseed at suboptimal
temperatures. J. Crop Sci. 14: 451454.
[6] Constable GA, Eveleigh R, Kay A, Marshall J. (1998). Replanting Guide - Cotton Seed Distributors Grower Information.
(Cotton Seed Distributors: Narrabri).
[7] Constable GA, Shaw A.J. (1988). Temperature requirements for cotton. New South Wales Department of Agriculture and
Fisheries P5.3.5.
[8] Davis TD, Curry EA. (1991). Chemical regulation of vegetative growth. Critical Rev. Plant Sci. 10: 151188.
[9] Duesterhaus B. (2000). Laboratory screening test for the evaluation of cold tolerance in cotton. Texas Tech University.
[10] Duesterhaus B, Hopper NW, Gannaway JR, Valco TD. (2000). A screening test for the evaluation of cold tolerance in
cottonseed germination and emergence. Proceedings of the Beltwide Cotton Conference. Memphis, Tennessee pp. 596599.
(National Cotton Council).
[11] Gavidia I, Zaragoza C, Segura J, Perez-Bermudez P. (1997). Plant regeneration from juvenile and adult Anthyllis
cytisoides, a multipurpose leguminous shrub. J. Plant Physiol., 150: 714718.
[12] Haughan PA, Lenton JR, Goad LJ. (1988). Sterol requirements and paclobutrazol inhibition of a celery cell-culture.
Phytochemistry 27, 24912500.
[13] Heckman NL, Elthon TE, Horst GL, Gaussoin RE. (2002). Influence of trinexapac-ethyl on respiration of isolated wheat
mitochondria. J. Crop Sci., 42: 423427.
[14] Lauterbach B, Krieg DR, Jividen GM. (1999). Fatty acid composition of lipid fractions in germinating cotton as affected by
temperature. Proceedings of the beltwide cotton conference. Memphis, Tennessee pp. 564565. (National Cotton Council of
America).
[15] Rademacher W. (2000). Growth retardants: effects on gibberellin biosynthesis and other metabolic pathways. Ann. Rev.
Plant Physiol. Plant Mol. Biol., 51: 501531.
[16] Tuck CA, Tan DKY, Bange MP, Stiller WN. (2010). Cold-tolerance screening for cotton cultivars using germination chill
protocols. Food Security from Sustainable Agriculture. Proceedings of 15th Agronomy Conference 2010. (Eds H Dove, RA
Culvenor). (Australian Society for Agronomy: Lincoln, New Zealand).
[17] Wanjura DF, Buxton DR. (1972). Water uptake and radicle emergence of cottonseed as affected by soil moisture and
temperature. Agronomy J., 46: 427431.
Increased Nutrient Uptake and Salinity Tolerance

in AhCMO Tansgenic Cotton
Huijun Zhang
1
, Jianlong Dai
2
and Hezhong Dong
2
1
Cotton Research Institute, Shanxi Academy of Agricultural Sciences, Yuncheng044000 China
2
Cotton Research Center, Shandong Key Lab for Cotton Culture and Physiology, Shandong
Academy of Agricultural Sciences, Jinan250100 China
E-mail: sjhuij@sohu.com, donghz@saas.ac.cn
AbstractMany previous studies have shown that salinity stress limits nutrient uptake in cotton (Gossypium
hirsutum L.), but the relationships between salt tolerance and nutrients uptake was not fully studied. The objective of
this study was to determine if improved nutrient uptake increases salinity tolerance of cotton. Two experiments with
either transgenic salt-tolerant cotton or a split-root system were conducted in the greenhouse. In the first experiment,
a transgenic AhCMO cotton line (CMO
4
) with increased salt tolerance and its wild line (SM
3
) were grown under
salinity stress in pots containing substrate mixture (seedling substrate: vermiculite = 1:1, v/v). In the second
experiment, cotton plants were cultured in hydroponics with a split-root system, in which one part of the root system
was non-stressed (C) and the other part was salt-stressed (S). Both sides of the root system were also fed with either
low (LN) or moderate level of nutrient solution (MN). Contents of essential nutrient elements (N, P, K, Ca, Mg, Fe,
Mn, Cu, Zn) and Na
+
in plant tissues, leaf photosynthesis (Pn) and plant biomass were determined after salinity
(NaCl) treatment in both experiments. In the first experiment, salinity stress with 150 mM NaCl reduced plant
biomass, leaf Chl, and Pn of both SM
3
and CMO
4
compared with their non-stressed controls. However, the CMO
4

suffered significantly lower reductions than the wild line, SM
3
, suggesting an increased salinity tolerance of CMO
4

relative to SM
3
. Total uptake and contents of main nutrient elements (N, P, K, Ca, Mg, Fe, Mn, Cu, Zn) in CMO
4

were higher than those in SM
3
. Also, less Na
+
accumulation and lower extreme ratios of Na/N, Na/P, Na/K, Na/Ca,
Na/Mg, Na/Fe, Na/Mn, Na/Cu and Na/Zn were observed in CMO
4
than in SM
3
30 days after salt stress (DAS).
Increased salt tolerance in transgenic AhCMO cotton may be attributed to its superior nutrient uptake compared with
SM
3
. In the second experiment, the non-stressed root half fed with moderate nutrient solution and salt-stressed half fed
with low nutrient solution (CMN/SLN) in the split-root system experienced increased dry weight (22.3%), leaf area
per plant (15.5%), leaf Pn (33.3%) and Chl content (35.3%), relative to those with salt-stressed root-part fed with
moderate level of nutrient solution and non-stressed root-part fed with low nutrient solution (CLN/SMN). Plants
absorbed more N, P, K, Ca, Mg, Fe, Mn, Cu and Zn, but less Na
+
under CMN/SLN than CMN/SMN. Moreover,
ratios of Na/N, Na/P, Na/K, Na/Ca, Na/Mg, Na/Fe, Na/Mn, Na/Cu and Na/Zn in CMN/SLN were lower than those
in CLN/SMN. Increased plant growth with high concentrations of nutrients in lower saline root part was also
attributed to enhanced uptake of nutrients. The overall results suggest that improved nutrient uptake increases
salinity tolerance of cotton.
Keywords: Cotton, Salt tolerance, Nutrient uptake, Split-root
INTRODUCTION
Soil salinity is one of the major abiotic stresses affecting plant productivity (Yildirim et al. 2009).
Although cotton (Gossypium hirsutum L.) is classified as a salt tolerant crop, it is negatively affected by
excessive salt in the soil, particularly in arid and semiarid regions. It is generally believed that soil
salinity affects plant growth and development by ways of osmotic stress, injurious effects of toxic ions
and the resulting nutrient imbalance (Sairam and Tyagi, 2004). Therefore, searching for ways or
techniques to reduce osmotic stress and ion toxicity under salinity has attracted great research attention
world-wide (Gorham et al., 2009; Munns and Tester, 2008).
Ways or techniques to reduce osmotic stress and ion toxicity have been explored by improving
agronomic practices. Seed-bed preparation (El-Swaify, 2000; Meiri and Plaut, 1985), plastic mulching
(Dong et al., 2008, 2009), and foliar application of plant growth regulators (Hoque et al., 2007) were
reported to reduce either osmotic stress or ion toxicity, and significantly decreased salt injury in the
greenhouse or saline field conditions. Glycine betaine is an osmoprotectant that is correlated with salt
tolerance (Sulpice et al., 2003; Sakamoto and Murata, 2001). Foliar application of glycine betaine
71
Increased Nutrient Uptake and Salinity Tolerance in AhCMO Tansgenic Cotton 421
improved salt tolerance of rice (Harinasut et al., 1996). Glycine-betaine is synthesized from choline
through two steps: cholinebetaine aldehydeglycine betaine, in which choline monooxygenase
(CMO) catalyzes the first step and betaine aldehyde dehydrogenase (BADH) catalyzes the second step
(Rhodes and Hanson, 1993). Transgenic cotton lines carrying CMO gene cloned from Atriplex hortensis
accumulated significantly more glycine betaine and less Na
+
than their wild line, and showed increased
salt tolerance in either greenhouse or field conditions (Zhang et al., 2007, 2009). Increased salt tolerance
of the AhCMO transgenic cotton lines was attributed to reduced osmotic stress and ion toxicity (Zhang
et al., 2009). However, it is still not clear if the increased salt tolerance is also related to nutrients uptake
under salinity stress.
Salinity levels are seldom uniform in saline fields. Hence, a part of the root system would be under
lower salinity than the other root part (Kaman et al., 2006; Zekri and Parsons, 1990). Such unequal salt
distribution can be accurately simulated with a split-root system, in which the root systems are divided
into two or more equal portions and each portion irrigated with varied concentrations of NaCl solution
(Shani et al., 1993; Zhu and Ito, 2000; Messedi et al., 2004; Lycoskoufis et al., 2005). Under the split-
root system, many studies indicated that unequal salt distribution improved plant growth and yield in
tomato (Tabatabai et al., 2004), cucumber (Sonneveld and De Kreij, 1999), halophytic shrub (Bazihizina
et al., 2009) and cotton (Dong et al., 2010) compared to equal salt distribution. However, the underlying
mechanism of plant growth enhancement under unequal salt distribution, particularly the relationship
between salt tolerance and nutrient uptake is not fully understood.
The relation between salt tolerance and nutrient uptake is an important basis for cotton fertilization in
saline soils. It was usually studied by using various fertilizer rates or forms (Elgharably et al., 2010;
Gimeno et al., 2009; Irshad et al., 2008). Previous studies with the traditional system have shown that
soil salinity inhibits uptake of nutrients in cotton, and proper fertilization alleviates the detrimental effects
of salinity under certain conditions (Rathert, 1983; Martinez and Lauchli, 1991; Chen et al., 2010).
Adequate N fertilization increased N uptake and plant growth at both low and medium soil salinities, but
had little effect at higher salinity even at an elevated N rate (Chen et al., 2010). As the amount of nutrient
uptake does not always change with fertilizer rate under moderate and high salinity, it is very difficult to
determine the relation between salt tolerance and nutrient uptake using the traditional system. In contrast,
varying nutrient uptake rates in cotton plants can be realized with unequal salt distribution under the
same salinity level in a split-root system, this helps to accurately study the tolerance-uptake relations.
In this paper, one experiment was conducted with an AhCMO transgenic cotton line to determine if
the improved salt tolerance of the transgenic cotton was related to increased nutrient uptake. The other
experiment was conducted in a split-root system to determine if plants with non-stressed root half in high
nutrient solution could uptake more nutrients and grow better than those with salt-stressed root half in
high nutrient solution. Our main objective was to determine if improved nutrient uptake increases salinity
tolerance of cotton.
Plant Culture and Treatment in Experiment 1
An elite Chinese cotton (Gossypium hirsutum L.) cv. Simian 3 (SM
3
, wild type) was transformed with the
AhCMO gene (gb: AF270651, patent no.: ZL00109 164.6) from Atriplex hortensis, mediated by
Agrobacterium tumefaciens in our previous study (Zhang et al., 2009). A transgenic line CMO
4
carrying
AhCMO gene at the T4 generation and wild line (non-transgenic) SM
3
were used in this experiment.
Seeds of CMO
4
and SM
3
were sown in 5 L plastic pots containing substrate mixture (seedling substrate:
vermiculite=1:1, v/v) and allowed to grow in the greenhouse at 32/24
o
C and relative humidity of 35
50%. Seedlings were thinned to five of uniform size per pot when the first true-leaf of most seedlings
fully expanded. Salinity treatment was imposed to seedlings at the 2 true-leaf stage by irrigating with
NaCl solution. This was added at 50 mM increments every 12h, until the final concentration of NaCl
reached 150 mM. The experiment was arranged into a completely randomized design with four
422
replicates
other day,
to avoid a
NaCl-free
Plant Cultu
Commerc
experimen
10 cm) co
of 16/8 h
leaves we
establish
blade on t
The top o
seedling.
parafilm.
with wate
topped up
grafted se
seedlings
3550% r
two unifor
Fig. 1: Establ
Accor
cotton see
nutrient so
The nutri
EDTA. Fe
adjusted t
Salini
solution (
was grad
combinati
split-root
NaCl + 1/
nutrient so
in the con
. Each replic
, and plants w
accumulation
e control (CK
ure and Treatm
cial high-yiel
nt. Seeds of
ontaining ste
, light intens
ere carefully
split-root sy
the hypocoty
of the rootsto
The sect
Grafted seed
er and imme
p with deion
eedling at tw
with two un
relative hum
rm split-root
ishment of a Split-
rding to our
edlings, and
olutions serv
tion solution
eNa; and (M
o pH 6.5 wit
ity stress trea
MN or LN),
dually increa
ions was thu
system, CMN
/8 concentra
olution, CLN
ntainer was r
World Cot
cate had 5 p
were irrigate
n of more sa
K). Seedlings
ment in Exper
lding cotton
SCRC28 we
rilized wet s
sity of 400
removed fro
ystems throug
yl 2 cm below
ock was cut t
tion was then
dlings were t
ediately cov
nized water a
wo weeks a
niform split-r
midity for 20
t systems we
-root System thro
Rootstock (). (
preliminary
1/8-strength
ved as moder
n consisted
M): 20 H
3
BO
th KOH.
atment was a
, and those w
ased at a 5
us formed by
N/SLN (CM
ated nutrient
N = 0 mM N
replaced at a
tton Research C
plants. Irriga
ed until a lar
alt than expe
s were sampl
riment 2
(Gossypium
ere sown at a
sand. Boxes
mol m
2
s
om the sand a
gh grafting (
w the two co
to form a de
n inserted in
transferred t
ered with p
as required a
after graftin
root systems
days. Nutrie
ere selected f
ough Grafting. (a) a
(c) A Well-grafted
trials, half-c
h nutrient sol
rate-level (M
of (mM): 2.
O
3
, 1.0 ZnSO
achieved by a
without NaC
0 mM incr
y adjusting
MN = 0 mM N
solution) an
NaCl + 1/8 c
a 3-day interv
Conference on T
ation was co
rge amount o
ected. Plants
led and analy
m hirsutum L
approximatel
were placed
1
PAR, and
and washed w
(Fig. 1). Bri
otyledons, le
eep at the
nto the
to plastic pot
lastic bags t
and renewed
g, the plast
were transfe
ent solution
for further ex
a Cotton Seedling
d Seedling with Two
concentrated
lution is quit
MN) and low-
.5 Ca(NO
3
)
2
O
4
, 0.2 CuSO
addition of 1
Cl addition se
rement per
concentratio
NaCl + half-
nd SMN/CLN
oncentrated
val. The amo
Technologies fo
onducted in
of saline wate
s irrigated w
yzed 30 days
L.) cultivar S
ly 3 cm depth
in growth c
temperature
with water. U
efly, a
aving about
e same posit
incision of t
ts containing
to prevent w
d weekly. W
tic bag and
erred to gree
was renewe
xperiment.
with Shaped
o Split-roots (Graf
nutrient solu
te inadequate
-level of nutr
2
, 0.5 NH
4
H
O
4
, 1.0 MnS
150 mM NaC
erved as non
day until re
ns of NaCl
concentrated
N (SMN = 1
nutrient solu
ount of evap
or Prosperity
the morning
er leaked thr
with non-salin
s after salt str
SCRC 28 w
h in plastic b
chambers und
e of 30. Se
Uniform seed
shaped inci
1/3 of the hy
tion of the h
the plant and
g aerated nut
wilting. The
When a new l
parafilm w
enhouse to gr
ed per day. H

Incision (). (b) P
fting Position ).
ution is good
e. Thus, half
rient (LN) tr
2
PO
4
, 2.5 K
SO
4
, 0.005 (N
Cl (S) to the
n-salt stresse
eached 150
and nutrient
d nutrient sol
150 mM NaC
ution). The w
porated water
g (7:007:30
rough the bo
ne water ser
ress (DAS).
was used in t
boxes (20cm
der a light/da
eedlings with
dlings were
ision was ma
ypocotyl tiss
hypocotyl fro
d closely wra
trient solutio
nutrient sol
leaf emerged
were removed
row under 32
Healthy seed
Part of a Seedling a
d for normal
f- and 1/8 co
eatments, res
KNO
3
, 1.0 M
NH
4
)
6
Mo
7
O
2
correspondin
ed control (C
mM. Two
ts in both si
lution, SLN
Cl + half-co
whole nutrien
r was replen
0) of every
ttom holes
rved as the
the second
m 15cm
ark regime
h two true
selected to
ade with a
sues intact.
om another
apped with
on, sprayed
lution was
d from the
d. Grafted
2/24 and
dlings with
as Shaped
growth of
oncentrated
spectively.
MgSO
4
, 0.1
24
, and was
ng nutrient
C). Salinity
treatment
ides of the
= 150 mM
oncentrated
nt solution
nished with
distilled water daily. Seedling plants were sampled and analyzed 42 days after treatment. The experiment
was arranged into a completely randomized design with four replicates. Each replicate had 3 plants.
Data Collection in Both Experiments
Data collected were fresh and dry weight of a whole plant and its organs (leaf, stem and root),
concentrations of mineral elements (N, P, K, Ca, Mg, Fe, Mn, Cu, Zn and Na
+
) in plant tissues, Pn rate
and Chl content in the 4
th
leaf on the main stem from growth terminal at 30 DAS in the 1
st
experiment
and 42 DAS in the 2
nd
experiment.
Determination of Growth Parameters
In the first experiment, plants were removed from pots carefully at 30 DAS. The remaining roots in the
substrate were also collected by washing the potted substrate through a 4 mm sieve. In both experiments,
plants were divided into leaves, stems and roots, and their respective fresh weight (FW) was weighed.
Leaf area was measured by passing the leaves through a LI-3000 area meter (LI-COR, Lincoln, NE,
USA). Dry weight (DW) was determined after oven drying at 75 until a constant weight was reached.
Determination of Physiological Parameters
Net photosynthetic (Pn) rate of the 4
th
fully expanded young leaf on the main-stem from terminal was
taken between 09:00 and 11:00 h on cloudless days when ambient photosynthetic photon flux density
exceeded 1500 mol m
2
s
1
, using a LI-6400 portable photosynthesis system (Li-Cor, Lincoln, NE,
USA). Leaf chlorophyll (Chl) contents were determined as described in He et al. (2002). Briefly, 0.20 g
fresh leaves were placed in a 100 mL test tube. The tissues were homogenized with a polytron after
adding 10-15 ml pure methanol. The homogenate was then filtered and made up to 100 mL with pure
methanol. The Chl concentration in the supernatant was spectrophotometrically determined by measuring
the absorbances at 652 and 665 nm for Chl a and Chl b, respectively.
Determination of Mineral Concentrations
Ashing was carried out by means of incineration in a muffle oven at 45025 until an ash was obtained.
The ash was acid digested (HCl) for the determination of micro and macro elements. Nitrogen was
determined by Kjeldahl procedure. Phosphorus was determined spectro-photometrically (TU-1901,
Beijing, China). The elements potassium (K), sodium (Na), calcium (Ca), magnesium (Mg), iron (Fe),
manganese (Mn), cuprum (Cu) and zinc (Zn) were measured by an atomic absorption spectrophotometer
(TAS-990, Beijing, China) equipped with hollow cathode lamps. Recovery of added known amounts of
standards to samples gave 98105% of expected values for all minerals.
Statistical Analysis
Data were statistically analyzed with DPS Data Processing System (Tang and Feng, 1997). Means were
separated using a t-test.
RESULTS
The 1
st
Experiment
Plant biomass
Salinity stress with 150 mM NaCl significantly reduced plant growth of both SM
3
and CMO
4
in terms of
their fresh weight and dry weight of root, stem and leaves (Table 1). Root, stem, leaf and total plant of
CMO
4
were decreased by 33.9, 45.8, 37.7 and 36.8% in fresh weight and 20.0, 28.4, 30.0 and 24.1% in
dry weight, respectively, while those of SM
3
were decreased by 58.8, 61.3, 65.5 and 62.7% in fresh
weight and 50.0, 56.6, 60.0 and 53.8% in dry weight, compared with their respective NaCl-free controls.
It was noted that the salt-induced decrement in both fresh and dry weights in CMO
4
was significantly
lower than that in SM
3
.
TABLE 1: FRESH AND DRY WEIGHTS OF TRANSGENIC AHCMO COTTON CMO
4
AND ITS WILD LINE SM
3
30 DAYS AFTER SALT (150 MM NACL) STRESS
Cultivar Treatment Fresh Weight (g. Plant
-1
) Dry Weight (g. Plant
-1
)
Root Stem Leaf Total Root Stem Leaf Total
SM
3
CK 6.86a 8.05a 11.09a 26.0a 1.23a 1.10a 1.63a 3.96a
Salt 2.80d 3.11c 3.80d 9.7d 0.64d 0.49c 0.68c 1.81d
CMO
4
CK 5.59b 5.91b 7.60b 19.1b 1.01b 0.70b 1.21b 2.92b
Salt 3.78c 3.27c 4.85c 12.0c 0.83c 0.53c 0.84c 2.20c
*Different letters within a column indicate significant differences at p=0.05.
Photosynthesis
Salt stress significantly reduced net photosynthetic (Pn) rate and chlorophyll (Chl) content in the 4
th
main
stem leaf from terminal for both SM
3
and CMO
4
at 30 DAS compared with their respective control
(Table 2). However, the decrement in Pn (26.9%) and Chl content (17.5%) in CMO
4
was lower than that
of Pn (36.3%) and Chl (24.4%) in SM
3
. Moreover, Na
+
concentration in roots, stems and leaves of CMO
4

was 6.1%, 11.6% and 26.4% lower than that of SM
3
under saline stress, respectively (Table 2). The result
indicated less accumulation of Na
+
in CMO
4
than in SM
3
tissues under salt stress.
TABLE 2: CHLOROPHYLL (CHL) CONTENT, PHOTOSYNTHETIC (PN) RATE OF MAIN-STEM LEAVES AND NA CONTENTS IN TRANSGENIC AHCMO COTTON CMO
4
3
30 DAYS
AFTER SALT (150 MM NACL) STRESS.
Cultivar Treatment Na content (mg.g
-1
) Chl
(mg.g
-1
FW)
Pn
(molCO
2
.m
-2
.s
-1
) Root Stem Leaf
SM
3
CK 7.53c 3.33b 4.00c 17.2a 21.5a
Salt 11.50a 13.80a 13.41a 13.0c 13.7c
CMO
4
CK 6.59d 3.03b 3.25c 17.7a 23.4a
Salt 10.81b 12.21a 9.86b 14.6b 17.1b
* Different letters within a column indicate significant differences at p=0.05.
Uptake of some mineral elements
Differences in mineral nutrient content (uptake) and the ratios of Na to other mineral nutrient elements
were observed between SM
3
and CMO
4
under salt stress. NaCl stress increased the content of N, P, K,
Ca, Mg, Fe, Mn, Cu and Zn in the leaves of CMO
4
, compared with the non-stressed control. Whereas in
the leaves of SM
3
, content of P, K, Ca, Fe, Mn and Zn decreased and only N, Mg and Cu concentrations
increased . Similar results were obtained in the stem and root. Moreover, the content of N, P, K, Ca, Mg,
Fe, Mn, Cu and Zn in leaf, stem and root in CMO
4
were higher than that in SM
3
under NaCl stress
(Table 3).
TABLE 3: NUTRIENT ELEMENTS CONTENTS OF TRANSGENIC AHCMO COTTON CMO
4
3
AT 30 DAYS AFTER SALT (150 MM NACL) STRESS
Cultivar Treatments Macronutrients (mg.g
-1
DW) Micronutrients (g.g
-1
DW)
N P K Ca Mg Fe Mn Cu Zn
-----------------------------------------------------------Root-----------------------------------------------------
SM
3
CK 7.4c 0.94c 5.56c 0.33a 0.22b 366a 40.0b 28.2c 49.0c
Salt 8.5b 0.97c 5.32d 0.26c 0.17c 174d 28.1c 32.1b 36.0d
CMO
4
CK 7.5c 1.37b 5.95b 0.34a 0.21b 323b 44.1b 23.3d 70.3b
Salt 10.4a 1.75a 7.21a 0.29b 0.25a 258c 53.0a 35.0a 125.0a
-----------------------------------------------------------Stem-----------------------------------------------------
SM
3
CK 8.5c 1.54b 8.60b 0.43d 0.19c 301b 24.2d 26.1c 18.2d
Salt 11.0b 1.24c 8.41b 0.46c 0.22b 225c 31.4c 30.0b 26.3c
CMO
4
CK 7.9c 1.01d 8.40b 0.48b 0.17c 289b 36.1b 23.0d 50.1b
Salt 13.1a 2.07a 9.80a 0.51a 0.26a 379a 43.8a 34.1a 81.0a
-----------------------------------------------------------Leaf------------------------------------------------------
SM
3
CK 15.5c 2.56a 5.50b 1.26c 0.56c 404b 136c 44.0b 83c
Salt 16.8b 1.66c 5.01c 1.22d 0.60b 328d 97d 46.1a 56d
CMO
4
CK 15.3c 2.28b 4.60d 1.31b 0.57bc 375c 213b 36.0d 109b
Salt 18.2a 2.48a 5.80a 1.46a 0.65a 537a 266a 40.0c 139a
*Different letters within a column about root, stem and leaf indicate significant differences at p=0.05.
Salt stress decreased total amount of mineral nutrients per plant in both SM
3
and CMO
4
compared
with their respective control (Fig 2). However, the amount of N, P, K, Ca, Mg, Fe, Mn, Cu and Zn in
CMO
4
was reduced by 4.4, 6.4, 7.6, 24.9, 13.8, 16.1, 12.6, 2.3 and 2.3%, while the amount of N, P, K,
Ca, Mg, Fe, Mn, Cu and Zn in SM
3
was decreased by 49.5, 65.7, 56.8, 58.2, 55.6, 68.9, 67.1, 50.9 and
65.3%, respectively. The results indicated that the decrement in CMO
4
was lower than in SM3.
Moreover, total amount of mineral nutrients per plant in transgenic AhCMO cotton were significantly
higher than in SM
3
under NaCl stress.

Fig. 2: Total Accumulation of Nutrient Elements Per Plant in Transgenic AhCMO Cotton CMO
4
and its Wild Line SM
3
30 Days after Salt (150 mM NaCl)
Stress. Means Followed by Different Letters Differ Significantly (p=0.05). Vertical Bars Show SD of Four Replicates with Five Plants Per Replicate
Salinity stress significantly increased the ratios of Na to main nutrient elements in different tissues
for both cultivars relative to their respective control, but the increment of Na/N, Na/P, Na/K, Na/Ca,
Na/Mg, Na/Fe, Na/Mn, Na/Cu and Na/Zn in CMO
4
was significantly lower than that in SM
3
. The ratios
of Na to main nutrient elements in leaf, stem and root of CMO
4
were also lower than that in SM
3
under
NaCl stress (Table 4).
TABLE 4: RATIOS OF NA TO NUTRIENT ELEMENTS IN TRANSGENIC AHCMO COTTON CMO
4
3
30 DAYS AFTER SALT (150 MM NACL) STRESS.
Cultivar Treatments Na/N Na/P Na/K Na/Ca Na/Mg Na/Fe Na/Mn Na/Cu Na/Zn
-----------------------------------------------------------Root--------------------------------------------------------
SM
3
CK 1.03b 8.0b 1.37c 23.0c 34.2c 21.1c 188c 271c 154b
Salt 1.16a 10.0a 1.85a 35.1b 59.0a 56.2a 356a 311b 275a
CMO
4
CK 0.88c 4.7c 1.11d 19.1c 32.2c 20.0c 151c 267c 99c
Salt 1.03b 7.3b 1.78b 47.0a 52.1b 50.1b 242b 370a 102c
-----------------------------------------------------------Stem-------------------------------------------------------
SM
3
CK 0.39c 2.27d 0.39c 7.8c 18.4c 11.3c 128c 128c 188b
Salt 1.28a 11.0a 1.64a 30.0a 62.0a 62.2a 442a 456a 537a
CMO
4
CK 0.38c 3.0c 0.36c 6.3d 17.1c 11.2c 84d 130c 61d
Salt 0.72b 6.3b 1.33b 26.1b 51.0b 35.0b 299b 389b 161c
-----------------------------------------------------------Leaf--------------------------------------------------------
SM
3
CK 0.26c 1.6c 0.72c 3.2c 7.1c 9.9c 29b 91c 48c
Salt 0.86a 8.7a 2.88a 12.1a 24.0a 44.1a 151a 316a 262a
CMO
4
CK 0.22c 1.4c 0.71c 2.5d 5.7d 8.1d 15c 91c 29d
Salt 0.47b 4.0b 1.69b 6.7b 15.3b 19.0b 37b 246b 75b
* Different letters within a column about root, stem and leaf indicate significant differences at p=0.05.
0
10
20
30
40
50
N K
A
c
c
u
m
u
l
a
t
i
o
n

(
m
g
.

P
l
a
n
t -
1
)

. SM3-CK SM3-Salt
CMO4-CK CMO4-Salt
0
2
4
6
8
P Ca Mg Fe
A
c
c
u
m
u
l
a
t
i
o
n

(
m
g

P
l
a
n
t -
1
)

.
0
100
200
300
400
Mn Cu Zn
A
c
c
u
m
u
l
a
t
i
o
n

(
g
.

P
l
a
n
t -
1
)
a
a
a
a
a
a
a
a
a
b
b
b
b
b
b
b
b
b
c
c
c
c
c
c
c
c
c
d
d
d
d
d
d
d
d
d
426
THE 2
ND
EX
Plant Biom
Treat
CMN
CLN
CMN
CLN
* Different lette
In the spl
plant. Th
concentra
plus mod
(CMN/SL
(22.3%), c
and salinit
Photosynth
The uptak
in CMN/S
Treatm
CMN/S
CLN/S
* Differ
Fi
XPERIMENT
mass
TABLE 5
tments
-------------
N/SLN
N/SMN
-----------
N/SLN
N/SMN
ers within a column a
lit-root syste
he root-half
ated (1/2-con
derate level
LN) significa
compared w
ty stress plus
hesis and Na
+

ke and partiti
SLN than in C
TABLE 6: LEAF AR
ments
Sp
SLN CMN
SLN
SMN CLN
SMN
rent letters within
g. 3: Cotton Plants
World Cot
5: FRESH AND DRY WEI
Split Roo
-------------------
CMN 2
SLN 1
CLN 1
SMN 1
------------------
CMN 4
SLN 3
CLN 3
SMN 2
about fresh weight a
m, one root-
was also fe
ncentrated) (M
of nutrition
antly increas
ith the treatm
s moderate n
Accumulation
ioning of Na
+
CLN/SMN,
REA, CHLOROPHYLL (CH
UNDER
Na Content
plit Root S
N 1.3d 5
N 8.1a
N 1.6c 5
N 7.7b
n a column indica
s (a) and the 4th M
tton Research C
IGHTS OF COTTON UNDE
Root
ot
------------Fresh
21.3a
13.3b
18.1a
10.9b
--------------Dry
4.13a
3.02b
3.41b
2.26c
and dry weight indica
-half was sa
ed in either
MN) nutrien
in one side
sed plant gr
ment combin
nutrient in the
n
+
varied betw
especially in
HL) CONTENT, PHOTOS
R UNEQUAL SALT AND N
(mg.g
-1
)
Stem Leaf
5.32b 11.8b
5.77a 13.5a
ate significant diff
Main-stem Leaves
Conference on T
ER UNEQUAL SALT AND
Total Weight
h Weight(g. pla
34.6a
29.0b
y weight (g. pla
7.17a
5.64b
ate significant differ
lt stressed (S
r low conce
nt solution. T
e and salini
rowth in ter
nation with n
e other side (
ween CMN/S
n leaf (12.6%
SYNTHETIC (PN) RATE O
NUTRIENT DISTRIBUTIO
Chl
(mg.g
-1
F f
b 9.2a
a 6.8b
ferences at p=0.0
(b) under Unequa
Technologies fo
D NUTRIENT DISTRIBUTI
Stem
ant
-1
)------------
49.1a
38.6b
ant
-1
)-------------
16.3a
13.3b
ences at p=0.05.
S) and the o
entrated (1/8
Treatment co
ity stress plu
rms of fresh
non-salinity s
(CLN/SMN)
SLN and CLN
%) and stem (
OF ON MAIN-STEM LEA
N 42 DAYS AFTER TRE
FW) (m
5.
l Salt and Nutrient
or Prosperity
ION 42 DAYS AFTER TR
Leaf
-------------------
31.3a
26.3b
-------------------
6.2a
5.3b
other was not
8-concentrate
ombination w
us low nutr
h weight (22
stress plus lo
) (Table 5).
N/SMN. Les
7.8%) (Tabl
AVES AND NA CONTENTS
EATMENT
Pn
molCO
2
.m
-2
.s
-1
12.8a
9.6b
t Distribution 42 D
REATMENT
Total w
--------------
115.
93.9
-------------
29.6
24.2
t-stressed (C
ed) (LN) or
with non-sali
rient in the
2.7%) and d
ow nutrient i
ss Na was ac
e 6).
S IN PLANT TISSUES
1
)
Lea
(cm
2
1
1
Days after Treatm
weight
.0a
9b
6a
2b
C) for each
r moderate
inity stress
other side
dry weight
in one side
ccumulated
af area
.plant
-1
)
345a
164b

ent
The leaf area, plant height, Chl content and Pn rate were higher in CMN/SLN than in CLN/SMN
(Table 6 and Fig. 3. a). Clear symptoms of nutrient deficiency were observed in the 4th main-stem leaf of
CLN/SMN, compared with that of CMN/SLN (Fig. 3. b). CMN/SLN had great advantage in
photosynthesis and plant growth over CLN/SMN, although both treatments were imposed with the same
level of salinity to their root systems.
Uptake of Main Mineral Nutrients
CMN/SLN treatment increased the concentration of N, P, K, Ca, Mg, Fe, Cu and Zn in leaf relative to
CLN/SMN (Table 7). Similar trends were also observed in their roots and stems. There was a significant
difference in concentrations of main mineral nutrients between the two sides of a split root for both
treatments. More N (139%), P (40.9%), K (13.0%), Ca (20.9%), Mg (9.2%), Fe (29.9%), Mn (20.5%),
Cu (7.7%) and Zn (25.6%) accumulated in CMN side than in CLN. Compared with SLN, concentrations
of N, P, K, Mn and Cu in SMN was only increased by 18.6, 9.1, 3.7, 16.5, 10.3%, respectively, and Ca,
Mg, Fe and Zn concentration even decreased.
TABLE 7: NUTRIENT CONTENTS IN DIFFERENT PLANT TISSUES UNDER UNEQUAL SALINE AND NUTRIENT DISTRIBUTION 42 DAYS AFTER TREATMENT
Elements Root Stem Leaf
CMN/SLN CLN/SMN CMN/SLN CLN/SMN CMN/SLN CLN/SMN
CMN SLN CLN SMN
M
a
c
r
o
n
u
t
r
i
e
n
t
s

(
m
g
.
g
-
1

D
W
)
N 16.0a 9.7c 6.7d 11.5b 3.5a 2.1b 13.3a 10.8b
P 3.1a 2.2c 2.2c 2.4b 1.5a 1.3b 1.5a 1.3b
K 7.8a 5.4b 6.9a 5.6b 5.3a 3.9b 7.6a 5.9b
Ca 5.2b 5.9a 4.3c 4.1c 4.8b 3.4a 8.5a 7.8b
Mg 3.8a 3.6b 3.48c 3.3d 3.1a 2.8b 4.6a 4.3b
M
i
c
r
o
n
u
t
r
i
e
n
t
s

(
g
.
g
-
1

D
W
)

Fe 291a 263b 224c 256b 104b 139a 290a 263b
Mn 40.0a 31.5d 33.2c 36.7b 30.4a 29.2b 57.3a 58.9a
Cu 33.7a 30.2c 31.3b 33.3a 25.2a 24.5a 29.5a 28.7a
Zn 108b 119a 86d 96c 85a 78b 110a 104b
* Different letters within a row indicate significant differences at p=0.05.
Total amounts of main nutrient elements per plant in CMN/SLN were significantly increased relative
to CLN/SMN (Fig. 4). The amounts of N, P, K, Ca, Mg, Fe, Mn, Cu and Zn per plant in CMN/SLN were
75.3, 43.4, 52.6, 54.1, 33.1, 19.7, 23.9, 24.9 and 63.2% more than in CLN/SMN. CMC/SLC significantly
decreased the ratios of Na
+
to most nutrient elements in different tissues, especially in leaf, compared to
CLN/SMN (Table 8).
TABLE 8: RATIOS OF NA TO NUTRIENT ELEMENTS IN ROOT, STEM AND LEAF OF COTTON PLANT UNDER UNEQUAL SALT AND NUTRIENT DISTRIBUTION 42 DAYS AFTER TREATMENT
Ratios Root Stem Leaf
CMN/SLN CLN/SMN CMN/SLN CLN/SMN CMN/SLN CLN/SMN
CMN SLN CLN SMN
Na/N 0.08d 0.83a 0.23c 0.66b 1.52b 2.75a 0.89b 1.25a
Na/P 0.43d 3.64a 0.70c 3.15b 3.55b 4.44a 7.89b 10.35a
Na/K 0.17d 1.48a 0.22c 1.35b 1.01b 1.48a 1.56b 2.28a
Na/Ca 0.25d 1.36b 0.36c 1.84a 1.11a 1.70b 1.39b 1.73a
Na/Mg 0.35b 2.22a 0.44b 2.29a 1.72b 2.06a 2.57b 3.13a
Na/Fe 4.53c 30.45a 6.84b 29.52a 51.16a 41.52b 40.83b 51.16a
Na/Mn 33d 254a 46c 206b 175b 198a 207b 228a
Na/Cu 39d 265a 49c 227b 211b 236a 401b 469a
Na/Zn 12.2d 67.3b 17.8c 78.7a 62.6b 74.0a 107.6b 129.4a
* Different letters within a row indicate significant differences at p=0.05.

Fig. 4: Total Accumulation of Nutrient Elements Per Plant under Unequal Salt and Nutrient Distribution in Cotton at 42 Days after Treatment. Means
Followed by Different Letters Differ Significantly (p=0.05). Vertical Bars Show SD of Four Replicates with Three Plants Per Replicate
DISCUSSION
Even as a salt-tolerant crop, cotton will be adversely affected in plant growth and development as well as
yield and fiber quality if soil salinity level beyond a threshold of 7.7 dS m
-1
(Chinnusamy, 2005; Maas,
1990). In the first experiment, we found a marked inhibition of fresh weight, dry weight, Pn and Chl
content in both CMO
4
and its wild line SM3 under salinity stress, but the inhibition effect of salinity on
CMO
4
was significantly lower than SM3. The result was in agreement with our previous reports that salt
tolerance of CMO
4
was greatly improved due to the introduction of AhCMO gene relative to its wild line
SM3 (Zhang et al., 2009). In the second experiment, a split-root system was established by grafting of
cotton seedlings, and unequal salt distribution in the root zone was thus constructed by irrigating one root
side with water and the other side with NaCl solution. It was shown that the fresh and dry weight, leaf
area, Pn and Chl content in CMN/SLN treatment were significantly higher than that in CLN/SMN. Since
salinity stress imposed to only half root also greatly reduced plant growth as we previously reported
(Dong et al., 2010), and both CMN/SLN and CLN/SMN were treated with same level of salinity stress
(150 mM NaCl) in the present experiment, CMN/SLN treatment improved salinity tolerance of cotton
compared to CLN/SMN.

0
50
100
150
200
250
N P K Ca Mg
A
c
c
u
m
u
l
a
t
i
o
n

(
m
g
.

P
l
a
n
t -
1
)

.
CMN/SLN CLN/SMN
a
a
a
a
a
b
b
b
b
b
0
1
2
3
4
5
6
Fe Mn Cu Zn
A
c
c
u
m
u
l
a
t
i
o
n

(
m
g
.

P
l
a
n
t -
1
)

.
a
a
a
a
b
b
b
b
Plant growth is affected by interactions of salt ions and availability or uptake of many essential
mineral nutrients (Greenway and Munns, 1980; Grattan and Grieve, 1992; Romero et al., 1994). Soil
salinity usually reduces uptake of both macronutrients like N (Dluzniewska et al., 2007), P (Rochester,
2010), K (Marschner, 1995), Ca (Yan et al., 2007) and Mg (Grattan and Grieve, 1999), and
micronutrients like Fe, Mn, Cu, Zn (Grattan and Grieve, 1999; Hirpara et al., 2005; Kholov et al.,
2009), and leads to nutrient imbalance in many plant species including cotton (Grattan and Grieve, 1999).
The ratios of Na to nutrient elements (Na/N, Na/K, Na/Ca and Na/Mg) have been used as an indication of
nutrient imbalance (Grattan and Grieve, 1999; Dluzniewska et al., 2007). In the present study, NaCl
stress significantly reduced nutrients (N, P, K, Ca, Mg, Fe, Mn, Cu and Zn) uptake in SM3 and CMO
4

relative to their respective non-saline treatment. However, the salt-induced decrements in nutrient uptake
in CMO
4
was significantly lower than that in its wild line SM
3
, and the absolute amounts of main nutrient
elements (N, P, K, Ca, Mg, Fe, Mn, Cu and Zn) in salt-stressed CMO
4
were significantly higher than in
salt-stressed SM
3
. Also less Na
+
accumulated in CMO
4
than in SM
3
as we reported previously (Zhang et
al., 2009). As a result, the ratios of Na/N, Na/P, Na/K, Na/Ca, Na/Mg, Na/Fe, Na/Mn, Na/Cu and Na/Zn
in CMO
4
was significantly reduced relative to its wild line SM
4
, and thus nutrient imbalance was
alleviated (Table 4). Although previous studies have shown that improved salinity tolerance of AhCMO
transgenic cotton was attributed to enhanced glycine betaine synthesis (Zhang et al., 2009), our results in
the present study further indicated that the increased salt tolerance was also associated with improved
nutrient uptake.
Split-root system provides an accurate system to simulate unequal salt distribution in the root-zone
(Bazihizina et al., 2009; Lycoskoufis et al., 2005). Many previous studies have shown that unequal salt
distribution improves plant growth or salt tolerance relative to equal salt distribution with the same level
of external salinity stress (Zekri and Parsons, 1990; Tabatabai et al., 2003; Bazihizina et al., 2009). In the
present study, treatment combination with non-salinity stress plus moderate nutrient level in one side and
salinity stress plus low nutrient level in the other side (CMN/SLN) significantly increased concentrations
of main nutrient elements (N, P, K, Ca, Mg, Fe, Mn, Cu, Zn) in different tissues, especially in leaf,
compared with the treatment combination with non-salinity stress plus low nutrient in one side and
salinity stress plus moderate nutrient in the other side (CLN/SMN). Similar trends were also observed in
total amount of nutrient elements per plant. The results affirmatively indicated that CMN/SLN
significantly improved nutrient uptake relative to CLN/SMN. Since both treatments were imposed the
same level of salinity stress, we suggest that the improved salt tolerance and plant growth in CMN/SLN
was attributed to the increased uptake of main nutrients. The results further showed that higher nutrients
uptake in CMN/SLN than in CLN/SMN was mainly attributed to much more nutrients uptake from the
CMN side than CLN side (Table 7). The content of Na
+
in CMN/SLN treatment is lower than that in
CLN/SMN, especially in leaf. The result was consistent with Sonneveld and Voogt (1990) who suggested
that unequal distribution of nutrients (EC=0.75/2.5) in the root environment increased N, P and K and
reduced Na
+
accumulation. More nutrients and less Na
+
uptake in CMN/SLN resulted in lower ratios of
Na
+
to the main nutrient elements (Table 8), which might alleviate nutrient imbalance of cotton.
In conclusion, introduction of AhCMO gene to cotton significantly increased salt tolerance of the
transgenic line CMO
4
. Such an improvement in salt tolerance was also associated with increased nutrient
uptake in CMO
4
. There was a clear interaction between unequal salt distribution and unequal nutrient
distribution on plant growth and salt tolerance of cotton plants in a split-root system. Plant growth and
salt tolerance as well as nutrient uptake were significantly improved in CMN/SLN relative to CLN/SMN.
Since levels of external salinity stress and nutrient supply were the same for both treatments, we suggest
that the improved salt tolerance and plant growth were attributed to increased nutrient uptake. It was thus
concluded that improved nutrient uptake can increase salinity tolerance of cotton under salinity.
ACKNOWLEDGEMENT
This work was supported by the earmarked fund for China Agricultural Research System (Cotton 2007
2011), the Major Project for Applied Agricultural Research (Cotton 20092011) and Seed Industry
(Cotton 20102012) of Shandong Province, and the National Natural Science Foundation of China
(30971720).
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Improvement of Partial Root-Zone Soil Environment

Increases Salinity Tolerance of Cotton
Hezhong Dong, L.I. Weijiang and L.I. Zhenhuai
Cotton Research Center/Shandong Provincial Key Lab for Cotton Culture and Physiology,
Shandong Academy of Agricultural Sciences, Jinan250100, P.R. China
E-mail: donghz@saas.ac.cn
AbstractSoil salinity is a major threat to cotton production worldwide. The objective of this study was to test
whether improvement at least in a part of the root-zone environment would enhance tolerance salinity and alleviate
salinity injury. Five experiments were conducted using a split-root system in the greenhouse or furrow seeding with
or without plastic mulching under salt-affected field conditions. The results showed that plastic mulching, furrow
seeding and early mulching or late-planting with short-season cotton either resulted in unequal salt distribution, or
increased moisture and temperature in the root-zone, and effectively reduced salt-injury to cotton. It is concluded that
improved partial root-zone environment increases salinity tolerance of cotton.
Keywords: Cotton, Salinity stress, Root-zone environment, Salinity tolerance, Plastic mulching
INTRODUCTION
Soil salinity has been a major concern to global agriculture throughout human history (Lobell et al.,,
2007). In recent times, it has become even more prevalent with intensification of land use (Meloni et al.,
2003, Egamberdieva et al., 2010). Cotton, though classified as one of the most salt-tolerant major crops,
its growth and development as well as yield and fiber quality are negatively affected by excessive salts in
the soil (Maas and Hoffman 1977, Qadir and Shams 1997, Higbie et al.,, 2010). Cotton is a pioneer crop
in reclamation of saline soils. But potential depends largely on ways and means to improve salt tolerance
of cotton. Although, some progress was made in salt-tolerance improvement, the development of salt-
tolerant cotton is not an easy job due to the complexity of the tolerance mechanisms and narrow
germplasm resource. There are a number of agronomic practices which can effectively control salt
damage through improvement in root-zone soil environment. Soil salinity expressed by ECe (electrical
conductivity of a saturated-paste extract), with values of 7.7, 12.5, and 17.1 dS m
1
are referred to as low,
moderate and high salinity level, respectively (Chen et al.,, 2009, Maas and Grattan, 1999). In general,
soil salinity delays and reduces germination and emergence, decreases cotton shoot growth, and finally
leads to reduced seed cotton yield and fiber quality characteristics at moderate to high salinity levels
(Khorsandi and Anagholi, 2009). Since soil salinity and the related stress originate from the root-zone
soil environment, it is hypothesized that improving at least part of the root-zone environment would
alleviate salt injury. The hypothesis was tested in greenhouse and field experiments.
MATERIALS AND MEDHODS
In a split-root experiment (EXP 1), potted cotton plants were grown in a split-root system in the
greenhouse and each root half was irrigated with either the same or two concentrations of NaCl (Fig. 1).
In a field experiment (EXP 2), we compared cotton grown on furrow-beds in saline fields with those
grown on flat beds as controls. A separate field experiment was conducted to evaluate the effect of
mulching with polyethylene film. Integration of plastic film with irrigation methods was also studied in a
separate set of experiment. Details of treatments provided in Dong et al. (2009, 2010 a, 2010 b).
72

RESULTS A
When the
leaf chlor
control. H
salinity o
significan
Furrow
earliness
unequal d
Row
water con
distributio
Further st
establishm
seeding al
Fig. 2
(a), C
Norm
establishm
cotton in a
cotton sig
reduced N
a
Improvem
Fig. 1: Effec
AND DISCUSSI
e entire root
rophyll (Chl
However, wh
on growth an
ntly increased
w-bed seedi
were signifi
distribution o
covering wi
nservation. I
on in the sal
tudy showed
ment, earline
lone (Fig. 2)
2: Effects of Se
Conventional Se
mal planting
ment, late m
a saline field
gnificantly im
Na+ concentr
ment of Partial R
cts of Unequal Sal
in cth
ON
system was
l) and photo
hen only half
nd yield wa
d compared t
ing induced
cantly impro
f salts in the
ith polyethy
In our field
line soil, in
d that the i
ss, yield, and
(Dong et al.
eding Modes on
eeding under M
of full-seaso
maturity, and
d of the Yello
mproved see
ration in cott
Root-Zone Soil E
t Distribution (100
he Root-Zone on Co
exposed to
osynthesis (P
f of the root s
as significan
to equal salt
unequal sa
oved. Such a
root zone (D
lene film en
experiment
which part o
ntegration o
d yield comp
., 2008).
n Stand Establi
Mulching (b) and
and Full Stand
on cotton in
increasing
ow River De
ed emergenc
on tissues re
b
Environment Inc
0/500 mM NaCl) a
otton Seedling Gro
the same co
Pn) were sig
system was e
ntly reduced
distribution
alt distributi
an improvem
Dong et al., 2
nhances plan
(EXP 3), p
of the root s
of plastic mu
ponents of co
shment in a Sa
d Furrowing Se
d Establishment
saline field
cost of inpu
elta (EXP 4),
ce and seedl
lative to norm
creases Salinity
and Equal Salt Dist
owth in the Green
oncentration
gnificantly r
exposed to lo
d. Plant biom
(Dong et al.,
on in saline
ment in yield
2010b).
nt growth by
plastic mulch
system devel
ulching with
otton more th
aline Field. Conv
eeding under M
t, Respectively
s in tempera
uts. After try
, we observe
ling growth
mal planting
c
y Tolerance of C

ribution (300/30
House.
of NaCl, lea
reduced rela
ow-salinity,
mass and se
, 2010b).
e fields; pla
d and earline
y increasing
hing also re
loped in rela
h furrow se
han effective
ventional Seedi
Mulching Resulte
y
ate areas is
ying late pla
ed that late p
due to incre
g ( Dong et a
Cotton
0 mM NaCl)
af area, plan
ative to the
the inhibitio
eed cotton y
ant growth,
ess was mai
g soil tempe
esulted in un
atively low-s
eeding enhan
ely mulching
ing without Mu
ed in Poor, Mod
faced with p
anting of sh
lanting of sh
eased tempe
l.,, 2010a).
433
nt biomass,
NaCl-free
on effect of
yield were
yield and
nly due to
rature and
nequal salt
saline soil.
nced stand
g or furrow

lching
derate
poor stand
hort-season
hort-season
erature and
Row mulching is conventionally applied after sowing, but pre-sowing evaporation in spring would
cause accumulation of salts and moisture loss in the surface layer of saline soils. Row mulching with
plastic film can be done 30 d before sowing (early mulching) in saline fields (EXP 5). Although both
conventional and early mulching could effectively improve stand establishment, plant growth, earliness
and lint yield of cotton, early mulching was more beneficial to stand establishment, plant growth and
yield. The increased benefits of early mulching were due mainly to the better control of root-zone soil
salinity, elevation of soil temperature and reduction of moisture loss (Dong et al., 2009).
CONCLUSION
About 23% of the worlds cultivated lands are saline. Plastic mulching, furrow seeding, late-planting of
short season-cotton, irrigation and fertilization either reduced salinity, increased temperature and
moisture, or increased supply of nutrients in the soil root-zone, thus reducing the salt damage in saline
fields. We conclude that improvement in a part of the root-zone environment increases salinity tolerance
of cotton.
REFERENCES
[1] Chen, W., Hou, Z., Wu, L., Liang, Y. and Wei, C. (2010) - Effects of salinity and nitrogen on cotton growth in arid
environment. Plant Soil 326, 6173.
[2] Dong, H., Kong, X., Luo, Z., Li, W. and Xin, C. (2010b) - Unequal salt distribution in the root zone increases growth and
yield of cotton. Eur. J. Agron. 33, 285292.
[3] Dong, H., Li, W., Tang, W. and Zhang, D. (2008) - Furrow seeding with plastic mulching increases stand establishment and
lint yield of cotton in a saline field. Agron. J. 100, 16401646.
[4] Dong, H., Li, W., Tang, W. and Zhang, D. (2009) - Early plastic mulching increases stand establishment and lint yield of
cotton in saline fields. Field Crop Res. 111, 269275.
[5] Dong, H., Li, W., Xin, C., Tang, W. and Zhang, D. (2010) - a: Late-planting of short-season cotton in saline fields of the
Yellow River Delta. Crop Sci. 50, 292300.
[6] Egamberdieva, D., Renella, G., Wirth, S. and Islam, R. (2010) - Secondary salinity effects on soil microbial biomass. Biol.
Fert. Soils 46, 445449.
[7] Higbie, S.M., Wang, F., Stewart, J. McD., Sterling, T.M., Lindemann, W.C. , Hughs, E., Zhang, J. (2010) - Physiological
response to salt (NaCl) stress in selected cultivated tetraploid cottons. Int. J. Agron. 1, 112.
[8] Khorsandi, F. and Anagholi, A. (2009) - Reproductive compensation of cotton after salt stress relief at different growth
stages. J. Agron. Crop Sci. 195, 278283
[9] Lobell, D.B., Ortiz-Monsterio, J.I., Gurrola, F.C. and Valenzuuela, L. (2007) - Identification of saline soils with multiyear
remote sensing of crop yields. Soil Sci. Soc. Am. J. 71, 777783.
[10] Maas, E.V., Hoffman, G.J. (1977) - Crop salt tolerancecurrent assessment. J. Irrig. Drain. Div. Am. Soc. Civ. Eng. 103,
115134.
[11] Maas, E.V., and Grattan, S.R. (1999) - Crop yields as affected by salinity. In: Skaggs RW, van Schilfgaarde J (eds)
Agricultural Drainage, Agron Monogr 38. ASA, CSSA, SSA, Madison, WI, pp 55108.
[12] Meloni, D.A., Oliva, M.A., Martinez, C.A., Cambraia, J. (2003) - Photosynthesis and activity of superoxide dismutase,
peroxidase and glutathione reductase in cotton under salt stress. Environ. Exp. Bot. 49, 6976.
[13] Qadir, M. and Shams, M. (1997) - Some agronomic and physiological aspects of salt tolerance in cotton (Gossypium
hirsutum L.). J. Agron. Crop Sci. 179, 101106.
Do Female-Led Farms Perform Less Well

in Cotton Production? Insight
from Hebei Province (China)
Michel Fok
1
and Guiyan Wang
2
1
CIRAD, UR SCA, Montpellier, FRANCE
2
Hebei Agricultural University, Baoding, Hebei Province, CHINA
AbstractMost of the emerging countries witnessed outmigration of farming community away from agriculture to
cities. This was mainly due to rapid industrialisation, fast rising service sectors growth and low profitability of
farming being agriculture is a gamble of rainfall. Generally, it is the men folk who leave their wives behind and
migrate to cities in search of better alternative jobs. This phenomenon gives rise to the feminization of agriculture and
raises the concern of lower productivity. A few studies in developing countries have indicated that female-led farms
lack access to production factors such as land, labour and capital.
In China, the migration of farmers to cities and the feminization of agriculture were well documented, but studies
on the impact of feminization on agricultural performance and productivity are rare. The impacts of feminization on
agriculture have been assessed on a regional or cropping system level, but not on a specific crop.
In light of the above, the present paper deals with the status and performance of the feminization on cotton
cropping in Hebei Province (Northern China). As cotton cropping is widely acknowledged to be both labour and
capital intensive it is most appropriate to assess not only the impacts of feminization on its productivity but also to
check the assumption whether female-led farms lack factors of production. Therefore, the study was conducted to
analyse the cotton cropping systems on identified farms run by women and compare their yield and gross income with
the alternative farms. The study stems its base on primary data collected through survey method for the period 2006 to
2009. Farms were considered to be female-led, when husbands or their spouse was away for more than five months per
year.
INTRODUCTION
Most of the emerging countries witnessed outmigration of farming community away from agriculture to
cities. This was mainly due to rapid industrialisation, fast rising service sectors growth and low
profitability of farming being agriculture is a gamble of rainfall. Generally, it is the men folk who leave
their wives behind and migrate to cities in search of better alternative jobs (Lastarria-Cornhiel, 2006).
This phenomenon gives rise to the feminization of agriculture in which three definitions are generalised:
women are taking over field tasks from men; Female participation rate in field work as compared to man
is increasing; women accounts for more than half of the labour force in fields.
In China, the feminization phenomenon has been particularly analysed and debated because of the
large extent of rural men migrating to cities. Several publications by Chinese scholars implicitly attribute
feminization of agriculture as the share of female in agricultural labour has increased from less than half
to more than half. Gao (1994) was among the first to point out the issue of feminization and to link it to
the migration of rural men to cities. Cheng (1998) pointed out outnumbering of females over males in as
the labour force in fields through the Fourth National Census, 1992 and that the trend was increasing
(Zhou and Sun, 2006). Chen and Xu (2008) reported that, at national level, women amounted to more
than 80% of the labour force in rural areas in 2000. Zhou et al. (2007) opined that this figure could reach
up to 90% in some provinces with the vibrant economic development there. Referring to the definition of
the female participation rate in field work, de Brauw et al. (2008) debated the reality of the phenomenon
of feminization in Chinese agriculture which is contrary to the findings of Zhou and Sun (2006). He et al.
(2010) found that in Jiangsu province, females were the major decision makers on farms.
73
Few studies in developing countries indicates that the feminization phenomenon raises the concern of
lower productivity in agriculture, as female-led farms lack access to factors of production such as land,
labour and capital (Chikwama, 2010). Sun and Zhou (2008) reported that most of the Chinese scholars
regarded feminization as a threat to agricultural productivity. Zhao and Zhao (2009) considered this
phenomenon as an impediment to further development of the rural economy in China. The pessimistic
view on productivity may arise from the association between feminization and aging that some scholars
highlighted in China (Zhou and Song, 2008).
Nevertheless, studies on the impact of feminization on agricultural performance remain rare. In
China, the impact of feminization have been assessed on a regional or cropping system level (de Brauw
et al., 2008), without confirming decline in the farming productivity. No study was undertaken to assess
the impact of feminization on productivity of a specific crop level.
The present paper deals with the status and performance of the feminization of cotton production in
China. As cotton cropping is acknowledged to be labour- and capital-intensive, it is appropriate to assess
the impacts of feminization on its productivity and to check the assumption that female-led farms lack
production factors.
This paper deals with cotton production in Hebei Province, located in the Yellow River Valley, northern
China. This province is one of the major cotton production locations in China. It was the first place where
Bt cotton was commercially released in China, both with Monsanto varieties incorporating its Cry1Ac
gene and Chinese varieties integrating the synthesized Cry1A gene (Guo and Cui, 2000; Guo and Cui,
2004). Favourable conditions for licensing
1
Chinese Bt gene to Chinese breeding organizations were set
to encourage the supply of new Chinese Bt cotton varieties and create market competition. This was in
line with the institutional shift through the application
2
of new laws on variety intellectual protection and
seed marketing whereby only authorized and registered varieties could be legally marketed (Xu and Fok,
2010).
Our study was based on primary data from surveys covering four successive years from 2006 to
2009. Data from farmers growing cotton in 38 villages of five major districts in Hebei Province (Handan,
Xingtai, Hengshui, Cangzhou and Shijiajiang) were collected.
To carry out the surveys, it was decided not to pass through local extension services, whose presence
might have influenced the farmers' answers. Enumerators were selected from students of Agricultural
University of Hebei whose families were doing farming in the cotton areas of Hebei province. Therefore,
all the enumerators were familiar with agriculture and cotton growing. As they belonged to the same
province, they gained confidence of native cotton farmers. Student-enumerators were trained on carrying
out the survey and the survey was done during the Spring Festival (end of January or beginning of
February). This period coincided with off-season when most of the farmers have already sold off their
products from the previous calendar year.
Enumerators were asked to survey 20-30 farmers randomly using recall enumeration techniques
through semi-directed questionnaires. The survey was conducted with the aim of determining farmers'
cultivation practices and income in connection with the structures of their farms. Farmers were asked to
answer on the basis of their memories. Villages and farmers surveyed were not the same from one year to
the next.
The institutional framework in China does not force farmers to buy seeds through merchants every
year (Fok and Xu, 2010b). Farmers can hold back seeds from one season to another, or mutually
1
Up to 2007, royalties due for using the Chinese Bt gene were paid once and for all at the time of the official registration of a
new Bt cotton variety and not related to the amount of seeds sold. The fee has fluctuated from US$35,000 to 40,000.
2
The laws were first issued in the first half of the 1990s, amended several times, but the decrees of application came into force in
late 1999 or early 2000.
Do Female-Led Farms Perform Less Well in Cotton Production? Insight from Hebei Province (China) 437
exchange seeds with or without monetary compensation. Farmers' practices in using seeds were
addressed in our surveys by asking whether they renewed seeds (instead of using seeds they held back
from the former season), how frequently and from whom (notably merchants).
Production costs were recorded for all the surveyed farms. As the valuation of family labour used is
debatable in rural areas, we did not integrate the family labour cost in total production costs. Cotton
production costs and yields were only obtained at farm level and not at plot level. This was not a
limitation when farmers used only one variety. Collected data were processed to analyse the structures of
farms and farm families, with emphasis on the share of female members and on the involvement of
family members in farm and off-farm activities, respectively.
The present study adopted a criterion that the farms were led by females when the farm heads (the
males in China) were engaged in off-farm activities for at least five months a year. Two groups of farms
namely male-led and female-led were compared with regard to the land areas cultivated, areas under
cotton production, cost of cultivation, and two performance indicators, namely cotton yield and gross
income.
A multi-regression analysis was done to check the extent to which yield was dependent on three
groups of factors. Group 1 was composed of farm structure factors regarding the gender of the farm
managers and the assistance of children in field work. A study on the same set of data revealed that farm
structural factors such as family size, total cultivated area, etc had no impact on yield. Group 2 consisted
of factors related to cotton areas and farmers' behaviour in using varieties and seeds (number of varieties
used, the extent of annual seed renewal, and the frequency of seed purchases from merchants). Group 3
pertained to cost of production (corresponding to seeds, plastic mulching, irrigation, fertilization, pest
control and disease control). Weed control by herbicide, soil preparation, growth regulators, etc were
integrated in "other costs".
RESULTS
For most of the variables related to farm families (Table 1), the data were consistent over the four years,
although the samples varied between years, in terms of both composition and size. In total, survey
covered 2492 family members living in farming families. Feminization, if any, was not due to females
outnumbering males in families. Female accounted for 45.4% of the total population, as against the sex
ratio of 120 (120 males for 100 females), which was well above the common sex ratio of 105-106
worldwide. The sex ratio was quite similar for family members at least 16 years old who could be
considered as potential active labour.
Farm heads were 47 years old in 2009, 0.8 years older than their wives. Globally, their children were
of working age, the sons being almost two years older than the daughters. In 2009, they were 21.5 and
18.6 years old, respectively.
The feminization of Chinese agriculture may be sustained by the higher frequency of females
involved in field work, even though the rate of all permanently involved family members was low.
Among all farming family members with at least 16 year old, only 56.7% were permanently involved in
field work. This is an indication of the specificity of China where agriculture is barely the main
occupation of farming families. As already pointed out, females were less numerous than males, but they
amounted to 55.8% of the permanent field workers. The wives of farm heads were the most permanent
workers in the fields. Over the four years, 91.1% of them were involved permanently in field work, much
more than their husbands (66.2%). The impression of feminization of agriculture was reinforced by the
low participation rate of farm children. Farmers' sons or daughters with at least 16 years old took part in
field work at a rate of 9.0% and 5.2%, respectively.

The observed feminization was the result of greater involvement of male in off-farm activities. Over
the four year period, 94.3% of farm families had at least one member involved in off-farm activities on a
long duration basis (at least five months a year). Males were more involved than females in off-farm
activities (40.8 vs 14.4%) but the large gap was due to the very marginal involvement of farm heads'
wives. Between boys and girls, the gap was far less (56.7 vs 38.3%). In the study, we assumed that farms
were female-led when their husbands were involved in off-farm activities on a long term basis. Over the
four-year period, for the 819 farms from which we obtained detailed product costs, 249 farms (30.4%)
were female-led (Table 2). Cotton farms were tiny in size (0.75 ha of cultivated land) and they were
smaller for female-led farms. 29.4% of farms managed to rent some land. This possibility was more
limited for female-led farms. Considering all farms per capita cotton area was 0.44 ha, but this area was
smaller for female-led farms and also for the cotton area per family member over the age of 16.
TABLE 1: CHARACTERISTICS OF FARM FAMILIES AND THEIR INVOLVEMENT IN OFF-FARM ACTIVITIES

Note: Figures in brackets are total numbers; figures in square brackets are standard deviations.
With regard to variety and seed use practices, there were no differences between farms led by males
or females. In case of all farms 31.1 % use non-Bt cotton varieties, 69.6% farms goes for annual seed
renewal (69.6%) and about 83% of farms source seeds through merchants (83.0%).
There were differences in performance between the two types of farms, but not in the expected
direction. Compared to male-led farms, female-led farms performed better in yield with a similar level of
total production costs. Female-led farms had higher costs to control diseases, but lower costs in a few
cases (irrigation and other costs encompassing, for example, herbicides, growth regulators, etc.). These
differences might be related to higher labour constraints (case of irrigation) or more routine practices
(spraying against cotton plant diseases).
The better performance of female-led farms was more balanced in economic terms. This better
performance was observed in case of cotton production value per area and gross income per area. Since
female-led farms had a smaller area, their total cotton production value and total gross income were
lower compared to male-led farms. This is the reason for lower labour productivity in terms of gross
income per family member over 16 years old in female-led farms.
Features of farm families 2006 2007 2008 2009 All
Number farms surveyed 119 207 360 175 861
Farm demography
% females among all family members 49.7 [366] 44.0 [621] 44.1 [1008] 46.7 [497] 45.4 [2492]
% females among family members 16 years old 49.7 [340] 44.1 [580] 44.1 [966] 45.9 [468] 45.2 [2345]
Number family members 3.1 (1.0) 3.0 (0.9) 3.0 (1.0) 3.1 (1.0) 3.0 (1.0)
Number of children living in farm families 1,0 1,0 1,0 1.2 1,0
Farm family members' ages
average age of farm holder 44.5 (9.2) 45.5 (9.2) 46.9 (8.7) 47.0 (8.9)
average age of farm holder's wife 44.5 (9.3) 44.6 (9.0) 45.8 (8.3) 46.2 (8.9)
average age of farmers' sons 20.8 (6.0) 20.8 (6.5) 22.2 (6.8) 21.5 (6.3)
average age of farmers' daughters 18.3 (5.7) 18.9 (5.7) 20.1 (6.6) 18.6 (6.5)
Permanent involvement in farming
% family members concerned, at least 16 years old 60.6 [340] 60.7 [580] 56.4 [966] 49.6 [468] 56.7 [2354]
% females among total permanently involved members 56.3 [206] 55.7 [352] 53.4 [545] 61.2 [232] 55.8 [1335]
% farm heads concerned 75.7 [115] 68.3 [205] 67.1 [343] 55.0 [160] 66.2 [823]
% farm heads' wives concerned 95.6 [113] 94.4 [197] 87.2 [313] 91.4 [151] 91.1 [774]
% farm children concerned
% heads' sons concerned, over 16 years old 4.4 [67] 10.1 [139] 12.6 [222] 2.8 [106] 9.0 [534]
% heads' daughters concerned, over 16 years old 1.8 [56] 7.4 [68] 65 [107] 3.8 [78] 5.2 [309]
% farms with permanent participation of children in field
works
3.4 [119] 7.2 [207] 8.8 [340] 3.8 [159] 6.7 [825]
Involvement in off-farm activities
% farms with at least one member off-farming 97.5 [119] 95.7 [207] 93.8 [340] 91.2 [159] 94.3 [825]
% males concerned, over 16 year old 38.0 [171] 38.3 [324] 40.6 [540] 44.7 [253] 40.5 [1288]
% females concerned, over 16 year old 19.5 [169] 10.2 [256] 14.6 [426] 14.9 [215] 14.4 [1066]
% farm heads concerned 22.6 [115] 25.4 [205] 27.4 [343] 33.8 [160] 27.5 [823]
% farm heads' wives concerned 4.4 [113] 2.5 [197] 6.7 [313] 2.6 [151] 4.5 [774]
% holders' sons concerned, over 16 years old 69.6 [56] 61.5 [117] 61.9 [202] 61.7 [94] 62.7 [469]
% holders' daughters concerned, over 16 years old 58.1 [43] 40.4 [52] 42.9 [91] 44.3 [61] 45.3 [247]
TABLE 2: COTTON CROPPING, COSTS AND RETURNS, COMPARISON BETWEEN MALE- AND FEMALE-LED FARMS

Note: Figures in brackets are total numbers; figures in square brackets are standard deviations. Comparisons are made between male- and
female-led farms * significant at 5%, ** significant at 1%, *** significant at less than 0.1%; Other costs pertain to soil preparation, sowing, weed
control by herbicides, growth regulation of cotton plants and occasional labour hiring.
TABLE 3: FACTORS AFFECTING YIELDS ON COTTON FARMS

Cotton cropping, costs, and returns All Farms Male-led farms Female-led farms
Number of farms 819 570 249
Total cultivated land (ha) 0.75 (0.54) 0.78 (0.62) 0.62 (0.39)**
Cultivated land per capita (ha) 0.26 (0.20) 0.28 (0.21) 0.23 (0.17)**
Cultivated land per capita, at least 16 years old (ha) 0.29 (0.23) 0.31 (0.24) 0.25 (0.19)**
Own land (ha) 0.61 (0.31) 0.62 (0.29) 0.51 (0.25)**
Own land per family member, at least 16 years old (ha) 0.24 (0.16) 0.25 (0.15) 0.21 (0.17)**
Land renting
% of farms concerned 29.4 [819] 29.2 [570] 29.6 [249]
average rented land (ha) 0.46 (0.71) 0.51 (0.81) 0.35 (0.44)*
Cotton area (ha) 0.44 (0.46) 0.49 (0.53) 0.36 (0.33)**
Cotton area per family member, of at least 16 year old (ha) 0.17 (0.17) 0.19 (0.19) 0.13 (0.12)**
Number of varieties cultivated 1.6 1.6 1.5
Percentage of farms with at least one non-Bt variety 31.1 [819] 33.2 [570] 26.5 [249]
Seed renewal, % of farms with systematic annual renewal 69.6 [806] 67.4 [530] 74.3 [276]
Seed source, % of farms with systematic purchase 83.0 [806] 81.8 [560] 85.8 [246]
Cotton yield, seedcotton (kg/ha) 3795 (840) 3735 (765) 3945 (945)**
Total cash-expenses production cost (US$/ha) 817 (181) 821 (190) 806 (152)
seed cost (US$/ha) 87 (58) 87 (59) 85 (52)
mulching plastic cost (US$/ha) 58 (23) 58 (23) 59 (21)
Irrigation cost (US$/ha) 54 (27) 56 (27) 50 (27)*
Fertilisation cost (US$/ha) 294 (106) 294 (110) 294 (92)
Pest control cost (US$/ha) 171 (79) 173 (79) 165 (75)
Disease control cost (US$/ha) 25 (33) 21 (25) 35 (42)**
Other costs (US$/ha) 127 (75) 131 (85) 121 (48)*
Total cotton production value (US$) 1178 (1120) 1270 (1214) 967 (832)**
production value per area (US$/ha) 2661 (754) 2596 (685) 2831 (875)**
production value per family member, at least 16 years old
(US$)
439 (417) 476 (451) 357 (312)**
Total cotton gross income (US$) 808 (774) 865 (836) 679 (592)**
Gross income per area (US$/ha) 1850 (792) 1175 (733) 2023 (886)**
Gross income per family member at least 16 years old (US$) 300 (287) 323 (311) 250 (217)**
Independent variales Value Std deviation t value Pr > t
Constant 3441,715 186,260 18,478 < 0,0001
Female-led farms 191,419 63,406 3,019 0,003
Help from children 331,142 114,373 2,895 0,004
Cotton area -12,528 4,432 -2,827 0,005
Number of varieties 96,063 40,142 2,393 0,017
Annual seed renewal 26,618 67,094 0,397 0,692
Seed sourcing from merchants -229,545 93,222 -2,462 0,014
Perception that diseases need to be controlled -7,179 105,288 -0,068 0,946
Seed cost -2,681 1,155 -2,321 0,021
Plastic mulching cost 3,714 2,610 1,423 0,155
Irrigation cost 8,590 2,062 4,166 < 0,0001
Fertilization cost -0,074 0,532 -0,139 0,890
Pest control cost -0,491 0,758 -0,647 0,518
Disease control cost 8,004 1,808 4,427 < 0,0001
Other costs 1,949 2,271 0,858 0,391
The better yield performance of female-led farms was confirmed by the multivariate regression
analysis (Table 3). The female-led farms have shown a positive effect on seed cotton yield. The positive
effect of assistance from the farm children was significant cotton being a labour intensive crop. The
number of varieties that farmers used had a positive impact on yield, contrary to seed costs and seed
provision from merchants. With regard to the costs of cultivation practices, only irrigation and disease
control exhibited positive effects.
DISCUSSION
The study covered cotton production in Hebei province a major cotton belt of China. Cotton is produced
on small farms where the average land holding is less than half a hectare (Fok et al., 2005; Pray et al.,
2001). China is probably the country where cotton farms are smallest. The results were obtained from
samples of farms whose representativeness was derived from the characteristics of the farm families.
Feminization was observed in cotton production where, females amounted for more than half of the
labour force in field work (55.8%) as females were essentially involved in farming (91.1%). These
figures are consistent with the empirical data analysed by Mao and Liu (2009) but lower than those of
Deng (2008), who carried out a large-scale survey and found that females accounted for 74.7% of the
field labour force in 2006. The differences observed might be due to variations between provinces.
Mainly wives of farm heads, who remained involved in farming were permanent workers in the fields,
apart from those who were engaged in off-farm activities on a long-term basis (about 30% of them).
It was found that about 31% of farms being led by females as their husbands were involved in off-
farm activities for more than five months a year. Our result was consistent with Deng (2008), who
observed in 2006 that 20.7% of farms were led by females alone, and even up to 31 to 33% in some
provinces.
The study confirmed the high frequency of farming family members involved in off-farm activities.
We found that 94.3% of farm families had at least one member engaged in off-farm activities. The figure
was higher than that found by Chen et al. (2010) where it was 74.3%. Further, it was found that
youngsters engaged in off-farm activities compare to farm heads.
Because of their lower involvement in off-farm activities, females were the permanent workers in the
fields, mainly wives of farm heads, but their husbands remained involved apart from those who were
engaged in off-farm activities on a long-term basis (about 30% of them). The smaller the size of the
farms, the higher was the frequency of farm heads involved in off-farm activities on a long-term basis,
and consequently the frequency of farms being run by females. The results confirm the relevance of
addressing the issue of the performance of female-led farms because these farms accounted for 30% of
all cotton farms in Hebei province. The relevance also derives from their structural differences with
reference to farms run by males. The female-led farms were smaller in size, indicating the probable
causality between farm size and long term migration of the farm heads. The smaller the farms are, the
more difficult it is to make ends meet without additional income from off-farm activities, thereby pushing
farms heads to engage for many months in off-farm activities, leaving their wives alone to run their
farms. This mechanism is indeed found by most of the studies addressing the feminization of agriculture
in China (Gao, 1994; He et al., 2010; Li and Fang, 1999; Zhou and Sun, 2006).
From a technical perspective, female-led farms did not perform less well because they achieved
higher yield. This result is contrary to what used to be claimed for developing countries (Lastarria-
Cornhiel, 2006) and feared in China (Li and Fang, 1999; Wu and Zhang, 2008) although they were
seldom based on empirical analyses. Our result is nevertheless close to the rare empirical studies in China
(de Brauw et al., 2008; Zhang et al., 2004), which did not find confirmation that female-led farms
performed less well. Li (2001) found in 1996 that the return on women's agricultural activities was higher
than that of men, probably a direct consequence of more involvement in field work by women. In the
study, female-led farms performed well because they entailed a similar level of total production costs,
and also because they probably took better care of their smaller cotton plots. Their capacity to invest in
production costs was opposite to what used to be reported in other countries. We believe that this
difference arises from the involvement of their husbands in off-farm activities and the resulting income.
We found no evidence that women were less technically skilled, which was quite consistent with the
results of Song (1999). Our result was contrary to the productivity decrease that was feared to result from
the fact that women were less educated or was less familiar with techniques and farm management
according to Li and Fang (1999) but that was at the early stage of feminization and may no longer hold
now. Nevertheless, in economic terms, female-led farms performed less well because they were smaller.
Their gross income per unit area was better, as a consequence of higher yield and similar production
costs, but at farm level, this performance was mitigated by the smaller size of their cotton plots.
Female-led farms accounted for about 30% of cotton production in Hebei province. They were
smaller but technically they performed well. The higher yield they achieved was due to their capacity to
invest in production costs similarly to other farms probably the income earned from the off-farming
activities by the farm heads was also invested in the farming which resulted in better care of smaller
cotton farms.
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of Eastern China: A Case Study of Yucheng City, Shandong Province. Chinese Geographical Sci. 20(6): 545553.
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Debunking the Myths

J. Reed, E. Barnes and P. O'Leary
Cotton Incorporated, USA
AbstractCotton and cotton textile industries are important to the economic well-being of more than 100 countries,
developed and developing, alike. In spite of its benefits, cotton production in particular, has been mischaracterized as
having negative impacts on the environment. Among those voicing claims against cotton are businesses seeking a
competitive advantage and non-governmental organizations. The most common of these allegations are that cotton
production requires excessive amounts of pesticides and water. This paper examines those and other claims against
cotton and presents data to negate these assertions.
INTRODUCTION
Claims based on outdated information continue to be perpetuated despite repeated efforts to set the record
straight. Although cottons past environmental history does provide some basis for the claims, significant
improvements have occurred in recent years yet these gains are rarely acknowledged as part of cottons
environmental story, focusing instead on cottons outdated history. This paper will address common
misconceptions about cotton production in hopes of enabling others to counter misrepresentations of
cotton productions environmental impact.
PESTICIDES
Synthetic pesticides are widely used in agriculture to control crop losses caused primarily by insect, weed
and disease pests. Cotton is no exception. According to Cropnosis (personal communication. 2010), plant
protection chemicals worth US$40 billion were used globally in agriculture in 2009. Almost half of these
were herbicides to control weeds and to facilitate conservation tillage. Plant protection chemicals worth
US$2.5 billion were used in cotton. It has been estimated that over 80% of the global cotton crop would
be lost in the absence of crop protection products. Arthropod and weed pests would be the primary
contributors to this loss (Oerke & Dehne, 2003 ).
Claim
Cotton uses 25% of the worlds pesticides
The Facts
In the 1990s, the use of pesticides on cotton peaked; accounting for one quarter of the global annual
value, equivalent to US$ 2 to 3 billion (Murray, 1994 as cited in de Blcourt, 2010). The global
insecticides share used on cotton had declined from 19% in 2000 to 14%. In the same year, cottons
pesticide consumption accounted for 6.2% of global use (Cropnosis, personal communication. 2010). A
comparison of pesticide usage globally on the major crop groups is given in Table 1.
TABLE 1: COMPARISON OF THE GLOBAL USAGE OF PESTICIDES ON MAJOR CROP GROUPS. 2007 (CROPNOSIS, PERSONAL COMMUNICATION.2008)
Crop % Global Pesticides Used
Cotton 6.8
Fruits & Vegetables 29.7
Cereals 17.0
Soy 9.6
Maize 9.3
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and species of birds were higher in conservation tillage fields than in the conventional fields, regardless
of the season. But in the winter the number and species of birds in conservation tillage fields far exceeded
the numbers observed in conventional fields (Cederbaum, 2004). In addition, arthropods that are an
important food source for birds are more abundant in conservation fields than conventional fields.
Research in North Carolina showed that quail chicks required more than five times as much time to
obtain their minimum daily requirement of insects in conventional fields as in conservation fields or
natural fallow areas believed to be ideal quail habitat (Palmer, 1995 as cited in Fawcett & Towery, 2003).
Studies in the USA and in Australia show that Bt cotton has a positive impact on beneficial insects in
cotton fields, thus promoting biodiversity (Naranjo, 2005; Head et al., 2005; Whitehouse et al., 2005;
Torres & Ruberson, 2005). Thus, with the expanding acres planted to Bt varieties worldwide and
corresponding reduction in pesticide usage we can expect arthropod biodiversity in cotton fields to
increase.
Claim
Pesticide residue left on cotton products could be harmful to ones health.
The Facts
Assumptions are often made that because pesticides are applied to the cotton plant then there must be
pesticides on the fiber and subsequently on the finished textiles. Since 1991, the Bremen Cotton
Exchange has tested raw cotton on an annual basis from a minimum of 16 countries for over 200 toxic
substances, including heavy metals and pesticides according to Eco-Tex Standard 100. In 18 years there
have been only four instances where substances were detected in quantities exceeding the most stringent
level of the standard: two by only 0.01 mg/kg, another by 0.25 mg/kg and another by 0.7 mg/kg
(Baumwollbrse, 2010). Therefore, one would expect the likelihood that cotton products are
contaminated with pesticides to be almost nil.
BIOTECH CROPS
Biotech cotton varieties were first introduced in 1996. Two types of biotech cotton traits are currently
available for commercial production. The first provides tolerance to herbicides and the second provides
resistance to bollworms. One or both of these traits can be found in a cotton variety. Global adoption of
biotech cotton has risen dramatically from 800,000 hectares in 1996 to 21 million hectares representing
14% of cotton area in 2010 (James, 1997; ISAAA, 2011). Indeed, this year marks the tenth year of Bt
cotton production in India. During this time, India has seen an unprecedented 188-fold increase in Bt
cotton acreage, from 50,000 hectares in 2002 to 9.4 million hectares in 2010. The increased yields,
reduction in insecticide applications and profitability of resource poor farmers attributed to Bt cotton has
transformed cotton production in India into the highly viable industry that it is today (James, 2010)
Claim
Bt cotton poses an environmental safety risk.
The Facts
Prior to commercialization, Bt cotton, like all genetically-enhanced products, are subjected to a rigorous
regulatory evaluation by the EPA, USDA and the FDA (EPA, 2011). This includes comprehensive
environmental and human safety studies to rule out the possibility of harm to existing plants, non-target
organisms and humans. Strict refugia requirements for growers using Bt cotton ensure the durability of
the technology and maintain ecological balance.

Debunking the Myths 445
Claim
Herbicide tolerant crops increase the use of pesticides and cause super weeds.
The Facts
While use of broad spectrum herbicides, notably glyphosate, has increased since the introduction of
herbicide tolerant crops, the use of less environmentally benign products has fallen substantially, leading
to net benefits to the environment (USDA, 1991 2011). Furthermore, herbicide-tolerant crops together
with the environmentally benign herbicide glyphosate, have enabled widespread adoption of conservation
tillage. This has led to significant enhancements in soil health and has contributed greatly to the reduction
of agricultures environmental footprint by reducing soil erosion, greenhouse gas emissions and water
and energy inputs (Fawcett & Towery, 2002).
One concern with biotech crops is the exchange of genes from the biotech plants to wild relatives
potentially producing new biota that might disrupt the ecological balances. If this should occur, the
resultant plants can be safely and easily controlled with herbicides other than glyphosate.
Herbicide resistance is a recurring agronomic problem, not just with cropping systems that include
biotech crop plants but with all cropping systems where herbicides are used. However, without
herbicides, every weed is a super weed. Therefore, herbicides in conjunction with established IPM and
other good management practices play an important role in sustainable crop production.
WATER
Water consumption for cotton production is often misunderstood. Cotton is a highly drought-tolerant
plant and can prosper in high salt soils. These attributes enable cotton to be grown in areas where it is
often not feasible to grow any other crop. And while a harvestable crop of cotton is possible from an
annual supply of only 250 millimeters of water, supplemental irrigation as for most crops ensures a
consistently high yielding crop. Irrigation water is used in cotton production throughout the world but the
amount and distribution varies widely depending on region, climate, available technologies and
governmental policies.
Claim
Cotton is a water intensive crop.
The facts
Cottons global water footprint is about 2.6% of the worlds agricultural water use, lower than many
other commodities and is proportional to cottons land use

(Hoekstra & Chapagain, 2007).

In the USA
only 11% of the cotton crop is fully irrigated, 25% receives supplemental irrigation; the remaining 64%
of U.S. acres receive no irrigation water at all, instead, relying totally on rainfall

(USDA, 2009).
Furthermore, compared to 25 years ago, US cotton growers today are producing over twice the amount of
cotton for the same amount of irrigation water

(USDA, 2004). According to a survey of USA growers in
2008, 72% of U.S. cotton growers list an adequate water supply as one of their top five concerns. This
helps explain why 81% of the cotton growers who do use supplemental irrigation upgraded their
irrigation systems within the last 10 years (Reed et al., 2009). And although abundant yields of high
quality cotton are a primary goal of U.S. cotton growers, water-use efficiency growing more using less
water is a common sense priority for them, as well. The same survey showed that US growers are
moving away from surface irrigation systems (where leaching and non-uniformity of water application
are a greater challenge) to the more uniform sprinkler systems. These include highly efficient
technologies such as subsurface drip irrigation (SDI), and Low Energy Precision Application (LEPA).
SDI is a system which as the name describes delivers water via underground tubes where it is not subject
to evaporation. LEPA is also energy efficient since it less pressurization is required and the water is
delivered precisely to the base of the plant where its needed. These and other irrigation systems are
made further efficient by various kinds of sensing or application technologies such as water-saving
nozzles, soil moisture probes, or infrared leaf thermometers that detect when a plant is becoming water-
stressed, in which case, the plants are only watered on an as needed basis. Remarkably, since 2002, the
number of cotton acres being grown under the more efficient drip irrigation methods increased by 400%
(Reed et al., 2009).
SOIL
Top soil depth and soil quality are integral to cotton producers livelihoods. U.S. growers recognize that
preserving soil resources is paramount. One key technique in maintaining soil quality is conservation
tillage.
Claim
Cotton harms the soil
The facts
Tillage is the primary source of soil quality loss and is the primary reason most conventional operations
now use erosion control measures like conservation tillage. The advantages of conservation tillage are
many. Included are: reduced erosion, increase populations of micro and macro flora and fauna, reduced
runoff, increased soil moisture and reduction in fuel usage (Fawcett & Towery, 2003), In the US 2/3 of
surveyed growers use some form of conservation tillage which decreases soil loss and improves the
quality of the soil

(Reed et al., 2009). Cotton can tolerate poor, infertile soils and its nutrient requirements
are less than those for most crops. Because of its deep tap root, cotton makes efficient use of soil
nutrients and fertilizers. Chemical fertilizers and pesticides do not reduce soil quality or soil organic
matter, nor does their use preclude the use of animal manures.
ENERGY
Claim
Cotton production requires a large amount of energy
The Facts
There are 1.5 pounds of cottonseed in every pound of cotton lint. When accounting for the embedded
energy contained in the seed it takes less energy to produce a cotton crop than the energy produced
by the cotton crop because of the energy embedded in this cottonseed (Matlock, 2009). There is
no doubt that energy resources will be limited in the future, and optimizing cottons energy use will
continue to be an important priority for agricultural research in the US.
GREENHOUSE GASES
Claim
Large amounts of greenhouse gases are released by the production of cotton.
The facts
Cotton, like all green plants, take carbon from the atmosphere and stores it in the leaves, stems, roots and
surrounding soil. One key technique to ensure maximum carbon storage is conservation tillage, a practice
that leaves plant residues on the soil surface for erosion control and moisture conservation. Growing a
hectare of reduced tillage cotton removes 3.4 MT of CO
2
from the atmosphere every year

(Causarano et
al., 2006). In addition, the amount of CO
2
removed by cotton plants worldwide is equivalent to taking
over 7 million cars off the highways

(Andy Jordan, personal communication, 2007) and the 5.1 MT of
Debunking the Myths 447
cotton consumed in the U.S. sequesters 7.7 billion kg of CO
2
into textile products per year

(Reed et al,
2009). In the US, more than two-thirds of growers have adopted some form of conservation tillage with
the greatest adoption occurring over the past decade (Reed et al., 2009).
COMPETITON WITH FOOD
Claim
Cotton production competes with food production.
The Facts
At the beginning of each crop season, the cotton grower need not make a choice between growing food or
growing fiber because one crop cycle produces both. The cotton plant produces fiber, cottonseed for oil
and feed, and gin by-products which have multiple industrial uses. The same inputs of energy, water and
nutrients used to produce a kg of cotton fiber also produce 1.5 kg of cotton seed. In the future, this
protein-rich cotton seed will become an even more economically important feed stock but will also be
edible by people, thanks to ongoing biotech research aimed at removing gossypol, a naturally-occurring
but inedible pigment (Sunilkumar et.al. 2006).
SUMMARY
It is often claimed that Cotton production has a very large environmental footprint. The fact is, in the
United States the environmental impact of producing a pound of cotton has fallen substantially over the
last 20 years (Figure 1). There has been a 25% reduction in land requirement, a 34% drop in soil loss, a
49% decline in irrigation water use, a 66% reduction in energy, and a 33% decrease in GHG emissions
(Field to Market, 2009). Cotton Incorporated continuously works to improve cottons environmental
footprint through its funding of agricultural research programs throughout the U.S. and by assisting
growers in adopting technologies and practices that conserve natural resources and enhance grower
efficiency. These programs are highly valued by growers and have contributed significantly to cottons
environmental gains over the past 40 years. It is hoped that by providing facts to dispel myths in venues
such as these that others will be empowered to communicate cottons positive environmental story as
well.
REFERENCES
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[8] Crumby, T.I., Mastrangelo, P., Sloan, C., Finlayson, B., Hosea, R., Trostle, M., Mitchell, T., Wells, S. and Karner, M.
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environment by reducing the need to plow. http://www.ctic.org/media/pdf/Biotech2003.pdf
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Outcomes of On-Farm Agricultural Production in the United States, First Report, January 2009.
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scale comparison of arthropod populations on commercially manged Bt and non-Bt cotton fields. Environ. Entomol. 34(5):
12571266.
[14] Hoekstra, A.Y. & Chapagain, A.K. (2007) - Water footprints of nations: water use by people as a function of their
consumption pattern. Water Resource Management. 21(1): 3548.
[15] (ISAAA) International Service for Acquisition of Agri-biotech Applications. 2011. Global Status of Commercialized
Biotech/GM Crops: 2010. Brief 42-2010: Executive Summary.
http://www.isaaa.org/resources/publications/briefs/42/executivesummary/default.asp.
[16] James, C. (1997) - Global Status of Transgenic Crops in 1997. ISAAA Briefs No. 5. ISAAA: Ithaca, NY. pp. 31.
[17] James, Clive. (2010) - Global status of commercialized biotech/GN crops: 2010. ISAAA: Ithaca, NY. ISAAA Brief No.42.
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[26] Sunilkumar, G., Campbell, L.M., Puckhaber, L., Stipanovic, R.D. and Rathore, K.S. (2006) - Engineering cottonseed for
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Outcomes of On-Farm Agricultural Production in the United States, First Report, January 2009
Analysis of Growth and Instability

of Cotton Production in India
Anuradha Narala
1
and A.R. Reddy
2
1
Scientist,
2
Senior Scientist Central Institute for Cotton Research, Nagpur
AbstractPolicy decisions are often made based on the growth rates which depend on the nature and structure of the
data and instability in farm production. The present paper analyzes the growth and instability in cotton area,
production and productivity during the period 1951-52 to 2010-11. Growth and instability of cotton during pre-
introduction (1993-94-2001-02) and post- introduction of Bt cotton periods (2002-03-2010-11) was also analyzed. For
this purpose compound growth rates were estimated by fitting the exponential function and coefficient of variation was
worked out to find out instability associated. It was found that growth of cotton area and production was significant
during 1950s, 1990s and 2001-10. Growth of cotton production was highest during the period 2001-10. Growth rate
of productivity was also high during 2001-10. Instability analysis indicated that cotton area was more stable than
production and productivity. Thus, policies should be made to reduce the risk in cotton production and to make it
profitable so as to sustain the high growth rate experienced during the past few years.
INTRODUCTION
Cotton is an important commercial crop of India and plays a key role in the national economy. About 60
million people get employment either directly or indirectly in the agricultural and industrial sectors of
cotton production, processing, textiles and related activities and by way exports, the foreign exchange
earnings of cotton amounts Rs.3837.33 crores (CAB, 2008). With economic liberalization and
globalization sweeping the world there is a scope for our country to play a leading role in the cotton
production and export. It is encouraging to note that over the last few years the cotton production had
shown a significant increase. In 2010-11 season, it touched a record production of 325 lakh bales with an
average productivity of 496 kg/ha (Cotton Corporation of India, 2010). Majority of cotton produced in
India is consumed domestically and hence, export of cotton from India is only 5.5 million bales. If this
increasing production trend continues in years to come, India can become a major exporter of cotton.
Researchers have shown that with the adoption of new technologies on farmer's fields, it is possible to
increase the average productivity beyond 600 kg lint per ha to meet the increasing cotton demand.
Growth rates are the measures of past performance of economic variables. They are commonly used as
summaries of trends in time series data. They are developed to describe the trends in a variable over time.
Policy decisions are often made based on such growth rates which depend on the nature and structure of
the data. Instability in farm production is causing serious shocks to supply and farm income and there is a
growing concern about increased volatility in farm production, prices and farm income (Ramesh Chand
and Raju, 2008). Indian cotton production has undergone a metaphoric changes from 2002-03, after Bt
cotton was introduced in the country, since then significant increase in area, production and yield was
witnessed. Bt cotton now occupies 95% of the total cotton area in the country. These dynamic changes
underline the importance of studying the growth performance and instability of cotton before and after Bt
cotton introduction. Therefore the present study was undertaken to analyze the growth and instability in
cotton area, production and productivity during 1951-2010.
METHODOLOGY
The present analysis is based on the secondary data sourced from Cotton Advisory Board, Ministry of
Agriculture, and Government of India. The period of analysis is 1951-52 to 2010-11. The entire period
was sub divided into six sub periods of ten years each. Further to study the growth before and after Bt
cotton introduction the period from 1993-94 to 2001-02 was taken as pre-introduction of Bt cotton period
and from 2002-03 to 2010-11 as post- introduction of Bt cotton period. Compound growth rates (CGR)
75
of area, production and productivity of cotton were worked for different periods as well as for entire
period of analysis by fitting exponential function as given below:
X
t
= ab
t

Log X
t
= Log a + t log b
b = (1 + r) / 100
Where,
X
t
= Area/production/productivity of pulse crops in the year 't'
t = time element which takes the value 1, 2, 3, . n
a = intercept
b = regression coefficient.
CGR was worked out as follows:
CGR (r) = (antilog b - 1) x 100
Student 't' test was used to test the significance of the CGR.
Instability was analyzed by estimating coefficient of variation (CV) using the formula.
CV= ( / ) * 100
Where
= Standard deviation
= Mean
COTTON PRODUCTION SCENARIO OVER THE YEARS
TABLE 1: AREA, PRODUCTION AND YIELD OF COTTON IN INDIA
Year Area (m ha) Production (m bales) Yield (kg/ha)
1951-52 6.56 3.28 85
1955-56 8.09 4.18 88
1960-61 7.61 5.6 125
1965-66 7.96 4.85 104
1970-71 7.61 4.76 106
1975-76 7.35 5.95 138
1980-81 7.82 7.01 152
1985-86 7.53 8.73 197
1990-91 7.44 9.84 225
1995-96 9.04 12.86 242
2000-01 8.58 14.00 278
2005-06 8.68 24.4 478
2010-11 11.14 32.5 496
Source: CAB, Ministry of Agriculture, GOI.
Over the years, country has achieved significant quantitative increase in cotton production (Table 1).
Till 1970s, country used to import massive quantities of cotton in the range of 0.8 to 0.9 m bales per
annum. However, after the Government launched special schemes like intensive cotton production
programmes through successive five-year plans, that cotton production received the necessary impetus.
As a result of initiation of systematic cotton improvement programmes and the launch of AICCIP on 1
st

April 1967 for cotton improvement involving genetic, production and protection technologies has
resulted in increasing countrys productivity by 7-8 folds from 88 kg to 553 kg lint ha
-1
. The realized raw
cotton production of nation during 2010-11 was 32.5 m bales. It is expected that this trend is going to
remain more or less the same.

Analysis of Growth and Instability of Cotton Production in India 451
If we examine cotton area over the years it is clear that area increase is less than that of production
and productivity. Cotton area increased from 6.56 m ha in 1951-52 to 11.14 m. ha in 2010-11. But cotton
production increased nearly 10 folds during the period of analysis. Cotton productivity increased to
nearly 7-8 folds during the same period. Cotton productivity was only 85 kg/ha in the year 1951-52 and
increased to 496 kg/ha in the year 2010-11. Increase in productivity and production was more prominent
after 2000-01.
GROWTH OF AREA, PRODUCTION AND PRODUCTIVITY
Decadal growth rates of cotton area, production and productivity were worked out and are presented in
Table 2. Growth rate of cotton was negative during two decades only. But t - values indicated that this
negative growth was statistically not significant. Growth rate was positive in other periods as well as
overall period. Cotton area increased significantly during 1951-60, 1991-00 and 2001-10, the growth
rates were 1.91, 2.21 and 3.43 percent respectively. During other periods, growth rate of cotton area was
not significant. Growth rate of cotton area was positive and significant during overall period.
Cotton production during 1960-1970 witnessed significant negative growth rate whereas in 1951-60,
1991-00 and 2001-10 cotton production increased significantly. Growth of cotton production was highest
during 2001-10. During overall period of analysis also cotton production increased significantly at an
annual rate of 3.29 %.
Cotton productivity also recorded positive growth during all the periods except during 1961-70
which has shown a negative growth rate. Among the decades showing positive growth rate all were
statistically significant except the 1971-80 period. Productivity growth was highest during the last period
i.e. 2001-10. During this period cotton productivity increased at a rate of 5.97% per annum, this may be
due to introduction of Bt cotton. The growth rate for overall period was also positive and significant. The
productivity increased at a rate of 2.94% per annum during the overall period.
INSTABILITY
CV for different periods was worked out for area, production and productivity and is given in Table 3. It
is clear from the analysis that instability in cotton area was less when compared to production and
productivity. Coefficient of variation for area was only 10.72 % where as it was 72.70% and 60.68% in
case of production and productivity, respectively. Instability in cotton area was the least during 1961-70
followed by 1971-80. It was highest during 2001-10 followed by 1991-00 and 1951-60. Instability of
cotton production was highest during 2001-10 and lowest during 1961-70. Similarly, CV of yield was
also highest during 2001-10 followed by 1981-90.
TABLE 2: TREND IN GROWTH RATE (%) OF AREA, PRODUCTION AND YIELD OF COTTON IN INDIA
Period Area Production Yield
1951-52 to 1960-61 1.91** 4.22** 2.30*
(2.4747) (2.7554) (1.9673)
1961-62 to 1970-71 -0.58 -0.27* -0.30
(-1.9056) (-0.2826) (0.3557)
1971-72 to 1980-81 0.50 1.10 1.50
0.9215 (1.7045) (1.8230)
1981-82 to 1990-91 -0.97 3.32 4.31***
(-1.4736) (1.7611) (3.1234)
1991-92 to 2000-01 2.21*** 4.63*** 2.37*
(3.1864) (3.5850) (1.9493)
2001-02 to 2010-11 3.43*** 9.63*** 5.97***
(5.0203) (6.3209) (3.8722)
Overall 0.34*** 3.29*** 2.94***
(5.4220) (21.4523) (25.2330)
***1% Significant level; ** 5% Significant level; * 10% Significant level Figures in the parenthesis indicates t-value

TABLE 3: DECADE-WISE INSTABILITY (CV IN %) OF AREA, PRODUCTION AND PRODUCTIVITY OF COTTON IN INDIA
1951-52 to 1960-61 8.45 17.56 12.41
1961-62 to 1970-71 3.20 8.11 7.17
1971-72 to 1980-81 4.81 11.49 8.14
1981-82 to 1990-91 6.22 19.21 17.64
1991-92 to 2000-01 8.71 17.58 12.34
2001-02 to 2010-11 11.84 27.10 19.45
Overall 10.72 72.70 60.69
PRE AND POST- BT COTTON INTRODUCTION
Growth & Instability
Bt cotton in India was introduced in the year 2002. Since then the area, production and yield of cotton in
India has undergone dynamic changes. A comparative analysis of growth before and after the
introduction of Bt cotton in India (Table 4) suggests significant increase in the growth in area, production
and yield during the post-Bt introduction than the pre-Bt period. The growth in area, production and yield
during post-Bt introduction were 4.45, 9.72 and 5.02 % respectively, as against the pre-Bt introduction of
1.53, 4.37 and 2.83 %.
TABLE 4: GROWTH RATE (%) OF AREA, PRODUCTION AND YIELD OF COTTON IN INDIA BEFORE AND AFTER INTRODUCTION OF BT COTTON
Pre-Bt Introduction 1.53 4.3738** 2.83*
(1993-94 to 2001-02) (1.7088) (2.7953) (1.9519)
Post-Bt Introduction 4.45*** 9.72*** 5.02**
(2002-03 to 2010-11) (9.5795) (5.0940) (2.7834)
Overall 1.18*** 6.10*** 5.75***
(1993-94 to 2010-11 (3.0459) (10.6514) (9.3339)
***1% Significant level; ** 5% Significant level; *10% Significant level Figures in the parenthesis indicates t-value
Instability in area, production and yield of cotton before and after introduction of Bt-cotton in India is
given in Table 5. CV which is the indicator of instability, for area production and yield of cotton was
higher in post-Bt cotton introduction than pre-Bt introduction. During post-Bt period, the instability in
the production of cotton was maximum (24.47%) followed by yield (16.28%) and area (12.41%).
TABLE 5: INSTABILITY IN AREA, PRODUCTION AND PRODUCTIVITY OF COTTON (CV IN %) IN INDIA- PRE AND POST BT INTRODUCTION
Particulars Area Production Yield
Pre-Bt Introduction (1993-94 to 2001-02) 7.36 15.81 12.45

Post-Bt Introduction (2002-03 to 2010-11) 12.41 24.47 16.28

Overall (1993-94 to 2010-11) 10.47 38.10 32.14
CONCLUSION
From the above analysis it is clear that during the last period and in the post- Bt period there was a
significant increase in area, production and productivity and registered high growth rate. This may be due
to introduction of Bt cotton in India. Instability was also high during this period indicating that cotton
production increased over the periods. Although the cotton production and productivity is following an
increasing trend, it is associated with many problems. Cost of production is escalating due to the rise in
the prices of inputs. The prices of cotton are fluctuating from place to place and year to year making the
production risky. Most of the cotton area is sown with Bt hybrids which have very high seed cost. The
nutrient requirement is also high. Similarly the labour cost for cotton picking is also increasing
exorbitantly. Since the Bt cotton matures early and because of synchronized bole bursting at one time the
labour demand for cotton-picking increases abruptly. Considering these facts policies should be made to
Analysis of Growth and Instability of Cotton Production in India 453
reduce the risk in cotton production and to make it profitable. Programs and policies such as
rehabilitation of irrigation systems, adoption of improved technologies, strengthening of extension,
reducing risks in cotton production can play a vital role in achieving stability and to sustain the high
growth rate experienced during the past few years.
REFERENCES
[1] Cotton Advisory Board (2008) - Ministry of Agriculture, Government of India.
[2] Cotton Corporation of India. www.cotcorp.gov.in.
Ramesh Chand and S.S.Raju (2008) - Instability in Andhra Pradesh Agriculture A Disaggregate Analysis Agri.
Economic Res. Rev., 21: 283288.
Total Factor Productivity of Cotton

in Gujarat (India)
A.R. Reddy
1
, S.M. Yelekar
2
, R.B. Petkar
2
and N. Anuradha
3
1
Senior Scientist,
2
Senior Research Fellow,
3
Scientist, Central Institute for Cotton Research, Nagpur
E-mail: ar.reddy@gmail.com
AbstractGujarat is one of the important cotton producing states of India, which ranks first in production and
second in area in the country. An analysis was carried out to study the total factor productivity (TFP) of cotton in the
state for the period 1981-82 to 2007-08. Tornqvist-Theil indexing procedure was adapted to workout total input, total
output and total factor productivity indices. These indices were worked out for first period (1981-1990), second period (1990-
2000) and last period (2000-2008) separately. Input factors considered to workout TFP includes human labour (days/ha),
bullock labour (days/ha), machine labour (days/ha), seeds (kg/ha), manure (q/ha), NPK (kg/ha), and insecticides (l/ha).
The results indicated that the growth of TFP was positive and significant during the period of analysis. TFP increased at the
rate of 5.71% per a. % per annum during the period of analysis. This indicates the sustainability of cotton production in the
state. The growth of total input index was not significant during the period of analysis. But total output index showed a
significant growth of 5.44 % per annum. This was due to the increase in the TFP during this period. TFP growth was positive
during all the three periods. It was highest (10.75 %) during 2000-07 while it was lowest (1.05 %) during 1981-1990. Growth
of input index was negative during first two periods while it was positive during the last period. Similarly growth of total
output index was negative during the first period and positive during last two periods. This analysis indicates that the output
growth of cotton in this state is sustainable as it is driven by the positive TFP growth. It also indicates that the growth of output
can further be strengthened by improving the input growth.
INTRODUCTION
In agriculture, generally, productivity is measured in terms quantity produced per unit area of land. This
was mainly because land is one of the scarce input and producer used to maximize production per unit
area of land. But productivity will not entirely depend upon land. Other inputs like seeds, fertilizers,
manure, labour, plant protection chemicals etc., play an important role. Inadequacies in the measurement
of single factor productivity led economists to devise methods to measure total factor productivity (TFP).
TFP measures the increase in total output which is not accounted for by increases in total inputs. The
total factor productivity index is computed as the ratio of an index of aggregate output to an index of
aggregate inputs. Therefore growth in TFP is equal to the growth rate in total output less the growth rate in
total inputs. Earlier Laspyeres arithmetic indices were used most commonly to measure TFP (Pandya and
Shiyani, 2002). But most recent literature on TFP (Kumar and Mruthyunjay, 1992; Kumar and
Rosegrant, 1994; Desai and Nambudri, 1997; Mittal and Lal, 2001) has advocated and employed
Tornqvist Theil or translog index in their study for drawing meaningful inferences.
Many of the studies conducted in the past considered agricultural sector as a whole and only a few
studies were done on specific crops. No study was conducted to workout TFP of cotton in India
especially in Gujarat. Gujarat is an important cotton growing state in the country. It ranked first in cotton
production and second in cotton area. In Gujarat, cotton is grown on 26.3 lakh ha producing 102 lakh
bales of lint with an average productivity of 658 kg/ha (2010-11). In this study, a critical analysis is done
on the partial as well as total productivity of cotton for the period 1981 - 2008.
DATA AND METHODOLOGY
Data on cost of cultivation of cotton in Gujarat was collected from the Commission for Agricultural
Costs and Prices, New Delhi. The data were collected for the period 1981-82 to 2007-08. The period was
divided into three sub periods, period I (1981-82 to 1989-90), period II (1990-91 to 1999-2000) and
period III (2000-01 to 2007-08).
76
Total Factor Productivity of Cotton in Gujarat (India) 455
COMPOUND GROWTH RATES
Compound growth rates were worked out for area, production and productivity of cotton, inputs used and
partial input productivities for the period of study. Compound growth rates were worked out by fitting
exponential function as given below:
Xt = ab
t

Log Xt = Log a + t log b
Compound growth rate (r) = (antilog b - 1) x 100
Where,
Xt = Value of the variable in the year 't'
t = time element which takes the value 1, 2, 3, . n
a = constant
b = regression coefficient.
Student 't' test was used to test the significance of the compound growth rate.
PRODUCTIVITY OF INDIVIDUAL INPUTS
Productivity of individual inputs was worked out as the quantity of seed cotton produced per unit of each
input
Partial productivity = quantity of seed cotton produced / quantity of input used
TOTAL FACTOR PRODUCTIVITY
The Tornqvist-Theil index is a superlative index which is exact for the linear homogeneous translog
production function (Diewert, 1976). A further advantage of the Tornqvist-Theil index is that it accounts
for changes in quality of inputs. Because current factor prices are used in constructing the weights, quality
improvements in inputs are incorporated, to the extent that these are reflected in higher prices (Antle and
Capalbo, 1988). The Tornqvist-Theil index provides consistent aggregation of inputs and outputs under
the assumptions of competitive behaviour, constant returns to scale, Hicks-neutral technical change, and
input-output separability. However, Caves et al. (1982) showed that Tornqvist-Theil indices are also
superlative under very general production structures, i.e., non-homogeneous and non-constant returns to
scale, so they provide consistent aggregation across a range of production structures. TFP indices were
worked out by using Tornqvist-Theil indexing procedure expressed in logarithemic form, which is given
by the following equation:
Ln (TFP
t
/ TFP
t-1
) = ln (Q
t
/Q
t-1
) - (C
it
+ C
it-1
) ln (X
it
/ X
it-1
)
Where
Q
t
= Output of cotton in year t
C
it
= Share of input i' in total input cost
X
it
= Input i' in year t
Specifying the index equal to 100 in a particular year and accumulating the measure based on above
equation provides the TFP index.
Input variables considered for constructing the indices were human labour (days/ha), bullock labour
(days/ha), machine labour (hours/ha), seeds (kg/ha), manure (q/ha), NPK (kg/ha) and insecticides (lit
/ha).
Area, Production and Productivity
Cotton area in Gujarat increased from 15.7 lakh ha in 1980-81 to 26.3 lakh ha in 2010-11 while
production increased from 17.1 lakh bales to 102 lakh bales in the same period (Table 1). This
spectacular increase in production was mainly due to an increase in cotton productivity from a mere
185.36 kg lint/ha to 658 kg lint per ha. The cotton acreage increased at the rate of 2.34 % per annum
while cotton production and productivity increased at the rate of 7.77 % and 5.30 % per annum.
TABLE 1: AREA, PRODUCTION AND PRODUCTIVITY OF COTTON IN GUJARAT
Year Area (lakh ha)
Production
(lakh bales) Yield (kg/ha)
1980-81 15.7 17.1 185
1985-86 14.0 19.9 240
1990-91 9.2 15.0 277
1995-96 14.1 31.3 377
2000-01 16.2 23.8 251
2001-02 16.9 32.5 327
2002-03 16.3 30.5 317
2003-04 16.5 50.0 515
2004-05 19.1 73.0 651
2005-06 19.1 89.0 794
2006-07 23.9 103.0 733
2007-08 24.2 110.0 772
2008-09 23.5 90.0 650
2009-10 26.3 98.0 635
2010-11 26.3 102.0 659
CGR

2.34**
(5.944)
7.77**
(8.394)
5.3**
(8.910)
Note: *1 bale = 170 kg
** Significant at 1 % level
Figures in the parentheses indicate t values
Input Utilization in Cotton Production
Human labour, animal labour, machine labour, seeds, fertilizers, and plant protection chemicals are
important inputs used in cotton production. Human labour accounts for nearly 40 % of working cost.
Human labour utilization fluctuated between 66.93 man days to 146.75 man days/ha during the period of
analysis (Table 2). These fluctuations in human labour utilization were mainly due to variations in labour
requirement for picking and weeding operations. Significant growth was not observed in human labour
utilization during the period of analysis. Animal labour utilization showed a negative growth of 3.30 %
per annum and reduced from 9.7 animal pair days per ha in 1981-82 to 5.32 animal pair days per ha in
2007-08. This may be due to the mechanization of some of the farm operations. This is evident as
machine labour showed a positive growth of 6.80 % per annum. Machine power is mainly used for tillage
and land preparation. Still many field operations are done using animal power.
Quantity of seeds used per ha showed a sharp decrease during the period of analysis. It reduced from
12.5 kg/ha in 1981-82 to 3.0 kg/ha in 2007-08. There was a negative growth of 3.17 %per annum in
quantity of seed used. This was mainly due to an increase in area under hybrids especially Bt hybrids.
Seed cost is not only high but seed requirement reduced as hybrids are grown at a wider spacing.
Fertilizers (NPK) are another important input in cotton production, which registered a positive growth of
0.28 % annum. Fertilizers (NPK) consumption increased from 98.01 kg/ha in 1981-82 to 149.48 kg/ha in
2010-11. Significant change was not observed in the quantity of manures used in cotton production.
Cotton is prone to many pests and diseases, which necessitates the use of chemicals. As the quantitative
data is not available an index was developed by using price index. Quantity of pesticides used in cotton
production increased at the rate of 3.44 % per annum.
TABLE 2: QUANTITIES OF INPUTS USED IN COTTON CULTIVATION
(per ha)
Year Human Labour
(Man days)
Animal Labour
(Pair days)
Machine labour
(hrs)
Seeds
(Kgs)
NPK
(kgs)
Manures
(Qtl)
Insecticides
(Index)
1981-82 123 9.7 1.42 12.52 98.01 27.00 602.29
1982-83 144 14.1 2.82 7.25 162.63 37.39 970.73
1983-84 121 10.5 1.67 10.20 79.58 23.94 517.90
1984-85 107 9.9 0.70 9.14 78.19 23.30 481.71
1985-86 93 9.4 0.69 8.09 76.81 22.66 445.53
1986-87 80 8.9 0.67 7.03 75.42 22.02 409.34
1987-88 122 12.5 2.33 5.07 103.09 83.80 328.40
1988-89 113 9.8 0.73 6.76 88.24 30.10 840.45
1989-90 105 8.6 0.78 5.89 97.29 31.32 680.45
1990-91 115 9.2 1.35 5.61 111.18 38.87 715.19
1991-92 125 9.9 1.29 5.34 125.06 46.41 749.94
1992-93 134 10.6 1.02 5.06 138.95 53.96 784.68
1993-94 94 8.3 1.36 9.58 83.61 20.43 579.36
1994-95 96 7.4 0.54 8.83 83.12 25.56 754.64
1995-96 97 6.3 1.41 7.98 88.88 32.40 843.22
1996-97 99 5.6 2.74 7.34 82.14 35.81 1105.21
1997-98 105 6.2 3.37 7.55 89.69 39.05 1356.91
1998-99 108 6.0 3.27 6.98 90.62 41.08 1058.62
1999-00 92 6.3 2.85 6.08 81.41 39.26 1287.03
2000-01 67 5.4 3.72 6.14 59.19 41.70 473.47
2001-02 105 6.7 3.54 4.35 78.97 41.41 1196.80
2002-03 100 5.2 3.79 4.97 68.03 34.27 492.95
2003-04 140 5.8 4.85 4.61 94.85 26.59 963.04
2004-05 134 5.3 4.34 3.49 102.86 33.52 981.86
2005-06 147 6.0 4.38 4.41 142.46 36.81 1424.54
2006-07 121 5.2 3.81 3.54 124.93 33.31 1304.03
2007-08 124 5.3 4.29 3.02 149.48 36.75 1505.26
CGR

0.11
(0.229)
-3.30**
(-9.967)
6.80**
(5.184)
-3.17**
(-5.461)
0.28
(0.446)
0.83
(1.094)
3.44**
(4.067)
** Significant at 1 % level,* = Significant at 5 % level
Partial Productivities of Inputs Used in Cotton Production
Analysis revealed that productivity of land, human labour, animal labour and seeds increased while the
productivity of machine labour decreased during the period of analysis (Table 3). Productivity of land
showed a spectacular increase from 699 kg/ ha in 1981-82 to 1932 kg/ha in 2007-08. Productivity of land
increased at the rate of 8.66% per annum. Similarly, productivity of human labour increased from 5.6
kg/man day in 1981-82 to 15.6 kg/man day recording a positive growth of 5.11% per annum. Similarly
productivity of animal labour manures and seed showed a positive growth. Productivity of seeds, an
important input, increased from 55.8 kg/kg seed in 1981-82 to 639.7 kg/kg seed in 2007-08. Significant
growth of 4.92% was observed in the case of fertilizers whereas machine labour showed a non-significant
growth.
Analysis of individual productivity has many limitations, as an increase in the productivity of
individual input is not the effect of that input only. Other inputs which are applied in the production
process have also contributed to it. Thus, attributing this to any single factor is not appropriate
necessitating TFP analysis.

TABLE 3: PARTIAL PRODUCTIVITY OF INDIVIDUAL INPUTS
Year
Human Labour
(kg Seed Cotton/
Manday)
Animal labour
(kg Seed Cotton/
pairday)
Machine
Labour
(kg Seed
Cotton/Hour)
NPK
(kg Seed
Cotton/kg
NPK)
Manures
(kg Seed
Cotton/Q)
Seeds (kg
Seed
Cotton/kg)
Land
(kg Seed
Cotton/ha)
1981-82 5.67 72.36 493.85 7.13 25.89 55.83 699
1982-83 3.68 37.42 187.28 3.25 14.12 72.83 528
1983-84 4.38 50.39 316.54 6.63 22.06 51.76 528
1984-85 7.13 76.68 1086.96 9.75 32.70 83.34 762
1985-86 7.76 76.94 1042.38 9.41 31.91 89.41 723
1986-87 5.13 46.07 612.34 5.41 18.53 58.04 408
1987-88 1.73 16.74 90.21 2.04 2.51 41.42 210
1988-89 6.10 70.13 945.94 7.79 22.82 101.63 687
1989-90 7.14 87.67 963.87 7.74 24.04 127.84 753
1990-91 7.23 89.88 617.20 7.47 21.38 148.04 831
1991-92 5.47 68.77 527.19 5.45 14.67 127.61 681
1992-93 6.88 87.50 909.41 6.65 17.12 182.61 924
1993-94 11.22 126.51 773.84 12.59 51.54 109.92 1053
1994-95 10.68 137.33 1898.65 12.27 39.91 115.47 1020
1995-96 11.68 179.88 802.70 12.73 34.91 141.73 1131
1996-97 11.61 203.37 417.88 13.95 32.00 156.13 1146
1997-98 13.99 236.10 436.45 16.39 37.64 194.70 1470
1998-99 12.53 226.68 413.98 14.93 32.94 193.84 1353
1999-00 12.79 185.56 413.34 14.45 29.95 193.42 1176
2000-01 13.00 160.78 233.69 14.70 20.86 141.69 870
2001-02 9.78 152.11 289.70 12.99 24.78 235.86 1026
2002-03 10.56 204.37 278.12 15.48 30.73 211.87 1053
2003-04 10.16 246.81 293.65 15.02 53.59 309.11 1425
2004-05 11.52 291.30 355.75 15.02 46.09 442.69 1545
2005-06 13.17 320.80 441.18 13.56 52.49 438.10 1932
2006-07 15.98 374.60 507.73 15.46 58.00 545.76 1932
2007-08 15.59 363.33 450.79 12.92 52.57 639.74 1932
CGR 5.11** 8.81** -1.37 4.92** 4.36** 8.66** 5.22**
T value 8.1341 10.940 -1.3042 7.0421 4.4908 7.6325 8.5700
** Significant at 1 % level
* = Significant at 5 % level
Total Factor Productivity
Indices of total input, total output and TFP of cotton in Gujarat are given in Table 4. TFP index showed a
positive growth rate of 5.81% per annum during the period 1981-2008. The growth in the total input
index was negative but not significant. Output index showed a positive growth of 5.59% per annum
which was contributed by TFP growth only. As the growth in output is backed by the growth in TFP, the
output growth is sustainable. The output growth can further be accelerated by the enhanced input growth.
If we examine these indices in different time periods it is clear that TFP growth was highest during
the period III. During this period TFP increased at the rate of 10.79% per annum. This period coincides
with the introduction of Bt cotton, new insecticides as well as improved crop production technologies.
All these contributed to the growth of TFP in this state. Input index also showed positive growth during
this period. All these resulted in 12.2% output growth during this period which was comparatively higher
than other two periods.
During period I, though total factor productivity growth was positive, output index showed negative
growth. This was due to the negative growth in input index. Positive growth of TFP was offset by the
negative growth in input index. In period II, output recorded a positive growth of 6.43% though there was
a negative growth in the input index. This was made possible due to the positive growth of total factor
productivity during this period.
TABLE 4: TOTAL FACTOR PRODUCTIVITY OF COTTON IN GUJARAT
Year Total Input Index Total Output Index Total Factor Productivity Index
1982-83 100 100 100
1983-84 81.20 100.00 123.15
1984-85 75.61 144.32 190.87
1985-86 70.50 136.93 194.24
1986-87 63.74 77.27 121.24
1987-88 91.24 39.77 43.59
1988-89 84.06 130.11 154.79
1989-90 73.51 142.61 194.02
1990-91 82.59 157.39 190.55
1991-92 86.55 128.98 149.02
1992-93 89.66 175.00 195.19
1993-94 70.27 199.43 283.81
1994-95 68.53 193.18 281.88
1995-96 70.32 214.20 304.61
1996-97 73.43 217.05 295.58
1997-98 76.89 278.41 362.10
1998-99 74.67 256.25 343.17
1999-00 72.50 222.73 307.23
2000-01 57.51 164.77 286.52
2001-02 76.83 194.32 252.91
2002-03 69.77 199.43 285.83
2003-04 83.00 269.89 325.15
2004-05 78.13 292.61 374.52
2005-06 86.67 365.91 422.20
2006-07 76.31 365.91 479.47
2007-08 87.97 365.91 415.94
Compound Growth Rate
Period I (1982-90)
-1.81
(-0.1726)
-0.76
(0.5865)
1.05
(0.5762)
Period I (1990-00)
-1.71
(-1.1493)
6.43
(0.3602)
8.14
(1.1120)
Period I (2000-08)
1.41
(0.9752)
12.20**
(4.5627)
10.79**
(6.2287)
Overall period
-0.27
(-0.8774)
5.44**
(6.9653)
5.71**
(6.7866)
** Significant at 1 % level; * Significant at 5 % level;
Figures in the parentheses indicate t values
CONCLUSION
Our results indicate that the growth of TFP was positive and significant during the period of analysis, and
increased at the rate of 5.7% per annum. Total output index showed a significant growth of 5.44 % per
annum. This entire growth was was contributed by the TFP growth only as the growth in the input index
was insignificant. This analysis indicates that the output growth of cotton in this state is sustainable as it
is driven by the positive TFP growth. It also indicates that the growth of output can be strengthened
further by improving input growth.
REFERENCES
[1] Antle, J.M. and Capalbo, S.M. (1988) - An Introduction to Recent Developments in Production Theory and
Productivity Measurement. In: S.M. Capalbo and J.M. Antle (eds)- Agricultural Productivity Measurement and
Explanation, Resource for the Future Washington D.C.
[2] Caves, D.W., Christensen, L.R. and Diewert, W.E. (1982) - Multilateral Comparisons of Output, Input and
Productivity Using Superlative Index Numbers- Economic Journal, 92: 7386.
[3] Diewert, W.E. (1976) - Exact and Superlative Index Numbers- Journal of Econometrics, 4 (2): 115146.
[4] Desai, B.M. and Namboodiri N.V. (1997) - Determinants of Total Factor Productivity in Indian Agriculture- Economic and
Political Weekly, 32: 165171.
[5] Kumar, P. and Rosegrant, M.W. (1994) - Productivity and Sources of Growth for Rice in India- Economic and Political
Weekly, 29: 183188.
[6] Kumar, P. and Mruthyunjaya, (1992) - Measurement and Analysis of Total Factor Productivity Growth in Wheat- Indian
Journal of Agricultural Economics, 47(3): 451458.
[7] Pandya, M. and Shiyani, R. L. (2002) - Analysis of total Factor Productivity Growth in Food Crops of Gujarat-Artha
Vijnana, 44(3-4): 367374.
[8] Mittal, S. and Lal, R.C. (2001) - Productivity and Sources of Growth for Wheat in India- Agricultural Economics Research
Review, 14(2): 109120.
Transfer of Technology Initiatives for Profitable

and Sustainable Cotton Farming in India
An Empirical Analysis
S. Usha Rani
1
and S.M. Wasnik
2

1
Scientist (Agrl. Extension), Central Institute for Cotton Research, Regional Station,
Coimbatore -641003, Tamil Nadu, India,
2
Principal Scientist (Agrl. Extension), Central Institute for Cotton Research,
P.B. No. 2, Shankarnagar, Nagpur10, Maharashtra, India
E-mail: ushajoshua@rediffmail.com,wasniksm2007@yahoo.co.in
AbstractTransfer of Technology (TOT) programs implemented in cotton have underlined the importance of
problem solving, creating effective scientists and farmers linkage and transferring the latest cotton production
technologies. Front Line Demonstrations, Farmers Field Schools, Contract Farming Approach are some of the TOT
programs which created remarkable impact on cotton production. Analysis on the laurels and let downs of those
initiatives revealed that they have high farmers acceptability due to its focus on problem solving and the practical
application of knowledge. They were effective in increasing the yields, sharing the knowledge but handicapped due to
lack of professional execution and non-availability of latest technological dissemination tools for ready transfer.
Exclusion of novel extension methods viz., cyber, market led, Farmer-led and environmental extension was observed in
many of the programs. Market intelligence surveys for commercializing our technologies and institutional
arrangements for freeing indebtedness and risk coverage had never found a significant place in those programs. Media
utilization and efforts to organize the cotton growers were the other areas where our cotton extension programs created
a meager impact. On the other side, Indian cotton sector is facing serious challenges posed by the changes viz.,
changing technology Bt cotton, changing demands of the textile industries and non woven sectors and changing
scenario of retaining the top position in acreage and second position in production at world level. All these changes
could not impact much on productivity which is a major setback. Reforms made in the technology delivery system in
other cotton growing countries raised the productivity of cotton. Therefore, criticisms about the present Indian cotton
extension system and the emerging challenges compel to review and restructure the existing cotton extension system.
A contemporary synergetic extension model with novel extension concepts and facility to forecast technology
considering the pros and cons of past methods and changing perspectives are suggested in this paper.
INTRODUCTION
Cotton is one of the major fibre crops of global significance, which is, cultivated by 20 million farmers
worldwide. The global cotton production in 2010 was 25.1 M tons (1476 lakh bales of 170 kg/bale) from
34.0 M hectares. India has the largest acreage accounting for 33.0% and contributes 21.1% of global
production currently ranking next to China. The production increased from a meager 2.3 M bales (170
kg lint/bale) in 1947-48 to a record production of 32.50 M bales during 2010-11. Both the accelerated
transfer of technology due to the sustained and joint research and developmental efforts by the
state/central institutions and the private sector agencies and the phenomenal spread of Bt cotton hybrids
seem to be the main contributory factors for the break through achieved in cotton production. In our
country, various Transfer of Technology (TOT) programs in cotton have underlined the importance of
problem solving, creating effective scientists and farmers linkage and transferring the latest cotton
production technologies. Front Line Demonstrations (FLD), Farmers Field Schools (FFS), Contract
Farming Approach are some of the TOT programs which created remarkable impact on cotton
production. The introduction of Bt cotton is a milestone in the history of cotton improvement, changing
scenario of Indian cotton in terms of acreage, production, productivity, quality, species composition and
export importance. As a result, new challenges for cotton extension in India emerged. In order to survive
in this era of economic liberalization and gene revolution, attaining excellent extension strategies is
inevitable for profitable and sustainable cotton farming in the country. This paper makes an attempt to
review the laurels and letdowns of past cotton extension programmes and suggest innovative strategies
for sustainable cotton production in the country.
77
TRANSFER OF TECHNOLOGY INITIATIVES FOR COTTON CROP IN THE COUNTRY
It is more often criticized that the results of cotton research do not reach the farmers in time. Often, it is
observed that the extension efforts are handicapped due to inadequate interaction with the research efforts
and non-availability of latest technological information for ready transfer. It is essential that the
developments that benefit the producer and consumer is translated into action, quick enough to meet the
growing challenges of the future. It is also vital to understand the socio-economic implications of new
technologies and understand the constraints limiting the transfer of technology. The Indian Council of
Agricultural Research (ICAR) has always underlined the importance of Scientist- Farmer linkage for an
effective transfer of latest agricultural technologies. Towards this goal, several programmes such as, Lab
to Land Programme, Operational Research Project (ORP), Front Line Demonstrations (FLD), Integrated
Pest Management (IPM), Integrated Resistance Management (IRM), Institute Village Linkage
Programme (IVLP), Intensive Cotton Development Programme (ICDP), Farmers Field Schools (FFS)
etc., were launched and implemented with active cooperation of the ICAR Institutes, State Agricultural
Universities and Extension personnel of the State Department of Agriculture. These programmes
ensured not only the quick dispersal of technologies by linking the Scientists, extension personnel and the
farmers but also helped scientists to get a feed back on the response of farmers to the latest technologies.
Among them, the three major cotton extension programmes which are currently prevailing in the cotton
sector are explained below.
Front Line Demonstrations on Cotton
TABLE 1: NUMBER OF COTTON FRONT LINE DEMONSTRATIONS CONDUCTED FROM 1996-97 TO 2010-11 BY AICCIP CENTRES WITH BUDGET PROVISIONS
Year
Number of Demonstrations Total amount Sanctioned
(Rs in Lakhs) North Central South Total
1996-97 200 170 181 551 15.00
1997-98 275 240 245 760 20.00
1998-99 270 235 255 760 20.00
1999-00 290 265 510 1065 30.50
2000-01 475 725 550 1750 87.50
2001-02 185 375 240 800 39.00
2002-03 115 215 150 480 25.00
2003-04 100 160 90 350 20.00
2004-05 217 495 143 855+55* 100.00
2005-06 277 588 285 1150+16*+20** 60.00
2006-07 250 750 350 1350+21*+23** 72.00
2007-08 255 795 350 1400+24*+22** 75.00
2008-09 500 1250 550 2300+30*+24** 100.00
2009-10 325 850 425 1600+19*+19** 70.00
2010-11 200 325 175 700+16*+16** 30.00
Total 3934 7438 4499 15871+126*+124** 764.00
* - Number of unit demonstrations on farm implements
** - Number of unit demonstrations on Integrated Pest Management
The programme Front Line Demonstration on cotton has been performing better than all other cotton
extension programmes prevailing in the country. Since 1996, the All India Coordinated Cotton
Improvement Project (AICCIP), as a nodal agency conducted FLDs. Funding for FLDs was from ICDP
that was later changed to the Technology Mission on Cotton (TMC) under Mini Mission II. The Project
Coordinator (Cotton) coordinates and monitors the implementation of the FLD programme with
headquarters at the Central Institute for Cotton Research, Regional Station, Coimbatore. The FLDs are
organized through AICCIP centres besides CICR, Nagpur and its regional stations. So far 15871
demonstrations on cotton production technology, 126 unit demonstrations on cotton IPM and 124 unit
demonstrations on cotton farm implements were conducted all over the country (Table 1). The FLDs are
conducted for transfer of improved cotton production technologies, IPM and demonstration of farm
implements. The main emphasis of the demonstrations includes enhancing the production of cotton in
low productivity areas / problematic areas. A list of beneficiaries and their plot numbers were selected in
consultation with local Agricultural Officers. A Bench mark survey was conducted before taking up the
Transfer of Technology Initiatives for Profitable and Sustainable Cotton Farming in IndiaAn Empirical Analysis 463
trial including information on the crops and cropping system of the area, inter cropping, the average
yields of cotton and the local practices adopted in terms of irrigation, use of fertilizer, plant protection
and information on the cost of production for the area as a whole. An impact analysis after the harvest
was done out in the light of reduction in insecticide use, reduction in cost of cultivation and awareness of
modern technology.
Impact of Front line Demonstration in Cotton
The demonstration of high yielding varieties and hybrids suited for various agro-climatic conditions,
Integrated Nutrient Management (INM) practices, IPM strategies, use of bio-fertilizers and bio-
pesticides, efficient water management techniques like drip irrigation and use of compatible intercrops
conducted in various FLDs through the cotton growing tracts of the country have helped the farmers
reduce pesticide input significantly and make cotton production more profitable. Over years yield
increased to the tune of 15 to 37 per cent over the check (Table 2) and cost benefit ratio was around
1:1.99 to 1:3.30. In total, FLDs were effective in increasing the yields, sharing the knowledge but
handicapped due to lack of professional execution and non-availability of latest technological
dissemination tools for ready transfer.
TABLE 2: IMPACT OF FRONT LINE DEMONSTRATIONS (FLD) CONDUCTED BY AICCIP SEED YIELD (KG/HA)
Period
FLD
FarmersPractices
Percentage Yield Increase Due to FLDs
1997-98 1326 965 37.40
1998-99 to 2001-02 1344 1071 25.60
2002-03 to 2005-06 1487 1265 17.57
2006-07 to 2009-10 2002 1737 15.28
Average 1540 1260 23.96
Farmers Field Schools (FFS) on Cotton
FFS is a participatory approach to adult education adopted by the Indian Government since the 1990s,
towards the achievement of an ecologically sound, profitable and socially sustainable small scale farming
(Mancini, 2006). In cotton, FFS was introduced as a component of TMC-MM-II in 2005-06 to provide
season long training to farmers, enable them to grow a healthy crop by adopting best management
practices. FFS is a non formal education method for cotton farmers with 20 sessions during the crop
season. Personnel from State Department of Agriculture who participated in the Season long Training
programme (ToF) on cotton were facilitators for the program. One FFS has a maximum of 30 farmers.
The main aim of the programme is to enable farmers to take quality decisions based on regular eco
system analysis. FFS also has a social goal to promote the empowerment of farmers by building human
and social capital (Gallagher, 2000). In FFS, farmers are no longer positioned as receivers of already
developed technological packages, but as field experts, who collaborate with the extension staff to find
solutions relevant to the local realities. FFS programmes emphasize farmers ownership, partnership and
group collaboration. So far more than 5000 farmers were trained under this programme.
Impact of FFS on Cotton Farming
Studies showed that the adoption of IPM by FFS had significantly reduced pesticide usage. It is expected
to mitigate the adverse effects of pesticides overuse on peoples health (Mancini et al., 2005),
biodiversity (Walter-Echols et al., 2005) and water quality (CSE, 2003). The strong correlation between
knowledge level and reduction in pesticide use proved that a skill-oriented, knowledge intensive and
hands-on education approach, as used during FFSs, is an efficient system to deliver the complex IPM
principles to farmers. The FFS approach focuses on the importance to judge the necessity for plant
protection interventions on the basis of actual field needs, which is essential to achieve a more
sustainable agriculture. Substituting pesticides with bio-control agents or other technologies such as
biotech cotton is unlikely to become a definitive solution to sustain agricultural productivity, if these new
technologies are not paired with educational programmes (Yang et al., 2005). FFSs are also effective in
disseminating the technical knowledge but market intelligence surveys for commercializing the
technologies and institutional arrangements for freeing indebtedness and risk coverage had never found a
significant place in these programs.
Contract Farming
Contract farming is another extension program implemented in this country for considerable close
cooperation between agricultural production and agribusiness and for vertical way of coordination.
Contract farming was introduced in cotton to reduce the load on central and state level procurement
system, to increase private sector investment and to promote processing and value addition. In this
approach, the farmer is contracted to plant the contractors cotton variety /hybrid on his land. He has to
harvest and deliver to the contractor, a quantum of kapas, based upon anticipated yield and contracted
average at a pre-agreed price. Towards these ends the contractor supplies the farmers selected inputs at
full cost. The Super Spinning mills, Tamil Nadu, the Appachi Cotton Mills, Tamil Nadu, CITI-Cotton
Development Research Association (CITI-CDRA) and Royal Classic Mills, Tamil Nadu are some of the
leading contract agencies in cotton contract farming.
Impact of Contract Farming
Cotton researchers are of the firm opinion that contract farming is the answer to the problems faced by
the cotton mill sector and the farming community. Studies showed that the contract farming system
effectively established a link between the consuming industry and the supplier of the produce. They
added that through contract farming, the integrated cotton crop management practices were propagated
much faster and spread of spurious seeds and other agro inputs were checked as the concerned industry
supplied them. Farmers also got a reasonable price for their produce. But in some cases either the
sponsor broke the contract by not giving the pre agreed price or the farmer sold the produce to others.
This program was also effective in increasing the yields and knowledge sharing. However, lack of
professional execution and non-availability of guidelines are bottlenecks for sustaining the contract.
LACUNAE OBSERVED IN THE CURRENT COTTON EXTENSION PROGRAMS
Analysis on all the cotton extension programs that are functioning in the country revealed that they were
effective in some aspects but handicapped due to inadequate interaction with the research efforts.
Although they were effective in increasing with yields, but handicapped due to lack of professional
execution and non-availability of latest technological dissemination tools for ready transfer. Many of
them excluded the novel extension methods viz., cyber extension, market led extension, Farmer-led
extension and environmental extension for a wider reach. Technology forecasting is another major area
where our cotton extension programs attempted very less initiatives. Many of the programs neglected the
inclusive growth and development. Market intelligence surveys for commercializing our technologies
and institutional arrangements for freeing indebtedness had never found a significant place in those
programs. Media utilization and efforts to organize the cotton growers were the other areas where our
cotton extension programs created a meager impact. This is one side of the countrys cotton sector in
terms of TOT initiatives.
CHANGING PERSPECTIVES AND EMERGING ISSUES IN COTTON
The countrys agricultural sector is facing serious challenges posed by the degradation of natural
resources viz., deforestation, salinity, soil erosion, water contamination and depletion. The agricultural
extension service in the country was ineffective at reversing these negative trends. Presently, extension
services are reorienting their development strategies towards supporting farmers empowerment. Among
the various changes in cotton sector, three major issues are discussed below in this article.
Changing Scenario
Combined efforts of millions of cotton farmers, talented scientists and technology providers, dynamic
seed production industry in the private sector and field extension agencies and favourable weather
conditions made India to be accountable for about 32% of the global cotton area and 21% of the global
cotton production, currently ranking second after China. Indias contribution to global cotton production
increased from 14% in 2002 to 20.5% in 2007. The production increased from a meager 2.3 M bales (170
kg lint/bale) in 1947-48 to a record production of 32.5 M bales during 2010-11. Though, India ranks
second in the world in cotton production after China, even its best productivity of 566 kg/ha, places it at
24th rank in the list of 80 cotton producing countries. Despite the good progress made by public and
private sector research and development, it is a matter of concern that productivity started to decline from
566 kg/ha in 2007 to 522 kg/ha in 2008, 486 kg/ha in 2009 and 475 kg/ha in 2010.
Changing Need
All along with the area, production and productivity of Indian cotton, the quality profile of Indian cotton
has also changed. Long staple cotton which constituted 38% prior to 2002, increased to an estimated 85%
of the total cotton produced in 2010, primarily because of the Bt cotton hybrids, most of which are of the
long staple category. However, the Confederation of Indian Textile Industries (CITI) estimates that in the
25.8 M bales utilization capacity, the current requirement of the Indian textile Industry is 37% long and
extra-long staple cotton, 53% medium staple and 10% short staple.
Changing Technology
Bt cotton is an attractive alternative and novel technology for controlling the bollworms in cotton. The
area under this Bt cotton increased phenomenally from a few thousand acres in 2002 to more than 1.01
million hectares in 2005 and it is estimated to be more than 9.5 million hectares in 2011. This tremendous
adoption rate of the technology could be due to the relative advantage of the technology and marketing
efforts of seed companies. Simultaneously, the area under public research bred varieties and hybrids
reduced significantly to less than 8% of the total cultivable area. The area under hybrid cotton increased
from 40%in 2002 to 92% in 2010. The area under G. hirsutum varieties was 33% in 2000 and now less
than 3%. Similarly, area under G. barbadense, G. arboreum and G. herbaceum which was 6.6%, 25%and
13% during 1995, declined to less than 7% in 2010 for all the three species combined. With intensive
selection pressure of Bt toxins used in more than 90% area of Bt crops, and less adoption of refugia and
declining area of inter crops in cotton cropping systems, development of bollworm resistance to the Bt
toxins is an emerging concern.
EXTENSION STRATEGIES TO FOSTER COTTON PRODUCTION IN THE COUNTRY

Fig. 1: Contemporary Synergetic Extension Model with Novel Extension Concepts for Profitable and Sustainable Cotton Farming in India
In order to survive in the current changing scenario of cotton sector, attaining excellent relevant and
effective extension strategies is inevitable for sustainable cotton production in the country. On the basis
of the analysis of issues, problems, opportunities of threats, a relevant and feasible model (Figure 1) has
been worked out for carrying out the extension activities. The future cotton extension programmes in the
country must include innovations like, cyber extension, market led extension, environmental extension
and farmer led extension. Along with improving the productivity and income of the existing cotton
growers, it must organize the clients, strengthen the linkage between various stakeholders in cotton and
provide genuine information for forthcoming policies and programmes.
Extension Innovations for Future Cotton TOT Programmes
Access to knowledge is the major factor most commonly cited by farmers as their major limitations in
cotton farming. The widespread availability and convergence of Information and Communication
Technologies (ICTs) in India in recent years have led to unprecedented capacity for dissemination of
knowledge and information to the rural population. Utilizing the ICTs for dissemination of knowledge is
inevitable for the future cotton extension programmes. The present cotton extension must address the
cotton market information along with the production technologies since price is another major constraint
in cotton production as cited by many of the farmers. Evolving a cotton extension programme that plays
a catalytic role for ushering market led agricultural extension, highly scalable, planned through bottom-
up process, and implemented through active involvement and collaboration of agricultural market
committees in India is the need of the hour.
Consequent indiscriminate use of inputs like fertilizers, plant protection chemicals, growth
promoters, herbicides and the recently introduced transgenics in cotton production system have started
causing alarms to natural resources viz., soil, water and environment. So, cotton extension has to reorient
itself to address these emerging concerns of natural resource degradation and environmental safety. In
totality, the changing scenario in cotton sector forces us to have a clear and more locally controlled and
managed extension approach. The approach should promote farmers and other rural people as the
principal agents of change in their communities. Such an approach is farmer led extension approach. In
this approach, farmers are not only key agents to access services provided by professional extension
specialists and researchers, but also make many of the management decisions and do much of the
extension works.
Concerns of Future Cotton Extension Programs
Even though India crowns the credits of owning first in cotton acreage and second in cotton production at
world level, its productivity level is very low. Disseminating novel cotton production technologies
evolved by cotton researchers through innovative extension approaches should be the top most agenda of
our future cotton extension programmes. With globalization there is a major change in the trade in
cotton. Selling to multinational companies needs larger quantities of seed cotton of a standardized
quality. This can be done by contract farming and by farmers organizations. Cotton extension has to
play a catalytic role in mobilization and organization of various cotton farmers groups depending upon
the local need and nature of enterprise. Building effective linkage among the cotton research (technology
development) system, extension (technology transfer) system and client (Technology user) system is a
strategic issue in management of cotton sector. Worldwide a great deal of attention is being given to
explore the intricacies involved in forging strong functional linkages among the various stakeholders of
cotton. The recently experimented approaches in cotton extension viz., IVLP, Strategic Research and
Extension Plan (SREP) in Agricultural Technology Management Agency (ATMA) are fairly successful
in addressing the issue of R-E-F linkage. However, given the diversity of conditions and contexts
prevailing in our cotton sector, linkage issue needs to be looked more as a major issue in cotton extension
programmes. In the context of rapidly changing global cotton scenario, cotton extension must emerge as
a major policy instrument to address the growing challenges in cotton sector. In addition it should
explore and provide information on the researchable problems to the cotton research system and
adoptable research solutions to various cotton stakeholders.
CONCLUSION
Worldwide, it is now widely recognized that agricultural extension needs to reform in ways that allow it
to fulfill a diverse set of objectives. The emerging paradigm of extension-plus ranges from better linking
of farmers to input and output markets, to reducing the vulnerability and enhancing voice of the rural
poor, development of micro-enterprises, poverty reduction and environmental conservation and
strengthening the farmers organizations (Sulaiman and Hall, 2004). Similarly, cotton extension needs to
embrace a broadened mandate as cotton farmers find themselves in an even more complex production
and market environment, with an expanding need for information and services. Hence, including the
modern extension innovations viz., cyber, market led, environmental and farmer-led extension
approaches in our future cotton extension programmes is inevitable.
REFERENCES
[1] CSE Report (2003) - Pesticide residues in bottled water, Centre for Science and Environment, New Delhi, India.
[2] Gallagher, K.D. (2000) - Community study programmes for integrated production and pest management: farmer field
schools. In: Human resources in agricultural and rural development. Food and Agriculture Organization (FAO, Rome,
Italy, pp. 6067.
[3] Mancini F., Van Bruggen A.H.C., Jiggins L.S.J., Ambatipudi A., Murphy, H., (2005) - Acute pesticide poisoning of cotton
growers in Andhra Pradesh, International journal of Occupational and Environmental Health; 11: 221232.
[4] Mancini, F. (2006) - Impact of Integrated Pest Management Farmer Field Schools on health, farming systems, the
environment, and livelihoods of cotton growers in Southern India. Doctoral theis, Biological Farming Systems Group,
Wageningen University, The Netherlands
[5] Sulaiman, R.V. and Hall Andy, (2004) - Towards Extension -Plus: Opportunities and Challenges. Policy Brief 17, NCAP,
ICAR, New Delhi.
[6] Walter-Echols G., Soomro M.H., (2005) - Impact of FFS training of the FAO-EU IPM Programme for cotton in Asia on the
Environment. In: Ooi P.A.C., Praneetvatakul S., Waibel H., Walter-Echols G., The impact of the FAO EU IPM Programme
for cotton in Asia, Pesticide Policy Project Publication Series No. 9, Hannover University.
[7] Yang P, Li K, Shi S, Xia J, Guo R, Li S, Wang L, (2005) - Impacts of transgenic Bt cotton and integrated pest management
education on smallholder cotton farmers, International J. Pest Management 51: 231244.
Post Harvest Processing

Enzyme/Zinc Chloride Pretreatment of Short-

Staple Cotton Fibres for Energy Reduction
during Nano-Fibrillation by Refining Process
N. Vigneshwaran
1*
, Vilas Karande
1
, G.B. Hadge
1
,
S.T. Mhaske
2
and A.K. Bharimalla
1

1
Nanotechnology Research Lab, Chemical and Biochemical Processing Division, Central Institute
for Research on Cotton Technology,
2
Department of Polymer and Surface Engineering, Institute of Chemical Technology,
Matunga, Mumbai400019, India
e-mail: nvw75@yahoo.com
AbstractCellulose is a renewable, biodegradable and the most abundant biopolymer available on the Earth. Natural
Cellulosic fibers are synthesized mainly in plants and cellulose constitutes 40-50% of wood, 80% of flax and 90% of
cotton fibers. Microfibrils are defined as the fibers of 0.1-1.0 m diameter, with high aspect ratio and nanofibrils are at
least one dimension in nanometer scale (1-100 nm). Nanofibrils of cellulose have potential use in high efficiency filters,
tissue scaffolds and as reinforcing agent in composites. In this work, we have processed short-staple cotton fibres
through refining process for the production of nanofibrils. The refining process, through shear force, pumps water into
the secondary layer and loosens the compactness of fibrillar structure by disrupting the hydrogen bonding. To enhance
the efficiency of refining process, pretreatments using enzyme / zinc chloride were developed to open up the primary
layer. Cellulase enzyme pretreatment hydrolyzed the surface molecules in cellulose while zinc chloride act as a swelling
agent thereby increasing the accessibility to secondary layer. Since the refining is a continuous process with very low
residence time, minimum of 30 passes were required for complete fibrillation resulting in huge energy consumption.
With pretreatments, the required number of passes for nano-fibrillation reduced drastically to fifteen only. Degree of
polymerization of cotton fibres (11188) significantly reduced to 8144 in the case of fibrillation without pretreatment
while it was 5147 and 6949 in the case of fibrillation with enzymatic and zinc chloride pretreatments, respectively.
Energy required (for 20 g of cotton fibres) for initial 5 passes for fibrillation without pretreatment and enzyme and
zinc chloride pretreatments are 1.346, 0.6764 and 0.8053 MJ, respectively. In subsequent passes, no significant
difference was noticed. The pretreated fibres showed more than 50% reduction in energy consumption during refining
process. The refining of the cotton fibers without pretreatment required at least 30 passes to achieve a fibril diameter of
400 nm whereas still smaller size (~100nm) could be achieved only in 15 passes using enzymatic / zinc chloride
pretreatments. Nanofibrils of cellulose thus produced are now being evaluated for their use as fillers in biopolymer
nanocomposites for use in food packaging.
INTRODUCTION
Cellulose is a renewable, biodegradable and most abundant biopolymer available in the biosphere (Lee et
al., 2009) and is produced in nature at an annual rate of 1011-1012 tons (Zhao et al., 2007). Cellulose is
the main constituent of the plants serving to maintain their structure. The properties of cellulose like good
tensile strength, low density, biodegradability etc. leads to rising research interest. Cellulose is the
structural material of the fibrous cells with high level of strength and stiffness per unit weight and has a
straight carbohydrate polymer chain consisting of -1-4 glucopyranose units and a degree of
polymerization of about 10,000 (Kamel, 2007). The molecules aggregate and are present in the form of
microfibrils (Hult et al., 2003). The hydroxyl (-OH) groups in the cellulose structure play a major role in
governing the reactivity and physical property of the cellulose. Natural Cellulosic fibers are synthesized
mainly in plants and cellulose constitutes 40-50% of wood, 80% of flax and 90% of cotton fiber. In
recent years, many researchers and manufacturers use natural fibers to replace man-made fibers as
reinforcement material and fillers to make environmentally safe products. Cellulose fibers can be
mechanically disintegrated to the structural nanoscale fibrils (Ahola et al., 2008).

78
The word fibril has been described by various researchers to describe relatively long and very thin
pieces of cellulosic material. Microfibrils are defined as the fibers of cellulose of 0.1-1m in diameter
(Chakraborty et al., 2005), with corresponding minimum length of 5-50 m and nanofibrils are at least
one dimension in nanometer scale (1-100 nm). The micro/nanofibrils isolated from the natural fibers have
much better mechanical properties (Cao and Tan, 2002). Therefore much attention has been given in the
last decade to study how to make micro/nano fibrils and how to combine them with the different
polymers to make composites.
In the present study, effect of enzymatic and zinc chloride pretreatments of cellulose on fibrillation of
cotton fibers by refining process have been studied. The purpose of these pretreatments is to loosen the
structure of the fiber either by reducing the secondary forces such as hydrogen bonding and van der
Waals forces or by swelling the fibers. Pretreatments will also be helpful in obtaining cellulose
nanofibrils in an energy efficient way. Nanofibrils from cotton fiber were prepared by top down approach
using the Lab-Disc Refiner. The refining is a pulping method in which the fibers are separated from the
matrix by means of mechanical forces. The main objective of the process is to loosen and separate the
fibers from the matrix, to break the fiber layer, to peel the fiber cell wall to some extent, and to fibrillate
fibers to the desired quality.
For enzyme pretreatment; about 20 g of cotton fibers were dispersed in 2 l of acetate buffer (pH 4.8)
along with 1% of cellulase enzyme and stirring was done using mechanical stirrer at 45C for 30 min.
After this, 10 ml of 1 M NaOH solution was added into the suspension to deactivate enzyme followed by
washing with distilled water. For zinc chloride pretreatment; 71.5% solution of zinc chloride in water was
prepared and allowed to cool as the exothermal reaction raised the temperature. After cooling, 20 g of
cotton fibers were added into it and stirred for 1 h at 35C. Finally, the fibres were washed four times
using distilled water to remove the zinc chloride completely. Cotton fibers before pretreatment and after
the enzymatic / zinc chloride pretreatments were subjected to the refining process for fibrillation in a lab
disc refiner. The output (fibrillated cotton fibers) from the fibrillation zone of refiner was collected in a
vessel and one such process was completed in 2 minutes and considered as one pass. The sample was
passed through the refiner up to 30 passes and characterization was done after every 5 passes. The
schematic of entire process is given in figure 1.

Fig. 1: Typical Process for Preparation of Cellulose Nanofibrils
The nanofibrils obtained by this process were analyzed by scanning electron microscopy, atomic
force microscopy, and their degree of polymerization (DP) was analyzed by viscometric method.
Simultaneously, the energy consumption was analyzed using the energy meter attached with the refiner.
Enzyme/Zinc Chloride Pretreatment of Short-Staple Cotton Fibres for Energy Reduction During Nano-Fibrillation 473
The product obtained by refining process was analyzed by scanning electron microscopy and the
obtained micrographs were subjected to image analysis as given in table 1. After the SEM analysis,
diameter of the cellulose fibril was measured at 15 different locations of various images and an average
diameter was reported.
TABLE1: DIAMETER OF THE FIBRILLATED COTTON (IN NM SD) FIBERS BEFORE PRETREATMENT AND AFTER ENZYMATIC, ZINC CHLORIDE PRETREATMENTS MEASURED FROM SEM IMAGES
No. of Passes Control Cotton Fibres Enzymatic Pretreated Cotton Fibres Zinc Chloride Pretreated Cotton Fibres
5 8090.53 3390.18 2720.014
10 7090.46 3340.20 2040.14
15 6170.50 1420.07 2060.22
20 5330.47 1460.14 1680.10
25 4730.38 1540.10 1700.12
30 4520.35 1520.09 1740.11
From table 1 it has been observed that the diameter of the fibrillated before pretreatment has been
reduced to 453 nm after 30 passes from an initial diameter of 21 m. It is also observed that the diameter
of the fibrillated cotton fibers after Enzymatic and Zinc Chloride pretreatments has been reduced down to
152 and 175 nm, respectively. The cotton fibers fibrillated even after 30 passes sample without any
pretreatment has an average diameter of ~ 453 nm whereas less than this was achieved after 5 passes
after enzymatic and zinc chloride pretreatments. Figure 2 shows the SEM micrographs of the fibrillated
cotton fibres (enzyme pretreated) after every 5 passes; a and b corresponds to initial fibre while the
figures from c to h represents stage after every 5 passes.

Fig. 2: SEM Micrographs of the Fibrillated Cotton Fibers After Enzymatic Pretreatment
Figure 3 shows the SEM micrographs of the fibrillated cotton fibres (zinc chloride pretreated) after
every 5 passes; a and b corresponds to initial fibre while the figures from c to h represents stage
after every 5 passes. Also, the swelling of fibrils due to zinc chloride treatment is clearly visible in the
SEM micrographs.

Fig. 3: SEM Micrographs of the Fibrillated Cotton Fibers After Zinc Chloride Pretreatment
The AFM analysis of the fibrillated fibrils after 30 passes was carried out using silicon tip in a
tapping mode. After the AFM analysis, diameter of the cellulose fibril was measured by image analysis
and reported in table 2.

Enzyme/Zinc Chloride Pretreatment of Short-Staple Cotton Fibres for Energy Reduction During Nano-Fibrillation 475
TABLE 2: DIAMETER OF THE FIBRILLATED COTTON FIBERS BEFORE PRETREATMENT AND AFTER ENZYMATIC, ZINC CHLORIDE PRETREATMENTS MEASURED FROM AFM IMAGES
Sample Avg. Diameter (nm)
Before Pretreatment 4320.11
Enzymatic Pretreatment 980.02
Zinc Chloride Pretreatment 1560.07
Degree of polymerization is defined as the number of repeating units present in a polymer.
Mechanical stresses generated due to shear, impact forces has great influence on chain scission and hence
on degree of polymerization. During the fibrillation process cotton fibers are subjected to the shearing
and impact forces therefore chain scission as well as fibrillation takes place which was resulted in
significant reduction of degree of polymerization and diameter of the cellulose fibril. Table 3 provides
the information about the DP of cotton fibres at different stages of fibrillation.
TABLE 3: DEGREE OF POLYMERIZATION OF THE FIBRILLATED COTTON FIBERS BEFORE PRETREATMENT AND AFTER ENZYMATIC/ ZINC CHLORIDE PRETREATMENTS
No. of Passes Control Cotton Fibres Enzymatic Pretreated Cotton Fibres Zinc Chloride Pretreated Cotton Fibres
Control 1118821.08 806223.41 1003221.92
5 918322.55 730723.99 874922.88
10 834240.19 674224.43 821023.30
15 822723.30 669024.48 812846.70
20 816123.33 552625.40 807823.41
25 811123.37 522031.43 792923.52
30 812823.37 516525.70 694924.27
From table 3, it is observed that the degree of polymerization was reduced to 5165 and 6949 after
enzymatic and zinc chloride pretreatments, respectively; while that of pristine fibres was 8128. The
degree of polymerization reduction may be attributed to the continuous exposure of cotton fibers to the
mechanical forces when subjected to the Lab disc refiner.
Energy consumption during the fibrillation process is a major prohibitive factor for carrying out
nanofibrillation. So, reduction in energy consumption will be a major boost for the production of
nanofibrils. Table 4 shows the energy requirement for fibrillation after different pretreatments.
TABLE 4: ENERGY REQUIRED FOR FIBRILLATION OF COTTON FIBERS BEFORE PRETREATMENT AND AFTER ENZYMATIC, ZINC CHLORIDE PRETREATMENTS
No of Passes Energy Required ( MJ)
Before Pretreatment After Enzymatic Pretreatment After Zinc Chloride Pretreatment
5 1.346 0.6764 0.8053
10 0.8425 0.6519 0.7139
15 0.747 0.6386 0.7448
20 0.805 0.6152 0.7038
25 0.8515 0.6091 0.6516
30 0.754 0.626 0.6368
Initially more energy was required for fibrillation and as the number of passes increased less energy
was required. Before pretreatment initial energy consumption was more and after the pretreatments, it
reduced significantly. After 5 passes, the energy required was almost 50% less for the enzyme pretreated
cotton fibres compared to that of pristine fibres.
CONCLUSION
The enzymatic and zinc chloride pretreatments have significant effect on the fibrillation of the cotton
fibers. The finest fibrils were obtained after fibrillation of the enzyme pretreated fibers and the diameter
of the fibril was reduced to ~98 nm from an initial value of ~ 21 m. The degree of polymerization has
been significantly decreased after the fibrillation of enzyme and zinc chloride pretreated fibres. It has also
observed that both enzymatic and zinc chloride pretreatments have significant effect on energy reduction
and among the pretreatments, enzymatic pretreatment performs better.

ACKNOWLEDGEMENT
Financial support for this work was provided by National Agricultural Innovation Project (NAIP), Indian
Council of Agricultural Research (ICAR) through a sub-project entitled Synthesis and characterization
nanocellulose and its applications biodegradable polymers composites to enhance their performance
properties (417101).
REFERENCES
[1] Ahola, S., Salmi, J., Johansson, L.S., Laine, J. and Osterberg, M. (2008) - Model films from native cellulose nanofibrils.
Preparation, swelling, and surface interactions. Biomacromolecules 9:1273-82.
[2] Cao, Y. and Tan, H. (2002) - Effects of cellulase on the modification of cellulose. Carbohydrate Research 337:1291-1296.
[3] Chakraborty, A., Sain, M. and Kortschot, M. (2005) - Cellulose microfibrils: A novel method of preparation using high
shear refining and cryocrushing. Holzforschung 59:102-107.
[4] Hult, E.L., Iversen, T. and Sugiyama, J. (2003) - Characterization of the supermolecular structure of cellulose in wood pulp
fibres. Cellulose 10:103-110.
[5] Kamel, S. (2007) - Nanotechnology and its applications in lignocellulosic composites, a mini review. eXPRESS Polymer
Letters 1:29.
[6] Lee, S.-Y., Mohan, D., Kang, I.-A., Doh, G.-H., Lee, S. and Han, S. (2009) - Nanocellulose reinforced PVA composite
films: Effects of acid treatment and filler loading. Fibers and Polymers 10:77-82.
[7] Zhao, H., Kwak, J.H., Conrad Zhang, Z., Brown, H.M., Arey, B.W. and Holladay, J.E. (2007) - Studying cellulose fiber
structure by SEM, XRD, NMR and acid hydrolysis. Carbohydrate Polymers 68:235-241.
Optimal Cotton Covered Jute, Nylon

and Metal Core Spun Yarns for Functional Textiles
Production and Characterization
S.K. Chattopadhyay
1
, A. Yadav
1
, V.V. Kadam
1
, Bindu V.
1
, D.L. Upadhye
1
,
V.D. Gotmare
2
and A.K. Jeengar
2

1
Central Institute for Research on Cotton Technology (ICAR), Matunga, Mumbai 400019 India
2
Veermata Jijabai Technological Institute, Matunga, Mumbai400019 India

AbstractIn the present exploration, cotton covered jute, nylon and stainless steel core spun yarns were developed
using a DREF-3000 Friction Spinning machine. In the core-sheath yarn, the cotton fibres were optimally used as
sheaths by just covering and twisting them around the core components. All the yarn samples were converted into
suitable technical fabrics and evaluated for functional characteristics.
Fabrics made from cotton-jute core yarn were used for making upholstery fabrics with aesthetic improvement in
feel, cover, color and design. Since the cost of jute is much lower than cotton, the product will be cost-effective
compared to 100% cotton fabric. The increased integrity and better mechanical properties of cotton-nylon core yarns
were found due to a positive linear relation between the core content and the yarn breaking tenacity further reinforced
by the wrapping sheath fibres. The interface produced by the cotton sheath helped anchoring the fabric with various
rubber matrices without the need of any chemical adhesive treatment. The composite thus produced resulted in an
optimum peel adhesion strength of 3.11 kN/m between the rubber and the fabric, and is suitable for application in
beltings. The cotton-stainless steel core yarn was used to prepare flexible shielding fabrics for protection from harmful
electromagnetic waves. It was found that the shielding efficiency of the fabrics could be tailored by the fabric design,
and the developed shields could mitigate electromagnetic waves to the extent of 55%.
INTRODUCTION
Blending natural and man-made staple fibres is well-known, and in vogue in the yarn spinning industry.
The objectives of blending of two or more staple fibres are to improve functional and aesthetic quality of
textiles, processing performance and economy of production. However, blending has a major limitation -
continuous man-made filament cannot be processed with staple fibres. Therefore, blending techniques are
mainly restricted to producing apparel textiles rather than technical textiles. Technical textiles are used
for their technical performance and functional properties mostly by other user industry. It has a vast
potential to grow and excel in a developing economy like India. It has shown a healthy growth of 18%
from 2001 to 2008. The current technical textile consumption in India is estimated at Rs. 41,756 crores,
and is expected to grow at the rate of 6-8% (without the Government intervention), but 12-15% with
stimulus from the Government (Ministry of Textiles, 2010). Therefore, the thrust is on to develop high-
tech and hybrid yarns for application in the manufacture of technical textiles.
The Friction spinning, a fairly new technology in the domain of yarn manufacture has asserted itself
as one of the most important system in technical yarn development. In the friction spinning, the fibers
from the sliver after individualization are deposited by an air current into the gap between the cylinders
(Brockmans 1984; Lord and Rust 1990, 1991) rotating in the same direction, wherein either one or both
the friction drums are perforated having an inside suction to restrain the deposited fibers, and consolidate
them (Lord et al. 1987; Karnon 1986; Salhotra et al. 1995).
The DREF friction spinning technology also produces core-spun multicomponent yarns by using
filaments covered by staple fibre as a sheath (Fehrer, 1987, 1989). The main aim of using core-spun
yarns is to take the advantage of different properties of core and sheath components. While the core
improves the yarn strength and machine productivity, the sheath fibers add to physical properties of the
surface and impart softness, bulkiness and an appearance similar to staple yarns.
79
PRESENT STUDY
A DREF-3000 friction spinning machine has been used to produce core-spun yarns. In this machine, first
the core part is false-twisted by the torque generated by the rotating friction drums, and then staple fibres
are deposited on it to make a sheath that covers the core. On emerging from the twisting zone, the false-
twist of the core is removed and only sheath fibres get real twisted in the reverse direction. The resultant
yarns have the character of an almost twistless core and helically wound sheath fibres of varying helix
angles. In the present study, the core and sheath materials have been chosen looking into predetermined
end-uses. Three different core yarns were produced, viz: cotton-jute, cotton-nylon and cotton-stainless
steel. The staple cotton fibre used as sheath just about covers the core part. Such optimal use of cotton as
sheath in a core yarn will limit the use of cotton in the yarn. This implies that, if the core fibre is cheaper
than cotton, like in cotton-jute core yarn, the resultant product will be cost-effective compared to those
made from 100% cotton yarns. All the yarn samples so developed in the study were converted into
suitable technical fabrics and evaluated for their functional properties. A fabric made from cotton-jute
core yarn was used for making dyed upholstery fabrics. The cotton-nylon core yarn was used for
preparing matrices for rubber composites, the cotton-stainless steel core yarn was converted into flexible
shieldings for protection against harmful electromagnetic waves.
On the DREF-3000 machine, the spinning and carding drum speeds were kept at 3000 and 5000 rpm
respectively. The yarn delivery speed was 150 m/min for jute and nylon core yarns, but 120 m/min for
stainless steel core yarn.
Cotton-jute Core Spun Yarn and Fabric
The cotton used for the sheath of the yarn had 2.5% span length of 32.5 mm, uniformity ratio of 0.54,
micronaire of 3.5, bundle strength (3.2 mm gauge) of 25.5 g/tex and breaking elongation of 5%. It was
converted into a second drawn sliver of 0.18 hank (3.3 ktex), and two of the same were used
simultaneously on the DREF machine. The jute yarn was of 4.8 pounds (166 tex), and made from jute
fibres with fineness of 1.89 tex and bundle strength, 26.6 g/tex. A 2s Ne (295.3 tex) DREF-yarn was spun
with a core to sheath ratio of 67:33. The same was converted into an upholstery fabric of plain weave
with 12 ends and picks per inch (4.7 per cm), and of fabric weight of 298 g/m
2
(GSM). The grey fabric
was subsequently scoured, bleached, dyed with reactive dyes and softened by exhaust finishing method.
Cotton-nylon Core Spun Yarn and Fabric
For production of cotton-nylon core yarn, a nylon 6 multifilament of 420 Denier, 48 monofilaments and 0
twist, with a breaking strength of 3.08 kgf and hot air shrinkage of 4.8% (at 180C for 15 min) was used
in the core. Four cotton slivers of 0.12 hank (4.9 ktex) each were fed to the machine to supply the sheath
fibres. DREF-yarns of 1.5s and 2s Ne (393.7 and 295.3 tex) were prepared by using different core to
sheath ratio, viz.,12:88, 24:76, 36:64 and 48:52. This was to find out the effect of cotton sheath on the
mechanical properties of the DREF-yarn. Another yarn sample of 1s

Ne (590.6 tex) with core to sheath
ratio of 77:23 was spun in enough quantity to convert it into a fabric of 480 GSM with mockleno weave
on a sample loom. The fabric was used as matrices in single and double layers with different rubber
compositions to make textile-rubber composite samples of 4.5 and 7 mm thickness respectively.
Cotton-stainless Steel Core Spun Yarn and Fabric
A steel wire of 0.145 mm diameter along with 840 denier polypropylene multifilament consisting of 84
monofilaments was used in the yarn core. Cotton fibres separated from four second drawn sliver with a
hank of 0.25 (2.3 ktex) each were used to supply fibres for the sheath to cover the core, and produce a
core-yarn of 3s Ne (196 tex). The core to sheath ratio was 40:60. The yarn was used as a weft in a warp
sett prepared from a 20s Ne (29.5 tex) normal cotton yarn. Fabric samples of plain, 2/2 twill and
honeycomb weave constructions were woven with a cover factor of 20 to 22. Repeat samples were also
produced for confirmation of the results.

Measureme
The yarn
MVIG 20
nylon yar
were mea
conditions
2259-96.
respective
Measureme
The tensil
5 cm rave
tensile str
The trapez
ASTM D
rigidity.
Measureme
The peel s
D-1876. A
mm/min.
UTM. As
specimen
Measureme
Any barri
as a shie
quantitativ
follows:
EMSE
EMSE
and P
in
is
Optima
ent of Yarn Prop
diameter an
005) and a S
rn was meas
asured on a
s according t
The testing
ely.
ent of Fabric Pr
le strength o
elled strips.
rength with
zoid tear stre
4833-88 res
ent of Fabric to
strength, i.e.
An Instron U
The strap, w
the machine
was recorde
ent of Electro-M
er between a
eld. The El
vely how mu
E=10 log
10
(P
E in the unit
the input pow
l Cotton Covere
operties
nd the surfac
Scanning Ele
ured accordi
Star Univer
to ASTM D
speed and g
roperties
f the fabric w
Since the fa
wide-width
ength and th
spectively. T
o Rubber Adhes
, the fabric t
Universal Te
with a test sp
e started the
ed.
Magnetic Shield
an emitter an
lectro-Magne
uch an electr
P
out
/P
in
)
of decibel (d
wer (watts).
ed Jute, Nylon a
ce of DREF-
ectron Micro
ing to ASTM
rsal Tensile
2256-02, an
gauge length
was measure
fabrics were
method was
e index punc
The IS 6490-7
sion Strength
to rubber adh
esting Machi
ecimen size
test piece re
ding Effectivene
nd a receiver
etic Shieldin
romagnetic fi
Fig. 1: E
dB) is expres

and Metal Core
-yarns were
oscope (SEM
M D 1059-9
Testing Ma
nd the shrinka
h of testing
ed according
to be used
s also measu
cture strength
71 standard t
hesion (bond
ine was cali
of (25 X 10
eached T sh
ess (EMSE)
r that decreas
ng Effective
ield is attenu
EMSE Measuremen
ssed as a pow
Spun Yarns for
studied by
M) (Model: P
7 test metho
achine (UTM
age percenta
on the UTM
to ASTM D
for technica
ured accordin
h were meas
test method
d) strength w
ibrated and t
00 mm) was
hape, and the
ses the streng
eness (EMS
uated because
nt Set-up
wer ratio, wh
Functional Tex
using an Im
Philips XL-3
od. The tens
M) following
age of nylon
M were 300
D 5035-95 w
al application
ng to ASTM
sured as per
was used to
was determine
the crosshea
fixed betwee
e force (kN/m
gth of an ele
SE) is a m
e of the barri

here P
out
is th
xtiles
mage Analyze
30). The den
sile propertie
g the standa
according to
mm/min and
ith a strip siz
n rather than
M D 4595-95
ASTM D 45
determine th
ed according
ad speed was
en the two g
m) required
ctromagnetic
measure that
ier, and is ex
he output pow
479
er (Model:
nier of the
es of yarns
ard testing
o ASTM D
d 500 mm
ze of 7.5 x
n apparels,
5 standard.
533-91 and
he flexural
g to ASTM
s set at 50
grips of the
to peel the
c field acts
expresses
xpressed as
(1)
wer (watts)
An RF Network Analyzer (VNA) with a 50 impedence, was used to generate EM waves in the
frequency range of 300 KHZ to 3 GHZ and passed through an anechoie circular co-axial fabric holder
fabricated in accordance to ASTM standard D4935-99 (ASTM, 1999). It simulates the far field shielding
behaviour. The measuring set-up is shown in Figure 1. The purpose of the test is to measure
quantitatively the insertion loss that results from introduction of the fabric as a shield, when the
electromagnetic plane wave is applied as normal to the surface of the material. The power from the
transmitter is coupled to a receiver, first without any barrier to set up a reference level, and then with the
fabric shield introduced using a two-port network (Fig. 2). The ratio of the two powers gives the insertion
loss (IL) from which EMSE was calculated using formula (1).

Fig. 2: Two-Port Network
Cotton-jute Core Spun Yarn and Fabric
The properties of the parent jute yarn vis--vis the cotton jute core yarn are presented in Table 1. The
DREF-yarn diameter was found to increase by 64% compared to that of the parent jute yarn. The core to
sheath ratio of 67:33 was verified by pulling out the sheath fibres from the core. The tenacity of DREF
spun core yarn was found to reduce by 49.6%, while the breaking elongation increased by 17% compared
to the parent 4.8 lbs jute yarn. This is due to unopening of the true twist in the parent jute yarn by the
false-twist during friction spinning. Once out of the twisting zone, the true twist tends to return to its
original state, but get partially blocked by the just laid sheath fibres. The observation of lower tenacity in
cotton-jute core DREF-yarns agrees with our earlier finding (Chattopadhyay et al, 2009). The tenacity-
elongation curves of the parent jute and cotton-jute core yarns from multiple spinning are shown in
Figure 3.
TABLE 1: YARN PROPERTIES OF JUTE AND COTTON-JUTE YARN
Sr. No. Test Parameter Jute yarn Cottonjute Core Yarn
1 Yarn diameter (mm) 0.7 1.1
2 Yarn size (tex) 15.9 32.4
3 Breaking load (kgf) 1.51 (22.9) 1.52 (20.8)
4 Breaking elongation (%) 1.41 (16.5) 1.65 (18.1)
5 Tenacity (gm/tex) 94.9 47.0
(Figures in parenthesis indicate CV %.)

The f
reactive d
bleached
agent, 3%
The mater
Orange-3R
room tem
was unifo
the jute fa
long stora
cotton, it h
Cotton-nyl
Multifil
Core
(Den
42
84
126
168
From the
strength, t
denier (Ta
yarns, nam
significan
Multifil
Core
(Den
42
84
126
168
Optima
fabric made
dyes using o
at a tempera
% hydrogen p
rial to liquor
R and Red-5
mperature for
orm. Further,
abric. The in
age could be
has little or n
on Core Spun
lament
Size
ier)
C
Str
(K
0
0
60
80 1
analysis of
tenacity and
able 2 and 3
mely 1.5s a
nce of 0.01%
lament
Size
ier)
C
Str
(K
0
0
60
80 1
l Cotton Covere
Fig. 3
from cotton
optimum reci
ature of 98
peroxide, 0.3%
r ratio was 1
5B) with a 2%
20 minutes
, the lightfas
nherent probl
e masked by
no chance to
n Yarn and Fab
Core
rength
Kgf)
Core
Elo
2.6
5.7
8.8
10.7
data on the
work of rup
). The relatio
and 2s Ne w
(P<0.001) (F
Core
rength
Kgf)
Core
Elo
2.6
5.7
8.8
10.7
ed Jute, Nylon a
3: Tenacity-elonga
n-jute core y
ipes as deter
C for 45 m
% peroxide s
1:10. Next, th
% shade dep
with materia
stness for co
lem associate
y the cotton
be exposed
bric
TABLE 2: TENSILE PR
e Breaking
ongation
(%)
C
Sh
R
18.7 1
18.9 2
18.5 3
20.0 4
e mechanical
pture, it can b
onship betw
was found to
Fig. 4).
TABLE 3: TENSILE PR
e Breaking
ongation
(%)
C
Sh
R
18.7 1
18.9 2
18.5 3
20.0 4
and Metal Core
ation Curve for Jut
yarn was sc
rmined by i
inutes in a b
stabilizer and
he fabric wa
pth. The fabr
al to liquor r
otton-jute fab
ed with jute
n sheath. As
to direct sun
ROPERTIES OF DREF-3
Core:
heath
Ratio
Ya
Brea
Stre
(k
12:88 5
24:76 7
36:64 10
48:52 13
l properties
be said that t
een the nylo
o be linear w
ROPERTIES OF DREF-3
Core:
heath
Ratio
Ya
Brea
Stre
(k
12:88 4
24:76 7
36:64 9
48:52 11
Spun Yarns for
te and Cotton-jute
coured, bleac
initial trials.
bath with 0.
d 2.5% caust
as dyed with
ric was final
ratio of 1:10
bric was foun
fibres e.g., c
jute yarn re
nlight.
000 YARNS (1.5SNE)
arn
aking
ength
kgf)
Yarn
Elong
5.5
7.9
0.6
3.1
of friction s
these parame
on multifilam
with high c
3000 YARNS (2S NE)
arn
aking
ength
kgf)
Yarn
Elong
4.4
7.2
9.8
1.9
Functional Tex
e Yarn
ched and su
First, the fa
.75% scourin
tic soda on th
h reactive dy
ly softened b
. It has been
nd qualitativ
colour fading
emained in t
)
n Breaking
gation (%) T
19.0
23.6
24.0
25.3
spun yarns,
eters increas
ment denier a
orrelation-co
)
n Breaking
gation (%) T
19.5
22.1
25.0
24.1
xtiles

ubsequently
abric was sc
ng agent, 1%
he weight of
es (Turquois
by exhaust f
n found that
vely better th
g and yellow
the core cov
Yarn
Tenacity
(g/tex)
W
R
(g.m
14.0
20.0
27.0
35.6
such as yarn
e with increa
and the stren
oefficient of
Yarn
Tenacity
(g/tex)
W
R
(g.m
14.9
24.2
33.1
40.6
481
dyed with
coured and
% desizing
f the fabric.
se blue-21,
finishing at
the dyeing
han that of
wing during
vered with
Work of
Rupture
mm) X 105
2.6
4.6
6.4
8.7
n breaking
ase in core
ngth of two
f 0.99 at a
Work of
Rupture
mm) X 105
1.7
3.9
6.1
7.2
482
Since
filament (
dependent
by 11-110
wrapped
providing
loading of
parent mu
is found
catastroph
yarns spun

, the strength
(plotted in th
t on the stren
0% compare
crosswise o
transverse f
f the yarn. T
ultifilament a
spasmodic,
hic in nature
n on the frict
World Cot
Fig. 4: Re
h curves of
he same figu
ngth of core
ed to the pa
over the long
force that bi
he same cou
as well as the
and therefo
with nil or r
tion spinning
(a)
Fig. 5.
tton Research C
elation between Ya
DREF-yarns
ure), it can b
filament yar
arent nylon
gitudinally l
inds the indi
uld be corrob
e DREF-yarn
ore, associat
reduced fibre
g machine.
a: Nylon Core Mul
Conference on T
arn Core Size (Den
s follow the
be inferred th
rn. However,
filament. It
laid multifil
ividual filam
borated from
ns (Fig. 6). W
ted with fib
e slippage. Th

ltifilament b) Cotto
Technologies fo
nier) and Breaking
similar linea
he strength o
DREF-yarn
t implies the
aments [com
ments and arr
the load ver
Whereas the b
bre slippage,
his shows im
on-nylon Core Spu
or Prosperity
g Strength
ar trend as th
of DREF-spu
n strength is a
e sheath cot
mpare (a) an
rest their slip
rsus displacem
breakage for
, the breaks
mproved stru
(b)
un Yarn

hat of the pa
un core yarn
also found to
tton fibres,
nd (b) in Fi
ppage during
ment plots fo
the multifila
s for DREF
uctural integr
arent core-
n is mainly
o be higher
which are
ig. 5], are
g the axial
or both the
ament yarn
-yarns are
ity of such

The b
which is a
the core (
found to i
The 1
breaking
mockleno
the fabric
The m
reinforcem
for examp
fabric and

Optima
breaking elo
also assigned
(Fig. 7). Fina
ncrease with
1s Ne (590.6
strength of
o fabric with
was 480.
mechanical p
ment for belt
ple, natural,
d various typ
l Cotton Covere
(a)
Fig. 6. Load vs
ngation of D
d to increase
ally, the wor
h the core con
Fig. 7: Rel
6 tex) produc
3.1 kgf, elo
13 ends and
properties of
ting fabric. F
polychlorop
es of rubbers
ed Jute, Nylon a
s. Displacement P
DREF-yarn w
ed integrity o
rk of rupture
ntent at a sig
lation between Yar
ced cotton-ny
ongation of
d 20 picks pe
f the fabric
For the same
prene and eth
s for both wa
and Metal Core

lot of (a) Parent M
was found h
of DREF-yar
e, that is, the
gnificance of
rn Core Size (Den
ylon core ya
22.5% and
er inch (5.1 X
(Table 4) w
e, the fabric
hylene-propy
arp and weft
Spun Yarns for
Multifilament and (
higher than
rn caused by
e energy to b
f 0.01% (p<0
ier) and Breaking
arn with core
tenacity of
X 7.9 per cm
were found s
sample was
ylene (EPDM
ways has be
Functional Tex
(b)
b) DREF-yarns
the parent m
y wrapping o
break a core
0.001) (Table
Elongation
e to sheath r
39.4 g/tex.
m) on a samp
atisfactory t
s composited
M). The peel
een presented
xtiles
multifilamen
of sheath fib
spun DREF
e 2 & 3).

ratio of 77:2
It was wov
ple loom. Th
to use the su
d with variou
l strength be
d in Table 5.
483

nt by 20%,
res around
F-yarn was
23, had the
ven into a
he GSM of
ubstrate as
us rubbers,
etween the

TABLE 4: FABRIC PROPERTIES OF COTTON-NYLON CORE SPUN YARN
Sr. No. Test Parameter Value
1 Tensile strength (Ravelled strip)
a. Breaking Strength (N) Warp 1931
Weft 1905
b. Elongation at break (%) Warp 36.1
Weft 37.3
2 Tensile Strength (Wide width)
a. Breaking Strength in kN/m Warp 39.0
Weft 36.4
b. Elongation at break (%) Warp 37.2
Weft 37.8
3 Tear Strength (Trapezoid Tear)
Peak Mean Load (N) Warp 480
Weft 430
4 Index Puncture strength Resistance (N) 620
5 Flexural rigidity (mg.cm) Warp way 15471
Weft way 164
TABLE 5: PEEL STRENGTH OF FABRICS MADE FROM CORE SPUN DREF YARN IN RUBBER COMPOSITE
Sample No. Type of Rubber Peel Strength (kN/m) Average Peel Strength
(kN/m) Warp Weft
1 Natural rubber (NR) 2.62 3.11 2.87
2 Polychloroprene 2.32 2.16 2.24
3 Ethylene- propylene (EPDM) 2.62 3.11 2.87
Such cotton covered nylon fabric does not require extra chemical adhesive treatment, such as
Resorcinol Formaldehyde Latex (RFL), which otherwise would have been necessary if the fabric was
made with only nylon fibre. Apart from general chemical affinity between rubber and cotton, it appears
the cotton fibres has provided more number of binding points for the rubber because of their fibrous and
irregular surface. The yarn surface was viewed under scanning electron microscope (Fig. 8). The
integrated yarn made from many nylon monofilaments wrapped tightly by cotton fibres with high surface
irregularity was clearly visible in the cross-sectional view (Fig. 8a). The peel strength is an important
parameter that decides bonding between the rubber and the textile. It is the ability of a material to resist
forces that can pull it apart and separate into two parts. It was found that resultant fabric could yield an
average peel bond strength of 2.87 kN/m and seems suitable for application in beltings. The cotton
covered nylon core fabric yielded good peel strength with all the rubber matrices viz., natural rubber,
polycholoroprene and ethylene-propylene (EPDM).

(a) (b)
Fig. 8. (a) Cross-sectional and (b) Longitudinal SEM Image of Cotton-nylon Core Yarn

EMSE of Fa
Figure 9
incident f
EMSE va
correspon
It can
increase in
to be high
opaque co
shields c
electroma
Weave Ty
Plain
Plain
2/2 Twill
2/2 Twill
Honeycom
Honeycom
CONCLUSIO
We have
spun usin
observatio
U
an
as
m
T
de
te
In
ob
Optima
abrics Made fr
shows the t
frequency of
alue optimiz
nding frequen
n be seen the
n stainless st
hest for the p
ompared to t
could mitiga
agnetic wave
ype Thread
mb
mb
ON
developed a
ng a DREF
ons are made
Upholstery fa
nd could be
ssociated wit
masked with t
he loss of y
etwisting of
ends to return
n cotton cove
bserved. The
l Cotton Covere
rom Stainless
typical varia
f the electrom
e at a partic
ncy for the va
Fig. 9:
optimum EM
teel content p
plain weave f
twill and ho
ate Very-Hi
s by 34-55%
d Density (Wa
28 x
28 x
28 x
28 x
28 x
28 x
and character
F-3000 Frict
e from the pr
abric develop
coloured wi
th jute fibre
the covering
yarn tenacity
the jute yar
n once out of
ered nylon c
e increased in
ed Jute, Nylon a
s Steel Core Y
ation in EMS
magnetic wav
cular freque
arious fabric
EMSE Plot at Diffe
MSE increas
per unit area
followed by
oneycomb, th
igh Frequen
%. Further res
TABLE 6: MAXIMUM E
arp x Weft) Per
30 (11 x 11.8)
32 (11 x 12.6)
30 (11 x 11.8)
34 (11 x 13.4)
34 (11 x 13.4)
x 38 (11x 15)
rised optima
tion spinnin
resent explor
ped from cot
th reactive d
s like colour
of cotton sh
y with gain
rn by the fal
f the torque z
core yarn, sin
ntegrity and
and Metal Core
arn
SE of a wov
ves, ranging
ncy. Table
cs developed
erent Frequencies
es with incre
of the fabric
twill and ho
hey offer bet
ncy (VHF)
search to des
EMSE VALUES OF VAR
r Inch (Per cm
al cotton cov
ng machine
ration:
tton covered
dyes normall
r fading and
heath fibres.
in elongatio
lse-twist gen
zone, it get p
nce the nylon
better mecha
Spun Yarns for
ven fabric s
from 300 kH
6 presents t
in the presen
of Electromagnet
easing pick d
c. Further, th
oneycomb (T
tter shielding
to Ultra-Hi
ign improve
RIOUS FABRIC SAMPLES
m) Cover Fa
19.7
20.6
19.7
21.5
21.5
23.3
vered jute, ny
and their
d jute core ya
ly like all co
d yellowing d
n in cotton-j
nerated in fri
partially block
n is zero twi
anical proper
Functional Tex
shield when
Hz to 1.6 GH
the optimum
nt study.
ic Wave
density which
e optimum E
Table 6). As
g efficiency.
igh Frequen
d textile shie
S
actor Freque
6.15
6.15
6.38
6.38
7.05
7.05
ylon and sta
technical fa
arn is of imp
otton fabrics.
during prolo
-jute core ya
iction spinni
ked by the ju
sted, the abo
rties of cotto
xtiles
subjected to
Hz. It can be
m EMSE val

h is mainly a
EMSE value
plain weave
The develo
ncy (UHF)
eld is in prog
ency (HZ) E
5 x 10
8

5 x 10
8

8 x 10
8

8 x 10
8

5 x 10
8

5 x 10
8

ainless steel
abrics. The
proved feel
The inheren
onged storage
arn is attribu
ing. Though
ust laid sheat
ove phenome
on-nylon core
485
o different
e seen that
lue and its
assigned to
was found
s are more
oped fabric
bands of
gress.
EMSE (dB)
50
55
42
52
34
42
core yarns
following
and cover,
nt problem
e could be
uted to the
true twist
th fibres.
enon is not
e yarns are
due to the core content contributing positively, further aided by the reinforcement of surface
sheath fibres that resist fibre slippage during the yarn loading.
The mockleno woven fabric made from cotton covered nylon core yarn could be bonded with
different rubbers without the need of any extra chemical adhesive treatment.
The peel adhesion strength of the fabric with the rubber was found to be 3.11 KN/m, which is
enough to be used as composites for belting purposes.
It is found the Electro-Magnetic Shielding Efficiency (EMSE) value for a fabric shield typically
optimises at a particular frequency of the electromagnetic wave.
EMSE increases with increasing pick density of the fabric attributed mainly because of increase
in the stainless steel content. It is also the highest for the plain weave, which is more opaque than
similarly woven twill and honeycomb fabric shields.
The developed fabric shields could mitigate Very-High Frequency (VHF) to Ultra-High
Frequency (UHF) bands of electromagnetic waves up to 55% effectiveness.
REFERENCES
[1] ASTM D 499599 (1999)Standard Test Method for Measuring the Electromagnetic Shielding Effectiveness of Plane
Materials.
[2] Brockmans, K.J. (1984) Friction spinning analyzedInternational Textile Bulletin, Yarn forming 2: 523.
[3] Chattopadhyay, S.K., Dey S.K., and Sreenivasan S. (2009)Composite yarns from natural fibres for production of technical
textiles International Conference on Emerging Trends in Production Processing and Utilization of Natural Fibres 2:
338346.
[4] Fehrer, E. (1987) Friction spinning: The state of the art Textile Month, 9: 115116.
[5] Fehrer, E. (1989) Latest DREF: medium counts at 300 m/min Textile Horizons, 2(6): 2021.
[6] Karnon I. (1986) Friction spinning the masterspinner , Textile Month, 3: 3437.
[7] Lord, P.R., & Rust, J.P. (1991) - Fiber assembly in friction spinning Journal of the Textile Institute, 82: 465478.
[8] Lord, P.R., & Rust, J.P (1991) Variations in yarn properties caused by a series of design changes in a friction spinning
machine Textile Research Journal,. 61(11): 645.
[9] Lord, P.R. & Rust, J.P. (1990) The surface of the tail in open-end friction spinning Journal of the Textile Institute,
81(4):, 100103.
[10] Lord, P.R., Joo, C.W., & Asbizaki, T. (1987)The mechanics of friction spinning Journal of the Textile Institute,
78: 234254.
[11] Ministry of Textile (Govt. of India) (2010) National Fibre Policy 201011.
[12] Salhotra, K.R., Behra, B.K. and Joshi, V.K. (1995) Friction spinning: pros and cons The Indian Texitle Journal,
105(4): 8690.
The Cotton Length Analysis

using the Lengthcontrol
Iwona Frydrych
1
, Anna Pabich
1
and Jerzy Andrysiak
2
1
Technical University of Lodz, Faculty of Material Technologies and Textile Design,
Department of Clothing Technology and Textronics, Lodz, Poland
2
Textile Research Institute, Lodz, Poland
E-mail: iwona.frydrych@p.lodz.pl.com

AbstractLengthcontrol is a fiber length measuring device produced by Trtzschler, which can quickly give
information about the distribution of fiber length, the short fiber content (SFC) and the content of fiber hooks. It can
also give some suggestions of changing spinning machine's parameters to optimize its work. By using this
measurement system, it is possible to measure all kinds of fibers, which length is not higher than 63 mm and which are
formed in a sliver from the carding, drawing or combing processes. It can be successfully used in the inter-operational
control. Just after 10 minutes (the measurement duration), it gives necessary data to assess the quality of the fiber
sliver and to optimize machine work. The possibility of quick adapting parameters of the machine work to the actually
processing fiber length is especially important in the case of cotton fibers, which are characterized by a big variability
of length characteristics. In this work, there are compared the length results of measurements made on the
Lengthcontrol with results from the AFIS. Such a comparison provides us information about the quality of the
measurements on the Lengthcontrol.
Keywords: Lengthcontrol, cotton, fiber length, inter-operational control
INTRODUCTION
Cotton, as a natural fiber, is characterised by a significant variability of parameters, especially length
parameters. The reason for this is the fact that main fiber properties (length, strength, maturity) are
dependent on many factors such as soil and weather conditions, irrigation method, harvesting and ginning
method etc. (Frydrych I., 2005; Wakelyn P., Chaudry R, 2010). This natural differentiation causes several
problems such as adjustment machine working parameters during the processing. Apart from the well
known cotton parameter measurement systems such as HVI and AFIS, there is also a new one -
Lengthcontrol (TC-LCT) proposed by Trtzschler (Trtzschler GmbH&Co, 2006). Below results
obtained from the Lengthcontrol device will be analyzed in a comparison with results of the same cotton
fibers from the AFIS. The comparison was done with the AFIS system results, because both of them
require the sample in the sliver form.
Advanced Fiber Information System (AFIS) is a well known integrated system, which is used to
obtain the fiber characteristics on the basis of measurement of single fibers separated in the air stream
(Frydrych I., 2005). The AFIS-L test provides detailed information regarding important fiber properties
including several length parameters (mean length by number l
m
(n) and by weight l
m
(w), upper quartile
length by number, 2.5 % etc.) (Frydrych I., 2005; Wakelyn P., Chaudry R, 2010). With the AFIS use, it is
possible to determine the average sample length properties, and also their variation. This system is quick,
purpose oriented and reproducible measuring the raw material in the sliver form in the majority of
process stages in the spinning mill. It is thus possible, based on the forecasted supervisory measures and
early warning information, to practically eliminate subsequent complaints with respect to the finished
product (Frydrych I., Matusiak M., 2002).
LENGTHCONTROL
Lengthcontrol (TC-LCT) is a new measurement device for the use in the spinning mill. The scheme of
Lengthcontrol is presented in the Fig. 1, whereas its photo in Fig.2.
80
488
It is u
measurem
operator,
just after
by the sliv
the results
The m
fiber tuft i
the distan
LENGTHCO
The result
I grou
Upper
LCT-L
using friendly
ment of all fib
who doesn't
10 minutes.
ver just from
s are not influ
measurement
is carefully c
ce to the clam
NTROL PARAM
ts from the T
up - basic tes
r length [mm
Length [mm]
World Cot
F
y and it does
ber slivers up
need to hav
There is no
m the card ca
uenced by th
t principle is
combed at bo
mp, so for th
METERS
TC-LCT mea
t results (Bre
m] the value
] the value
tton Research C
Fig.1: A Scheme of
n't have any
p to 63 mm (
ve any specia
time-consum
an, draw fram
he calibration
Fig. 2:
based on a m
oth sides. An
his reason its
asurement ca
euer J., 2005
e is the uppe
is roughly e
Conference on T
the Lengthcontro
demands on
(but especial
al qualificatio
ming sample
me or combin
n or settings.
A Photo of Length
modified fibr
n optical sen
acting princ
n be divided
5; Breuer J., 2
er length or th
equal to the m
Technologies fo
l (Breuer J., 2005
n the room cl
lly cotton). T
ons. The resu
e preparation
ng machine.

hcontrol
rograph princ
sor measures
ciple is more
d into two gro
2005a):
he 2, 5 % spa
mean fiber le
or Prosperity
5)
limate. TC-L
This device i
ults of meas
n, because th
It is fully au

ciple. The sli
s the fiber m
similar to th
oups:
an length of
ength of the A
LCT is design
s operated on
surements are
he Lengthcon
utomatic and
iver is clamp
mass with a re
he HVI.
the HVI,
AFIS,

ned for the
nly by one
e available
ntrol is fed
d objective,
ped and the
eference to
The Cotton Length Analysis using the Lengthcontrol 489
Fiber hooks [%] - the value gives a measurement of fiber parallelization. The lower the value is, the
less fiber hooks in the sliver are.
II group additional results and parameters (Breuer J., 2005; Breuer J., 2005a):
LCT-length x hooks [mm] this value links the information regarding hooks and length. Fibers are
only really long if the length is long and the number of hooks is low,
Short fiber length [mm] this value shows that 10 % of fibers are shorter than this length,
Short fiber amount [%] - this value describes the percentage ratio of short and long fibers,
Short fiber content [%] - this value indicates the fiber content at 12,7 mm fiber length,
Staple gradient [%] - this value describes the ratio of short and long fibers.
Just after the measurement all the parameters are displayed on the screen and, moreover, a graphical
representation of the fiber length gradient is also possible. The way how the Lengthcontrol shows the
results is presented in Tables 1, 2.
TABLE 1: THE BASE TEST RESULTS

TABLE 2: ADDITIONAL TEST RESULTS

For the further analysis two length parameters measured by the Lengthcontrol have been chosen:
upper length (which according to the producer corresponds to the 2.5 % span length value from the HVI)
and LCT-length (which is comparable to the mean fiber length from the AFIS). There will be also
compared short fiber content (SFC) values.
SAMPLE PREPARATION
The subject of measurement were 15 American standard cottons differed by the length. Because the
Lengthcontrol can be fed only by the sliver, fifteen types of fibers had to be formed in such a structure.
For this purpose, Microdust and Trash Analyzer (MDTA) (presented in the Fig. 3) from the Textile
Research Institute (Lodz) was used. MDTA is a device, which can provide the cotton in the sliver form
and therefore, it was used for the sample preparation for the measurements on the Lengthcontrol
(Frydrych I., 2005; Wakelyn P., Chaudry R, 2010).
490
RESULTS A
For the le
weight L(
Upper len
analysis w
whole pop
The o
Lp.
m
1 2
2 2
3 2
4 2
5 2
6 1
7 1
8 2
9 1
10 2
11 2
12 2
13 2
14 2
15 2
The c
presented
Pa
AND THEIR ANA

ength analysi
(w), the mean
ngth. Moreo
was carried o
pulation (Gn
obtained mea
L(w) [mm]
mean S
21.4 0
21.3 0
25.6 0
24.4 0
25.1 0
19.7 0
18.8 0
28.1 0
19.9 0
26.0 0
23.5 0
25.3 0
23.6 0
24.5 0
23.6 0
coefficient o
in Table 4.
TABLE 4: THE VA
rameter from
L(w)
L(n)
World Cot
Fig. 3: A View o
ALYSIS
is the follow
n length by n
over, on the
out, the corr
niotek K., Ku
asurement res
TABLE 3: RESUL
AFIS

.D. m
0.3
0.1
0.2
0.2
0.3
0.2
0.2
0.2
0.3
0.1
0.2
0.2
0.2
0.2
0.4
of linear corr
ALUES OF LINEAR CORR
the AFIS
tton Research C
f the Microdust an
wing length p
number L(n)
basis of ob
relation coef
ucharska-Kot
sults from th
LTS OF CHOSEN COTTO
L(n) [mm]
mean
17.4
16.8
20.8
20.1
20.2
15.2
14.1
24.2
16.1
21.4
19.0
20.8
19.1
20.3
18.5
relation for
RELATION COEFFICIENT
Par
Conference on T
nd Trash Analyzer
arameters fro
) and two pa
btained resul
fficient was
t J., 2004).
he AFIS and L
N LENGTH PARAMETER
]
S.D.
0.4
0.3
0.3
0.3
0.4
0.1
0.3
0.3
0.4
0.2
0.3
0.2
0.3
0.3
0.4
two pairs o
S FOR THE CHOSEN LEN
rameter from t
Upper lengt
LCT length
Technologies fo
and the Sliver (Fr
om the AFIS
rameters fro
ts Rusing th
determined
Lengthcontro
RS FROM THE LENGTHCO
Upper Lengt
mean
23.42
23.48
27.52
27.05
27.53
22.53
22.88
29.43
21.28
27.20
25.91
27.40
25.91
26.56
26.17
f results fro
NGTH PAIRS OF RESULT
the LCT
th
h
or Prosperity
rydrych I.,2005)
S were chose
m the Lengt
he PQStat so
as well as i
ol are set in T
ONTROL AND AFIS
Lengthcon
th [mm]
S.D.
0.7
0.1
0.3
1.1
0.2
0.6
0.5
0.4
0.8
0.8
0.7
0.4
0.2
0.4
0.4
om both mea
TS FROM THE AFIS AND
Corr

en: the mean
thcontrol: LC
oftware, the
its significan
Table 3.
ntrol
LCT Length
mean
14.98
15.31
18.90
21.52
18.46
13.08
16.09
23.81
13.94
18.41
17.11
19.34
18.54
19.14
17.66
asurement sy
D LENGTHCONTROL
relation Coeffic
0.97
0.88
n length by
CT length i
e statistical
nce for the
h [mm]
n
8
1
0
2
6
8
9
1
4
1
1
4
4
4
6
ystems are
cient
The Cotton Length Analysis using the Lengthcontrol 491

Fig. 4: The Relationship between Parameters L(w) and Upper Length

Fig. 5: The Relationship between Parameters L(n) and LCT Length
As a result of carried out test of correlation coefficient significance for the whole cotton fiber
population the following values of probability were obtained:
- for parameters L(w) and Upper length p < 0.000001,
- for parameters L(n) and LCT length p = 0.000018.
Comparing these values with the assumed level of test significance =0.05, it can be stated that in
both cases there exist the linear correlation between the cotton length parameters for the whole
population. In Figures 4 and 5, there are presented the mentioned above relationships between
appropriate fiber length parameters and the linear regression equation is given.
SHORT FIBER CONTENT
Except the length parameters the AFIS and Lengthcontrol give us also information about the short fiber
content (SFC). This parameter demonstrates the percentage of fibers that are shorter than 12.7 mm. The
relationship between measurements made on the AFIS and Lengthcontrol is shown in Fig. 6 (Pabich A.,
Frydrych I., Raczyska M., Andrysiak J., 2010).
492
In Fig
by weight
Lengthcon
points, wh
CONCLUSIO
There we
Lengthcon
parameter
have been
On th
correlation
Presen
competitiv
product o
make an a
ACKNOWLE
This
Developm
industry
Action 2.1
REFERENCE
[1] Breuer
[2] Breuer
Sales In
[3] Frydry
10 Nr 3
[4] Frydry
(in Poli
[5] Gniotek
FIBER
[6] Pabich
parame
[7] Trtzsc
German
[8] Wakely
g. 6., there is
t from the AF
ntrol are the
here the diffe
ON
ere compared
ntrol with ap
rs from the L
n compared.
he basis of m
n between L
nted results
ve method f
f machines s
additional op
EDGEMENT
work is (p
ment of rese
CLO 2IN
1.
ES
J., (2005), Len
J., (2005)a, Le
nfo.
ch I., Matusiak
3(39), 2002.
ch I., (2005),
ish).
k K.. Kuchars
RS & TEXTILE
A., Frydrych I
eters. TEXSCI 2
chler GmbH &
ny.
yn P.J., Chaudr
World Cot
Fig. 6: Th
s presented a
FIS and SFC
most correla
erences are s
d the results
ppropriate L(
Lengthcontro
easurement a
(w) from the
show that a
for the older
such as the c
peration to pr
partially) sup
earch infrast
N TEX, fin
ngthcontrol LCT
engthcontrol LC
k M, Trends of
Cotton asse
ska-Kot J. (200
ES in Eastern Eu
I., Raczyska M
2010. Liberec.
Co. KG, Moen
ry M.R. (2010),
tton Research C
he Values of SFC P
a graph with
C from the L
ated with the
een, the maj
s of two len
(w) and L(n)
ol. Additiona
and statistica
e AFIS and U
a new system
ones, espec
card, draw fr
repare specia
pported by
tructure of i
nanced by O
T Far more th
CT The decis
AFIS Applicat
essment system
04), Commensu
urope, Vol. 12.
M., Andrysiak J
nchengladbach
Cotton: Techn
Conference on T
Parameters from t
three sets of
Lengthcontrol
e results of S
ority of resu
ngth paramet
from the AF
aly, the SFC
al result anal
Upper length
m of measur
cially in the
rame or com
al samples fo
Structural
nnovative te
Operational
han just a fiber l
ive step toward
tion in Researc
ms and method
urability of M
No. 2 (46)
J., (2010). The
(2006), TC-LC
nology for the 2
Technologies fo
the Lengthcontrol
f points relat
l. There is sh
FC by numb
lts is almost
ters (Upper
FIS, which se
C parameters
lysis it can b
h from the Le
ring the fiber
case of inter
mbing machin
or the measur
Founds in
echniques an
Programme
length measurin
ds quality assur
h and Industry,
ds. Editor of t
Measuring Instru
Lengthcontrol
CT fiber length
1
st
Century, ICA
or Prosperity
and AFIS
ted to the SF
hown that th
ber from the
equal.
length and L
eemed to be
from the Le
e stated that
engthcontrol.
r length L
roperation co
ne is the sliv
rement of fib
the frame
nd technolog
Innovative
ng device, Trtz
rance and qualit
, Fibres &Texti
the Technical
uments for Tex
a comparativ
measuring dev
AC.
FC by numbe
he SFC result
AFIS. In spi
LCT length)
the most sim
engthcontrol
there is a str

Lengthcontro
ontrol, wher
ver, there is n
ber length par
of the pro
gies of textil
Economy, 2
zschler Sales In
ty optimization
iles for Eastern
University of
xtile Science a
ve analysis of c
vice, Operating
er and SFC
ts from the
ite of some
) from the
milar to the
and AFIS
rong linear
ol can be a
e the half-
no need to
rameters.
oject titled
le clothing
2007-2013,
nfo.
n, Trtzschler
n Europe vol.
Lodz, Lodz
and Practice,
cotton length
instructions,

An Innovative Bio-chemical Approach

for Low Energy and Less Polluting Scouring
of Cotton Textiles
P.V. Varadarajan, R.H. Balsubramanya, Nayana D. Nachane,
Sheela Raj and R.R. Mahangade
Central Institute for Research on Cotton Technology, Mumbai
AbstractThe Indian textile scene is dominated by small and medium scale processing units. The scenario has not
undergone drastic changes even under the new economic order as witnessed in other sectors. The main requirement of
the Wet processing sector is low energy and less polluting processing technique. In this direction, CIRCOT has
successfully initiated and developed a new bio-chemical scouring technique for cotton goods. The new method is low
energy and least polluting one.
The newly developed method employs a mixed microflora developed and maintained at CIRCOT. Under this
treatment the fabric is subjected to anaerobic treatment at room temperature. In the above study, 100% cotton woven
fabric of low weight was employed. The fabric was subjected to anaerobic treatment at room temperature for various
length of time followed by peroxide bleach and dyed with hot brand reactive dye. The fabric samples at different stages
were evaluated for weight loss, whiteness index, C.V. of whiteness index, absorbency, viscosity, colour strength, fabric
strength and elongation at break. All the treatments were compared with conventionally kiered and bleached fabric as
control.
To determine the usefulness of the fabric for apparel applications, impact of the bio-scouring process on the low
stress mechanical characteristics were also investigated. The KES-FB system was employed and properties such as
Tensile, Bending, Compression, Shear, Surface friction and Roughness were evaluated. Bio-scoured fabric showed
better extensibility as compared to the conventional treatment. No change was observed in Bending rigidity on
bioscouring treatment. An increase in fabric surface smoothness was indicated in the case of bioscoured fabric.
Primary handvalues displayed an improvement in the case of bioscoured fabric as against the conventionally scoured
fabric. The results of the extensive studies show that the properties of the treated fabric are on par with the
conventionally processed ones. It is further observed that the colour value of the treated and dyed samples are in fact
higher than the conventionally processed ones.
The above process can be easily coupled to existing Hand Processing Units leading to considerable reduction of
pollution load along with appreciable saving of energy.
INTRODUCTION
Scouring is an important pretreatment operation in the processing of cotton and cotton blended
materials.The main objective of the above operation is to remove the non-cellulosic constituents of cotton
fibre which make the fibre non-absorbent posing serious technical problems in the subsequent wet
processing operations. In fact, the scouring operation determines the ultimate quality of the finished
product.
The scouring operation consists of treating the cotton goods with 1-2% of NaOH solution at high
pressure and temperature for 4-5 hours. The above operation is not only energy intensive but also
leads to environmental pollution. It is estimated that scouring operation consumes about 1% of the total
water used, contributes 54% of the total BOD and is responsible for 10-25% of the total pollution load of
the entire textile processing operations. It is pertinent to observe here that in view of the ever widening
gap between the demand and supply position of energy, serious effects are on, in almost every field of
activity either to cut down the un-necessary expenditure of energy or to adopt a low energy process.
In the light of the above observations it is not surprising that a number of studies
1
have been initiated
over the years to make scouring operation less energy intensive and more effective one. A survey of the
literature shows that but for the development of a number of chemical additives
2
-
6
, and use of certain pure
81
enzymes
7
-
9
, the basic operation of scouring remains essentially energy intensive and polluting in nature.
The present study therefore attempts a novel technique to produce a mixture of enzymes in-situ to make
the scouring operation less polluting and less energy intensive. The anaerobic technique developed at
CIRCOT
10
for the degradation of the cellulosic waste is employed in the present study to carry out
scouring operations on 100% cotton fabric.
Initially, quality and characteristics of apparel fabrics were evaluated by touching and feeling by
hand, leading to a subjective assessment of them. Around 1972, Kawabata, Niwa and their colleagues
15
-
16

developed an objective evaluation system based on the precise measurements of certain mechanical and
surface properties. The instruments used for these measurements was known as KES-FB system, wherein
instead of feeling fabric by hand, this instrument system touch fabrics to measure their mechanical
properties and surface properties under low load conditions. The mechanical property parameters are
converted to assess the Handle Value of the fabric. These elementary fabric mechanical properties are
generally believed to relate to important fabric characteristics such as drape, handle, tailorability,
wrinkling, creasing, shape-retention properties and other aesthetic characteristics. The present study
involves scouring through mixed microbial culture created in-situ followed by alkaline treatment and the
impact of such novel treatment on the low stress mechanical properties of bioscoured cotton have not
been studied till now.
In the above study 100% cotton woven fabric of low weight (78g/sqm) was employed. The fabric was in
grey state. The required quantity of fabrics was subjected to anaerobic digestion for 10h and 20 h
respectively. Microbial consortium was used to treat the fabric. The consortium comprised both aerobic
and anaerobic-types. Species belonging to Bacillus and Micrococcus sp. from Gram positive group and
Beijerinckia, Pseudomonas, Xanthomonas and Flavobacterium were from Gram negative group.
Aspergillus, Penicillium and Mucor were from fungi and Streptomyces was the alone Actinomycete.
All these were from the aerobic ones surviving under anaerobic conditions. As and when the system was
disturbed, these were acting as scavengers of oxygen and setting anaerobiosis. Among anaerobic groups,
species of Methanomicrobium, Desulfoto-maculum, Clostridium, Chlorobium, Ectothiorhodo-spire,
Thiodictyon and Rhodospirilhim were predominant. The presence of Chlorella as green alga and Anacystis
as blue green alga was found to grow profusely under anaerobic-conditions. One species of protozoan
belonging to the genus Monocercononas was also present
The anaerobic digestion was carried out in sealed glass jar employing a 100% mixed flora developed
and maintained at CIRCOT. The digestion was carried out at room temperature of approximately 32C.
At the end of the digestion period the samples were boiled with 0.5% and 1% NaOH solutions (owf) for
15 minutes, washed and air dried. The above treated samples were bleached with peroxide employing a
M:L ratio 1:20 with 3g/l peroxide,1.5 g/l Na-silicate and 1g/l NaOH (owf) at boil maintaining the pH at
10 to 11 for one hour. The bleached samples were dyed with hot brand reactive dye. One set of grey
sample was subjected to conventional kiering consisting of boiling with 1% NaOH under 15lh/inch squire
pressure for 4 hours followed by bleaching and dyeing. The fabric samples at different stages were
evaluated
11
-
13
for weight loss, wax content.whiteness index, uniformity of whiteness index, water
absorbency, viscosity, colour strength, fabric strength and elongation at break. All the treatments were
compared with conventionally kiered and bleached sample as control. The reflectance measurements of
all the samples were carried out using Jaypak - 4802 computerized colour matching system. From the
reflectance values, colour strength, expressed as K/S values, were calculated at the wavelength of
maximum absorption (X max) using the Kubelka - Munk equation.
The low stress mechanical properties were measured on the Kawabata Fabric evaluation System
(KES-FB) under standard conditions. The fabrics were tested on the five modules of the KES-FB system
viz KES-F1 Tensile and Shear Tester, KES-FB2 Pure Bending Tester, KES-FB3 Compression Tester and
KES-FB4 Surface Tester to measure the Tensile and Shear, Bending, Compression and Surface
properties respectively. The fabric hand value was evaluated using the Kawabata System of equations.
An Innovative Bio-chemical Approach for Low Energy and Less Polluting Scouring of Cotton Textiles 495
Tables l and 2 depict the comparative behavior of the fabric samples subjected to conventional and
anaerobic digestion for 10h and 20h, under different experimental conditions in respect of weight loss,
whiteness index, fabric strength and elongation, fluidity and, colour strength. It can be seen from the
above Tables that the anaerobic digestion carried out for 10 h under different experimental conditions
have in general shown lower weight loss as compared to the conventional kiering. Similar trend is
observed in respect of the bleached samples too. Table 2 which depicts the trend of weight loss under
20 h anaerobic digestion also shows a similar behavior but the weight loss is higher than those of the
samples subjected to 10 h digestion. It is interesting to observe an increasing trend in the whiteness index
of the anaerobically kiered sample followed by boiling with 0.5% and 1% NaOH (owf) respectively. The
treatment of anaerobic digestion followed by boiling with 1% NaOH confers the same whiteness index as
that of the conventionally kiered ones. In respect of the samples anaerobically treated followed by alkali
boiling and bleaching clearly show that with the increase in the concentration of NaOH used for
boiling, the whiteness index also increases. The whiteness index of the anaerobically treated boiled
with 1% NaOH and bleached samples almost compare with those of the conventionally kiered and
bleached samples. Similar trend is witnessed in the case of samples subjected to 20h anaerobic
digestion as depicted in Table 2.
The study also showed that the extent of removal of wax through anaerobic digestion followed by
0.5% and 1% open alkali boil is as efficient as that achieved through conventional kier boil. In fact
the wax content of the sample subjected to 10 h anaerobic treatment and 1 % alkali boil followed by
bleaching was lesser than that of the conventionally kiered and bleached fabric.
The water absorbency of the fabric is an important functional parameter. It can be seen from Table 1
that the fabric subjected to only anaerobic treatment for 10h is not absorbent but when the anaerobic
treatment followed by alkali boiling makes the fabric absorbent. It is further noted that an increase in
NaOH concentration employed for boiling after anaerobic treatment does not appear to have any
influence on water absorbancy property. The overall results of absorbancy as shown in Table 2
indicates that the samples become more absorbent with increase in the duration of digestion.
Table 1 presents an interesting picture of the behaviour of samples in respect of strength and
elongation properties. In general it is observed that both the conventional and the anaerobic kiering
treatments lower the fabric strength. It is evident from the Table 1 that the samples subjected to
anaerobic treatment followed by alkali boiling possess an improved strength retention as compared to
the conventionally kiered samples. Similar trend is observed in the case of the samples subjected to 20h
anaerobic treatment as depicted in Table 2. It can also be inferred that the longer duration of anaerobic
treatment leads to a lower strength retention. It is further noted that the strength reduction trend of the
bleached samples differ between the treated and control samples. It is seen that the extent of strength
reduction of anaerobically treated and bleached samples is much lower than conventionally kiered and
bleached samples. This could possibly be attributed to the lower degradation of anaerobically treated
samples as compared to the conventionally kiered samples as reflected in the fluidity values shown in the
Table 1 and 2. This could possibly be attributed to the reported fibrillar agglomarisation in the case of
cotton fibre samples subjected to anaerobic treatment as observed by Bhatawdekar et.al
14
The ends
and picks values of the treated samples indicate that a possible fabric structural differences have taken
place similar to that noticed during fabric shrinkage and hence to some extent the observed higher
strength retention could possibly be attributed to the changes in the ends and picks values. But the results
also show that the anaerobic digestion process also appears to be less degradative as reflected in the
lower fluidity values. It could therefore be safely observed that the anaerobic digestion process does
confer higher strength retention as compared to the conventional kiering process.
It is further noted that in all the samples whether conventionally kiered or subjected to anaerobic
digestion a general strength reduction is noted subsequent to dyeing. In respect of the elongation retention,
the anaerobically treated samples as shown in Table 1 show an entirely different trend to that of the
conventionally kiered samples. In comparison to the conventionally kiered samples where, a general
reduction in elongation is noted,the anaerobically treated samples that are subjected to alkali boiling on
the contrary show an increase in the elongation retention. Once again the observed increase in the
elongation may possibly be attributed to a relatively higher shrinkage factor of the treated fabrics as
compared to the conventionally kiered ones. Such anomalous behavior is not noticed in the case of
samples subjected to 20h anaerobic treatment.
Tables 1 and 2 also depict the whiteness index of the control and the anaerobically kiered samples.
Though the whiteness index of the samples subjected to only anaerobic kiering are much lower to that of
the conventionally kiered sample, the whiteness improves when the anaerobically treated samples are
given an alkali boiling. It can be seen, that the whiteness index of anaerboically treated samples followed
by 1 % alkali boiling is almost on par with that of the conventionally kiered samples. However, it is
interesting to note that in all the cases the anaerobically treated and bleached samples show superior
whiteness index values as compared to the conventionally kiered and bleached samples. It is also noted
that the whiteness index of bleached samples increase when anaerobic treatment is followed by alkali
boiling. In order to study the uniformity of whiteness achieved through anaerobic treatment, eight
reflectance measurements were undertaken on each of the samples and the C.V. of the whiteness index
was considered as a measure of the uniformity of whiteness. It can be seen from Table1 that the C.V.
value of the whiteness index had shown a drop from 2.57 for conventionally kiered to 1.27 and 0.99 for
the anaerobically treated alone and alkali boiled samples respectively. Though the C.V. of the sample
boiled with 1% NaOH is higher still it is much lower than the conventionally kiered sample. Thus on
the whole anaerobic treatment followed by alkali boiling appears to impart more uniform whiteness as
compared to the conventionally kiered samples. A similar trend is observed with the bleached
samples also.
In order to study the response of such anaerobically treated and bleached samples to dyeing, all the
samples were dyed to 2% shade employing a hot brand reactive dye. It can be seen from Table1 that the
anaerobically kiered samples in general show higher K/S values as compared to the conventionally kiered
samples. However,in the case of samples depicted in Table2, K/S values of treated samples are almost on
par with the control. The colour characteristics of the anaerobically treated sample match with that
of the conventionally kiered control sample. The L*a*b* values indicate that the anaerboically treated
dyed samples have a relatively higher colour strength with a slightly higher yellowish tinge.
The tensile properties of the conventionally scoured and bioscoured samples are measured using
tensile tester KES-FBI. The values EMT, WT and RT of the three samples are depicted in Table 3. EMT
is extensibility under a load of 500g/cm. It is a measure of fabric ability to be stretched under tensile load.
Bioscoured samples B and C showed better extensibility compared to conventional kiering. The values of
WT give a measure of work done during extension. WT is higher for samples B and C as compared to
conventional kiering. This suggests that the bioscoured samples are easily stretchable when subjected to
tensile deformation than that subjected to conventional treatment. RT is a measure of tensile resilience.
This represents the recovery of the fabric from tensile deformation. Values of RT were observed to be
less for bioscoured fabrics as compared to the conventionally scoured.
Table3 depicts the bending properties of the conventionally scoured and bioscoured samples. The
term B denotes the bending rigidity and 2HB denotes the hysteresis of bending moment. The
bending rigidity does not vary among the samples. Whereas, 2HB values of the bioscoured samples are
lower than the conventionally scoured ones indicating lower hysteresis losses in bending deformation as
compared to the conventionally kiered. In short the bioscoured ones showed better recovery from
bending deformation.
The compression parameters WC( compression energy),RC (compression resilience)and LC(linearity
of compression )is measured using KES-FB3 Compression Tester. Conventionally kiered sample showed
higher WC value indicating that the fabric is more compressable than the bioscored fabrics. Compression
resilience is the percentage of the extent of recovery or regain in fabric thickness when applied force is
removed. No significant difference was observed in RC values between the conventional kiered and
bioscored fabrics.
Surface properties of conventionally scoured and bioscoured samples are shown in Table 3.Values of
MIU is a measure of the mean values of coefficient of friction between fabric surface and metallic piano
modelsurface detector, whose surface is simulated to the finger surface. Sample C showed significant
decrease in MIU, MMD and SMD values indicating a much smoother surface for bioscoured fabric
compared to conventional kiered sample A. The increase in fabric smoothness is indicated in the value of
Numeri too which is 5.93 for sample C as against 5.39 for conventional kiering.
Shear tester KES-FBI was used to determine the G, 2HG and 2HG5 values of the fabric samples.
These values in brief is a measure of a fabrics ability to deform in its plane. The overall picture of
Table3 which show the shear property of the samples, do not show any change in its shear property
whether it is conventionally kiered or bioscoured under different conditions.
The Primary Hand Values of the three samples present interesting picture. Sample C showed an
improvement in fabric smoothness ( Numeri ). It also showed a marginal improvement in fabric softness
and fullness ( Fukuremi ) as compared to conventional kiering. This improvement in fabric smoothness
and softness resulted in a higher value for THV ( 2.89 ) for sample C as compared to the value of THV (
2.46 ) for conventional kiering.
CONCLUSION
The overall results of the above study seems to show that the anaerobic digestion followed by mild alkali
boiling could offer a simple, low energy, less polluting, eco-friendly kiering technique for 100% cotton
fabric. The quality of the kiered samples in terms of fabric strength, elongation whiteness index and
the uniformity of the whiteness is on par with that obtained through conventional kiering process. The
colour strength of treated and dyed samples is slightly higher than that of the control. In brief, the
properties of the fabric subjected to biochemical technique are comparable to that obtained in
conventional treatment. Impact of the bio-scouring process on the low stress mechanical characteristics
such as Tensile, Bending, Shear, Surface friction and Roughness of the Bio-scoured fabrics showed better
extensibility as compared to the conventional treatment. No change was observed in Bending and Shear
rigidity on bioscouring treatment. An increase in fabric surface smoothness was indicated in the case of
bioscoured fabric. Primary hand values displayed an improvement in the case of bioscoured fabric as
against the conventionally scoured fabric. The results of the extensive studies show that the properties of
the treated fabric are on par with the conventionally processed ones.
TABLE 1: FABRIC PROPERTIES OF 10 HOUR ANAEROBIC TREATMENT
S
l
.

N
o

T
r
e
a
t
m
e
n
t

W
e
i
g
h
t

L
o
s
s
%

A
b
s
o
r
b
a
n
c
y

W
h
i
t
e
n
e
s
s

I
n
d
e
x

C
.
V
.

o
f

W
h
i
t
e
n
e
s
s

I
n
d
e
x

F
l
u
i
d
i
t
y

S
t
r
e
n
g
t
h

R
e
t
e
n
t
i
o
n

%

E
l
o
n
g
a
t
i
o
n

R
e
t
e
n
t
i
o
n

%

E
n
d
s
/
p
i
c
k
s

K
/
S

1 0 As such - > lOmin 54.76 0.72 - 1 100 100 94/80 -
2 E0 Conv.Kier 13.2 Instant 70.81 2.57 2.2 77.4 95.5 96/90 -
3 El Con v. Kiered+B leached 13.6 Instant 85.99 1.32 6.8 73.8 89.4 99/89 -
4 E2 Conv,Kiered+Bleached+Dyed ~ Instant ~ - " 72.2 87.4 97/91 10.51
5 A0 Anaerobic Kiered 6.2 >2min 55.72 1.27 2.6 74.6 95.5 96/87 -
6 Al An.Kiered+Bleached 10.4 l-2min 79.16 0.28 3.4 89.3 124.0 98/88 -
7 A2 An. Kiered+Bleached+Dyed - Instant - - 78.2 115.4 100/87 11.77
8 A3 An.Kiered+0.5%NaOH 12.1 l-5sec 66.75 0.99 2.3 82.5 111.4 101/87 -
9 A4 An.Kiered-
K).5%NaOH+Bleached
12.1 l-5sec 82.60 0.61 2.9 88.9 124.0 102/90 -
10 A5 An.Kiered+0.5%NaOH+Bleach
ed+Dyed
- Instant - - - 79.4 97.2 101/90 11.35
11 A6 An.Kiered+l%NaOH boil 12.2 l-5sec 69.32 1.41 2.4 84.1 121.5 101/88 -
12 A7 An.Kiered+l%NaOH+Bleached 12.1 l-5sec 83.73 0.79 3.0 81.7 104.1 96/90 -
13 A8 AnKiered+1
%NaOH+Bleached+Dyed
- Instant - - - 77.0 94.7 100/90 11.24

TABLE 2: FABRIC PROPERTIES OF 20 HOUR ANAEROBIC TREATMENT
S
l
.

N
o
.

T
r
e
a
t
m
e
n
t

W
e
i
g
h
t
.

L
o
s
s
%

A
b
s
o
r
b
a
n
c
y

W
h
i
t
e
n
e
s
s

I
n
d
e
x

C
.
V
.

o
f

W
h
i
t
e
n
e
s
s

I
n
d
e
x

F
l
u
i
d
i
t
y

S
t
r
e
n
g
t
h

R
e
t
e
n
t
i
o
n

%

E
l
o
n
g
a
t
i
o
n

R
e
t
e
n
t
i
o
n

%

E
n
d
s
/

p
i
c
k
s

K
/
S

1 0 As such - > lOmin 54.76 0.72 - 100 100 94/80 -
2 EO Conv.Kier 13.0 Instant 70.81 2.57 2.2 77.4 95.5 96/90 -
3 El Conv.Kiered+Bleached 13.6 Instant 85.99 1.32 6.8 73.8 89.4. 99/89 -
4 E2 Conv,Kiered+Bleached+D
yed
- Instant - - - 72.2 87.4 97/91 10.51
5 BO Anaerobic Kiered 9.7 >2min 56.71 1.28 1.9 67.1 80.5 97/87 -
6 Bl An.Kiered+Bleached 11.0 lOsec 81.70 0.90 43 67.5 80.9 98/89 -
7 B2 An.
Kiered+Bleached+Dyed
- Instant - - - 55.6 67.9 100/88 10.27
8 B3 An.Kiered+0.5%NaOH
boil
12.4 Instant 66.42 2.04 2.1 69.4 93.1 100/90 -
9 B4 An.Kiered+0.5%NaOH+B
leached
12.7 Instant 86.23 0.47 3.8 67.5 91.5 98/89 -
10 B5 An.Kiered+0.5%NaOH+B
leached+Dy ed
- Instant - - - 69.8 91.1 99/90 10.67
11 B6 An.Kiered+l%NaOH boil 12.8 Instant 70.45 1.06 2.2 72.6 98.0 99/89 -
12 B7 An.Kiered+1
%NaOH+Bleached
13.2 Instant 88.69 0.42 6.0 72.2 92.3 96/88 -
13 B8 AnKiered+1
%NaOH+Bleached+Dyed
- Instant - - - 66.7 82.9 99/90 10.38
TABLE 3: LOW STRESS MECHANICAL PROPERTIES OF TREATED SAMPLES
Sr. No. Properties Sample A Sample B Sample C
1 Tensile Property
EMT % 9.97 11.44 10.92
WT (g.cm/cm2) 17.96 19.28 19.31
RT % 26.87 24.74 24.18
2 Bending Property
B(gf cm/cm) 0.021 0.020 0.020
2HB (gf cm/cm) 0.025 0.020 0.020
3 Surface Property
MIU 0.115 0.112 0.095
NMD 0.028 0.029 0.023
SMD( micron) 7.174 7.401 4.781
4 Shear Property
G (g/cm.deg) 0.56 0.55 0.56
2HG (g/cm) 1.59 1.66 1.59
2HG5(g/cm) 2.54 2.58 2.53
5 Compression Property
LC 0.387 0.232 0.254
WC (g.cm/cm2 ) 0.293 0.217 0.231
RC % 56.65 52.20 53.27
T (mm) 0.558 0.646 0.656
A - Conventially kiered sample
B Anaerobic scouring and 0.5 % NaOH boil
C - Anaerobic scouring and 1.0 % NaOH boil
REFERENCES
[1] Harmaker, S.R.- Colourage Annual 1998, p.18.
[2] Sanakari, V.D., Text Dyers & Printers, May 11, 1983.
[3] Burkitt, F.H. Am. Dyest. Rep. March 1978 p.51
[4] Text. Dyers & Printers.May 1983.
[5] Meyer Jim. Textile Horizon, April 1983
[6] Garrett, C.,J. Soc. Chem.& Col. 71,1955, p.83O
[7] Etters, J.N. and Annis, P.A. Am. Dyest Rep., 87, No.5, 1998, p.18.
[8] Etters. J.N. -colourage Annual 1998, p.87
[9] Hardin, I.R. and Kim J.-Book of Papers AATCC International Conference and Exhibition 1998, p. 319
[10] Khandeparkar. V.G., Balasubramanya, R.H. and Shaikh, A.J. -Process for the preparation of paper grade pulp from cotton
plant stalk by anaerobic digestion. Ind. Pat. No. 176891, July 1993.
[11] 1.S. 23491963.Method for determination of wettability of cotton fabrics.
[12] Handbook of Methods of Tests.,CIRCOT
[13] Handbook of Methods of Tests., CIRCOT
[14] Bhatawdekar, S.P. Sreenivasan, S., Balasubramanya. R.H. & Paralikar, K.M., Text Res. J., 62 (5), 290292 (1992)
[15] Kawabata,S., The standardization and Analysis of Hand Evaluation, 2
nd
ed. The Textile Machinery Society of
Japan, 1980.
[16] Kawabata, S., and Niwa, Masako, Fabric Performance in clothing and Clothing Manufacture, J. Textile Inst. 80,
1950 ( 1989)
The Within Bale Repeatability of Standardized

InstrumentS for Testing Cotton Fiber
Produced in Africa
E. Lukonge
1
,

M. Aboe
2, 4
, Gourlot
3
, J.P. Goz
3
and E. Hubl
3

1
Lzardi, Mwanza, Tanzania
2
Association Interprofessionnelle du Coton, Parakou, Bnin
3
Cirad, UPR SCA, F34398 Montpellier, France
3
Institut d'Administration des Entreprises, Montpellier, France
4
Universit de Haute Alsace, LPMT-EAC 7189 CNRS-UHA, Mulhouse, France
AbstractFiber length, fiber strength, micronaire, uniformity, reflectance and yellowness measured on standardized
instrument for testing cotton (SITC) are often used on cotton bales produced in the world for trading purposes with
full respect of agreed commercial tolerances in order to limit the frequency of claims. In Africa, almost no trading on
SITC data is made because we lack the study of within-bale variability of the given characteristics to deduce sampling
and testing protocols insuring the respect of the same agreed commercial tolerances. We then conducted this study of
the within-bale variability of fiber length and its uniformity, fiber strength, micronaire, reflectance and yellowness. We
took eight samples per bale within 455 cotton bales produced in 14 African countries during two crop seasons. Our
representative sample is then composed of over 3600 fiber samples which were analyzed in controlled conditions by
SITC in a laboratory fully respecting the international recommendations. We then achieved an estimation of the
within-bale variability of cotton fiber technological characteristics in most of the African cotton producing countries.
The results indicated the variability per country, per bale in some situations and it was noted that even the gins (saw
and roller) have also some effects in relation to within bale variability.
Keywords: cotton, fiber, within-bale variability, sampling, testing, repeatability, classification
INTRODUCTION
The issue of cotton fibre characterization is important for the productivity and quality of the products
obtained during processing operations such as spinning, weaving, etc. [2]. This explains why cotton
classification has been based on fiber characterization.
Sasser [3] and Knowlton [4] described the steps in fiber classing and testing that have gradually
impacted cotton classification and worldwide trade in cotton: the most recent, hereafter referred to as
Standardized Instruments for Testing Cotton (SITC), combines manual and visual classing in addition
to a fully automated instrument testing.
SITC have been increasingly used worldwide and the International Cotton Advisory Committee
(ICAC) estimates that 50% of the cotton traded in the world is classed thanks to SITC, either in addition
to or instead of manual and visual classing [5]. These instruments measure Micronaire, length, length
uniformity, Strength and color (Reflectance and Yellowness) at least.
In Africa, almost no bale is sold with instrumental result. As the within-bale variability and the
sampling and measurements procedures determine the precision of bale evaluations for those
characteristics, which in turn determine the risk of discrepancies exceeding commercial tolerances and
ultimately litigations, United State Department of Agriculture, Agricultural Marketing Services (USDA-
AMS) periodically performs variability studies in order to warrant a limited litigation risk in their given
conditions.
Only one publication was found focusing on the impact of the production conditions onto the within-
bale variability, however without any formal treatments for production conditions [6].
82
The Within Bale Repeatability of Standardized Instruments for Testing Cotton Fiber Produced in Africa 501
The assumption is that the variability of SITC measurements for any given cotton bale can differ
from one ginning mill to another. Indeed, equipment used, or the ginning conditions and/or the cropping
system used in the supply area of the ginning mill as well as seed-cotton management practices have
impacts on fiber characterizations.
This question is particularly important in developing countries, in particular in the fourteen African
countries that produce cotton (Benin, Burkina Faso, Cameroun, Ivory Coast, Mali, Mozambique,
Senegal, Sudan, Tanzania, Chad, Togo, Uganda, Zambia, Zimbabwe) considered in this study. In these
African countries, production conditions differ considerably from prevailing conditions in the USA. The
cotton farms are smaller, on average 0.6 ha [7] and the cropping system is largely manual [8].
Consequently, each cotton fiber bale includes fiber produced on a larger number of farms under
different field conditions and that a higher litigation risk may arise between some cotton companies from
this area and their customers.
In this publication, we checked the hypothesis that the application of the USA method provides
results precise enough for the trading of cotton in Africa, In the opposite case, we would develop other
sampling and testing modalities in order to insure that SITC methods match both the needs of SSA
producers and the agreed worldwide expectations in terms of reliability, precision and the trueness of the
results.
In general, the within-bale variability of fiber quality depends on the agricultural production
conditions and on the equipment used in the ginning mills and is affected by four main scales: 1) scale of
the cotton plant, [9] [10] where fibers from different cotton bolls vary; 2) scale of the cotton field, where
cropping conditions (agronomical impacts, climate, variety, cultivation practices) may differ [11, 12]; 3)
scale of the supply area of the ginning mills, as seed cotton from different farms is combined before
being transported to the ginning mill [13]; and 4) scale of the ginning mill and of their equipment
including the management of seed cotton [14] [15].
In this publication, the main focus was on supply area of the ginning mills and ginning equipment as
the main variability sources to find the level of within bale variability of the fiber characteristics as
measured by SITC for the bales produced in the African countries and the most appropriate sampling and
testing procedures for African countries to respect international repeatability requirements.
Since nothing has been done for African cotton, it is time for African countries to adapt the USA
methodology to avoid claims according to cotton quality methods and analysis procedures Therefore,
through CFC/ICAC/33 project funded by the Common Fund for Commodities and the European Union,
SITC tests on samples taken from the bales in the fourteen African producing countries for the two
cropping seasons were done to measure the sampling variance due to the operational sampling
conditions and testing using a SITC.
Two experiments on measurement of within-bale variability were conducted in two seasons: (2008-2009
crop season 1 and 2009-2010 crop season 2). Given the large number of the ginning mills in these
African countries, we chose twenty-two situations, representative of these countries, according to their
seed-cotton supply areas, their ginning equipment (roller vs saw) and the presence or absence of lint
cleaners. In crop season 1, 28 situations were sampled though it was half season and 35 situations were
sampled during crop season 2. Some situations remained the same in both seasons to allow us to repeat
the measurement in the same situations, and others were added in the second season to extend the sample
of the situations. For reasons of confidentiality, all countries and situations were encoded.
Sampling Cotton Fiber for the Characterization of Fiber Properties
We assumed that seed cotton transported in different trucks came from various villages, and would thus
induce different levels of variability when the seed cotton differed from one village or another. In our
experiment, we assumed that eighteen 225 kg bales of fibres can be produced from every seed-cotton
truck. So, to insure that each sampled bale comes from a different village, we decided to select one bale
out of every 20 in each situation,
Eight samples per bale from eight different layers were collected from every sampled bale. In each
situation, a total of 10 bales were sampled in crop season 1 and limited to 5 bales in crop season 2.
Including all selected ginning mills, the total numbers of bales and samples collected and tested were
respectively 280 bales and 2239 samples in crop season 1 and 175 bales and 1400 samples in crop season
2 (Table 1).
TABLE 1: LIST OF SITUATIONS, NUMBER OF BALES AND SAMPLES TESTED
Situations Crop 1 Crop 2
No. of Bales No. of Samples No. of Bales No. of Samples
C1G1 10 80 5 40
C1G2 10 80
C1G3 5 40
C2G1 10 80 5 40
C2G2 10 80
C2G3 10 80
C2G4 5 40
C3G1 10 80
C3G2 10 80 5 40
C3G3 10 80
C3G4 5 40
C4G1 10 80
C4G2 5 40
C4G3 5 40
C5G1 10 80
C5G2 10 79
C5G3 10 80 5 40
C5G4 5 40
C6G1 5 40
C6G2 5 40
C6G3 5 40
C7G1 5 40
C7G2 5 40
C7G3 5 40
C8G1 5 40
C8G2 5 40
C8G3 5 40
C9G1 10 80 5 40
C9G2 10 80 5 40
C9G3 10 80 5 40
C10G1 5 40
C10G2 10 80
C10G3 10 80
C10G4 5 40
C10G5 10 80 5 40
C11G1 10 80 5 40
C11G2 10 80 5 40
C11G3 10 80
C11G4 5 40
C12G1 10 80
C12G2 10 80 5 40
C12G3 10 80 5 40
C12G4 5 40
C13G1 10 80 5 40
C14G1 10 80
C14G2 10 80
C14G3 10 80
C14G4 5 40
C14G5 5 40
C14G6 5 40
Total number of bales 280 175
Total number of samples 2239 1400
Total number of situations 28 35
In crop season 1, the collection was done at the end of the ginning season, whereas in crop season 2,
the collection was done in the middle of the ginning season while, ideally, the samples should be
randomly selected throughout the ginning season.
Sample Testing
The six technological characteristics recommended by the CSITC Task Force of the ICAC [5] for testing;
Micronaire (Mic; Micronaire unit); Upper Half Mean Length (UHML, mm); Length Uniformity Index
(UI, %); Strength (Str, g/tex = 0.981 cN/tex); Reflectance (Rd, %); Yellowness (+b, Yellowness unit)
were measured.
For these quantitative variables assumption: when making a measurement on one sample of a bale,
two additive errors are experienced:
The sampling error: the sample mean differs from the bale mean
The measurement error: due to the re-sampling of a specimen within the sample, and to the
imperfection of the instruments.
One bale is the result of stacking successive layers in a continuous production process leading to the
assumption that within-bale variability results essentially from differences between the layers. Then we
estimated the variances of the two error components with a standard two-stage sampling method. One
sample from each of the eight layers was evenly distributed in each bale to be measured twice (total of
two replicates). The six technological characteristics were measured centrally in a controlled laboratory
using a SITC device, USTER Technologies model HVI 1000. Each replicate was carried out according to
ASTM 5867 requirements [16] with one measurement of Micronaire and two measurements of the
Length/ Uniformity Index, Strength, Color Rd and Yellowness.
All required precautions were taken to avoid any calibration drift or, if any drift occurred, to measure
it. The reference materials used for calibration were Universal Micronaire Calibration Cottons, Universal
High Volume Instrument Calibration Cotton Standards for length and Strength parameters and the colour
tiles delivered by the manufacturers. The reference material was also tested for every after 16 samples,
and the testing conditions were recorded. All test results were grouped together in a database for
statistical analysis using R software version 2.11.1 and SAS Institute software version 9.2.
Model of Exploration of the Variances
We used the following model for exploring the acquired results: result = (bale fixed effect) + (layer in the
bale random effect) + (replicate or measurement effect random effect) block effect [17, 18]. The indicial
of this model is as:
,],k
= m
+ A
,]
+ B
,k
+ E
,],k
(1)
Where: Y is the response variable
m
i
is the mean of the bale i
A is the random effect of the layer j in the bale i~ N(u, o
A
2
)
B
i, k :
is the effect of the block k in the bale i
E
i, j, k
is the error of measurement of the replicate k of the layer j of the bale i, residual effect linked to
the replicate in the layers ~ N(u, o
L
2
), independent from A
i is 1I bales
j is 1J layers in the bale
k is 1K replicates in each layer.
The two retained random effects retained as variability sources (A and E) are assumed to be
independent:
o
A
corresponds to the standard deviation of the random layer effect,
o
L
corresponds to the standard deviation of the residual effect.
Our goal is to evaluate o
A
and o
L
for each retained situation and to estimate the variance error as a
function of practical sampling (J sampled layers in a bale) and testing conditions (K tests per sample) or
N if one decides to mix cotton fibers from J layers to analyze them all together:
o
M
2
=
c
A
2
]i
+
c
E
2
]iKi
or respectively o
M
2
=
c
A
2
]i
+
c
E
2
Ni
(2)
The full paper exactly explaining the statistical theory of this experiment is published in Textile
Research Journal [1]. (3)
DEFINITION OF THE LITIGATION RISK AND CALCULATION MODALITIES
In this experiment, we made all steps necessary for evaluating the litigation risk that a given tolerance
(Table 2) is exceeded for each measured characteristic.
TABLE 2: TOLERANCES USED FOR CALCULATION OF THE LITIGATION RISK [16]
Characteristic Commercial tolerances
Micronaire +/- 0.1 unit
UHML +/- 0.508 mm
UI +/- 1 %
STR +/- 1.5 cN/tex
Rd +/- 1 %
+b (Yellowness) +/- 0.5 unit
For our numerical calculations, we chose a litigation risk of 10%, and the standard deviation of the
difference is the commercial tolerance divided by the quintile (100-5)% of the normal distribution =
1.65.
-1
(1-u)
=
-1
(0.95)
= 1.6S. The standard deviation of the mean of the bale is then:
o
M
=
1oI
(1.65.2)
(5)
Three ways of decreasing the sampling variances are
Increasing the number of replicates. Then J=1 and K>1. This is pertinent if the contribution of
the measurement error is larger than that of the between layer error.
Increasing the number of layers. Then J > 1 and K = 1. This is pertinent if the contributions of
the between layer and within layer variances are well balanced.
Increasing the number of layers and replicates that are parts of the combined sample without
increasing the total number of replicates. Then J>1 and N=1. This is pertinent when both
variances are comparable in level..
Bales from the ginning mills within the bold circle will show less than 10% litigation risk with 1
layer sampled and 1 replicate. Bales from the ginning mills within the dotted ellipses will show less than
10% litigation risk with J layers sampled and K replicates.
In practice, the number of layers J should be limited to two as it is only easy to remove samples of
fiber (cutter method) from the top and bottom outside layers of the bale. Then according to the position in
this chart, one can deduce the sampling and testing protocoles to e applied in the commercial
classification process.
From results obtained on reference materials tested in the sets of samples, we did not observe any hour
effect or day of testing effect in the analysis, thus proving that external conditions did not affect the
technological measurements.

Fig.
Explorator
The cont
characteri
characteri
situations
could be i
that, the n
the situati
replicate.
layers sam
marked w
In all
cleaner in
ginning m
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right uppe
In the
situations
When
samples to
agreed tol
both Sigm
cotton ma
As in
Micronair
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The Within B
1: Example of Rela
ry Analysis of
tribution of
istic concern
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are illustrate
included in t
number of la
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Bales from
mpled and K
with arrows ar
cases, the e
ndicated som
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er corner of t
e case of on
ensure a liti
n all data wi
o grab per b
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maA and Sigm
anagement fr
dicated in S
re and two
ss.
Bale Repeatabil
tionship Linking th
the Acquired
the factors
ned. To illu
2). That sho
ed by a point
the J=2 and
yers per bale
the bold circ
the situation
K=1 replica
re those show
effect of the
me variations
here the situa
seed cotton m
the Figure 2)
ne sample tak
gation risk lo
ill be availa
ale and the n
the lowest l
maE values
rom field to g
Sample testin
measuremen
lity of Standardi
he Standard Devia
Results
s layer an
ustrate these
ows the Sigm
t. Many poin
d N=J.K=1
e needs to be
cle showed
ns inside the
ate in the c
wn in the pre
crop season
s among the
ations with r
management
) we took our
ken from on
ower than 10
able for all c
number of te
litigation risk
are high and
gin, and of fi
ng, we rem
nts of Leng
zed Instruments
tion between Laye
nd replicate
e results, we
maA vs Sigm
nts are includ
1 ellipse tha
e increased i
less than 10
dotted ellips
omposite te
evious box pl
n (crop seaso
e situations.
roller gins w
t at the ginni
r fiber sampl
ne layer with
0% (those ins
characteristic
ests to be per
k as possible
d that some a
ibers from th
mind that one
gth/ Uniform
s for Testing Co
ers (SigmaA) vs. w
e differed
e display on
maE relation
ded in the J=
n in the J=
in the compo
0% litigation
se showed le
sting possib
lots.
on 1 or 2) an
There was
were often fa
ng mill at th
les and the sa
h one replica
side the limit
cs, we will
rformedper s
e. However,
adjustments
he gin stand t
e replicate co
mity Index, o
otton Fiber Prod
within Layer Stand
depending
ne example
nship for Mi
=1 and K=1
1 and K=2
osite testing p
n risk with 1
ess than 10%
bility. The tw
nd the prese
also a prob
ar from the o
he time (exam
aws in those
ate, we obse
t curve in bo
be able to d
sample in or
for some situ
will also be
to the pressed
orresponds t
of Strength,
duced in Africa
ard Deviation (Sig
on the tec
of the tec
icronaire wh
circle and m
ellipse. We
possibility. B
1 layer samp
% litigation ri
wo extreme
ence or absen
blem of diff
origin of the
mple of the p
gins.
erved that on
old).
deduce the n
rder to respec
uations, it ap
necessary in
d bale.
o one measu
of Color R
505

gmaE)
chnological
hnological
here the 65
more points
concluded
Bales from
pled and 1
isk with J
situations
nce of lint
ferences in
e chart, we
point in the
nly certain
number of
ct both the
ppears that
n the seed-
urement of
Rd and of

Fig. 2: Example of Micronaire: Standard Deviations between Layers (SigmaA) vs within Layer (SigmaE) for all Available Situations
Actual Limitations of the Study
In this study, we did not consider reproducibility conditions that could appear when results are
different from one classing laboratory to the next.
It will be also necessary to periodically quantify the within-bale variability for each situation in
order to ensure the litigation risk for any given situation.
Finally, we did limit the litigation risk to 10% for any single bale while commercial agreements
and contracts generally concern lots of several bales and the General Rules of Cotton
Associations; the lot litigation risk will have to be evaluated as well.
CONCLUSION
The measurement of within bale variability for six technological characteristics of cotton fibers produced
in fourteen cotton producing countries in Africa was done. Technological characterizations of these
samples were achieved in operating conditions that ensured the repeatability of the characterizations.
Within bale variance was categorized as sampling variance on one hand and as replicate variance on
the other. The goal is to propose sampling and testing procedures. Using the procedures proposed based
on African conditions; Africa will be able to start the instrumental classification of their cotton fibers
with a sampling and a testing procedure that respects international standards and with a low litigation
risk. This could also lead to improvements of seed cotton management and limit the risk of litigation. The
final aim is to give confidence in SITC results to final users in Africa as well as in any other place on
Earth.
ACKNOWLEDGMENT
The authors wish to thank the financial contributors who made this study possible. The study was
undertaken as part of project CFC/ICAC/33 Commercial Standardization of Instrument Testing of
Cotton, which was funded by the Common Fund for Commodities, an intergovernmental financial
institution established within the framework of the United Nations, headquartered in Amsterdam, the
Netherlands, and by the European Union in the framework of its "All ACP Agricultural Commodities
Programme under the sponsorship of the International Cotton Advisory Committee (ICAC) Washington
(USA) and implemented by the Faserinstitut Bremen (FIBRE), Germany.
The authors would like to thank the personnel of the Tanzania Bureau of Standards and Tanzania
Cotton Board Dar es Salaam, Tanzania on one side and the one from CERFITEX, Sgou, Mali who
performed all the fiber characterizations for the present study.
Finally, the authors want thank the African Cotton Companies who allowed us to take samples in
their facilities for running this experiment.
DISCLAIMER
This report was prepared within the project CFC/ICAC/33. The views expressed are not necessarily
shared by the Common Fund for Commodities and/or the European Commission and/or the International
Cotton Advisory Committee. The designation employed and the representation of material in this report
do not imply the expression of any opinion whatsoever on the part of the Common Fund for
Commodities and/or the European Commission or the International Cotton Advisory Committee
concerning the legal status of any country, territory, city or area or its authorities, or concerning the
delineation of its frontier or boundaries.
REFERENCES
[1] Abo M, Gourlot J-P, Goz E, Hubl P And Sinoimeri A. New Findings On Within Bale Repeatability Of Standardized
Instruments For Testing Cotton Measurements On Cotton Fiber Produced In West And Central Africa. Textile Research
Journal (Under Press). 2011.
[2] Sasser Pe And Smith Cb. High Volume Instrument Test System A Tool For Textile Manufacturing. 1984.
[3] Sasser Pe And Moore Jf. A Historical Perspective Of High Volume Instrument Developments In The U. S. Itmf-Icctm.
Brme1992, P. 215.
[4] Knowlton J. International Developments In Cotton Classification. Beltwide Cotton Conferences. New Orleans, Usa:
National Cotton Council Of America, 2005, P. 22547.
[5] Icac-Secretariat. Instrument Testing Of Cotton At The Producer Level For Trading Purposes. Washington D.C., Usa2011.
[6] Thibodeaux Dp, Senter H And Cui X. Within Bale Variations Of Cotton Fiber Properties. Beltwide Cotton Conferences.
Nashville, Usa: National Cotton Council Of America, 2008, P. 1542.
[7] Levrat R. Culture Commerciale Et Dveloppement Rural, L'exemple Du Coton Au Nord-Cameroun Depuis 1950.
Paris2010, P.292 P.
[8] Levrat R. La Culture Du Coton Dans La Zone Franc. Paris, France2010, P.264 P.
[9] Davidonis Gh, Johnson A, Landivar Ja And Hood Kb. The Cotton Fiber Property Variability Continuum From Motes
Through Seeds. Textile Research Journal. 1999; 69: 7549.
[10] Davidonis Gh, Johnson As And Johnson Rm. Quantification Of Within-Plant And Within-Field Yield And Fiber
Variability. Crop Management. 2004; Http://Ddr.Nal.Usda.Gov/Bitstream/10113/11887/1/Ind43806179.Pdf.
[11] Clouvel P, Goz E, Sequeira R, Dusserre J And Crtenet M. Variability Of Cotton Fiber Quality. New Frontiers In Cotton
Research: Proceedings - Washington: Icac, 2000. 2000.
[12] Meyer V.G. And Meyer Jr. Some Sources Of Variability In Boll And Fiber Properties Of Cotton (Gossypium Hirsutum L.).
Crop Science. 1970; Vol. 10: Pp. 699702
[13] Dimitrova L. And Bozhinov M. Variability Of Cotton Fiber Properties Caused By Genotype And Some Environmental
Factors. Plant Science. 1988; Vol.25: N9: 2734.
[14] Gourlot J-P. Les Tendances Dans La Standardisation Du Coton Sur Le March Mondial. Rle Et Place De La Recherche
Pour Le Dveloppement Des Filires Cotonnires En volution En Afrique: Actes - Montpellier: Cirad, 2000. 2000,
P. 1136.
[15] Usda. Cotton Ginners Handbook. Agricultural Handbook N 503. Washington, D.C., Usa1977, P. 110.
[16] D5867 A. Standard Test Methods For Measurement Of Physical Properties Of Cotton Fibers By High Volume Instruments.
In: Materials Asfta, (Ed.). Annual Book Of Astm Standards. Philadelphia, Pa (Usa) 2005, P. 88693.
[17] Saporta G. Probabilit, Analyse Des Donnes Et Statistique. 2 dition Ed. 2006, P.P. 1542.
[18] Philippeau G. Thorie Des Plans D'exprience, Application L'agronomie. 1989: 205.

A Vision for Technical Textiles in this Decade

A. Subramaniam
Retd-Scientist, Madupra Coats, Consultant, Coimbatore, India
INTRODUCTION
Technical Textiles can be defined in simple term As Textiles any other than Apparel or Furnishing
Textiles. This could be Medical Textile, Agri, Building and Geo Textiles etc.
COTTON FABRICS
Woven, Non-woven, Knitted or any other manufacture items were from immemorial days for their usage
in Medical and Agricultural areas. During last 20
th
century, Cotton through yarn and fabrics contributed
substantially to Industrial / Technical Textiles in terms of Tyre cord for tyres from Egyptian cottons and
from short staple Indian cotton of 1 staple (25.4mm) to produce well known Beltings of World
standards for transmission and conveyor usage and fabrics for Tents to Defense and Tarpaulin for
domestic purposes. But during the later part of the last Century, Synthetic and other Man-made Glass
filament gradually replaced cotton due to their superior strength and desirable qualities for Technical
Textiles. Still Cotton yarn was useful to cover Synthetic for adhesion purpose with Rubber for example
Cotton/Nylon Beltings for conveyor. Cotton fibre and yarn are still in use without any Physical and
Chemical treatment in western countries blending with Synthetics at 3 to 5% to manufacture parts for
Auto Industry of Bonnet and Doors and expected to increase further by 10% during this decade.
CONTRIBUTION BY INDIAN COTTON BREEDERS
During the last century, tremendous progress had been achieved by the Indian Breeders adopting new
techniques in their breeding work. As a result, new varieties of cotton, such as, Shankar 4 & 6, then
Varalaxmi, all in the range of long staple contributing high productivity. Later, SUVIN the highest extra
long staple of 38-40mm came into production recording very high fibre strength, better than Egyptian
cotton of KARNAK and MINOFI. All these Indian variety cottons served in the Apparel Industrys
requirement very well.
This writer was very much involved in consuming these varieties for Sewing Thread and high quality
fabrics like Poplin, spinning 100s to 120s yarn from SUVIN.
REQUIREMENT OF TECHNICAL TEXTILES
The first and foremost quality requirement is fibre strength and fibre strength alone for Technical
Textiles. Count range require of Technical Textiles are from 6s, 10s, 12s, 16s and 20s count maximum
and do not require long staple cotton, as they are unnecessarily for their FITNESS OF PURPOSE. Hence,
this writer suggest that our Breeders should concentrate in developing a short staple cotton with a
maximum staple length of 1 (25.4mm) and of short duration, preferably from Rain-fed areas with high
yield. Breeders should attempt fibre strength of 25 gms/tex and above. If this can be achieved by
sustained effort and thrust is given on an emergency basis during this decade, then India can be Leader in
developing such short staple cotton. Formers will enthusiastically welcome such high yield varieties and
get a better price, as the Industry will benefit to utilize them in High Tech areas, not only at 100% level
but also in components with Synthetic fibre/filament.
83
A Vision for Technical Textiles in this Decade 509
PRESENT WORK BY THE WRITER
This writer had done some development work to prove a point that a short staple of high strength cotton
recording 23-25 gms/tex instead 20 gms/tex achieved today. This is done by using the by-products of
long staple cotton while it is manufactured in the yarn and spinning them to Coarser count from 6s to 20s
and woven into fabric. Interesting results will be shown during the World Cotton Conference to
convince the Cotton Breeders that a short staple high yield fibre can revolute with high strength fibre of
23 to 25 gms / tex to the prosperity of Formers and Industrialists of Textiles.
Cotton Stalk: An Additional Raw Material

to Board Industry
R.M. Gurjar, P.G. Patil, A.J. Shaikh and R.H. Balasubramanya
Central Institute for Research on Cotton Technology, Mumbai, India
AbstractA supply chain model comprising collection, cleaning and chipping of cotton stalks and their
transportation to board manufacturing factories has been evolved. Pilot plant trials and large scale industrial trials
have been conducted for making particle board and hardboard from cotton stalk. The techno-economic feasibility of
manufacturing boards from cotton stalk has been demonstrated to board industry which is now convinced about its
potential. In the not-very-distant future, many particle board manufacturers are likely to start using cotton stalk as an
alternative raw material in place of sugarcane bagasse and hardwood. Once the industrial use of this agrowaste picks
up, the farmer would be able to earn additional income from the sale of cotton stalk to the particle board manufacturers.
Growth of industrial activity, rural employment generation and conservation of forest resources are other national
benefits that are bound to follow.
INTRODUCTION
Environmental concerns have, in recent years stimulated researches in the exploitation of renewable
resources. Such researches make better economic sense when they relate to utilization of waste materials
like cotton stalk. With its vast area under cotton cultivation, India is undoubtedly the largest producer of
cotton stalk among world countries and stands to benefit immensely from commercial exploitation of this
putative agrowaste. Profiled in this paper is CIRCOTs saga of R & D efforts that have revealed the
economic potential of cotton plant biomass abundant in many Afro-Asian countries.
King cotton rules the world of textiles despite inroads made by synthetic fibres. The economy of
about 90 cotton growing counties is greatly influenced by cotton. Cultivated in over 30 million ha, the
annual world cotton production is about 22 million tonnes constituting 36% of the total fibre production
and consumption.
In recent years, India has emerged as the second largest producer of cotton next to China. The cotton
production in 2009-10 in India stands at 5.1 million tonnes as against Chinas 6.8 million tonnes. Other
major producers are USA, Pakistan, Brazil, Uzbekistan, Turkey, Australia, Turkmenistan, Greece, Syria
and Egypt.
In area under cotton cultivation, India tops the world list (10.1 Mha) with China (5.4 Mha) and USA
(3.1 Mha) closely following. In productivity, however, India lags behind most other countries though in
recent years, there has been substantial improvement.
DISTRIBUTION OF COTTON IN INDIA
TABLE 1: COTTON CROP IN DIFFERENT STATES IN INDIA (2010-11)
State Production of Cotton (In Lakh Bales) Area (Million Ha)
1 Gujarat 90.00 2.422
2 Maharashtra 62.00 3.194
3 Andhra Pradesh 53.00 1.138
4 Punjab 17.50 0.604
5 Haryana 14.00 0.483
6 Madhya Pradesh 18.00 0.630
7 Karnataka 9.00 0.402
8 Rajasthan 7.50 0.339
Table 1 (Contd.)
84
Cotton Stalk: An Additional Raw Material to Board Industry 511
Table 1 Contd.
9 Tamil Nadu 5.00 0.119
10 Orissa 1.50 0.050
11 North 39.00 1.426
12 Central 170.00 6.246
13 Southern Region 67.00 1.659
14 Others 0.50 0.058
15 Loose 12.00
Total 290.00 9.436
Two leading cotton growing States in India are Gujarat and Maharashtra which respectively account for
22% and 32% of total area under cotton in the country (Table 1). The average yield is much higher in
Gujarat than in Maharashtra on account of better irrigation in the former. These two States together
contribute over 50% of Indias cotton crop.
CROPPING SEASON
In the northern States of Punjab, Haryana and Rajasthan which are largely irrigated, cotton is harvested in
the two months of October & November. Farmers in these areas cut away cotton plants even if some
green bolls are still left, so as to clear the land early for the ensuing wheat crop. Stalks are removed in
about a months time. In other regions, which are mostly rainfed, harvesting of cotton takes place from
October to February. Cotton stalks are uprooted from around FebruaryMarch and the process goes on
till May-June since there is no pressure to vacate the field for any other crop.
LOW INCOME FROM COTTON FARMING
Relatively low yield in rainfed areas has rendered cotton farming somewhat unremunerative in India.
Most farmers are unable to make a living out of cotton cultivation. Ways and means to increase the
returns from cotton farming, therefore, need to be explored.
RESEARCH ON BY-PRODUCT UTILISATION
The diverse products available from cotton crop after the harvest of seed-cotton and ginning include seed,
linters, hulls, oil and meal which are classified under the broad head by-produce. Cotton stalk is the
other biomass available in the field after the harvest of seed-cotton. In general, there is lack of focus on
judicious utilisation of cotton by-produce not only in India but also in all the Afro-Asian countries whose
economies are influenced by cotton. The bulk of cotton seed is subjected to what is known as whole seed
crushing for extraction of oil and cotton stalks are disposed of by burning in the field itself as otherwise
they would harbour several insects and pests which would be harmful for the future crop. A small
fraction of seeds is consumed as cattle feed while some of the stalks is used as domestic fuel. In whole
seed crushing valuable components like linters, hulls, protein and large fractions of oil go unutilised
instead of fetching the much-needed additional returns to farmers. Equally unacceptable is the burning
off of cotton stalks that have in recent years proved to be of immense economic potential.
COTTON STALK
It is estimated that about 25 million tonnes of cotton stalk is generated in India every year. Most of the
stalk produced is treated as waste though a part of it is used as fuel by rural masses. The bulk of the stalk
is burnt off in the field after the harvest of the cotton crop as pointed out earlier. Cotton stalk contains
about 69% holocellulose, 27% lignin and 7% ash.
In contrast to other agricultural crop residues, cotton stalk is comparable to the most common species
of hardwood in respect of fibrous structure
3
and hence it can be used for the manufacture of particle
boards, preparation of pulp and paper, hard boards, corrugated boards & boxes, microcrystalline
cellulose, cellulose derivatives and as substrate for growing edible mushrooms.
TECHNOLOGY FOR PARTICLE BOARDS FROM COTTON STALK
Research work on the preparation of particle boards in CIRCOT dates back to 1979-80 when cotton stalk
chips were used for the first time (Pandey and Mehta, 1979). Detailed studies have since been made to
arrive at the appropriate process sequence and to identify process parameters that would ensure the
required qualities for the particle board (Pandey and Mehta, 1980; Gurjar, 1994). The process involves
the following steps:
Chipping of stalks to 1.5 - 2.0 cm size;
Rechipping to particles of 20 mesh size and 8 mesh size;
Mixing of chips with synthetic binder such as urea formaldehyde or phenol formaldehyde;
Preparation of a three-layered mat comprising coarser particles for the core layer and finer ones
for the top and bottom layers;
Pressing the mat between heated platens of a hydraulic press for specific time and pressure.
The board thus made is cooled to attain dimensional stability and then cut to the desired size. By
using different chemicals and additives, the boards can be made water proof, fire proof, termite resistant,
etc. These boards have been found to meet BIS specifications in respect of quality characteristics. Due to
the lower cost of raw material and reduced power required for its conversion into finished product, the
cost of particle board made from cotton stalk will be much lower than that of boards made from wood.
The data presented in Table 2 clearly show that the particle boards from cotton stalks possess all the
desirable properties to be used for internal as well as external applications such as false ceiling,
partitioning, paneling, etc.
TABLE 2: PROPERTIES OF THREE-LAYERED PARTICLE BOARDS FROM COTTON STALK
Sr. No. Properties Unit Flat Pressed Three-layer/
Multilayer Particle Board IS
3087-1985
Cotton Stalk
Particle Board
Type I Type II
1 Density Kg/m3 500 900 ---- 750
2 Average Moisture % 5-15 --- 11
3 Water Absorption 2 h soaking 24 h soaking % 10-20 40-80 20-40
4 Swelling Thickness % 8 12 9
5 Swelling due to surface Absorption % 6 9 6
6 Modulus of Rupture (MOR) Up to 20 mm
Above 20 mm
N/mm2 15.0-12.5 11.0-11.0 17.6
7 Internal bond strength Up to 20 mm Above 20 mm N/mm2 0.45-0.40 0.3-0.3 0.51
8 Screw withdrawal strength Face Edge N 1250-850 1250-700 1400-860
9 Nail withdrawal strength N 1250 --- 1300
Process Parameters and Product Quality
TABLE 3: PROPERTIES OF PARTICLE BOARDS FROM COTTON STALKS WITH UREA FORMALDEHYDE AS BINDER
Resin Content
(%)
Thickness (mm) Density (kg/m3) Modulus of Rupture
(N/mm2)
Water
Absorption (%)
Swelling due to Surface
Absorption (%)
0 7.1 700 6.1 77 28
3 7.4 720 7.4 57 22
5 7.6 760 9.3 42 18
8 8.0 780 12.4 33 12
10 8.2 820 13.3 31 10
12 8.4 840 13.9 28 9
13.5 8.5 840 17.4 25 8
15 8.9 880 18.6 22 6
Increase in resin content results in improvement of product performance. Density of boards and modulus
of rupture are found to increase while water absorption and surface swelling show progressive decline as
the resin content is increased. Data in Table 3 demonstrate that by altering the process variables, it is
possible to get particle boards of any desired quality.
Uses of Particle Boards
The applications of particle boards are many. The application areas identified include door panel inserts,
partitions, wall panels, pelmets, furniture items, floor and ceiling tiles, etc. for residential houses,
commercial buildings, schools, hotels, theatres, etc. In recent years, particle board is being used
increasingly in place of commercial plywood in the preparation of printer blocks.
In all the above applications, substitute materials for particle boards are timber, commercial plywood,
marine plywood and block board in general and for false ceilings in place of plaster of Paris. The
advantages of particle board are many:
It is free from natural defects of wood, like tendency for warping.
It is easier to fix. For instance, the factory-made panel doors from particle board are available in
a ready-to-fix form. Similarly, for wall panelling, false ceilings, table tops, etc., pre-laminated or
pre-veneered particle boards can be used with advantage.
It is cheaper than substitute materials.
With proper protective surface coating and edge covering, particle board can be made termite
proof and fire resistant. It can take a variety of surface finishes, like laminations, veneers, paint,
varnish, polish, etc. Attractive wall paper can also be used as surface finish for particle boards.
Even though the process for the preparation of particle boards from cotton stalks was developed a
decade ago, it was not accepted by the boards industry for commercial adoption. Some of the important
reasons being
Absence of cost effective supply chain mechanism
Absence of pilot plant facility for fine tuning of technology and demonstration
Non availability of data on techno-economic feasibility of the process.
To address some of the above issues CIRCOT had submitted a project proposal to the Common Fund
for Commodities (CFC), Netherlands, through the International Cotton Advisory Committee, USA for
seeking financial assistance. The CFC realized the importance and potential of the project and sanctioned
an amount of US$ 918,886 for undertaking the above study. Some of the important achievements made
in the project are described below:
ESTIMATION OF AVAILABILITY OF COTTON STALK
In Maharashtra, where cotton crop is grown mostly under rainfed conditions, stalks yield was as low as 1
to 1.5 tonnes per ha. The stalks yield from Karnataka ranged from 1 to 2.5 tonnes/ha in case of rainfed
crop and up to 4 tonnes/ha from irrigated fields. The data clearly showed that the yield of cotton stalks
from Maharashtra is the lowest. It was also noted that cotton type (variety/hybrid) plays a very important
role besides conditions of growth. The information on State-wise availability of cotton stalks is given in
Table 4.
The survey revealed that in the North, the major part of cotton stalks is used as domestic fuel.
However, farmers were ready to part with at least 50% the cotton stalk available with them for a payment
of Rs. 400 to 500 (US $ 8 to 10 ) per tonne. In Gujarat the stalks are mostly burnt in the field itself. In
Maharashtra, though farmers are using the stalks as household fuel, they were willing to sell it for as low
as Rs. 300 to as high as Rs. 500 per tonne. In Karnataka, the survey showed that most of the farmers use
cotton stalk as fuel. Some of them exchange the stalks for farm yard manure. They expressed their
willingness to sell the stalks for a price of Rs. 500 per tone (1 US$ = Rs. 45/-)

TABLE 4: AREA UNDER CULTIVATION, PRODUCTION OF INDIAN COTTON & AVAILABILITY OF COTTON STALKS IN INDIA (2010-11)
State Production of Cotton
(In Lakh Bales)
Area (Million Ha) Availability of Stalks
(Million Tonnes)
1. Gujarat
2. Maharashtra
3. Andhra
Pradesh
4. Punjab
5. Haryana
6. Madhya
Pradesh
7. Karnataka
8. Rajasthan
9. Tamil Nadu
10. Orissa
11. North
12. Central
13. Southern
14. Region
15. Others
16. Loose
90.00
62.00
53.00
17.50
14.00
18.00
9.00
7.50
5.00
1.50
39.00
170.00
67.00
0.50
12.00
2.422
3.194
1.138
0.604
0.483
0.630
0.402
0.339
0.119
0.050
1.426
6.246
1.659
0.058
9.69
9.58
3.41
3.00
2.42
1.89
1.21
1.70
0.36
0.15
7.12
21.16
4.98
0.17

Total 290.00 9.436 33.58
COLLECTION AND CLEANING OF COTTON STALK
Cotton is a seasonal crop harvested in India from October to February. Cotton stalks which are available
only between December and May will require storage over several months to ensure adequate raw
material supplies to board manufacturers for the entire year's production.
Cotton stalks are bushy in nature and have very low bulk density. Collection and transportation are,
therefore, expensive. Further, on storage in stick form, cotton stalks get degraded by insect attack.
Success of cotton stalks as an industrial raw material would depend on the establishment of a sustainable
supply chain to reach them to the industry.
The entire cost economics of board manufacturing technology and the acceptance of cotton stalk by
industry will depend to a large extent on the cost of raw material in a readily usable form made available
at the factory gate. Therefore, the logistics of economic collection of cotton stalks, chipping and
transportation from field to industry, and its proper storage in different forms at various centres are
crucial factors that decide the economic viability of this raw material.
Trials were conducted in three successive seasons to arrive at the most economic mode of cotton
stalk collection in and around Nagpur where the crop is raised under rainfed conditions. In this region
after the picking of seed cotton, the stalks are not cleared from the field immediately since there is no
subsequent crop. The study started right from the stage of sensitizing farmers about the utility of cotton
plant stalks.
It is comparatively easier to uproot the stalks immediately after the picking is over since the moisture
still present in the soil facilitates uprooting. A metallic device available locally helped in uprooting the
stalks effortlessly as compared to manual pulling. It has been observed that as many as 7-8 labourers
could clear the stalk from one hectare of land in a day of 8 hours. Although there was no significant
difference between manual pulling and uprooting with the help of the mechanical device in respect of
speed, the drudgery involved in bending and pulling with hands could be avoided in the latter case. In
view of this, the simple mechanical device is recommended for uprooting the stalks. The stalks thus
collected should be left for 4 to 5 days in the sun during which time, the leaves are shed. The boll rinds
can be removed by gently beating the stalks on a wooden mallet. The cleaned stalks could then be
subjected to chipping at a nearby chipping centre.
Three different models were attempted initially for economic collection and transportation of cotton
stalks at three different locations near Nagpur:
Among the various models attempted, the most suitable model is the third model which
comprises uprooting of stalk, storage, manual cleaning, chipping by use of a tractor-driven
chipper at a centralized chipping centre not farther than 5 km from the field and transportation of
chips to the factory within 50 km distance by a truck.
Transporting cotton stalks beyond 5 km before chipping and beyond 50 km after chipping so as
to make it available in an appropriate form to the industry would not be an economically feasible
endeavour.
On an average, the cost of cleaned and chipped stalks to be made available at the factory gate
situated within 50 km from the production centre (farm) would workout to about Rs.1500-2000
(US $ 30.0 40.0) per tonne of the raw material with 10% of moisture.
COST OF READY-TO-USE COTTON STALK CHIPS
Using the large volume of data on collection, chipping and transportation of cotton stalks, it has been
possible to work out the economics of each of these operations. From this analysis the cost of cotton stalk
chips made available at the particle board factory has been arrived at. Details are given in Tables 5 and 6.
The raw material cost for particle board manufacture from cotton stalk thus works out to Rs. 1960 (US $
39.2) per tonne of the ready-to-use material.
TABLE 5: COST OF COLLECTION AND CHIPPING OF COTTON STALKS
Operations Cost Per Tonne
Rupees US$
Uprooting and cleaning 500 10.0
Chipping 230 4.6
Tractor hiring 360 7.2
Total 1090 21.8
TABLE 6: TOTAL COST OF READY-TO-USE COTTON STALK CHIPS DELIVERED AT FACTORY SITE
Operation Cost Per Tonne
Rupees US$
Labour charges for uprooting, cleaning and
chipping
1090 21.8
Transportation charges 320 6.4
Loading and unloading charges 50 1.0
Raw material cost 500 10.0
Total 1960 39.2
TRANSPORTATION OF STALKS AND CHIPS
A critical study of the logistics of cotton stalk collection, chipping and transportation has revealed the
following facts:
Transporting chipped cotton stalk is more economical than transporting the stalk as such.
It would be appropriate to employ bullock carts and tractor trolleys to carry cotton stalk to the
chipping centre and use lorries to deliver the chips at the factory.
Transporting distance plays a major role in deciding the effective cost of the raw material.
Fifty kilometers should be considered as the maximum permissible distance for economic
transportation of chipped material.
Storage Trials
In order to find out the shelf life of cotton stalks, a large quantity of the unchipped material was stored in
the open, on a stone platform. Similarly, two lots of chipped cotton stalks packed in gunny bags were
also stacked, one lot in the open and the other inside a godown. Observations were made every month for
colour and insect attack, and chemical analysis was done once in a month to find out the changes if any in
chemical composition (Table 7). The following facts emerged from this trial:
Insect attack is rampant in unchipped stalks kept in shade or in the open.
No significant change in the chemical composition occurs in the case of chips stored in godown.
Chipped stalks are not susceptible to insect attack.
A marginal reduction in holocellulose content is noticeable in chips stored in the open.
TABLE 7: DATA FROM CHEMICAL ANALYSIS OF STORED COTTON STALK CHIPS
Sl. No. Month Moisture (%) Lignin (%) Holo Cellulose (%) Ether Extractives (%)
A B A B A B A B
1 July 14.2 16.0 26.6 25.4 82.1 77.1 7.1 7.2
2 August 14.0 15.9 26.1 25.0 81.4 76.4 7.0 7.1
3 September 12.0 11.9 26.0 24.8 81.2 75.7 7.0 7.2
4 October 11.2 12.9 25.9 24.5 81.1 75.4 6.5 6.8
5 November 11.8 11.4 25.8 24.2 80.9 75.2 6.8 7.1
6 December 11.1 11.2 25.5 24.1 80.7 75.2 6.7 6.5
7 January 11.3 11.4 25.2 24.1 80.5 75.1 6.7 6.8
8 February 11.3 11.4 25.5 24.7 81.1 75.0 6.5 6.3
9 March 11.2 11.4 25.5 24.6 80.9 74.7 6.9 4.5
10 April 11.0 11.1 25.6 24.5 80.5 75.1 6.6 6.4
11 May 10.1 11.5 25.3 24.7 80.4 74.9 6.5 6.4
12 June 13.2 14.1 25.2 24.4 81.0 74.8 6.4 6.3
A: Stored in shed; B: Stored in the open
A MODEL COTTON STALK SUPPLY CHAIN FOR A 20 TPD PARTICLE BOARD PLANT
It is known that for producing 1 tonne of boards, about 1.5 tonnes of chips are required. Therefore for
running a 20 TPD particle board plant, 30 tonnes of chips would be required each day. Our studies have
shown that it is possible to get about 1.5 tonnes of ready-to-use chips from one hectare land around
Nagpur. Hence if a factory is to run only on cotton stalks it is necessary to get the material from 6000 ha
of farm land which will provide 9000 tonnes of chips for board production in a 20 TPD plant working for
300 days in a year.
Storage
CIRCOT study has shown that cotton stalks are normally uprooted in Nagpur area when the plant is
almost dry (devoid of leaves). If such plants are uprooted and left in the field for three days and manually
cleaned to remove the boll rinds before being subjected to chipping, the chipped material would be left
with a moisture of around 12%. During transportation to the factory, the percentage of moisture stabilizes
at about 10%.
Considering that the chips have to be stored in the factory premises for at least one month, about 900
tonnes are to be stacked in its premises and the rest to be stored in 9 decentralised places by groups of
farmers (say in 9 villages connected well with transport services).
Chipping Stations
It has been estimated that about ten chipping stations are required to be set up. Each chipping station
must be provided with one tractor-driven mobile chipper (outsourcing). Each chipper has an output rate
of about 500 kg/h and can provide about 3-4 tonnes per day and in a month it is possible to generate
about 90-120 tonnes of chips. Each chipping station will have to store about 1000 tonnes of chips. These
chips will be stored in three stockpiles of 3 metres height and each pile is to be covered by polythene
sheets to prevent spoilage during rainy season. The stock piles will be adequately separated from one
other so as to facilitate loading of chips in trucks.
The bulk density of cotton stalk chips is about 0.14 g/cc. The average area occupied by a 3-metre
high stockpile would be around 70 m
2
. The space required in each chipping centre would, therefore, be
around of an acre.
Cotton Stalk Collection for Each Chipping Station
As said earlier, about 1000 tonnes of chips are to be generated and stored in each chipping centre. For
this, stalks must be collected from 600-700 hectares of land. Based on earlier trials at CIRCOT, four
labourers can uproot and collect stalks from one acre in a day. This means, 10 persons are required to
uproot the stalk available in 1 hectare. This also means that 10 persons would get employment for one
week, only for uprooting. The same number of persons are required for cleaning the material as well.
Chipping will employ four persons daily for one month.
Supply of Cotton Stalk Chips
The chips will have to be transported to the factory under the direct supervision of the factory itself to
ensure that supply takes place at the required rate.
ESTABLISHMENT OF 1 TPD PILOT PLANT
One-tonne-per-day (1 TPD) pilot plant was commissioned for preparation of particle boards. Accordingly
a pilot plant of indigenous design was procured and installed at the Ginning Training Centre of CIRCOT
at Nagpur (India). The layout of the pilot plant is shown in Fig. 5 while the process sequence is illustrated
in the flow-chart in Fig. 6. The pilot plant comprises an array of several machines :
Hammer Mill
Drum Chipper
Rechipper
Rotary Dryer
Glue Blender
Mat Former
Pre Press (cold)
Hydraulic Hot Press
Cutting Machine
Sanding Machine

Fig. 5: Layout of Pilot Plant
The component machines except the last two are linked by conveyors that transfer the material from
one stage to the next till the boards are formed.
Pilot Plant at GTC Nagpur

Fig. 6: Flow-Chart of Particle Board Pilot Plant

Fig. 7
Material Balance
After optimizing the process parameters on the pilot plant, regular production trials were undertaken and
boards of various thicknesses, densities, etc. were made. On the basis of these production trials and the
systematic data thus collected, a material balance for preparation of particle board from cotton stalks was
worked out, as shown in the following chart.
Material Balance for the Preparation of Particle Boards from Cotton Plant Stalks

Fig. 8
The most significant facts that emerge from the chart are the following:
One tonne of cleaned cotton stalk chips with 10% moisture yields 0.7 tonne of plain boards
with 6% moisture.
To prepare 1 tonne of plain boards with 6% moisture, about 1.4 tonnes of cleaned cotton
stalk chips with 10% moisture are required.

Fig. 9: Particle Boards Produced on Pilot Plant
COMMERCIAL TRIALS
CIRCOTs particle board technology subjected to refinement on the pilot plant was tried on an industrial
scale in large board manufacturing units. In the first instance, about 30 tonnes of ready-to-use cotton stalk
chips were supplied to M/s Ecoboard Industries Ltd. at Velapur near Pandharpur in central India followed
by a second lot of 50 tonnes. The chips were delivered from Nagpur, and boards of 13.5 x 6 size were
prepared with different thicknesses (9 mm, 12 mm & 18 mm) and surface finishes. Laminated boards
thus manufactured were used for making different furniture items and also for panelling some rooms in
CIRCOT, Mumbai, GTC of CIRCOT, Nagpur, DOCD, Mumbai and ICAR Headquarters, New Delhi.
Tests results shown in Table 8 indicate that it is possible to prepare good quality boards in the existing
industry without any modification.
TABLE 8: PROPERTIES OF COTTON STALK BOARDS MADE IN INDUSTRIAL TRIALS
Properties Interior Grade Boards BIS Specification
12 mm 18 mm
Density (kg/m3) 713.1 699 500-900
Modulus of Rupure (N/mm2) 15.4 12.0 11.0
Tensile Strength (N/mm2) 0.9 0.5 0.3
Water Absorption (%) 2 hours 20.4 56 40
Water Absorption (%) 24 hours 42.2 95 80
Screw Withdrawal (N)Face 1762 1610 1250
TECHNO-ECONOMIC FEASIBILITY OF PARTICLE BOARD PLANTS
The technical feasibility of particle board manufacture from cotton stalk was examined on the basis of
industrial trials conducted at M/s Eco-board Industries Pvt. Ltd., Pune and Archid Ply Industries, Mysore.
In both the industries, the trials were successful and good quality boards suitable for lamination were
manufactured. No technical problems were encountered during the processing and no changes or
modifications in the existing plant were found necessary.
For an examination of the economic viability of cotton stalk as raw material for particle board
manufacture, a 30-tonne trial was undertaken at M/s Eco-board Industries Pvt. Ltd., Pune having installed
capacity of 200 tonnes/day but running at 60% capacity utilization. In these trial boards of 9 mm, 12 mm
and 18 mm were prepared. The data in Table would substantiate the fact that particle board production
from cotton stalks in an established board manufacturing unit is indeed an economically viable
proposition.
Thickness Density (g/cc) Production Cost Per sq. ft. Selling Price Per sq. ft. Profitability %
Rs. US $ Rs. US $
9 mm 0.73 13.94 0.28 17.1 0.34 22. 7
12 mm 0.72 15.00 0.30 18.63 0.37 24.2
18 mm 0.72 19.25 0.38 24.53 0.49 27.4
COST ESTIMATION FOR A PARTICLE BOARD PLANT OF 10 TPD CAPACITY
On the basis of extensive information gathered from wide ranging R & D efforts under the project, it has
been possible to work out the profitability of particle board plants. For a 10 TPD plant the capital
investment comprising land, buildings and plant & machinery will work out to over Rs. 58 million (US $
1.16 million) while the production cost after duly considering depreciation, interest on investment etc
would be around Rs. 38.76 million (US $ 0.78 million). The cost of production per tonne of particle
board would be about Rs. 12,920 (US $ 258). At current selling price levels for particle boards, the return
on investment will work out to about 19%. Details are given below in tabular form.
Production Highlights
Production capacity: 10 TPD (384 boards of 8x4x12 mm),
Raw material used:15 TPD of cleaned chips/day
No. of days of production in a year : 300
No. of shifts per day: 3
Total production in a year: 3000 tonnes
TABLE 9: PROJECT COST AND PRODUCTION COST IN CASE OF A 10 TPD PLANT
A Capital Investment Rs. (Million) US $
1. Land & Building
Land : about 1 hectare 2.50 50000
Building : (Area : About 10,000 sq.ft., raw material storage) 4.50 90000
2. Plant and Equipment 42.20 844000
Table 9 (Contd.)
Table 9 Contd.
3. Auxiliary and service Equipments, Margin money for working capital 90.00 180000
Total Project Cost 58.20 1164000
B Cost of Production
1. Raw Material & Utility 20.50 410000
2. Labour & Supervision 3.51 70200
3. Repairs , Maintenance & Overheads 2.80 56000
(I) Total Manufacturing Cost 26.81 536200
(II) General Expenses 1.75 35000
(III) Depreciation & Interest 10.20 204000
Total cost of production B (I+II+III) 38.76 775200
Cost of production per tonne of board Rs.12,920 258
TABLE 10: PROFITABILITY OF A 10 TPD PARTICLE BOARD PLANT
Rs. (Million) US $
1. Gross Annual Income 49.77 995400
2. Annual Cost of Production 38.76 775200
3. Annual Return (2-3) 11.01 220200
4. Return on Investment (ROI) 19% 19%
*Selling price per unit (8x4x12 mm) @ Rs. 13.5 per sq. ft. = Rs. 432.00 (US $ 8.64)
COST ESTIMATION FOR A PARTICLE BOARD PLANT OF 20 TPD CAPACITY
A similar exercise has been done in respect of a 20 TPD plant for which the capital investment including
cost of land, buildings and plant & machinery works out to over Rs. 75 million (US $ 1.50 million). The
production cost, after taking into consideration depreciation, interest on investment etc., works out to
about Rs. 74.60 million (US $ 1.49 million). The production cost per tone of particle board would be
about Rs. 12430 (US $ 249). The return on investment would be about 33.5% which is significantly
higher than for a 10 TPD plant. Details are shown in tabular form.
Production Highlights
Production capacity: 20 TPD (770 boards of 8x4x12 mm)
Raw material used: 29 TPD of cleaned chips/day
No of days of production in a year: 300
No. of shifts per day: 3
Total production in a year: 6000 tonnes
TABLE 11: PROJECT COST AND PRODUCTION COST IN CASE OF A 20 TPD PLANT
A Capital Investment Rs. (Million) US $
1. Land & Building
Land : about 1 hectare 2.50 50000
Building: 25,000 sq.ft. 9.05 181000
2. Plant and Equipment 50.00 1000000
3. Auxiliary and service Equipment 2.50 50000
4. Margin money for working capital, pre-operative expenses,
contingency etc.
11.17 223400
Total Project Cost 75.22 1504400
B Cost of Production (80% capacity utilization)
1. Raw Material & Utility 41.50 830000
2. Labour & Supervision 5.50 110000
3. Repairs & Maintenance 2.70 54000
4. Plant overheads 1.00 20000
(I) Total Manufacturing Cost 50.70 1014000
(II) General Expenses 3.10 62000
(III) Depreciation & Interest 20.80 416000
Total cost of production B (I+II+III) 74.60 1492000
Cost of production per tonne board Rs. 12430 249

TABLE 12: PROFITABILITY OF A 20 TPD PARTICLE BOARD PLANT
Rs. (Million) US$
1. Gross Annual Income 99.79 1995800
2. Annual Cost of Production 74.60 1492000
3. Annual Return (2-3) 25.19 503800
4. Return on Investment (ROI) 33.5% 33.5%
*Selling price per unit (8x4x12 mm) @ Rs. 13.5 per sq. ft. : Rs. 432.00 (US $ 8.64)
CONCLUSIONS FROM THE TECHNO-ECONOMIC STUDY
A particle board plant with an installed capacity of 10 tonnes per day and involving a capital
investment of about Rs. 60 million (US $1.6 million) can ensure a profitability of about 20%
with cotton stalk used as the raw material.
A plant with a higher capacity of 20 TPD can bring higher returns of up to 33%.
For sustainable supply of raw material, an agency should be identified for organizing collection
and chipping of cotton stalks and delivering the chips at the particle board factory.
Existing particle board plants manufacturing boards from hardwood, bagasse, etc. can use cotton
stalk as an additional raw material.
POST PROJECT SCENARIO
The successful completion of the project in 2009 encouraged many farmers, entrepreneurs and board
industries particularly in Maharashtra and Gujarat to collect stalks for use in board making.
a) M/s. Godavari Particle Board Palnt (10 TPD) near Nanded in Maharashtra have been using
cotton stalks for the preparation of particle boards. They are preparing blended boards also
(cotton stalks and bagasse). They are able to collect about 500-1000 tonnes of cotton stalks
through an organized collection mechanism. This exercise provided employment specifically to
landless labourers. About 100 farmers are involved daily in collection, chipping and
transportation.
b) A 100 TPD particle board plant based on cotton stalks was commissioned in 2009 by M/s. Rushil
Decor Ltd. In Dhrangadhra village in Surendranagar District, Gujarat. Their annual requirement
of cotton stalks is 50,000 tonnes. They have identified six cotton stalk collection centres, each
centre covering an area of 8000 ha to 25,000 ha in a distance of about 15-20 km. They identified
contractors, panchayat representatives, self help women groups and Rural Development
Agencies to undertake this job. This plant with innovation collection mechanism generated
4,00,000 man days employment annually benefitting 1000 farmers and 5,000 landless labourers.
c) Based on CIRCOT technology, CIRCOT in collaboration with MITCON consultancy services
prepared a bankable project proposal for setting up of a 10 TPD particle board manufacturing
plant from 100% cotton stalk at Washim, Maharashtra. Ministry of Social Welfare, Govt of
Maharashtra sanctioned a loan of 680 lakhs to Tulsai Magasvargiy Audyogik Sahakari Sanstha
(TMASS), Washim. The construction of the plant is in progress and expected to be over by June,
2011. About 100 farmers will be involved in the procurement and supply of cotton stalks and
about 150 persons will be involved in direct and indirect employment.
d) M/s. Bajaj Steel Industries, Nagpur have already started procuring cotton stalks by mechanically
uprooting, baling and transporting the baled stalks to the centralized chipping centres. Their
interest is to supply the stalks as a fuel to boilers.
e) Many NGOs in and around Akola, Jalgaon and other places in Maharashtra have started
collecting cotton stalks and supply to many user industries.
f) Apart from this, M/s. Aurobindo Laminations in Nagpur are using cotton stalks in making
particle boards.

ACKNOWLEDGEMENT
The authors are grateful to:
1. CFC, Netherlands for the financial assistance.
2. ICAC, USA for constant help, guidance, supervision and encouragement.
3. ICAR, for providing all the infrastructural facilities for effective monitoring , guidance and
permission to participate in the meeting and presentation of data.
4. To all my project colleagues, specially to Dr. S. Sreenivasan, Former, Director, CIRCOT for
constant and continuous support and guidance.
5. To. M/s. Eco-Boards, M/s. Jolly Boards, M/s. Archid Ply and other industries for permitting to
undertake commercial trials.
REFERENCES
[1] Balasubramanya R. H., Shaikh A. J., Paralikar K. M. and Sundaram V., Spoilage of Cotton Stalks During Storage and
Suggestions for its Prevention, J. Indian Society for Cotton Improvement, 15:34-39, (1990).
[2] Sundaram .V., Balasubramanya R. H., Shaikh A. J., Bhatta I. G. and Sitaram M. S., Utilisation of Cotton Stalks, J. Indian
Society for Cotton Improvement, 14(1):94-99 (1989).
[3] Pandey S. N. and Shaikh A. J., A study on Chemical Composition of Cotton Plant Stalk of Different Species, Indian Pulp
and Paper, 41: 10-13 (1986).
[4] Pandey S. N. & Mehta A. K., Industrial Utilisation of Agril. Products : Cotton Plant Stalk, Research and Industry, 24(2):75,
(1979).
[5] Pandey S. N. & Mehta A. K., Particle Boards from Cotton Stalks, Research and Industry, 25:67-70, (1980).
[6] Gurjar R. M., Cotton Stalk Particle Boards A Timber Substitute, Research and Industry, 39(9):153-155, (1994).
[7] Mahanta D., Particle Board and Hardboard from Cotton Stalk, Indian Chemical Manufacturer, 22(8):15-21, 1984.
[8] Guler C. & Ozen R., Some Properties of Particleboards made from Cotton Stalks (Gossypium hirsitum L.), European
Journal of Wood and Wood Products, 62(1), 40-43, March, 2004.
[9] Negi J. S. and Chawla J. S., Composite Boards from Cotton Stalks, Research and Industry, 40(12):267-271, (1995).
[10] Narayanamurti D., Fibre Boards from Indian Timbers, Indian Forester, 86(1): 5-15, (1960).
[11] Pandey S. N., Das R. N. and Day A., Particle Board from Jute & Its Lamination A new Process, Research and Industry,
35:227-229, (1990).
[12] Negi J. S., Utilisation of Lantana camera Laminated composite Boards, Research and Industry, 31(1):22, (1994).
[13] Fadl N. A. and Sefain M., Hardboard from Retted Rice Straw and Cotton Stalk, Research and Industry, 28(8):95, (1983-84).
[14] Balasubramanya R. H., Shaikh A. J. and Sreenivasan S., Cotton Crop and Industry Waste , in Environment and Agriculture
Edited by Chadha K. L. and Swaminathan M. S., Malhotra Publishing House, New Delhi: 2008.
[15] Khandeparkar, V. G., Balasubramanya, R. H. and Shaikh, A. J. (1993), A Process for the Preparation of Paper Grade Pulp
from Cotton Plant Stalk by Anaerobic Digestion. (Indian Patent No. 176891, July, 1993).
[16] Fadl N. A., Sefain Z. and Rekha M., Hardening of Cotton Stalk Hardboards, Indian Pulp and Paper, 33(2):3, August (1978).
[17] Fadl N. A., Heikal S. O., El-Shinnawy and Moussa, M. A., Hardboard from Cooked Rice Straw Blended with Cotton
Stalks, Indian Pulp and Paper, 35(1):19-22, (1980).
[18] Pandey, S. N. and Shaikh, A. J., Utilisation of Cotton Plant Stalks for Production of Pulp and Paper, Biol. Wastes, 21, 63-
70, (1987).
[19] Shaikh, A. J., Blending of Cotton Stalk Pulp with Bagasse Pulp for Paper Making, Biol. Wastes, 31, 37-43, (1990).
[20] Pandey S. N., Ghosh I. N. & Day A., Utilisation of Non-wood Fibrous Ra material for Pulp, Paper and Board, Research and
Industry, 40(12):285-288, (1995).
[21] Pandey S. N. and Shaikh A. J., Production of Various Grades of Paper from Cotton Plant Stalk, Indian Pulp and Paper, 40:
14-18 (1985).
[22] Balasubramanya, R. H., (1981), An Edible Mushroom Crop on Cotton Stalks, J. Indian Soc. Cotton Improv., 6, 104-105.
[23] Balasubramanya R. H. and Khandeparkar V. G., Mushroom Crop on Spent Cotton Stalks, Indian Society for Cotton
Improvement J., 14:85-86, (1989).
Differential Speed Setting Facility for Roller

and Beater in Gins for Higher Ginning Rates
S.B. Jadhav and K.R.K. Iyer
Former Senior Scientist and Former Director, CIRCOT, Mumbai, India
AbstractIn the present-day double roller (DR) gin, being used in many cotton-growing countries of the world, the
rollers rotate with a speed of about 90 rpm while the reciprocating knives (beaters) make about 1000 oscillations per
minute. Studies at CIRCOT have shown that different speed combinations of the rollers and the beaters can produce
spectacular changes in ginning rate (kg/hr) in cottons of different staple classes. In the conventional DR gin,
differential speed variation is not possible because both rollers and beaters are operated by the same drive mechanism.
As a result, when the rollers are made to rotate faster, the beaters too would get faster making more number of
oscillations.
In the modified DR gin designed at CIRCOT, two independent drives are provided for the rollers and the beaters
whereby differential speed adjustment is rendered possible. It has been shown that higher roller speeds such as 110 to
160 rpm coupled with a beater speed of 1000 oscillations per minute can increase the ginning rate by 50-140 per cent.
The increase is found to be more in the case of cottons of longer staple lengths. Interestingly, higher rates of processing
do not cause fibre damage. The new gin design holds promise for a substantial reduction in processing cost.
INTRODUCTION
Ginning rate in the double roller (DR) gins is far lower than that in the saw gin
1
. Though slower, the DR
gin is gentler to the fibre and preserves the quality of lint as compared to saw gin
2
. The Indian ginning
industry processes 85% of cotton with roller gin
3
whereas in the world as a whole roller ginning accounts
for only 15%. However, the slow ginning rate of the roller gin has made it expensive to maintain and
operate. In this gin, the roller and beater are driven with a single drive such that the ratio of the beater
frequency to roller speed remains fixed. There is no provision for altering this ratio, which researches
have revealed,
4-6
controls the efficiency of the process while ginning different varieties of cotton.
Attempts to increase the ginning rate have been made by Gallium and Armojo
7
and Chellamani et al
8
. These researchers used rotary knife system instead of oscillating knife for beating the seeds to separate
them from the fibre.
The present exercise was aimed at designing an improved version of the double roller gin by
providing separate drives for the roller and the beater to increase the ginning rate. This provision is not
available in any of the existing commercial models of DR gin.
THE REDESIGNED GIN
The gear box in the DR gin was suitably redesigned and installed on the gin stand. The modification
allows independent drives for the roller and the beater whereby any desired speed of the roller and beater
is possible. The redesigned gin is known as variable speed (VS) gin.
The machine employed for modification was a commercial Platts DR gin. The modified gin retains
all the machine elements of the DR gin except the gear box. In the conventional DR gin the gears
controlling the motion of the roller and beater are linked to a single shaft and thus have fixed angular
frequency, (generally in the ratio of 1:10). In the VS gin there are two independent shafts, one for the
roller and the other to effect the reciprocating motion of the beater by means of a crank, each provided by
a different motor. The machine diagram of VS gin is shown in Fig.1.
85
Fig. 1: M
Machine Sp
The moto
diameter o
pulley con
and 110 rp
The o
diameters
with frequ
oscillates
experimen
fulcrum o
fixed kniv
mm to sui
Procedure
Experime
beater fre
short fibre
have been
The f
coupled w
respective
shifted to
each varie
The s
speeds (16
are design
The t
manufactu
This set o
C in all
Table
3 & 4 sho
and AFIS
average o
conditions
Differen
Modified DR Gin [G-
pecifications
or that drives
of the step p
nnecting the
pm.
other motor,
of 6 and 4.5
uencies of 1
with 950 op
nt. The slidi
of the lever a
ves. The over
it the varietie
e
nts were con
equency in r
e content in
n considered
first experim
with roller s
ely in Table
the appropr
ety was proce
econd set of
60,130, &11
nated as S1, S
third and fin
urers recom
of tests was u
the four Tab
es 1 and 2 su
ow importan
S machines.
f three value
s of humidity
ntial Speed Sett
-Gear Box, W-Gear
s the roller ro
pulley varies
gear box is k
, which driv
5 inch. The b
1000 and 75
pm. This com
ng weights,
at a pre-dete
rlap width be
es processed
nducted with
relation to va
the resultan
in the optim
ment was con
speeds of 16
1 as well a
riate pulley f
essed thrice
f tests was co
10 rpm) usin
S2 & S3 in T
nal set of te
mmended spee
used as the c
bles.
ummarize res
nt fibre prope
Three tests w
es for each qu
y and temper
ting Facility for
r Wheel, P-Pul-Ley
otates with th
in steps of 7
kept at 6 inc
ves the beat
beater driver
50 opm. In
mbination wa
which exert
ermined posi
etween fixed
in the tests.
h the variable
arious chara
nt lint, incide
mization study
nducted with
60, 130, and
s Table 3. F
fixed on the
at each comb
onducted with
g another pu
Table 2 as we
est was con
ed of roller (
control while
sults of expe
erties of lint
were carried
uality param
rature (65 + 2
Roller and Beat
y R-Roller, M-Movin
he speed of
7.5, 6.0 and
ch. This pulle
ter assembly
r with a pulle
conventiona
as kept as the
t force on th
ition for mai
d knives and
e speed (VS)
acteristics of
ence of neps
y. The experi
h a constant
d 110 rpm.
For the reduc
motor shaft.
bination of ro
h the beater
ulley combin
ell as in Tabl
nducted on t
(95 rpm) and
e critically an
riments toge
processed in
d out for eac
meter was rec
2% RH & 27
ter in Gins for H
ng Knife, O-Beater
1420 rpm an
4.5 inch whi
ey drives the
y, is also fi
ey of 9 inch
al DR gin, r
e control for
he roller, we
intaining con
moving kniv
gin to determ
f seed cotton
s, seed coat f
iment was ca
t beater freq
The results
ced speeds o
In all exper
oller and bea
frequency of
nation for the
le 4.
the same gin
d oscillation
nalyzing the
ether with oth
n various tri
ch of the con
corded. All th
7 + 2
0 C
)
Higher Ginning R

r Shaft, F-Fixed Kn
nd is fitted w
ile the diame
e rollers with
itted with th
diameter thu
roller speed
r all the eight
ere kept at 3
nstant pressu
ves was varie
mine the opt
n being ginn
fragments, a
arried out in
quency of 1
are designa
of 130 & 11
riments, 10 k
ater speeds.
f 750 opm fo
e beater driv
nning mach
frequency (9
results. The
her performa
ials. Fibre te
ntrol and tre
he tests were
Rates
nife, and C-Conecti
with a step p
eter of the ro
h the speeds o
he step pull
us oscillates
is 95 rpm a
t cottons in t
30 inch away
ure between
ed between 9
timum roller
ned. The stap
and ginning p
the followin
000 opm su
ated as S1,
10 rpm the V
kg of seed co
or the same t
ve. Here too
hine operated
950 opm) of
data are des
ance data wh
ests were don
eated sample
e conducted a
525
ing Arm]
pulley. The
oller driver
of 160,130
ley having
the beater
and beater
the present
y from the
roller and
9 mm to 11
r speed and
ple length,
percentage
ng steps.
uccessively
S.2 & S3
V-belt was
otton from
three roller
the results
d with the
the beater.
signated as
hile Tables
ne on HVI
es, and the
at standard
Influence of Machine Parameters on Lint Out-turn
Two important machine parameters that influence the ginning rate and lint quality in ginning are the
speed of the roller and the frequency of the oscillating beater. The effect of the roller speed and the beater
frequency on ginning rate (GOT) and related parameters will be clear from data in Tables 1 and 2.
TABLE 1: GINNING OUT-TURN AT DIFFERENT ROLLER SPEEDS WITH A BEATER FREQUENCY OF 1000 OPM
Variety Treatment Ginning Rate
Kg/hr
Lint Out- turn
Kg/hr
Ginning
(%)
% Increase in
Ginning Rate
% Increase in
Lint Out-turn
G Cot DHy 7 S1
S2
S3
C
180
180
180
105
72
72
72
40
33.7
33.8
33.6
34.0
71.0
71.0
71.0
-
80.0
80.0
80.0
-
K.2 SI
S2
S3
C
163
143
100
80
60
45
38
29
35.9
35.7
35.9
36.0
103.7
78.7
25.0
-
106.8
55.1
31.0
-
Jayadhar S1
S2
S3
C
254
171
240
150
82
60
73
51
31.8
31.6
31.8
32.0
69.3
14.0
60.0
-
60 .7
17.6
43.1
-
H.8 S1
S2
S3
C
170
189
160
98
60
69
52
35
35.0
35.8
35.8
35.0
73.4
92.8
63.3
-
71.4
97.1
48.5
-
JKHy.1 S1
S2
S3
C
200
138
137
103
65
53
53
37
35.4
35.5
35.4
35.6
94.1
33.1
33.0
-
75.6
43.2
43.2
-
G Cot 10 S1
S2
S3
C
160
100
112
96
58
36
41
36
32.6
32.4
32.8
33.0
66.7
4.1
16.6
-
61.1
0.0
13.8
-
DHB.105 S1
S2
S3
C
166
175
172
115
60
62
62
40
35.6
35.2
35.5
35.8
44.3
52.5
49.5
-
50.0
55.0
55.0
-
DCH.32 S1
S2
S3
C
220
151
167
85
72
58
53
30
33.3
33.2
33.5
33.5
158.8
77.6
96.4
-
140.0
76.6
93.0
-
Sl: 160:1000, S2: 130:1000, S3: 110:1000 and C: 95:950
TABLE 2: GINNING PERFORMANCE AT DIFFERENT ROLLER SPEEDS WITH A BEATER FREQUENCY OF 750 OPM
Cotton Treatment Ginning Rate
Kg/hr
Lint Out- turn
Kg/hr
Ginning
(%)
% Increase
In Ginning Rate
% Increase
in Lint
Out-turn

G Cot Hy.7
S1
S2
S3
C
145
140
120
105
57
56
48
40
33.7
33.9
33.6
34.0
38.0
33.3
14.3
-
42.5
40.0
20.0
-

K.2

SI
S2
S3
C
109
82
82
75
42
35
31
29
35.9
35.7
35.9
36.0
45.3
9.3
9.3
-
44.8
20,6
6.8
-

Jayadhar
S1
S2
S3
C
180
160
164
150
66
58
57
51
31.8
31.6
31.8
31.0
20.0
6.7
9.3
-
29.4
13.7
15.7
-
Table2(Contd.)
Differential Speed Setting Facility for Roller and Beater in Gins for Higher Ginning Rates 527
Table2Contd.

H.8
S1
S2
S3
C
120
116
113
98
46
45
45
35
35.0
35.8
35.8
35.0
22.4
18.4
15.3
-
31.4
28.5
28.5
-

JKHy.1
S1
S2
S3
C
130
100
100
90
52
41
41
38
35.4
35.5
35.4
35.6
44.4
11.1
11.1
-
36.8
7.8
7.8
-
G Cot 10 S1
S2
S3
C
113
105
105
96
40
37
38
36
35.6
35.4
33.8
33.0
17.7
9.4
9.4
-
111
2.8
5.5
-

DHB.105
S1
S2
S3
C
125
124
120
115
50
50
48
40
35.6
35.2
35.5
35.8
8.6
7.8
4.3
-
25.0
25.0
20.0
-

DCH.32
S1
S2
S3
C
150
150
120
83
48
48
46
30
33.3
33.2
33.5
33.5
80.7
80.7
44.6
60.0
60.0
53.3
TABLE 3: QUALITY PARAMETERS OF LINT AT DIFFERENT ROLLER SPEEDS WITH A BEATER FREQUENCY OF 1000 OPM
Cotton Treatment 2.5% Span
Length (mm)
Short Fibre
%
Mic (10-6g/in) Mat.
Ratio
Fibre Strength
g/tex
Neps/g SCF/g
G Cot DHy
7
S1
S2
S3
C
22.3
23.3
23.2
20.8
13.5
13.4
14.1
11.1
52
5.3
5.3
5.3
0.89
0.90
0.87
0.87
20.8
21.6
194
21.5
95
128
139
118
11
20
27
16
K.2 S1
S2
S3
C
24.4
24.2
22.6
23.8
20.0
19.7
18.3
15.2
4.3
4.2
4.2
4.0
0.79
0.83
0.79
0.77
20.8
20.4
22.5
21.6
261
250
280
243
48
55
40
44
Jayadhar S1
S2
S3
C
24.7
23.3
22.7
24.7
15.0
15.8
13.7
10.7
4.3
4.4
4.3
4.4
0.90
0.88
0.89
0.91
19.1
16.1
15.7
15.4
193
182
195
161
19
16
18
16
H.8 S1
S2
S3
C
25.5
25.3
24.7
25.8
14.1
13.7
15.0
10.3
4.4
4.4
4.5
4.5
0.79
0.94
0.89
0.86
22.5
20.8
22.5
22.8
156
115
92
91
17
11
15
12
JKHy.1 S1
S2
S3
C
28.2
26.1
25.6
25.5
15.6
18.5
10.9
12.7
3.8
3.6
3.7
3.7
0.80
0.79
0.78
0.86
23,7
21,6
23.6
23.6
185
192
157
144
24
31
17
26
G.Cot 10 S1
S2
S3
C
25.6
24.8
25.0
26,4
13.9
14.7
13.4
10.2
4.3
4.4
4.0
4.3
0.86
0.85
0.89
0.86
20.9
19.9
17.9
18.9
167
138
155
124
48
38
51
24
DHB.105 S1
S2
S3
C
28.8
27,4
27.0
27.9
9.9
13.4
16.6
10.0
2.9
3.2
3.3
3.4
0.87
0.80
0.87
0.78
23.5
21.6
19.9
20,8
185
170
175
159
22
23
21
15
DCH.32 S1
S2
S3
C
32.7
31.8
30.9
33.1
13.6
13.1
12.3
7.5
2.9
3.2
3.3
3.4
0.78
0.78
0.77
0.77
25.5
21.6
21.4
21.5
212
190
200
170
19
18
19
16

TABLE 4: QUALITY PARAMETERS OF LINT AT DIFFERENT ROLLER SPEEDS WITH A BEATER FREQUENCY OF 750/MIN
Cotton Treatment 2.5% Span
Length
(mm)
Short
Fibre %
Mic
(10-6g/in)
Mat.
ratio
Fibre
Strength
g/tex
Neps/g SCF/g
G Cot DHy.10 S1
S2
S3
C
22.8
22.7
23.2
20.8
7.8
13.5
11.1
11.1
5.0
4.9
5.5
5.3
0.93
0.91
0.94
0.97
21.0
21.0
21.6
21.5
121
147
165
118
14
19
30
16
K.2 S1
S2
S3
C
25.8
23.1
23.2
23.8
11.5
18.9
18.9
15.2
4.1
3.9
3.4
4.0.
0.87
0.78
0.78
0.79
22.4
19.2
19.9
21.6
205
305
410
243
35
50
50
44
Jayadhar S1
S2
S3
C
24.4
24.5
24.4
24.7
9.8
11.5
13.6
6.7
4.3
4.8
4.1
4.4
0,89
0.88
0.87
0.91
19.6
19.9
18.4
15.4
140
229
202
161
12
13
23
11
H.8 S1
S2
S3
C
24.0
25.2
25.3
25.8
16.9
15.2
12.7
10.3
3.4
4.1
4.3
4.5
0.79
0.84
0.84
0.86
18.3
22.9
20.8
22.8
135
137
138
91
35
15
17
12
JKHy.1 S1
S2
S3
C
27.3
24.5
26.4
25.5
13.6
17.9
10.6
14.4
3.3
3.2
3.2
3.7
0.79
0.75
0.81
0.86
24.8
24.7
24.4
23.6
196
259
326
144
42
38
48
26
G Cot 10 S1
S2
S3
C
25.2
23.7
23.6
26.4
14.8
13.6
16.9
12.4
3.5
3,2
3.0
4.0
0.80
0.78
0.82
0.86
23,7
25.2
22.4
18.9.
116
171
194
124
27
17
31
24
DHB.105 S1
S2
S3
C
27.7
28.3
27/6
27.9
12..9
12.1
14.7
10.0
3.1
2.7
2.6
3.4
0,75
0.75
0.75
0,78
21.5
24.8
22.9
20.8
179
164
200
159
30
21
41
15
DCH.32 S1
S2
S3
C
35.4
33.4
33,3
33.1
11.4
12.6
9.8
7.5
2.8
2.8
2.5
2.9
0,79
0.77
0.78
0.77
28.2
27.1
23.8
21.5
202
190
200
170
18
17
16
16
From Table 1 it will be clear that the ginning rate and the lint out-turn in almost all the experiments
with changed speed ratios are much higher than with the normal (control) speed of roller and beater.
Among the three roller speed ratios (S1-160: 1000, S2-130: 1000 & S3-110: 1000), the highest ratio
(160:1000) gave the maximum ginning rate in all the cottons. The maximum increase in ginning out-turn
for the longest staple cotton DCH.32 was observed to be 158.8%. The increase in the lint out-turn ranges
from 50% for DHB.105 to a maximum of 140% for DCH.32. The speed ratio of 130:1000 gave the
highest ginning out-turn for medium staple like G.Cot.DHy.7 (80%) followed by DHB.105 (55%)
Table 5 gives the increases in ginning rate and lint out-turn averaged for for all the eight cottons
tested.The averages are found to be the highest for a roller speed of 160 rpkm coupled with the beater
frequency of 1000 opm. The numerical difference between percentage increase in ginning rate and lint
out turn for a given variety is dictated by its ginning percentage.
TABLE 5: AVERAGE INCREASE IN THE GINNING RATE AND LINT OUT-TURN OVER THE CONTROL
Sr. No Speed Ratio % Increase in Ginning Rate % Increase in Lint Out-Turn
Experiment-1
1 S1 (160: 1000) 84.8 80
2 S2 (130:1000) 52.5 49.8
3 S3 (110:1000) 51.5 50.8
Experiment-2
1 S1 (160:750) 27.4 29.7
2 S2 (130:750) 20.3 22.5
3 S3 (110:750) 12.1 13.2
It is well known that for a given variety with an average staple length of say, 24 mm,, there would be
some fibres with a length of 30 mm. The tips of such fibres are most likely to be picked up and gripped
first. Other shorter fibres are entangled with them and are dragged forward near the ginning point along
with the seed and then get ginned. The different speeds have given different out-turns for cottons and the
reasons for the same are discussed.
Differential Speed Setting Facility for Roller and Beater in Gins for Higher Ginning Rates 529
The rollers employed in the conventional double roller gin are of 16.5 cm diameter rotating at a speed
of (n) 95 rpm such that the surface speed (V= n d/2) lies between 48 and 50 m/min. The beater
oscillates with a frequency of 950 opm. Since the beater oscillates symmetrically with respect to the edge
of the fixed knife, one half of the time is utilized for feeding seed-cotton and the other half is for ginning.
Since the beater frequency is 950 opm, during the feeding interval of 1/30 sec, the rollers move a distance
of 26 mm. If it is assumed that during the feeding interval, the probability of a fibre of average length
being picked up by the roller is p (0<p<1), the length L of the fibre picked and pulled would be 26p mm.
It may be noted that for efficient ginning the fibres must be fully drawn out so as to ensure firm grip
and this requires that the staple length of the cotton is equal to the parameter L. Thus it is clear that
probability p must be as high as possible for ginning cottons with nearly 24 mm staple length. To
improve the value of the parameter L it is suggested that the roller speeds may be increased if the staple
length is more than 26 mm.
One representative example will reveal how a roller speed of 160 rpm in combination with an
oscillation frequency of 1000 opm is better suited for extra long staple cottons like DCH.32. The surface
speed (V) of the roller is given by
V = . d /2
where w is the angular velocity and d the diameter of the roller. If n is the number of revolutions per
minute,
V=2.n.d/2.
If the roller has a diameter of 15 cm, and it rotates with 160 rpm,
V = 2 . 160. 15/2 =. 125.68 cm/sec.
Similarly for roller speeds of 130, 110 and 95 rpm, the corresponding surface speeds are 102.1, 96.4
and 74.6 cm/sec.
In the first set of experiments the beater frequency was kept at 1000/min, the time period (time per
oscillation) being given by
t = 0.06 sec.
Since the feeding time per gin cycle (t
f
) is half of this time t
t
f
=0.03 sec.
The total movement of the roller during the time t
f
is 125.6 x 0.03 =3.768cms =37.7mm. Similarly
for 130, 110 and 95 rpm of roller speeds the corresponding movement of the roller during the same time
would be 30.6, 25.9 and 22.3 mm. respectively.
The time needed by a fibre on a seed to be caught between the roller and the knife-edge is a statistical
parameter and can be taken as
p x t
f
where p stands for the probability factor.
At the roller speed of 160 rpm, the fibres of average length (p x .37.7 mm) are caught between the
roller and the knife-edge and for them ginning starts in the first cycle itself. Thus for ginning long and
extra long cottons the highest speed of 160 rpm is justified and is expected to yield higher lint out-turn
than by employing the lower speeds.
It may be noted that the experiments performed with a roller speed of 130 rpm always results in
higher yield for medium staple cottons where the staple length varies between 24 and 30 mm. It is thus
concluded that the higher speed has a positive impact on the probability factor.
Most of the Indian cottons are of medium staple category and for them a roller speed of 130 rpm
coupled with a beater frequency of 1000 opm is found to be optimum for high lint yield. The same
condition cannot be taken for long staple cottons. In such cases, even if the grip is strong enough to
prevent fibreslippage, the seed may fail to get ginned in the first cycle. Nevertheless, it will utilize
subsequent cycles to get ginned. This would mean that ginning will be delayed and the out-turn will be
lower.
The other experiment with the same roller speeds combined with the lower oscillation frequency of
the beater (750 opm) also gives comparatively higher output (Table 2) than in the control experiment
with higher beater frequency (1000 opm). In this case, the feeding rate of seed cotton is more but the
beater frequency is less, thereby decreasing the ginning rate.
CONCLUSION
The VS Gin is an excellent replacement for the conventional DR gin presently being employed in Indian
ginning industry. It has the flexibility to permit optimization of settings to increase the productivity for
all types of cotton, short, medium and long staples.
For efficient ginning of cotton, the fibre must be dragged toward the ginning point i.e. at the edge of
the fixed knife. This can be achieved by setting different roller speeds relative to staple length of cotton
under the ginning process. At the recommended speed for the roller, the oscillating beater separates the
seed from the fibre with minimum number of strokes, at the same time increasing the ginning out-turn
and preserving fibre length, which is the most important quality parameter of the lint. The following are
the salient features of the performance of the VS. Gin:
In most cottons the ginning out-turn has recorded increase, ranging from 60% to 100% over the
production possible with the old model gin.
The roller speed of 160 rpm combined with an oscillation frequency 1000 cycles per minute for
the beater is the best for ginning extra-long Indian cottons.
For medium staple cottons in the length range of 24-28 mm, a roller speed of 130 rpm in
combination with 1000 oscillations per minute for the beater is the most appropriate.
The combination of 110 rpm for the roller and 1000 cycles per minute for the beater is ideal for
the cottons having length 20 to 23 mm.
Important fibre parameters such as 2.5% span length, fibre tenacity and other characters are
preserved.
ACKNOWLEDGEMENT
The authors are grateful to Dr. A. J. Shaikh, Director CIRCOT for according permission to present this
paper at WCRC-5.
REFERENCES
[1] Comparative Performance of Different Types of Gins, A report submitted by CTRL, ATIRA & AIFCOSPIN to IDA/IBRD,
Sponsored by NCDC, India (1984).
[2] Sundaram, V., "Contribution of Cotton Technological Research Laboratory to the Improvement of Cotton Ginning in
India", CTRL Publication, New Series, No. 144 (1980).
[3] Chaudhary, M Rafiq, "Harvesting and Ginning of Cotton in the World", Proceedings of Beltwide Cotton Conferences,
Vol.2 of 2 pp 1617-1619 (1997).
[4] Johnson, F., Mayfield, W., Lalor, W. F. and Hughes, S. F. "Ginning Developments", Textile Asia, 23(10), 49-52(1992).
[5] Vizia, N.C. and Iyer K.R.K.; "Ginning Research in India and Future Prospects", National Seminar, Mumbai (1999)..
[6] Nanjundayya, C. and Iyengar, R. L. N., A Resume of Various Investigations Carried Out on the Ginning Of Indian
Cottons, Tech. Bull. Series, B No.50, Tech. Laboratory, Matunga, Bombay (Oct 1955).
[7] Gillium, M. W. and Armojo, C.B., American Society for Agriculture Engineers, Vol.43 (4) 809-817 (2000).
[8] Chellamani, K.P. Parthasarathy, N. and Jaikumar, V., "Design and Development of High Production Gin and Lint Cleaner",
Asian textile Journal, (June 2000).
Influence of Quality Attributes

of Individual Bales on Yarn Quality
R.P. Nachane
CIRCOT, Mumbai, India
AbstractCotton being a commercial crop of great economic importance, there exists a value chain in the sense that
the seed cotton is converted into lint and through the yarn and fabric route into garments and made ups for both
internal consumption and export. However, in this conventional value chain there are several weak as well as missing
links. The crucial unit operation involved in the value chain is ginning, i.e., conversion of seed cotton into lint, is still
considered to be one of the weakest links characterized by excessive use of energy, low productivity, absence of cleaning
and lack of facilities for quality assessment of the lint that this sector produces.
Even though the Spinning Industry in India is modernized, it mainly depends upon the ginning industry for raw
material. Cotton bales prepared in ginning industry are not individually tagged with fibre characteristics. Therefore,
spinning units are procuring cotton bales on the basis of fiber characteristic values got from random sampling of bales.
This special study has been carried out to find out the effect of segregation of bales, mainly based on fineness, after
tagging of individual bales according to fibre properties, on the yarns so produced. Results indicate that for high
micronaire of above 4.5 for a given variety grown in one location, no statistically significant differences in yarn
properties are observed. However, for yarns produced from finer cottons of less than four micronaire, many of the yarn
properties are highly influenced by the fineness. It is therefore recommended that individual bale tagging with
segregation of bales based on fineness can result in better quality yarns, leading to better quality fabrics and garments.
INTRODUCTION
The Indian Textile Industry uses about 60% cotton as its raw material, unlike the global textile industry
that has a 40:60 mix of cotton and manmade fibres. There has been a phenomenal increase in cotton
production in India in recent years.
Cotton being a commercial crop, there exists a value chain in the sense that seed cotton is converted
into lint and through the yarn and fabric route into garments and made-up for both internal consumption
and export. However, in the conventional value chain there exist a few missing as well as weak links.
Ginning is considered to be one of the weakest links characterized by excessive use of energy,
presence of contaminants & trash and lack of facilities for quality assessment of individual bales.
Although spinning sector is performing better with modern facilities, weaving/knitting sector still needs
to improve in quality and product up gradation to meet the international standards. Further, processing
such as preparatory chemical treatments of yarns and fabrics, eco-friendliness, energy use efficiency, and
its treatment are factors that need immediate attention. In the handloom sector, workers are exposed to
harmful chemicals and environment is also vitiated with chemical effluents in rural areas.
Some of these weak links have been addressed under a World Bank funded project entitled, A
Value Chain for Cotton Fibre, Seed and Stalk: An Innovation For Higher Economic Returns to Farmers
and Allied Stake Holder sanctioned by National Agricultural Innovation Project (NAIP) under
Component 2 in consortium mode with Central Institute for Research on Cotton Technology (CIRCOT),
Mumbai as lead centre and Central Institute for Cotton Research (CICR), Nagpur & Super Spinning Mill
Ltd., Coimbatore, as consortium partners. This value chain is shown in the figure below.
As regards status of research and technology in the production system, although cost and resource
conservation technologies like IPM, IRM and INM have been perfected at the field level and
demonstrated in several farms across the country, their benefits have not percolated to small scale
growers. Also, quality seeds and inputs have remained scarce to these farmers. It is felt that an
Integrated Cultivation approach by bringing together small holding farmers and ensuring quality inputs
86
at competitive price would not only bring down the cost of cultivation but also result in better quality
cotton with low levels of contaminants.
CIRCOTsCottonValueChain
Cotton
Production
CleanCotton
picking
Ginning
Baleswith
FibreQuality
YarnProduction
Delinteringofseeds
&separationtoHull
&Kernel
Collectionof
Stalks&Chipping
Bioscouring&
Dyeingwith
NaturalDyes
Oilrecovery
fromKernel
Edible
Oyster
Mushroom
Hulls
Bio
enrichment &
cattlefeed
trial
Cottonseed
oil,seed
meal&
edible
protein
Fabric
production
, garment
making&
marketing
Weaving
home
furnishing&
Marketing
Supplyto
board
making
factory&
Utilisation
Linters

CICR with its expertise on production technologies executed this job in the current project and
CIRCOT did the post harvest on-farm and off-farm management to produce clean seed cotton. For this,
certified cotton seeds, fertilizers, pesticides, etc., were procured for farmers of the adopted villages near
Nagpur, Coimbatore and Sirsa by CICR. Sowing and related activities were taken up along with crop
growth monitoring in the farmers fields under supervision of CICR scientists. The integrated production
technologies developed by CICR, were adopted for raising medium, long and extra-long cottons in the
adopted villages. Details about cotton produced are as given below:
Adopted Villages Area in
Acres
Number of
Farmers
Quality of Seed General Sowing Period
Nandura & Loni villages at Nagpur 60 30 Bunny Bt In the month of June
Vadapaddur village at Coimbatore 92 110 RCHB 708 Bt In the month of August
Nejadela Kalan & Jhopra villages at Sirsa 82.75 34 Bioseed -6488 BG I In the month of April
Kapas produced by these farmers was procured by CIRCOT under the project and was ginned in
modern ginning factories in the respective area and pressed into bales. While pressing into bale,
representative fibre samples were collected and evaluated using HVI. The fibre attributes so obtained
were assigned to these individual bales.
In the work presented here, these bales were segregated using fibre fineness and spun into yarns
separately for each segregated group. Influence of fibre properties on yarn parameters is studied and
reported here.
LABELING OF BALES WITH FIBRE QUALITY
In this project, individual bales were tagged with their fibre properties. For spinning purpose bales were
segregated in groups based on their fineness expressed as micronaire value. Yarn was manufactured in
different lots from these segregated bales. This particular exercise has been carried out to study effect on
yarn properties after segregation of bales based on fibre properties of the same variety of cotton.
STATISTICAL METHOD USED
When yarn is manufactured from different lots of cotton bales identified on the basis of micronaire value
two sources of variation may exist in yarn properties.
Between group variation, and
Influence of Quality Attributes of Individual Bales on Yarn Quality 533
Within group variation
In this case each group of bales segregated on the basis of micronaire value is identified with
particular code. From the lot of each code, equal numbers of yarn cones were drawn at random by using
random number table. This type of sampling is called as
Representative or Stratified Sampling
Number of samples to be tested was decided on the basis of coefficient of variation percentage and 95%
confidence interval for the mean value. To determine whether significant variation exists or not among
the yarns prepared from different groups, we decided to carry out analysis of variance (ANOVA). It is
essentially an arithmetic process for portioning total sum of squares into components associated with
recognized sources of variation. It has been used to advantage in all fields of research where data are
measured quantitatively.
In analysis of variance one finds the ratio F of two variances to be compared. In the present case
variation in yarn properties among the group of bales and variation in yarn properties within the group of
bales is analyzed.
The calculated F value is compared with the tabular F value for N
1
and N
2
degrees of freedom to
decide whether to accept the null hypothesis of no difference between population means or the alternative
hypothesis of a difference. The interpretation of F depends upon the particular problem, but good general
rule is
a) If the calculated value of F is less than the 5% probability level in tabular value, then the
difference is not statistically significant.
b) If the calculated value of F falls between the 5% and the 1% probability level tabular value, then
there is some evidence of a difference but that this evidence by no means is conclusive.
c) If the calculated value of F is greater than the 1% probability level tabular value then a real
difference certainly exists.
Degrees of freedom N
2
is based on total number of observations and N
1
is based on total number
groups. Degrees of freedom refers to independent comparison available in estimating variances.
Results are presented and analysed below centre wise.
Sirsa
At Sirsa, Haryana Bioseed-6488 BG was used. Total 285 quintals of kapas was procured. After ginning
63 cotton bales were prepared.
TABLE 1: AVERAGE FIBRE PROPERTIES OF BALES
2.5% SL (mm) UR% Micronaire (g/inch) Tenacity (g/tex)
29.0 45 4.9 22.6
Yarn was spun to 24s count in five different groups as shown below. Each lot consisted of about
eight bales amounting to approximately one and a half tonne. From each lot five yarn cones were drawn
at random to test yarn properties.
TABLE 2: GROUP OF BALES SEGREGATION BASED ON MICRONAIRE
A B C D E
4.7 4.8 4.9 5.0 4.7 5.0

TABLE 3: YARN TEST RESULTS (LEA TEST AV.)
Group Count Strength CSP
A 23.7 102.5 2431
B 24.1 97.2 2334
C 23.9 100.0 2388
D 23.6 106.5 2511
E 23.7 102.3 2417
Grand 23.8 101.7 2416
TABLE 4: SINGLE YARN STRENGTH TEST RESULTS
Group Breaking Strength Tenacity Elongation %
A 353.5 14.2 5.2
B 348.7 14.2 5.3
C 344.7 14.0 5.3
D 349.5 14.0 5.5
E 353.2 14.1 5.2
Grand 349.9 14.1 5.3
TABLE 5: USTER EVENNESS TEST RESULTS
Group U% Thin Thick Neps
A 13.0 20 348 253
B 13.3 19 363 294
C 12.8 15 332 261
D 12.4 11 250 209
E 12.6 13 287 216
Grand 12.6 16 316 346
F Values Within and Between (Combined)
TABLE 6: WITHIN TOTAL 25 READINGS AND BETWEEN 5 SETS OF MICRONAIRE VALUES
Mic. X Lea
Count
Mic. X
Lea
Strength
Mic. X
CSP
Mic. X Single
Thread Str
Mic. X
Elongation
%
Mic X
Tenacity
Mic. X
U%
Mic. X
thin
places
Mic. X
thick
Places
Mic. X
neps
1.14 1.99 2.21 0.23 1.23 0.2 4.05 1.44 3.39 2.23
For D.F. N
1
= 4 and N
2
= 20
F value at 5% level of significance = 2.71
Result indicates that there is no significant difference for yarn properties between groups for 24s
count yarn after bale segregation based on fineness.
Nagpur
At Nagpur, Maharashtra, Bunny Bt was used. Total 349 quintal of kapas was procured. After ginning 67
cotton bales were made ready.
TABLE 6: AVERAGE FIBRE PROPERTIES OF BALES
2.5% SL (mm) UR (%) Micronaire (g/inch) Tenacity (g/tex)
30.5 46 3.2 24.4
Yarn was spun to 30s count combed in five different groups as shown below. Each group consisted
of about eight bales amounting to approximately 1.5 tonne. From each lot twenty two yarn cones were
drawn in random to test yarn properties.
TABLE 7: GROUP OF BALES SEGREGATION BASED ON MICRONAIRE
A B C D E
3.1 3.2 3.3 3.1-3.6 Mix

Influence of Quality Attributes of Individual Bales on Yarn Quality 535
A 29.9 94.8 2836
B 30.4 91.3 2775
C 29.8 93.2 2775
D 29.7 94.4 2801
E 29.4 97.2 2858
Grand 29.9 94.2 2809
A 326.6 6.5 16.5
B 321.5 6.9 16.5
C 323.1 6.6 16.5
D 319 6.4 16.2
E 335.2 6.4 16.8
Grand 325.1 6.6 16.5
A 11.5 6 167 298
B 11.9 9 226 330
C 12.2 11 330 571
D 12 9 256 425
E 12 5 220 407
Grand 11.9 8 240 426
F Values Within and Between (Combined)
Mic. X Lea
Count
Mic. X Lea
Strength
Mic. X
CSP
Mic. X Single
Thread STR
Mic. X
Elongation
%
Mic X
Tenacity
MicX
U%
Mic. X
Thin
places
Mic. X
Thick
Places
Mic. X
NEPS
7.69 8.68 4.63 4.23 2.8 1.56 8.86 7.33 19.5 19.22
For D.F. N
1
= 4 and N
2
= 105
Results indicate that there is significant difference between groups for 30s count yarn after bale
segregation in the case of Lea Count, Lea and Single Thread Strength, Evenness of yarn and Thin
places. Difference in case of Thick place and Neps is highly significant. For single yarn tenacity
and breaking elongation there is no significant difference.
Coimbatore
At Coimbatore, Tamilnadu, RCBH-708 Bt was used. Total 687 quintal of kapas was procured. After
ginning cotton 142 bales were ready.
TABLE 12: AVERAGE FIBRE PROPERTIES OF BALES OF COIMBATORE COTTON
2.5% SL (mm) UR (%) Micronaire (g/inch) Tenacity (g/tex)
28.3 48 4.5 21.5
Yarn was spun to 80s count in seven different groups as shown below. Each group consisted of about
eight bales amounting to approximately one and a half tonne. From each lot twenty yarn cones were
drawn at random to test yarn properties.

TABLE 13: BALE SEGREGATION BASED ON MICRONAIRE
A B C D E F G
3.2-3.6 3.6 3.7-3.8 3.4-3.8 2.9-3.0 3.1-3.2 2.9-3.2
A 81.1 39.8 3228
B 81.7 36.6 2988
C 81.6 37.9 3091
D 81.2 37.9 3073
E 80.8 40.1 3238
F 81 39.8 3225
G 80.9 40 3237
Grand 81.2 38.9 3154
A 147.5 5.5 20.3
B 136.3 5.1 18.8
C 138.9 5.2 19.2
D 140.0 5.1 19.2
E 143.9 5.8 19.7
F 144.3 5.9 19.8
G 144.4 6.0 19.4
Grand 142.2 5.5 19.5
A 11.7 30 84 262
B 12.9 88 198 415
C 12.7 80 169 373
D 12.6 69 154 301
E 12.1 48 147 367
F 12.0 35 136 313
G 12.0 38 135 310
Grand 12.3 55 146 334
F Values within and between (Combined)
Mic. X Lea
Count
Mic. X Lea
Strength
Mic. X
CSP
Mic. X
Single
Thread Str
Mic. X
Elongation
%
Mic X
Tenacity
Mic. X
U%
Mic. X thin
places
Mic. X
thick places
Mic. X
neps
1.31 17.41 23.44 8.8 18.75 4.33 31.83 17.68 21.02 7.68
For D.F.N
1
= 6 and N
2
= 133
F-table value at 5% level of significance = 2.17
F-table value at 1% level of significance = 2.96
Results indicate that there is highly significant difference between groups for 30s count yarn after
bale segregation in case of Lea Strength, Lea CSP, Single thread strength and elongation, and all
the evenness properties of yarn. Even for tenacity of single yarn, difference is significant.
CONCLUSION
Statistical analysis of results indicates that bale segregation based on fibre properties has significant
effect on properties of yarn spun from cotton of fine and extra fine characteristics. It does not seem to
have significant effect on the properties yarn spun from coarse cotton. It is therefore recommended that
individual bale tagging with segregation of bales based on fineness can result in better quality yarns,
leading to better quality fabrics and garments, particularly for cotton from varieties giving medium and
fine fibres.
Development of an Automatic Roller Grooving

Machine for Making Helical Grooves on Rollers Used
in Roller Ginning Machines
T.S. Manojkumar
1
V.G. Arude
2
and S.K. Shukla
2

1
Programme Co-ordinator, KVK, CPCRI, Kasargod, (India)
2
Scientist (Sr. Scale), Ginning Training Centre, Central Institute for Research on Cotton
Technology, Amaravati Road, P.O. Wadi, Nagpur-440 023 (India)
E-mail: dr.tsmanojkumar@gmail.com
AbstractDouble roller gins are the most popular ginning machines used in India. The rollers used in double roller
gins become smooth due to friction between the roller and knife during continuous operation. Helical grooves are made
on the rollers after every 16 to 20 working hours to make the roller surface rough to facilitate the lint to adhere to the
roller at a faster rate. In the present practice of manual grooving with hacksaw, the depth, breadth and spacing of the
grooves are not uniform and is a laborious process requiring an hour to groove roller. This results in lower
productivity of ginning machines and damages the roller affecting its durability. To overcome these problems an
automatic roller grooving machine was developed to make helical grooves on the roller. It consists of main frame, head
stock, tail stock and a cutter assembly mounted on a movable trolley. The rotary motion of the roller and forward
motion of the trolley was synchronized together with suitable drive mechanism. The rotary motion of the saw blade
cutter was achieved with the appropriate mechanism. With this mechanism roller completes the revolution of 270
o
and
the cutter assembly moves forward by the length of the roller. A mechanism consisting of a screw shaft and handle
arrangement is used for adjusting the depth of groove. A mechanism consisting of a chuck on a spindle with an
indexing arrangement is developed for making consecutive helical grooves at a specified distance parallel to each other.
The rotary cutter saw blade was mounted at an angle of 17
0
to the roller axis on a vertical plane. The machine was
successfully tested and its performance was found to be satisfactory. The machine can groove the roller in ten minutes
with 18 equally spaced grooves of uniform depth and breadth of 2 mm with accuracy. The re-grooving on the same
impression after reduced roller diameter was also successfully carried out. The automatic roller grooving machine can
successfully replace the existing manual method of grooving and will avoid the drudgery involved in this operation.
INTRODUCTION
Ginning machines are devices which separates mechanically the fibres and seed of the seed cotton. Roller
gins are the most popular ginning machines used in India. Normally four types of roller gins, namely
laboratory model gins, single roller (SR) gins, double roller (DR) gins, and rotary knife gins are used for
ginning. The roller gins work on the principle of Ma-carthy's gin. Ginneries are presently using different
materials for rollers viz., chrome composite leather, newspapers, walrus leather, rubber-canvas composite
material, cotton woven fabrics, etc. Among these materials the chrome composite leather is the most
widely used roller material. The roller of a given length is made of chrome composite leather washers,
which are compressed at a pressure of about 14 kg/cm
2
on a steel shaft. The shape of steel shaft inside the
discs can be square, rectangular, pentagonal, etc. The washers are made of partly finished chrome tanned
leather sheet, each 0.75 to 1.25 mm in thickness, which are glued together. Each washer consists of 10 to
20 numbers of leather flaps. The sheet is finally cut with the help of a die in circular shapes to make the
washers. Individual washers are perfectly stitched with single cotton thread. Each washer is about 13 to
19 mm in thickness. The ginning roller is made by inserting required number of washers over a steel
shaft. By applying optimum pressure on the inserted washers, the washers are compressed together,
which forms a ginning roller. This roller is finished on a lathe machine and grooved.
The surface of the chrome composite leather roller gets polished very fast while running. During the
continuous operation of the DR gin, due to friction between the roller and knife, the rollers become
smooth and the surface of leather roller needs to be roughened by cutting grooves to get optimum ginning
output. Due to this the ginning out put of the machine drops continuously with running time. To
87
overcome this, the chrome leather roller is required to be removed from the machine and then the surface
is roughened by cutting grooves. The surface of the roller is given a smooth finish and helical grooves of
2 mm width and depth are to be cut on it such that the spacing between the consecutive grooves is
uniform throughout the length of the roller. The purpose of the groove is two-fold namely, they prevent
abortive seeds and motes from sticking under the knife; and they provide place for the fibres to enter
readily under the knife, assuring a constant flow of fibres over the roller, there by increasing the
production. The helix makes a half to three fourth revolution about the length of the roller. Rollers
should be grooved every day before the start of the ginning or once in every 16 20 working hours. The
grooving of the rollers is necessary to increase the rough surface for the lint to adhere to the roller at a
faster rate. The grooves should not be appreciably wide or distinctly V-shaped.
This process is very labour intensive job and in India is done preferably manually using saws. For
carrying out grooving the roller is mounted on a stand. Two men pull a saw in a reciprocating manner by
standing on either side of the roller. The roller has to be rotated by hand while cutting to give a helical
groove. Rotating the roller with hand while cutting is not at all uniform and hence the successive grooves
cut on the roller are never parallel to each other and sometimes overlap each other. Since it is a manual
operation, the uniformity in depth and width of groove can not be maintained.
Some ginneries use marble cutting tool and grooving is done by rotating the roller with one hand
while moving the motor along with the tool on the other hand on the surface of the roller in a helical
direction. This operation is very dangerous as the motor has to be held in hand with the cutting tool
rotating at very high rpm and may cause serious injury to the operator. The risk of getting electric shock
also is very high which makes the grooving very risky job. In this operation also the grooves made are
never parallel to each other and the depth of grooving is also varying which causes damage to the fibre
during ginning.
Some ginneries also use conventional grooving machines in which the roller has to be rotated by
hand as well as the trolley carrying the cutting tool has to be moved in the forward direction by hand.
Due to these manual operations, operator does not have any control on the forward speed and the angle of
rotation of the roller which results in grooves with non uniform spacing. Also the non uniform grooving
has a adverse effect on the fibre and seed quality and on the production capacity of the machines. There
should not be any damage to the properties of the ginned lint and quality of seed obtained while
processing in gins. Besides, maintaining the seed quality and ginning percentage, preservation of fibre
properties viz. 2.5% span length, length uniformity ratio, micronaire value, tenacity are equally important
for cotton.
To overcome these difficulties an auto-grooving machine has been developed to make helical
grooves with precise depth control and with uniform spacing between two grooves.
An auto groovier machine has been designed. The 2D and 3D drawings of the designed machine were
prepared in high end software. A prototype of an autogroover machine has been developed and fabricated
at Ginning Training Centre of Central Institute for Research on Cotton Technology (CIRCOT), Nagpur.
Design and Fabrication
Autogroover consist of main frame, head stock, tail stock and a cutter assembly mounted on a movable
trolley developed for making helical grooves on leather rollers. The rotary motion of the roller and
forward motion of the trolley was synchronized together with suitable drive mechanism. The rotary
motion of the saw blade cutter was achieved with the appropriate mechanism to cut precise groove on the
roller. The machine is provided with an indexing mechanism for making consecutive helical grooves at a
specified distance parallel to each other on the roller. Adjustments viz. change of angle of helical grove,
number of grooves, distance between successive grooves, depth and width of groove, speed of roller,
cutting blade and trolley could be made easily for carrying out smooth grooving of the roller.
Development of an Automatic Roller Grooving Machine for Making Helical Grooves on Rollers Used in Roller Ginning 539
Base Frame
The roller grooving machine base frame was designed to 2.4 m length, 0.6 m width and 0.66 m height
with channel section frame of 100 x 50 mm. The frame was supported at the four corners by four legs
made of the same section with the total height of the frame at 0.66 m. The head stock and tail stock were
mounted on either end of the frame to hold the roller firmly in a horizontal direction parallel to the length
of the frame. Two rails were provided on the top of the frame to move a cutting blade assembly
consisting of a cutting tool with driving motor from one end to the other. Accurate machining was done
to obtain the geometric centre of chuck fixed on the headstock and revolving centre of the tailstock
concentric in one line. The rails for guiding the movement of the trolley was made with 50mm square bar
fixed on the top of the main frame on either sides.
TABLE 1: SPECIFICATIONS OF THE AUTOGROOVER MACHINE
S.N. Particulars Values
1 Cutter blade diameter (mm) 100
2 Cutter blade thickness (mm) 1.5
3 Cutter blade speed (rpm) 1400
4 Power required to drive cutter blade (HP) 1
5 Power required to drive the roller and trolley (HP) 1
6 Trolley size (mm x mm) 600 x 350
7 Number of groves 18
8 Overall size of the machine (m x mm x mm) 2000 x 600 x 660
Head Stock
The head stock consisted of a cast iron box having size, 270 mm length, 300 mm width and 350mm
height. The opposite faces were bored and fitted with a spindle having 50 mm diameter and 500 mm
length. On the inner side of the spindle a 150 mm true chuck was fitted to hold the roller. On the outer
side of the spindle a bevel gear was mounted on bush bearing which could be rotated independent of the
chuck. The spindle and the bevel gear are locked together by an indexing mechanism. This consists of an
arm fixed on the spindle perpendicular to it and having a spring loaded nail which locks it to a hole made
on the bevel gear. At the same radius a total of eighteen holes are made at 20
0
angles apart. The bevel
gear was attached to a pinion gear which is mounted on a shaft fitted vertically on the on the base frame.
The movement of the chuck and the cutter assembly are synchronized for uniform forward and backward
motion.
Tail Stock
Screw type tail stock was made with 50 mm screw shaft and 4 TP threading attached to a square nut
connected to the top sliding block. The forward and backward movement was obtained by rotating a
handle attached to the screw. The roller was secured between the chuck and the tail stock horizontally for
grooving by moving the tail stock towards the chuck by rotating a handle provided on the screw attached
to the tailstock. Gripping of the roller is provided by tightening the chuck on the head stock.
Cutter Assembly
A 600 mm X 350 mm horizontal trolley was fabricated with 45 x 45 x 5 mm L section material and
mounted on 4 pulleys to slide on the rails provided on the main frame. A rotary saw blade and motor
assembly was mounted assembly was mounted on a sliding spindle mounted on the trolley. This
assembly could be lifted or lowered to adjust the depth of groove by rotating a wheel attached to a gear
box on which a spindle is mounted. The rotary saw cutter was fixed at an angle of 17
o
to the direction of
forward movement of the trolley. . The cutter blade was positioned exactly above the centre line of the
roller. This trolley was synchronized with the rotation of the chuck.

Fig. 1: Schematic Diagram of Autogroover Machine (Front Elevation)

Fig. 2: Schematic Diagram of Autogroover Machine (Top View)
The performance of the autogroover machine was evaluated and the capacity, power requirement and
energy consumption of the machine was studied. The machine performance was compared with the
manual method of grooving. The precision and accuracy of the grooving was also noticed.
The roller was mounted horizontally between chuck and revolving centre. The cutting circular saw
was adjusted to just touch the roller by bringing the tool post carrier near to the chuck. Then the cutter
saw was rotated by starting the motor and was slowly brought down by rotating the handle attached to the
spindle of the gear box mounted on tool post carrier. By this method the depth of cut was suitably
adjusted to 2 mm. Then the trolley was moved forward by starting the motor for drive assembly. As the
Development of an Automatic Roller Grooving Machine for Making Helical Grooves on Rollers Used in Roller Ginning 541
trolley with cutter saw moves forward, the roller rotates in clockwise direction as it is attached to the
same shaft through a pinion and a bevel gear mechanism. This way the synchronization between the
forward movement of the trolley and clockwise rotation of the roller was made in such a way that when
the saw cutter moves from the head stock end to the tail stock end, the roller makes a rotation of 270
0
. A
helical groove was made on the surface of the roller when the trolley was moved forward. The indexing
for making further grooves was done manually by adjusting the pin on the head stock spindle to lock with
the successive holes on the bevel gear which aligns the roller de-linking from the trolley movement. A
total of eighteen grooves were thus made on the roller.
Performance Evaluation
The developed machine was successfully tested at Ginning Training Centre of Central Institute for
Research on Cotton Technology (CIRCOT), Nagpur. The chrome composite leather rollers used in cotton
ginneries were used for the trials. Roller was mounted on the autogroover machine and grooving was
carried out. The necessary adjustments of indexing mechanism and the cutter assembly were carried out
before starting the machine. The performance of the machine was found to be satisfactorily. The precise
and accurate helical grooves were made on the roller with the help the machine. The equally spaced
grooves of uniform depth and breadth of 2 mm were cut with precision and accuracy. The successive
grooves at the designed spacing between two grooves were also carried out. The uniform spacing
between the two successive grooves throughout the length of the roller was noticed. The re-grooving on
the same impression after reduced roller diameter was also successfully carried out.
The actual time required for cutting a grove was found to be 30 seconds which includes the time
required for forward and return movement of the trolley. The total time required for grooving the
complete roller with 18 grooves was found to be 10 minutes. Six rollers were grooved with the
autogroover machine in an hour.

Fig. 3: Pictorial View of Autogroover Machine
Grooving on the similar roller was carried out with conventional manual method by a saw pulled by
two men. Manually it takes an hour to groove the complete roller with 18 grooves. The depth, width and
spacing of the grooves were not uniform throughout the length of the roller. Non uniform spacing and
improper depth and width of the groves adversely affect the fibre quality and the ginning output. The
autogroover machine can successfully replace the existing manual method of grooving and will avoid the
drudgery involved in this operation.

Fig. 4: Pictorial View of Manual Grooving of Leather Roller
CONCLUSION
1. Autogroover machine was designed and developed to make helical grooves on the roller used in
cotton ginneries. The rotary motion of the roller and forward motion of the trolley was
synchronized together with suitable drive mechanism. The rotary motion of the saw blade cutter
was achieved with the appropriate mechanism.
2. The machine was successfully tested and its performance was found to be satisfactorily. The
machine was found to groove the roller in ten minutes with 18 grooves on it. The equally spaced
grooves of uniform depth and breadth of 2 mm were cut with precision and accuracy.
3. The re-grooving on the same impression after reduced roller diameter was also successfully
carried out.
4. The autogroover machine can successfully replace the existing manual method of grooving and
will avoid the drudgery involved in this operation.
REFERENCES
[1] Antony, W.S., 1990. Performance characteristics of cotton ginning machinery Transactions of the ASAE, Vol. 33(4): 1089-
1097.
[2] Arude V. G, 2004. Roller replacement for DR gins at appropriate time: A surer way to preserve quality and earn more.
CIRCOT ginning bulletin, Vol (4) Issue (1): 2-3.
[3] Arude V. G., Shukla S.K., Patil P.G., 2004. Effect of leather roller wear on ginning output in roller gins. Journal of the
Indian Society for Cotton Improvement. Vol. 29 No.2: 106-115.
[4] Arude V. G., Shukla S.K., Manojkumar T. S., 2008. Cotton Ginning: Technology, Trouble shooting and Maintenance. A
book published by CIRCOT, (ICAR) Mumbai. PP: 1-240.
[5] Lal, M.B., 2001. Role of TMC in the production of trash and contamination free cotton in India, Paper presented in
National seminar on Relevance of Ginning in the Production of trash free cotton held at GTC of CIRCOT, Nagpur. Book
of papers: 3-8.
[6] Sharma, M.K., 1999. Current Scenario of Ginning Industry. Paper presented in International seminar on Cotton and its
utilisation in the 21
st
century held at CIRCOT, Mumbai, Book of papers: 161-168.
The Effect of Quarantine Treatments

on the Physical Properties of Cotton Fibres
and Their Subsequent Textile
Processing Performance
M.H.J. Van, Der Sluijs
1
, F. Berthold
2
and V. Bulone
3
1
CSIRO Materials Science and Engineering, Geelong, Australia

2
Innventia AB, Stockholm, Sweden

3
Royal Institute of Technology (KTH), School of Biotechnology
and Swedish Centre for Biomimetic Fibre Engineering, Stockholm, Sweden

Despite its declining market share of the world fibre market, cotton still remains an important fibre,
not only for the textile and clothing industry but also as a valuable source of income to a large number of
countries. Cotton is currently grown in over 60 countries worldwide, with around 31% exported annually
for processing in textile mills. With this comes the risk of exotic pests and diseases entering a country
which can seriously affect its unique environment and native flora and fauna. Quarantine thus plays a
critical role to ensure that a country remains free from serious pests, weeds and diseases present in other
parts of the world. Most countries that import cotton fibre insist on a phytosanitary certificate which may
require that bales of cotton are treated to ensure that the consignment is free of live insects, soil and other
debris. The quarantine treatments used are generally chemical (fumigation) and in some instances
radiation (gamma irradiation). This paper examines the effects of these various quarantine treatments on
the physical properties of cotton fibres and the subsequent effect on textile processing performance.
METHODS AND MATERIALS
Standard Upland cotton and Extra Long Staple (ELS) cotton were used to determine the effect of the
various quarantine treatments on the physical properties of cotton fibre. Commercially grown Australian
Upland saw ginned (Sicala V2R) as well as ELS roller ginned (Pima A8) cotton grown during the
2006/07 season under commercial growing conditions and machine harvested were used for this study.
The Sicala V2R cotton was grown at Trangie in the Macintyre Valley of New South Wales (NSW) and
the Pima A8 was grown at Moree in the Gwydir Valley of NSW. Samples were gathered from various
parts of the bales and sent for fibre testing.
FIBRE TESTING
Fibre testing was carried out on the cotton fibre to determine an initial profile for comparison and is
described as untreated (NT).
Bale (raw fibre) samples were conditioned under standard conditions of 20C +/-2C and 65% +/-3%
relative humidity for 24 hours as per ISO 139. The samples were then tested on an Uster Technologies
1000 High Volume Instrument (HVI), as per American Society for Testing and Materials Standard
(ASTM) D5867, for Micronaire, staple length, length uniformity, staple strength and elongation (Table
1). The visual grade, based on colour (colour grade) and visible trash (leaf grade) of the fibre was
determined by classification of cotton according to the United States Department of Agriculture (USDA)
established standard grade boxes.
Fibre fineness was determined using the CSIRO Cottonscan instrument, which determines fibre
fineness (linear density) by measuring the length of fibre in an accurately weighed specimen of fibre
snippets. Combined with an independently measured Micronaire value from the HVI, the average fibre
maturity was also calculated using Lords empirical relationship between Micronaire, maturity ratio and
fineness (Table 2).
88
Bale samples were also tested for nep, seed-coat neps (SCN) and short fibre content (SFC) by an
Uster Technologies Advanced Fibre Information System (AFIS PRO), as per ASTM D5866 (Table 3).
QUARANTINE TREATMENTS
In order to determine the effects of the various quarantine treatments on the physical fibre properties,
samples were treated as per the standard methods prescribed by the Australian Quarantine and Inspection
Services. Samples of cotton were subjected to irradiation and fumigation at Steritech Pty Ltd in
Melbourne, Victoria. For irradiation, samples were gamma irradiated at various dosages (21, 29, 57 and
74 kGy) using the radioactive isotope Cobalt 60. Samples for fumigation were exposed to ethylene oxide
(ET) at a rate of 1200g/m for 5 hours at a minimum temperature of 50C. Samples subjected to
fumigation by methyl bromide (MB) were treated at ISS Australia in Sydney, New South Wales, at a rate
of 32g/m for 24 hours.
Following treatment all the samples were retested on the same instruments and under the same
conditions to determine their physical properties, as described earlier.
PHYSICAL ANALYSIS
Physical analysis was also conducted on samples both before and after quarantine treatment in order to
explain any changes in the mechanical properties of the cotton fibre. Cotton fibres (only NT and 74 kGy)
(2nm Chromium coated) were imaged using a Hitachi S4300 scanning electron microscope (SEM), to
detect any physical changes.
Cotton samples (only NT and 21, 29, 57 & 74 kGy) were prepared for molecular weight
determination of cellulose as described by Evtuguin et al. (2003). Samples (1 g) of cotton fibres from
each irradiated line and the corresponding untreated controls were extracted for six hours in
toluene/ethanol (2/1) using a Soxhlet extractor. Husks and impurities were removed and 200 mg of the
remaining material was transferred to a beaker and mixed with 10 ml of a 10% peracetic acid solution
whose pH had been preliminarily adjusted to 4.8 with NaOH. The reaction was performed for 30 minutes
at 75C and the solution cooled on ice and filtered. The cotton samples were then washed three times
with 100 ml deionised water and once with 100 ml acetone/ethanol (1/1). The samples were then air dried
at ambient temperature.
Size-exclusion chromatography (SEC), a proven and effective technique for measuring molecular
weights (MW) of cellulose, was then performed on 15 mg of each dried cellulose sample treated with
peracetic acid. Cellulose was swollen in water and solvent exchanged three times with dry N, N-
dimethylacetamide (DMAc) before the addition of 2.1 ml of an 8% LiCl/DMAc solution.
Ethylisocyanate (0.22 ml) was then added to the mixture to fully dissolve the cellulose during 5 days
incubation at room temperature. The cellulose solutions were then analysed by SEC using a 2690
Separation Module, equipped with 4 x Mini-Mix-A (20m; 4.6 x 250 mm) columns, as described by
Berthold et al. (2004).
TEXTILE PERFORMANCE
In this part of the study we determined what the effects are of the various quarantine treatments (only NT,
21, 29, 57 & 74 kGy) on the textile processing performance and quality of the Upland and ELS cotton.
Moisture Regain
Moisture regain was tested according to ASTM 1576. Samples were wet out in water with small amount
of wetting agent, squeezed out and left to dry under standard laboratory conditions for 2 weeks prior to
testing.

The Effect of Quarantine Treatments on the Physical Properties of Cotton Fibres 545
Dyeing
Ten grams of each sample was put into a dyeing basket and placed in an Ahiba Turbomat dyeing
machine. Samples were then scoured and dyed with Cibacron red LS6G (1%) reactive dye using a
standard recipe. Upon completion, the samples were removed, hydro extracted (using a centrifuge to
remove excess water) and air dried. Each sample was then put in the viewport of a Gretag Macbeth Color
Eye 7000A spectrophotometer. Each sample was measured 5 times and the readings averaged.
Yarn Production
In order to determine textile performance of the cotton samples, a hybrid system was used which consists
of a re-furbished Shirley miniature spinning plant manufactured by the Platt Company in the UK. This
system consists of using the miniature card and draw frame to produce slivers, which are then transferred
to a full-scale spinning system for further processing through a further draw frame passage and the
production of roving, prior to spinning, as described by van der Sluijs et al. (2009). Draft and twist was
optimized for each sample to deliver a 30 Ne carded yarn with a twist factor of e 3.7 (792 turns per
meter). One yarn bobbin per irradiation dosage was tested for quality parameters
Spun yarns were conditioned under standard conditions of 20C +/-2C and 65% +/-3% RH for 24
hours and tested for linear density (count) as per Australian Standard (AS) 2001.2.23, twist as per AS
2001.2.14, evenness, hairiness and imperfections using an Uster Technologies 4-SX evenness tester as
per ASTM D1425. Tensile properties were determined using the Uster Technologies Tensorapid 3, as per
ISO 2062:2009.
Fabric Production and Dyeing
After yarn testing, the left over yarns were waxed and wound but not cleared using a Schlafhorst 238RM
winder. Wound yarns were then knitted on a Lawson Hemphill 25.4cm 10inch F.A.K. circular knitting
machine, using a cover factor of 1.37 to produce a fabric weight of 153 g/m
2
.

The dyed fabric was conditioned under standard conditions of 20C +/-2C and 65% +/-3% RH for
24 hours and tested for mass per unit area by AS 2001.2.13, for bursting strength as per ISO 13938-
2:1999 and abrasion resistance using the Martindale instrument as per AS2001.2.25-06.
Physical Results
By any measure, the fibre properties of the two cottons used in this study can be considered as good
quality and above the Australian base grade
1
. As expected, the ELS cotton had much higher bundle
tenacity and was inherently finer than the Upland cotton (Table 2), which would have positively affected
the bundle tenacity result.
TABLE 1: FIBRE RESULTS BY THE HVI 1000
1*
AND VISUAL GRADE
Variety Tenacity
g/tex
Elongation
%
Length
Inch
SFI
%
UI
%
Micronaire
g/Inch
Visual
Grade
Upland 29.8 6.1 1.161 9.1 82 4.08 11~3
ELS 44.5 6.4 1.358 5.9 86 4.01 1
1
Calibrated using HVI ICC Upland and Pima Calibration Cottons
*
Average of 10 tests
TABLE 2: FINENESS
2
BY COTTONSCAN AND CALCULATED MATURITY RESULTS

Variety Fineness (MTEX) Maturity Ratio
Upland 187 0.71
ELS 167 0.75
2
Average of 5 Tests
1
Australian Upland base grade quality is staple length > 1.130 inch, strength > 29 g/tex and Micronaire in the range of 3.5-4.9.
ELS base grade quality is staple length > 1.350 inch, strength > 44 g/tex and Micronaire in the range of 3.5-4.1.
546
Variety
Upland
ELS
3
Avera
The a
with y-er
treatments
fibre.How
the fibre, w
In the
21 kGy, w
in fibre str
In the
reducing b
each subs
strength c
did increa
decrease i
The e
graphicall
age of 5 Tests
average resul
rror bars of
s by either M
wever gamm
with these ef
e case of Upl
with the fibre
rength was a
e case of the E
by almost 5
sequent radia
corresponded
ase at a dos
in elongation
xtent of the
ly illustrated
World Cot
TA
Nep
lts after the
one standar
MB or ET ha
ma irradiation
ffects becom
and cotton, t
e strength sig
a gradual, but
ELS cotton,
g/tex (11%)
ation dosage
d with a grad
sage of 21
n with a dosa
decrease in f
in Figures 1
F
Fig
tton Research C
ABLE 3: NEP, SEED-CO
s/ gram
326
167
various qua
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ad little or no
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ming more ap
the strength o
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the strength
from 44.5 g
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dual but signi
and 74 kGy
age of 29 kGy
fibre strength
and 2.
ig. 1: Effect of Qua
g. 2: Effect of Quar
Conference on T
OAT NEP AND SFC RES
rantine treat
n, giving an
o significant
e lower dosa
parent and si
of the fibre r
educing with
decrease in f
was affected
g/tex to 39.4
strength was
ificant decrea
y. Notwithst
y and 57 kGy
h and elonga
rantine Treatment
rantine Treatment
Technologies fo
SULTS BY THE AFIS PR
SCN/ gram
21
10
tments are sh
n indication
t effect on th
ages, has an e
ignificant as
reduced signi
h each dosage
fibre elongat
d more signif
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s significantl
ase in fibre e
anding this
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ation due to t
ts on Fibre Streng
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or Prosperity
RO
3
hown in Fig
of the varia
he physical p
effect on the
the dosage s
ificantly afte
e. Correspon
tion with eac
ficantly than
gamma irradi
ly affected.
elongation, a
anomaly, th
the various q

gth
tion
SFC(w)%
8.9
2.9
gures 1 to 3,
ation. The f
properties of
e physical pr
strength incr
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nding with th
ch dosage.
n for the Upla
iation of 21
This decreas
although the
here was a
quarantine tre

%
expressed
fumigation
f the cotton
operties of
eased.
adiation of
he decrease
and cotton,
kGy. With
se in fibre
elongation
significant
eatments is

For b
were not
short fibr
subsequen
determine
decrease
decreasing
There
colour de
Spotted G
on the co
irradiation
The e
Figure 3.
STRUCTUR
In order t
irradiation
weight de
As ca
fibres wer
2
The propor
Th
both the Upla
affected by
re index (S
nt higher do
ed by the AF
at the 29 kG
g from 86%
e was a slight
eteriorated fr
Good Middlin
olour of the
n did not seem
extent of the
AL ANALYSIS
o determine
n, the sampl
etermination.
an be seen by
re found betw
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e Effect of Qua
and and EL
gamma irrad
FI)
2
increas
osages. This
FIS PRO. Th
Gy dosage,
to 83% with
t impact on t
from Good M
ng (13~3) co
e cotton as
m to make a
increase in S
Fig.
OF IRRADIATE
the reason f
les were ana

y the SEM im
ween the untr
of fibres shorter
rantine Treatme
S cotton, M
diation. Fibr
sed significa
s trend was
his in turn als
with Upland
h subsequent
the colour of
Middling W
otton as the g
Pima cotton
any differenc
SFI due to th
3: Effect of Quara
ED COTTON SA
for the signif
alysed by a
mages shown
reated and tr
r than one half i
ents on the Phy
Micronaire, m
re length wa
antly at the
also noted
so had an ef
d cotton dec
higher dosag
f the Upland
White (11~3)
gamma irrad
n is naturall
e to the colo
he various qu
ntine Treatments
AMPLES
ficant change
range of tec
n in Figure 4
reated sample
inch, measured
sical Properties
maturity, fibre
as also not si
29 kGy d
in the shor
ffect on lengt
creasing from
ges.
d cotton, altho
to Good M
diation dosag
ly creamier
ur.
uarantine trea
on Short Fibre Ind
es in the phy
chniques inc
, no significa
es.
by the HVI.
s of Cotton Fibre
e fineness a
ignificantly
dosage, grad
rt fibre cont
th uniformity
m 82 to 80%
ough the effe
Middling Lig
ges increased
than Uplan
atments is gr
dex %
ysical fibre p
cluding SEM
ant differenc
es
and nep cont
affected; how
dually increa
tent %, by w
y which also
% and the E
fect was mini
ght Spotted
d. There was
nd cotton an
raphically ill

properties aft
M, and SEC
ces to the sur
547
tent values
wever, the
asing with
weight, as
o tended to
ELS cotton
imal as the
(12~1) to
no impact
nd gamma
lustrated in
fter gamma
molecular
rface of the

548
DETERMINA
The estim
cotton, wh
These
Upland co
parameter
actual rea
lines from
irradiation
progressiv
apparent M
irradiation
cellulose
fibrillar st
the more
the observ
data, sam
irradiation
fibres is n

Fig. 4: Represent
ATION OF CELL
mated Molecu
hile the Upla
Fig. 5
e MW corres
otton. This di
rs have never
ason for the o
m their origin
n doses were
ve than that
MW of all ce
n dose of 74
chains at the
tructures, wh
crystalline d
ved alteration
mples from w
n doses com
not significan
World Cot
tative SEM Images
LULOSE MOLEC
ular weight (
and cotton ex
5: Estimated Molec
spond to a de
ifference ma
r been repor
observed dif
nal value (10
e accompanie
observed be
ellulose samp
kGy. These
e most sensi
hile higher d
domains of th
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wood have b
mpared to the
ntly modified
tton Research C
of Untreated (Lef
CULAR WEIGH
(MW) of the
xhibited a sig
cular Weight of Ce
egree of poly
ay reflect gen
rted to affect
fferences rem
0
6
or 7 x 10
5
ed by a conti
etween the n
ples decreas
results sugg
tive points p
doses are req
he fibres. Th
chanical prop
been shown
e untreated s
d upon irradia
Conference on T
ft) and 74 kGy Irra
T
e non irradia
gnificantly lo
ellulose from Untre
ymerization
netic variatio
the MW of
mains unclea
Da) to ca 2.
inued decrea
non irradiate
ed from 2.5
gest that a 21
probably rep
quired to furt
he decrease i
perties of th
to exhibit a
samples, wh
ation (Aoki e
Technologies fo
adiated (Right) Cot
ated cellulose
ower apparen
eated and Gamma
(DP) of ca 6
on or differen
cellulose fro
ar. The obser
5 - 3 x 10
5
D
se in MW, b
ed samples a
- 3 x 10
5
Da
kGy irradiat
resenting do
ther decrease
in molecular
e cotton fibr
a decreased m
hile the degre
et al. 1977).
or Prosperity
tton: Top Upland; B
e samples w
nt MW (7 x 1
Irradiated Cotton
6200 for the
nt growth con
om different
rved apparen
Da under a d
but the observ
and those exp
a to 10
5
Da w
tion dose pro
omains of low
e the size of
size is most
res upon irra
molecular w
ee of crystal

Bottom ELS Cotton
was 10
6
Da fo
10
5
Da) (Figu

n Fibre
ELS and 43
nditions, alth
cotton lines
nt MW dropp
dose of 21 kG
ved decrease
posed to 21
when exposed
ovokes the b
wer crystalli
f cellulose ch
t likely respo
adiation. Sim
weight upon
llinity of the

n
or the ELS
ure 5).
300 for the
hough such
. Thus, the
ped for all
Gy. Higher
e was more
kGy. The
d to a total
breakage of
inity in the
hains from
onsible for
milar to our
increasing
e cellulose

TEXTILE PE
Moisture R
Figure 6 s
fumigatio
ELS cotto
decreasing
change.
Dyeing
Figure 7 s
shade and
decreases
Th
ERFORMANCE
Regain
shows the mo
n with either
on fibre. Irrad
g as the dos
shows a pho
d hence dye
and hence th
F
e Effect of Qua
oisture regain
r MB or ET
diation with
sage increase
Fig. 6: Moisture R
otograph of th
e uptake is
he shade of t
Fig. 7: Dye Uptake o
rantine Treatme
n of the untr
has a signif
a 21 kGy do
es to 57 kG
Regain of Upland a
he untreated
clearly visib
the samples d
of Untreated and T
ents on the Phy
reated and tre
ficant effect
osage gives s
Gy, after whi
nd ELS Cotton Fibr
d and treated
ble. As the
decrease.
Treated Cotton Fib
sical Properties
eated fibre sa
on the moist
similar result
ich the mois
re at Various Quar
fibre sampl
irradiation
bre at Various Qua
s of Cotton Fibre
amples. It is
ture regain o
ts with the m
sture regain
rantine Treatment
les after dyei
dosages inc
arantine Treatmen
es
interesting t
of both the U
moisture rega
does not sig

ts
ing. The diff
crease, the d
nts
549
to note that
Upland and
ain steadily
gnificantly
ferences in
dye uptake

550
Each
Each sam
were mea
Difference
difference
values ne
significan
Yarn Resul
There wer
treated fib
the irradi
elongation
The y
NT cotton
with an ir
into fabric
the yarn s
strength r
impossibl
74 kGy.
sample was
mple was me
asured using
es between
e from the u
ear or greate
nt on the basi
lts
re no signific
bre samples.
ation dosage
n results.
yarn strength
n and the 21
rradiation do
c by the wea
strength dec
reduced to
e. The yarn
World Cot
then put in t
asured 5 tim
g the CIELA
the samples
untreated fibr
er than one
is of the mon
TABLE 4: FIBRE
Upla
N
M
E
2
2
5
74
EL
N
M
E
2
2
5
74
cant differen
However, a
e strength in
h, for both th
kGy irradia
osage of 29 k
aving and als
reased even
18.7cN/tex,
strength was
tton Research C
the viewport
mes and the r
AB colour s
s were asses
re samples a
between th
nochromatic n
E APPEARANCE REPORT
and L*
NT 49.0
MB -4.3
T -2.2
1 -3.3
9 1.2
7 1.7
4 5.74
LS L*
NT 48.1
MB 1.3
T -1.2
1 4.52
9 3.7
7 7.4
4 7.8
nces in the ev
as can be see
ncreases, wi
he Upland an
ted cotton. H
kGy, which w
so the knittin
further to b
which wou
s further redu
Fig. 8: Yarn Stren
Conference on T
of a Gretag
readings ave
scale, using
ssed in term
and takes in
he untreated
nature of the
TED IN LIGHTNESS (L),
* a*
05 54.19
34 -0.47
20 -0.61
38 -3.97
7 -3.86
7 -4.85
4 -7.71
* a*
10 52.51
5 0.36
27 1.23
2 -1.75
1 -3.36
6 -5.95
8 -7.83
venness and
en in Figure
ith the resul
nd ELS cotto
However, the
would make
ng processes
below 10 cN
uld make p
uced to 14.8
ngth of Untreated a
Technologies fo
Macbeth Co
eraged. The
L (lightnes
ms of delta E
nto account d
and treated
e dyed sampl
, REDNESS (A), BLUEN
b*
20.16
0.26
-0.53
-4.05
-4.55
-5.13
-6.95
b*
20.08
-3.17
1.68
-3.17
-3.59
-6.09
-7.04
imperfection
8, the yarn
lts typically
on yarns, did
ere was a sig
e the Upland
. With an irr
N/tex. In the
processing o
cN/tex with
and Treated Cotto
or Prosperity
olor Eye 7000
appearance
ss), a (redne
E (E), whi
differences i
d samples w
les.
ESS (B), AND E
E
0
2.0
1.07
2.9
2.7
3.2
5.1
E
0
0.78
1.13
2.8
2.8
5.1
5.8
n values betw
strength sign
following t
d not reduce
gnificant redu
d cotton yarn
radiation dos
case of the
f fabric on
h irradiation d

n
0A spectroph
of dyed fibr
ess) and b (
ich it the to
n L, a and b
were deemed
ween the unt
nificantly de
the fibre str
significantly
uction in yar
n impossible
sage of 57 an
e ELS cotton
high speed
dosages of 5
hotometer.
re samples
(blueness).
otal colour
b. Delta E
d as being
treated and
ecreases as
rength and
y from the
rn strength
to process
nd 74 kGy
n, the yarn
d weaving
7 kGy and

As ca
elongation
between N
reduction
case of th
and a dos
to 3.7%).
Fabric Res
As can be
increases,
following
irradiation
strength w
although s
The a
that abras
6000 rubs
CONCLUSIO
The fumig
on the phy
have an e
Th
an be seen in
n. As was th
NT, Upland
in yarn elon
he Upland co
age of 74 kG
In the case o
sults
e seen in Fig
indicating
the yarn str
n dosage. Fo
with a dosage
significant, th
abrasion resis
sion resistanc
s as compare
ON
gation treatm
ysical proper
effect on the
e Effect of Qua
n Figure 9, th
e case with t
and ELS cot
ngation betwe
otton, the yar
Gy almost re
of the ELS co
F
gure 10, the
that the fab
rength result
or both the
e of 21 kGy.
he reduction
Fig. 10: Fab
stance was d
ce of both th
d to NT cotto
ments by eith
rties of the c
e physical pr
rantine Treatme
his decrease
the yarn stren
tton and the
een NT Upla
rn elongation
educed the ya
otton, the yar
Fig. 9: Yarn Elonga
fabric burst
bric strength
ts in Figure
Upland and
. The fabric
n in strength w
bric Bursting Stre
determined on
he Upland an
on failing at
her ethylene
otton fibre. H
roperties of
ents on the Phy
in yarn stren
ngth, there w
21 kGy irrad
and and ELS
n at 57 kGy w
arn elongatio
rn elongation
ation of Untreated
ting strength
decreases a
1. Fabrics b
d ELS cotto
strength con
was not as la
ength of the Untrea
nly for NT a
nd ELS cotto
~ 50 000 rub
oxide or me
However gam
the fibre, wi
sical Properties
ngth coincid
was no signif
diated cotton
cotton and t
was similar t
on by half w
n continued t
and Treated Cotto
significantly
as the irradi
became sign
on, there wa
ntinues to dec
arge as betwe
ated (NT) and Trea
and 74 kGy i
on was signi
bs.
ethyl bromid
mma irradiat
ith these eff
s of Cotton Fibre
ded with a gr
ficant differe
n. There was
the 29 kGy i
to the results
when compar
to steadily dr

on
y decreases
iation dosag
nificantly we
as a significa
crease with t
een the NT a

ated Cotton
irradiated fab
ficantly affe
de had little o
tion, even at
fects becomi
es
radual decrea
ence in yarn
s however a
rradiated cot
s of the 29 k
ed to NT cot
rop with each
as the dosag
es increases
eaker with ea
ant decrease
the higher do
and the 21 kG
brics. The re
cted, both fa
or no signifi
the lower do
ing more app
551
ase in yarn
elongation
significant
tton. In the
Gy dosage
tton (6.6%
h dosage.
ge strength
s, typically
ach higher
e in fabric
osages but,
Gy dosage.
sults show
ailing at 5-
cant effect
osages, did
parent and
significant as the dosage strength increased. Our results show that the physical fibre properties of the
cotton such as; strength, elongation, length uniformity, short fibre and to a lesser extent length and colour
are affected by gamma irradiation. Moisture regain and dye uptake were also significantly affected. In
addition, there were differences between the Upland and ELS cotton varieties. Analysis by various
microscopy methods revealed no noticeable surface damage. However, the apparent molecular weight of
the cellulose present in cotton fibre decreased even at low gamma irradiation dosage (21 kGy),
irrespective of variety. The decrease in molecular size is in all likelihood responsible for the changes in
the physical properties of the cotton fibers upon irradiation.
From a textile processing performance point of view, yarn results show that the various irradiation
dosages did not have a significant impact on the evenness and imperfection values. However, gamma
irradiation had a significant effect on the yarn strength and elongation results of both the Upland and ELS
cotton, with the results typically following the fibre strength and elongation results. The fabric results
also show that gamma irradiation does have a significant effect on fabric strength and abrasion resistance
for both the Upland and ELS cotton, with the results typically following the yarn strength results.
The salient message from this study is that if any cotton lint needs to be treated by quarantine one
must insist on chemical (fumigation) by either ethylene oxide or methyl bromide as gamma irradiation,
even at low dosages, severely damages the physical properties of cotton lint. These physical damages can
cause major issues if the cotton lint is to be used to either calibrate fibre testing instruments, to
benchmark various cottons or for commercial processing.
ACKNOWLEDGEMENT
The authors gratefully acknowledge the financial support of the Cotton Research and Development
Corporation and CSIRO Materials Science and Engineering. They also acknowledge the assistance of
Jeff Church, Colin Veitch and Bernadette Lipson in compiling this report. The support of Steritech and
the Auscott Classing facility is also acknowledged. The authors also acknowledge the assistance of Fred
Horne, Mark Freijah, Phil Henry, Lisa O Brien, Peter Herwig and Colin Brackley for processing the
fibre through to fabric as well as Susan Miller and Liz Coles for testing the samples.
REFERENCES
[1] Aoki, T., Norimoto, M. and Yamada. T., (1977) Some Physical Properties of Wood and Cellulose Irradiated with Gamma
Rays. Wood Res. 62, pg 1928F.
[2] Berhold, F., Gustavsson, K., Berggren, R., Sjholm, E. and Lindstrm, M., (2004) Dissolution of Softwood Kraft Pulps by
Direct derivatization in Lithium Chloride/ N, N-Dimetylacetamide. J. Appl. Polym. Sci., 94: 424431
[3] Evtuguin, D.V. Tomas, J.L. Silva, A.M.S and Neto, C.P. (2003) Characterization of an acetylated heteroxylane from
Eucalyptus globulus Labill. Carbohydr. Res. 338: 597604.
[4] Lord, E., (1956) Airflow through plugs of textile fibres. Part II. The Micronaire Test of Cotton. J. Text. Inst.
47: T16 T47
[5] International Cotton Advisory Committee, (2010) Cotton: World Statistics.
[6] Van Der Sluijs, M.H.J, Long, R. and Gordon, S., (2009) An Alternative Miniature Cotton Spinning System, Beltwide
Cotton Conference, pg 14461452
The Impact of Cotton Fibre Maturity

on Dye Uptake & Low Stress Mechanical Properties
of the Fabric
S. Venkatakrishnan and R.P. Nachane
CIRCOT, Coimbatore

AbstractIn the Present study True Maturity has been evaluated in Premier ART-2, after testing with reference
standard Maturity Samples, supplied by the instrument manufacturer, whose maturity values measured and labeled
using Image processing technology from the renowned FASER INSTITUT, Bremen, Germany and the line of best fit
for Maturity values has been observed in ART-2 and the linear regressed line is 99%, which indicates the
measurement principle is an excellent way of measuring Maturity. Further In the present study, in order to know the
impact of cotton fibre Maturity on Dye uptake and low stress mechanical properties, two groups of cotton samples has
been collected (long staple with 33mm and extra long staple with 36mm), from different growth areas in order to have
broad ranges of Maturity.The above groups of cotton is tested in PREMIER ART 2 instrument for True Maturity.
Yarn and knitted samples were produced from each cotton sample and all the process parameters for producing such
samples were kept constant. The k/s value of grey and dyed fabrics produced from different growth regions of cotton
sample dye differently due to the differences in Maturity. The knitted fabric produced from the yarn, spun from high
matured cotton showed higher compressional recovery, tensile resilience and smoother surface compared to the fabric
produced from the yarn spun from low matured cotton samples.
OBJECTIVES
This research paper deal with the newly introduced fibre parameter called True Maturity measurement
from PREMIER ART 2 Cotton Fibre Testing Instrument.
The objective of the research paper is to find out
Traceability and co relation of True Maturity measurement.
Repeatability of the instrument
Usefulness of TRUE MATURITY from PREMIER ART 2 on dye uptake and low stress
mechanical properties.
Traceability
The 4 reference materials used are namely RSM 1, 2,3 and 4 having the maturity values measured and
labeled using Image processing technology from the renowned FASER INSTITUT, Bremen, Germany
and supplied by the instrument manufacturer. These materials cover the wide range of maturity values
from 0.72 to 1.06
Repeatability of the Instrument
The reference materials from FASER INSTITUT, Germany (namely RSM 1, 2,3 and 4) were tested in
PREMIER ART 2 for True Maturity measurement on various days to ascertain the REPEATABILITY of
the instrument.

89
554
Usefulness
In order to
prepared.
second gr
cottons w
groups of
were prod
kept cons
stress mec
IMPORTANC
The world
select cott
also impo
establishe
finishing
problems
out any sa
several gr
procured
difference
fibres in a
lowering o
In 10
variation a
shows the
The F
Fluoresce
properties
type of sy
enough to
successful
but gener
together, t
s of True Matu
o find out th
First group
roup contain
were taken fro
f cottons wer
duced from e
tant. The dif
chanical prop
CE OF MATURI
d wide expan
ton and it he
osed new ch
ed varieties a
problems w
witnessed a
avings in raw
rowth region
from differe
es in Maturit
a sample is u
of appearanc
0 % cotton,
and hairiness
e % influence
Fibre Propert
nce. Therefo
s in the lay d
ystem to cate
o control to
lly to contro
rally it never
then addition
World Cot
urity from Pre
he impact of
containing 4
ning 2 cotton
om different
re tested in P
each cotton s
fferences in
perties of eac
ITY
nsion of cott
elped in redu
hallenges fo
and growing
were controll
and is eviden
w material co
ns within th
ent growing
ty of the fib
undesirable a
ce grade due
, the cause
s arises durin
e of each par
ties that hav
fore it is ess
down to get c
gorize the co
o eliminate
ol Barre, if
r happens in
nal testing a
tton Research C
emier Art 2 on
True Maturi
4 cottons of
ns of extra l
growth area
PREMIER A
sample and a
k/s value be
ch cotton sam
ton market h
ucing cost an
or the cotton
g regions, so
ed to some
nt from the r
osts obtained
he country.
regions and
bre. Earlier s
s this causes
to formation
of Fabric B
ng the prepar
rameter in rel
ve major infl
sential for t
consistent dy
otton bales in
the Barre
f the cotton
n a Mill. If
and Maturity
Conference on T
n Dye Uptake
ity values on
long staple
ong staple v
as which cov
ART 2 for Tr
all the proces
etween sampl
mple were te
has opened u
nd improving
n buyers. Ea
o by control
extent. In r
rejection of
d by purchas
Unfortunate
seed varieti
studies repor
s excessive w
n of neps, un
Barre is as
ration proces
lation to the
luence in the
the Spinning
yeing and fin
n an effort to
, especially
being proces
many cotton
y information
Technologies fo
and Low Stre
n fabric prope
variety with
variety with
ver a wide ra
rue Maturity
ss parameter
les of each g
sted using K
up wider pos
g quality and
arlier cotton
ling microna
recent years,
100% cotton
sing different
ely cotton w
ies/ hybrids,
rted that, th
waste in proc
even dyeing
sociated wit
ss in the spin
dyeing defec
e causes of
g Mill to co
ishing of kni
o control the
in Knitted
ssed is from
n varieties o
n is necessar
or Prosperity
ss Mechanica
erties, two g
h staple lengt
staple length
ange of matu
y values. Yar
s for produc
group were a
KAWABATA
ssibilities for
d at the same
n purchase w
aire of the c
, there were
n knitted fab
t varieties / h
with similar
dye differen
e presence o
cessing, lappi
, etc (1,2,3).
th raw mate
nning Mill. T
cts particular

barre are F
ontrol the a
itted fabric. M
average Mic
Fabric. Mi
m same variet
or growing a
ry.
al Properties
groups of sam
th of about 3
h of about 3
urity values.
rn and knitte
ing such sam
analysed. Al
A evaluation
r the spinnin
time this sit
was restricte
cotton, the d
e lot of dyei
brics and it e
hybrids of co
Micronaire
ntly mainly
of excessive
ing of fibres
erial, yarn co
The followin
rly Fabric B
Fineness, Ma
above mentio
Many Mills h
cronaire, and
icronaire can
ty and growt
areas are be

mples were
33mm and
36mm. All
The above
ed samples
mples were
so the low
system.
ng Mills to
tuation has
ed to well
dyeing and
ing related
easily wipe
otton from
that have
due to the
e immature
on rollers,
ount, twist
g pie chart
Barrre.
aturity and
oned fibre
have some
d this is not
n be used
th regions,
eing mixed
The Impact of Cotton Fibre Maturity on Dye Uptake & Low Stress Mechanical 555
Maturity: Literally, cotton fibre maturity is defined based on the cell wall thickness of the fibre. If
the fibre wall is thicker and the hollow portion called lumen is very thin, then the fibre is called as
Matured Fibre. On contrary, if the fibre wall is very thin and the hollow portion is very large then the
fibre is treated as Immatured Fibre. Any fibre lying between these two limits can be treated as Half
Matured.
COTTON FIBRE MATURITYMEASUREMENT METHODOLOGIES
The following are the most important methodologies available at present for measuring cotton fibre
maturity.
Direct Method of Measurement
Caustic soda swelling method
Polarized light method
Image based cross section measurement
Direct measurement involve direct estimation of cotton maturity, In case of sodium hydroxide
swelling technique, the cotton fibres are immersed in NaOH and the appearance of fibres are analysed.
Ribbon like fibres and convoluted fibres are treated as Immatured fibres and rod like fibres are treated
as Matured fibres. In India, three methods have been recognized as standard for the determination of
Maturity using of cotton fibres by Caustic soda technique (4). Method I is followed at CIRCOT. Method
II is based on the ASTM (5) and Method III on the British standards (6)
In case of Polarized light technique, the classification of Maturity is made depending upon the
interference colours produced by the fibres (7, 8).
Image based maturity is the globally accepted methodology of measuring maturity since it is most
accurate and free from human errors. In image based maturity measurement, the cross section of the fibre
is analysed to determine Maturity values. The cross- section of a cotton fibre contains measurable
information directly related to to fibre Maturity. Much research has been conducted using the image
analysis technology to measure cotton Maturity and other parameters from cotton fibre cross-sections (9,
10, and 11).
Indirect Method of Measurement
Double compression methods used in High Volume Instruments manufactured by reputed firms are one
of the indirect methods of measuring maturity. In spite of the importance of maturity, there is no direct or
indirect measurement method that is both fast and reliable. The lack of standards of reference for
maturity has made it impossible to calibrate the existing instruments (airflow instruments with double
compression), further it cant be refined without the reference standard for Maturity. In the present study,
the PREMIER ART-2, Instrument has been tested with reference samples supplied by the instrument
manufacturer, whose maturity values are determined by image analysis technology at Bremen Institute.
TRACEABILITY AND REPEATABILITY
PREMIER ART 2 cotton testing instrument was tested using the following, four different Reference
standard maturity cotton samples.
TABLE 1: MICRONAIRE AND MATURITY COMBINATION OF REFERENCE STANDARD COTTON SAMPLE
High Maturity RSM 3
Maturity: 0.95
Micronaire: 3.20
RSM 4
Maturity: 1.06
Micronaire: 4.02
Low Maturity RSM 1
Maturity: 0.72
Micronaire: 2.76
RSM 2
Maturity: 0.81
Micronaire: 3.48
Low Micronaire High Micronaire
556
Matur
manufactu
correlation
measuring
To de
the referen
SD is low
deviation
0.01(show
The f
PREMIER
DETAILS OF
In the pre
groups of
growth ar
PREMIER
operator c
reported in
rity Testing h
urer and the
n coefficient
g maturity.
etermine the
nce standard
wer for highe
of 0.02 bet
wn in Fig.2)
following F
R ART 2 and
F MATERIALS U
sent study, in
f cotton sam
reas in orde
R ART 2 ins
carried out
n Table-1, gi
World Cot
has been carr
line of best f
t (R) is 0.99
between day
d cottons and
er Maturity s
tween readin
igures show
d their standa
USED
n order to kn
mples (long s
er to have b
trument for T
10 tests and
iven below:
tton Research C
ried out usin
fit for MR va
909, which in
ys repeatabili
d calculated th
samples. The
ngs, where
w the correla
ard values m
now the influ
staple with
broad range
True Maturit
d the average

Conference on T
ng the four re
alues measur
ndicates the
ity, 10 readin
he Mean and
e reference s
as RSM-3
ation of Tru
measured usin
Fig. 2
Fig. 3
uence of mat
33mm and e
es of maturi
ty. Samples
e of two res
Technologies fo
eference stan
red in PREM
measuremen
ngs were tak
d standard de
sample RSM
and RSM-4
ue Maturity
ng image ana
turity on fini
extra long s
ity.The abov
are tested by
sults given b
or Prosperity
ndard cotton
MIER ART 2
nt principle
ken from each
eviation for 5
M-1 and RSM
4 showed a
y values for
alysis technol

ished fabric,
staple with 3
ve groups o
y two operato
by the opera
supplied by
is shown in
is an excelle
h sample in a
5 days. It app
M-2 showed
standard de
r cottons me
logy.

we have col
36mm), from
of cottons is
ors separately
ator will be
instrument
Fig.1. The
ent way of
a subset of
peared that
a standard
eviation of
easured in
llected two
m different
s tested in
y and each
taken and

KNITTING A
X and Y s
for 40s N
factor of 1
obtained f
shade. Co
standard c
low stres
evaluation
having di
spinning s
standard k
reactive d
source at
RESULTS A
Influence o
Fig. 3a: K/S
In the pre
between X
samples is
K/S value
The s
to control

T
T
Group
Long Staple
Extra Long
AND DYEING
samples havi
e using a mi
1.32 and tigh
from these tw
olour measur
colour measu
ss mechanic
n system. In
ifferent Micr
system. Yarn
knitting mac
dyes at 0.5%
10
0
observer
AND DISCUSSI
of Fibre Matur
S Vs. Wave Length
esent study,
X and Y sam
s that the X a
es of grey and
hift in K/S v
and maintai
The Impact of Co
TABLE 1: MEASURED FI
Sa
e
g Staple
ing a Mic va
iniature spin
htness factor
wo samples
rements were
uring instrum
al propertie
the second g
ronaire and
n samples ar
chine. Fibre,
% shade. Co
rs in a standa
ONS
rity on the K/
h for Sample X and
it has been
mples even th
and Y sampl
d dyed fabric
value is main
in the maturit
otton Fibre Mat
BRE PROPERTIES OF LO
ample ID 2.
X
Y
A
B
C
D
alue of 2.9 an
nning system
of 15.4 on a
of this grou
e taken on gr
ment. The gr
s viz. comp
group of extr
Maturity va
re knitted wi
, Yarn and
lour measur
ard colour me
S Value of Dy
d Sample Y at Grey
observed th
ough the Mi
les are prepa
cs are shown
nly due to th
ty for each lo
urity on Dye Up
ONG AND EXTRA LONG
.5% Span Leng
32 to 33 mm
36 to 38 mm
nd having tw
. Yarn samp
a standard kn
up are scoure
rey and dyed
rey fabrics o
pression, ten
ra long stapl
alues, are sp
ith a cover f
knitted fabr
rements were
easuring inst
yed Fabric Mad

y Stage Fig. 3b
at there is a
cronnaire va
ared from cot
n in Figure 3
he difference
ot to control
ptake & Low Stre
STAPLE COTTON USED
gth Micron
2.9
2.9
3.5
3.5
3.7
3.7
wo different m
ples from eac
nitting machi
ed and then
d fabric usin
obtained from
nsile and sh
le cotton, fou
pun separate
factor of 1.32
rics obtained
e taken for
trument.
de from X a
b: K/S Vs. Wave Le
a difference
alues are sam
tton having d
and 4.
in maturity
shade variat
ess Mechanical
D IN THE PRESENT STUD
nnaire True
9
9
5
5
7
7
maturity valu
ch group wer
ine. Fibre, Y
dyed with r
g D65 sourc
m X and Y
hear propert
ur different s
ely for 60s
2 and tightne
d are then sc
grey and dy
and Y Cotton
ength for Sample X
in dye uptak
me. The only
different Tru
values. So i
tions apart fr
l
DY
e Maturity
0.77
0.82
0.87
0.78
0.79
0.83
ues are spun
re knitted wi
Yarn and knit
eactive dye
ce at 10
0
obs
samples are
ties in KAW
samples A, B
Ne using a
ess factor of
coured and
yed fabrics u
n Samples
X and Sample Y at
ke and shade
difference be
ue Maturity v
it is essential
rom micronai
557
separately
ith a cover
tted fabrics
with 0.5%
ervers in a
e tested for
WABATA
B, C and D
miniature
f 15.4 on a
dyed with
using D65

Dyed Stage
e variation
etween the
values. The
l for a mill
ire.
558
Influence o
of the Grey
The Low
has been
from high
from high
The fabric
Numeri (6
in the coe
between t
(41.3%) t
overall rat
Influence o
from Extra
Similar d
samples A
C&D are
of Fibre Matur
y Fabric Made
stress mech
clearly obse
h matured fib
h matured fib
c Y exhibite
6.06). This w
efficient of f
the two fabri
o tensile def
ting of fabric
TA
of Fibre Matur
a Long Staple
ifference ha
A, B, C and
shown separ
World Cot
rity on the Low
e from X and
hanical prope
erved that, th
bre compared
bre Y show
ed much smo
was a result o
friction MM
ics. The fabr
formation co
c Y as knitted
BLE 2: LOW STRESS M
KAWA
Compre
Extensi
Tensile
Geome
Coeffic
Koshi
Numeri
Fukuram
Total H
rity on the K/S
Cotton A,B
as been obse
D. The K/S
rately in Figu
tton Research C
w Stress Mech
d Y Cotton S
erties evalua
here is a 17
d to fabric X
wed higher fle
oother surfac
of lower valu
MD. No signi
ric Y showe
ompared to f
d inner wear
MECHANICAL PROPERTIE
ABATA proper
ession Recover
ibility (EMT %
e Resiliency (RT
trical roughnes
cient of friction
i
mi
Hand Value
S Value of Dye
B, C & D S
erved in dye
S values of g
ure 4.
Conference on T
hanical Prope
Samples
ted in KAW
% increase i
X, which is m
exibility than
ce than the fa
ues of geome
ificant differ
ed higher ext
fabric X. By
r garment wa
ES OF FABRICS X AND Y
rty
ry (RC %)
)
T %)
s (SMD) in m
(MMD)
ed Fabric Mad
Samples
e uptake and
grey and dye
Technologies fo
erties
WABATA sy
in compressi
made from lo
n the fabric m
fabric X as is
etrical rough
rence was ob
tensibility EM
virtue of hig
as higher than
Y MEASURED IN KAWA
Fabric X F
44.91
21.88
36.99
m 10.33
0.02
6.23
4.21
5.72
3.24
de
d shade varia
ed fabrics of
or Prosperity
stem are sho
ion recovery
ow matured
made from lo
s evident fro
hness SMD (
bserved in th
MT (24.2%)
gher Numeri
n fabric X.
BATA EVALUATION SYS
Fabric Y
52.54
24.20
41.34
4.76
0.01
5.39
6.06
5.50
3.39
ation among
f A&B, A&C
F
own in Tabl
y of the fabri
fibre. The fa
ow matured f
om the highe
4.76) and th
he fullness (
) and higher
i and lower
STEM
g the extra l
C, A&D, B&

Fig. 4 (Contd.)
e-2, and It
ic Y made
abric made
fibres X.
er rating of
e variation
(Fukurami)
r resiliency
Koshi, the
ong staple
&C, B&D,

The w
the sampl
difference
find any s
True Matu
there is no
within the
T
Fig. 4 Contd
Fig
wide differen
les A & C
e in True Ma
significant d
urity of cott
o difference
e limit.
The Impact of Co
d.
g. 4: K/S vs. Wave
nce in True M
C are highl
aturity value
difference in
ton than Mic
in k/s value
otton Fibre Mat
Length Graph for
Maturity valu
ly reflected i
e even thoug
K/S values.
cronaire. In c
s of grey and
urity on Dye Up
Samples A, B, C an
ue of cotton
in the K/S v
gh there is a
. It clearly in
case of C &
d dyed fabric
ptake & Low Stre
nd D by Comparing
(0.09) in the
alue. In B
slight differ
ndicates that
& D Samples
c, since micr
ess Mechanical
g between each Ot
e samples A
& C samp
rence in Mic
t dye uptake
, It has been
ronaire and m
l
ther
A& B and
ples, there is
cronnaire and
e depends m
n clearly obs
maturity are
559
d (0.08) for
s not much
d we dont
more on the
served that
controlled
CONCLUSION
The True Maturity measurement obtained from PREMIER ART 2 are traceable to image analysis
based maturity values measured at internationally recognized FASER INSTITUT located at
Bremen, Germany. The Repeatability of True Maturity measurement verified and is within the
acceptable limit.
Cotton with similar Micronaire,which are produced from different growing regions and seed
varieties/ hybrids have varying dye uptake mainly due to the differences in Maturity of the fibre.
Such Maturity variations reflected clearly in True Maturity values measured by the instrument.
Knitted Samples produced from different maturity values show clearly the dye uptake
differences. Such differences correlate well with the True Maturity values provided by the ART 2
Fibre testing instrument and also, the low stress mechanical properties measured in
KAWABATA evaluation system clearly correlates well with the True Maturity values of those
samples.
Yarn spun from high matured cotton can be preferred to produce wrinkle free fabric and
garments, as is evident from the low stress mechanical properties.
It is essential for a Spinning Mill, to control and maintain the maturity of cotton in each lay down
to control shade variations in the finished fabric apart from micronaire.
ACKNOWLEDGEMENT
Thanks are due to Mr.M.Sampath Kumaar, Research Assistant, CIRCOT, Coimbatore, for evaluating the
fibre properties in ART-2 instrument.
REFERENCES
[1] Gulati, A.N., and Ahmad, N., J.Text. Inst., 26, T26191935)
[2] Gulati, A.N., Indian Text. J., 60,65,140(1949)
[3] Smith, B., Text.Res.J., 61, 137 (1991)
[4] IS 236-1968: Determination of CottonFibre Maturity ( by sodium hydroxide swelling method) ISI Hand book of Textile
Testing, Bureau of Indian Standards, New Delhi, 1982 Edn., P.86
[5] ASTM Designation: D 144293. Standard Test Method for Maturity of Cotton Fibres (Sodium hydroxide Swelling and
Polarized Light Procedures) 1998 Annual Book of ASTM standards, Vol.07.01., p 377.
[6] BS 30851968: Determination of Cotton Fibre Maturity (Estimation by Classifications of Fibres swollen in Sodium
hydroxide Solution), BS Handbook, No.11, Methods of Tests for Textiles, British Standards Institution, London, 1974
Edn.,p.2/44.
[7] Schwarz, E.R. and Hotte, G.H., Text. Res. J., 5, 370(1935)
[8] Schwarz, E.R., and Shapiro, L., Rayon Textile Monthly, 19, 371, 421, 480, 570(1938)
[9] Thibodeaux, D.P and evans, J.P., Cotton fibre Maturity by Image Analysis, Textile Res.J., 56, 130139, 1986
[10] Xu, B., Pourdeyhimi, B. and Sobus, J., Fibre cross-sectional shape Analysis Using Imaging Techniques, Textile Research
Journal, 63, 717730,1993
[11] Xu, B. and Ting, Y., Fibre Image Analysis, Part I: Fibre Image Enhancement, J. of Textile Institute, 87, 274283, 1996.
Exploration of Residual Hazardous Compounds

on Cotton Fibers
Syed Zameer Ul Hassan and Jiri Militky
Dept. of Textile Materials, Technical University of Liberec, Czech Republic
AbstractA new method based on measurement of bio-electrical signals caused by enzymatic inhibition of
acetylcholinesterase (AChE) has been performed in this study for the detection of organophosphorous pesticides and
carbamates being the strong inhibitor of AChE and prevents its normal function of the rapid removal of acetylcholine
(Ach). Biosensor Toxicity Analyzer (BTA) equipped with electrochemical sensors was used for the testing and enzyme
activity was determined by acetylthiocholine chloride (ATCCl) as enzyme substrate. The monitoring of changes in bio-
electrical signals caused by the interaction of biological substances and residues were evaluated. Two samples of cotton
Giza 86 from Egypt of the crop 2009/2010 were analyzed. One of them was the classical conventional cotton and the
other was organic cotton without utilizing the synthetic pesticides. Cryogenic homogenization was carried out for
sample pretreatment and Soxhlet extraction method (SOX) was used with two different solvents; hexane and
dichloromethane for both of the samples, respectively. The resulted extracts were concentrated and then injected in the
BTA and the results were also compared with Gas Chromatography equipped with mass spectrometry (GC-MS).
Keywords: Acetylcholinesterase, Acetylcholine, Acetylthiocholine chloride, Biosensor Toxicity Analyzer,
Cryogenic homogenization, Soxhlet extraction method, Gas Chromatography.
INTRODUCTION
Cotton is the most important natural textile fiber in the world, used to produce apparel, home furnishings
and industrial products (Philip J. Wakelyn, 2007). Cotton has always been a major part of the textile
industry and today provides almost 38% of the world textile consumption, second only to polyester,
which recently took the lead (MYERS, 1999). Cotton production is highly technical and difficult because
of pest pressures and environment, e.g. drought, temperature and soil nutritional conditions. The total
area dedicated to cotton production accounts approximately 2.4% of arable land globally and cotton
accounts for an estimated 16% of the worlds pesticide consumption (Blackburn R.S, 2009).
Pesticides are widely used for the control of weeds, diseases, and pests all over the world, mainly
since after Second World War, with the discovery of some organic compounds with good insecticide or
herbicide activity. At present, around 2.5 million tons of pesticides are used annually and the number of
registered active substances is higher than 500 (Turiel Esther, 2008). Excessive use of pesticides causes
severe environmental degradation and research efforts are being made all over the world to find
alternatives for these harmful chemicals (El-Nagar, 2004). Humans can be exposed to pesticides by direct
or indirect means. Direct or primary exposure normally occurs during the application of these compounds
and indirect or secondary exposure can take place through the environment or the ingestion of food
(Turiel Esther, 2008). This is why development of natural biological methods of insect control was
initiated. Cotton grown without the use of any synthetically compounded chemicals (i.e. pesticides,
fertilizers, defoliants, etc.) is considered as organic cotton. It is produced under a system of production
and processing that seeks to maintain soil fertility and the ecological environment of the crop (S. Gordon,
2007).
Pesticides are toxic compounds that may cause adverse effects on the human and the environment.
Benzoylureas, carbamates, organophosphorous compounds, pyrethroids, sulfonylureas and triazines are
the most important groups (Alder Lutz, 2006). Organophosphorous pesticides (OP) are a class of
compounds that includes derivatives of phosphoric, phosphorous, thiono-phosphoric, and thion-thiolo
phosporic acids esterified with methyl or ethyl and different alcohol groups (Muccio, A.D, 2006).
Organophosphorous (OP) compounds are among the most toxic substances and are thus commonly used
as pesticides, insecticides and chemical warfare agents (Deo, 2005). The organophosphates and
90
562
carbamate
acetylchol
resulting
and in vi
Inhibition
pesticides
As the
pesticide r
the last fe
gas chrom
spectrome
are time c
laboratori
Biose
detection
and achie
from a ce
response,
(Buerk, 1
inhibition
Adina, 20
In thi
pesticides
Biosensor
this meth
limit.
PRINCIPLE
The targe
Acetylcho
synapses
neurotran
inhibitors
blocks the
es are power
linesterase (A
in the buildu
ital organs
n of AChE b
s such as orga
e pesticide r
residue play
ew years for
matography (
etry (GC-MS
consuming, e
es (Mulchan
ensors based
of OP comp
evable oppor
entralized la
low cost of
1993). Elect
n of AChE a
006).
s paper, we
s based on ac
r Toxicity A
od is simple
OF BTA
et for many
olinesterases
within the
smitter acety
of AChE. T
e nerves to pr
World Cot
rful inhibitor
AChE) whic
up of the ne
produce ser
by any xenob
anophosphat
residue is a p
ys a very imp
the detection
(GC), high-p
S), immune a
expensive an
ndani Priti, 19
on the inhib
pounds. (Deo
rtunity to pe
aboratory du
fabrication, p
trochemical
and the inhib
report anoth
cetylcholines
Analyzer (BT
e, fast and m
insecticides
s (AChE) bi
nervous s
ylcholine. Pe
This results i
rocess the sig
tton Research C
rs of acetylch
ch is essentia
eurotransmitt
rious sympt
biotic compo
tes and carba
potentially se
portant role i
n of organop
performance
assay and flu
nd require hi
999).
bition of ace
o, 2005). Ele
erform biom
ue to their a
possibility o
biosensors
bition degre
her method f
sterase inhib
TA). Compar
more sensitiv
s is an enzy
ological role
ystem of t
esticides bloc
in the accum
gnals proper
Fig. 1: Biosensor T
Conference on T
holinesterase
al for the func
ter acetylcho
oms and ev
ound is used
amates (ML H
erious hazard
in minimizin
phosphorous
liquid chrom
uorescence (
ighly trained
etylcholine e
ectro analytic
medical, envi
advantages s
f miniaturiza
for measure
ee is proport
for the determ
bition using
red with oth
ve for pestic
yme called
e is the termi
the insects
ck the cataly
mulation of a
rly (Mulchan
Toxicity Analyzer W
Technologies fo
e (Naggar, 2
ction of the c
oline which i
ventually de
d as a tool f
Hannam, 200
d to human h
ng risk. Many
pesticides. T
matography (
(Hu, 2010).
d personnel,
sterase (ACh
cal sensors a
ronmental, f
such as high
ation and eas
ement of th
tional to the
mination of
AC1.W2.R1
er kinds of
cide determin
acetylcholin
ination of im
and mamm
ytic activity
acetylcholine
ndani Ashok,
With Microflow Uni
or Prosperity
009). They c
central nervo
interferes wi
eath (Mulcha
for assessme
08).
health, the co
y methods h
The most wi
(HPLC), gas
However, th
are available
hE) have bee
and biosenso
food and ind
h selectivity
sy to integrat
hese pesticid
e pesticide c
Oganophosp
/ACCHE se
electrochem
nation with
nesterase (A
mpulse transm
mals by rap
of the active
e in the syna
2011).
it
can irreversi
ous system (H
ith muscular
andani Asho
nt of toxicit
ontrol and de
have been de
idely used m
chromatogr
hese techniqu
e only in sop
en widely us
ors provide a
dustrial anal
and specifi
te in automat
des are base
concentration
phorus and c
ensors with t
ical AChE b
much lower
ChE) (Jos
missions at c
pid hydrolys
e center, thus
aptic membra
ibly inhibit
Hu, 2010),
r responses
ok, 2001).
ty of some
etection of
veloped in
methods are
aphy-mass
ues, which
phisticated
sed for the
an exciting
lysis away
city, rapid
tic devices
ed on the
n. (Arvinte
carbamates
the help of
biosensors,
r detection
L, 2008).
cholinergic
sis of the
s acting as
ane, which

Exploration of Residual Hazardous Compounds on Cotton Fibers 563
Biosensor toxicity analyzer (BTA) works on the above mentioned principle and monitors the activity
of the inhibition of AChE with the help of sensors which are equipped with an enzymatic membrane of
AChE enzyme which is immobilized. It consists of two major parts, one of which is the Microflow unit
and the other is Bioanalyzer. The microflow unit has the capillary arrangement which allows precise and
constant flow of the liquid onto the active surface of the AChE sensor for a high level of repeatability and
sensitivity in the measurements. The module has an integrated chamber in which the sensor can easily be
placed or replaced (Fig 1).
EXPERIMENTAL
Materials
Two samples of Egyptian cotton variety namely Giza 86 (G86) were collected from the cultivation
season 2009/2010. This variety is one of the most long staple Egyptian cotton exports. One of them was
the classical conventional cotton and the other was organic cotton without utilizing the synthetic
pesticides.
Reagents and Apparatus
HPLC grades Hexane and dichloromethane solvents were used for the extraction procedure. Anhydrous
and granular sodium sulfate was used as dehydrating agent throughout the extraction process. Glass wool
was used as supporting material in soxhlet extraction after cleaned with acetone. A round flask (250ml)
with soxhlet and condenser glassware was used to conduct the soxhlet extraction. MOPSO sodium salt
was used for the preparation of buffer solution in BTA, where as Acetylthiocholine chloride (ATCCl) as
enzyme substrate and Neostigmine methyl sulfate as enzyme inhibitor. AC1.W2.R1/ACCHE Sensors
were used for the monitoring of AChE inhibition.
Gas Chromatograph in an analytical system was equipped with a temperature programming
capability, splitless injector, and capillary column MS-VF-5. It is equipped with an automatic sample
injector. The operating condition of the gas chromatograph analytical system were adjusted as
mentioned: Helium carrier gas with a flow rate of 1.0 ml/min, injector temperature 280
0
C, column
temperature 280
0
C, started at 80
0
C and increased for the first 08 min with the rate 10
0
C/ min and after
this initial phase with the rate 4
0
C/ min, holding time 10 min and FID detector temperature 120
0
C.
All the above experiments were done at the Technical University of Liberec, University of Pardubice
and Bvt technologies, Czech Republic.
Sample Preparation
The determination of pesticides in samples at low concentrations is always a challenge. The main aim of
any extraction process is the isolation of analytes of interest from the selected sample by using an
appropriate extracting phase. The development of an appropriate sample preparation procedure involving
extraction, enrichment, and cleanup steps becomes mandatory to obtain a nal extract concentrated on
target analytes (Turiel Esther, 2008). It is always necessary to carry out some pretreatments to get a
homogeneous and representative subsample. Even if the sample is apparently homogeneous, that is, an
aqueous sample, it will be at least necessary to perform a ltration step to remove suspended particles,
which could affect the nal determination of target analytes. However, solid samples need to be more
extensively pretreated to get a homogeneous subsample (Jos L, 2008).
Cryogenic Homogenization
It is necessary for the determination of the residual pesticides in cotton samples to turn the sample into
finely chopped or ground powder. This grinding procedure should be done carefully to avoid heat
generation (El-Nagar, 2004).
564

Both
pre-chilled
liquid nitr
min for gr
samples w
nitrogen v
After each
2 shows t
and blend
Soxhlet Ext
Soxhlet E
hexane an
was transf
wool laye
separately
stored for
conventio
RESULTS A
All the ab
For the c
process.
Hexane
samples of c
d Teflon mil
rogen surrou
rinding and 1
which had b
vapor. Only
h successful
the different
ded, the powd
traction (SOX
Extraction me
nd dichlorom
ferred to the
er. The thimb
y. Samples w
r further ana
onal cotton w
AND DISCUSSI
bove mention
convenience
World Cot
Fig.
classical conv
ll in the form
unding. Each
1 min for coo
been transfer
2 - 3 gm of
homogeniza
steps for the
der was samp
X)
ethod was us
methane were
e soxhlet thim
ble was place
were extracte
alysis. At th
with hexane &
ON
ned four extr
we will des
tton Research C

2(a): Raw Cotton (
ventional co
m of pallets w
h sample is m
oling. After t
rred to a cle
the raw cott
ation, the res
e cryogenic h
pled, cleaned
sed for the e
e used for ea
mble in betw
ed in the extr
ed for overni
he end of th
& dichlorome
racts were in
scribe the re
Fig. 3: Classi
Conference on T
(a) (b) (c)
(b) Freezer Mill (c
otton and org
which contai
milled for ap
the milling th
ean Teflon b
ton would fit
ulting powd
homogenizat
d and stored
extraction fro
ach of the sa
ween two lay
raction appar
ght. The extr
he extraction
ethane and o
njected in BT
esults with r
ical Cotton Sample
Technologies fo

c) Homogenized Sa
ganic cotton a
ns a concent
pproximately
he resulting p
bag and sea
t into the mi
er was poole
tion. Once th
for analysis.
om both of th
ample. A tot
yers of dehy
ratus charged
ract then con
n process w
rganic cotton
TA and then
respect to th
e with Hexane
or Prosperity

ample
are arranged
tric Teflon ri
y 10 minutes
powder is sa
led were he
ll at a time a
ed in another
he entire sam

he samples.
tal of 0.5 gm
drated sodiu
d with 230 m
ncentrated by
e have four
n with hexan
n analyzed in
he solvent u
d around the
ing and Teflo
s with an int
ampled. The
eld overnigh
and grind su
r clean Teflo
mple was hom
Two differen
m homogeniz
um sulfate ov
ml of both th
y turbo evap
r extracts i.e
ne & dichloro
n Gas chrom
used for the

inside of a
on puck in
terval of 2
raw cotton
t in liquid
uccessfully.
on bag. Fig
mogenized
nt solvents
zed sample
ver a glass
he solvents,
porator and
e. classical
omethane.
atography.
extraction

Both the s
the slot an
the sampl
compare
activity ar

It can
this respo
an inhibit
inhibitor o
Dichlorome
The same
dichlorom
that there
the standa

samples of c
nd starting th
le is added
the inhibitio
re shown in F
n be seen in t
nse (I) and
or to the enz
or sample ad
ethane
procedure a
methane and t
is a clear res
ard inhibitor.
Exploratio
classical cott
he pump, the
and then th
on of the sa
Fig 3 and Fig
the above gra
d also the rela
zyme. Inhibit
ddition. The c
as mentioned
the resultant
sponse on the

n of Residual H
ton and organ
buffer solut
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Fig. 4: Organ
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ting effect is
calculated va
d in the case
graphs are s
e addition of
Fig. 5: Classical C
Hazardous Comp
nic cotton w
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neostigmin
the standard
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i
), whi
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f both the sam
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nt hexane is
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6. It is quite
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After some st
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566
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TABLE 1:
Classica
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1.416
3.208
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Fig 7 shows
ane and Fig
nt dichlorom
Fig. 7
Fig. 8: Com
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al conventional
A)] Relative
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7: Comparison of C
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Conference on T
otton Sample with
and relative
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ponse and mo
TIVE INHIBITION FOR CL
l cotton
e Inhibition [R
0.0002855
0.0004602
the samples
arison of clas
e comparison
Classical and Orga
ical and Organic S
Technologies fo
h Dichloromethane
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show the cha
ore inhibition
LASSICAL AND ORGANIC
R
i
] Response
6.5
3.5
with hexane
ssical (red) a
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anic Sample with H
Sample with Dichlo
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n with each s
C COTTON
Organi
e [I(nA)]
527
524
e and dichlor
and organic
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Hexane
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amples. It ca
intensity of t
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ic cotton
Relative Inhib
-0.00007
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an be seen
the current
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i
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441
btained by
ple (green)
ton sample
Exploration of Residual Hazardous Compounds on Cotton Fibers 567
Although there are chemical compounds and pesticides detected such as 4,4- DDT, 4,4-DDE but
organophosphorous and carbamates pesticides and their derivatives are not detected in all the four
extracts.
CONCLUSION
This study demonstrates a method based on AChE inhibition. Contrary to other sophisticated methods,
this is an easier, faster and cheaper method. It is a method that offers to different investigators an easy
way to detect the presence of organophosphorous pesticide. Further research must be needed to verify the
usefulness of the method presented here for the screening of pesticides on some more varieties of cotton
of different regions.
ACKNOWLEDGMENT
The Authors would like to thank Dr. Jan Krejci, CEO of Bvt Technologies, for his support. This work
was supported under Student Grant Scheme (SGS) by Technical University of Liberec, Czech Republic.
REFERENCES
[1] Alder, L. (2006). RESIDUE ANALYSIS OF 500 HIGH PRIORITY PESTICIDES: BETTER BY GC MS OR LC
MS/MS? Wiley InterScience , 25, 838 865.
[2] ARVINTE, A. (2006). DEVELOPMENT OF A PESTICIDES BIOSENSOR USING CARBON-BASED ELECTRODE
SYSTEMS. In L.S. Chirila, & L.S. Chirila (Ed.), Chemicals as Intentional and Accidental Global Environmental Threats
(pp. 337343). Springer.
[3] Blackburn, R.S. (2009). LIFE CYCLE AND ENVIRONMENTAL IMPACT. In L. GROSE (Ed.), Sustainable Cotton
Production. Cambridge : Woodhead Publishing Limited.
[4] Buerk, D.G. (1993). Biosensors: Theory and Applications. USA: Technomic Publishing Company,Inc.
[5] Deo, R.P. (2005). Determination of organophosphate pesticides at a carbon nanotube/organophosphorus hydrolase
electrochemical biosensor. Analytica Chimica Acta , 530, 185189.
[6] El-Nagar, K. (2004). Extraction of Residual Chlorinated Pesticides from Cotton Matrix. Journal of Textile and Apparel,
Technology and Management , 04 (02).
[7] Hu, H. (2010). A novel chemiluminescence assay of organophosphorous pesticide quinalphos residue in vegetable with
luminol detection. Chemistry Central Journal.
[8] Jos L. Tadeo. (2008). Pesticides : Classi cation and Properties. In J.L. Tadeo (Ed.), Analysis of Pesticides in Food and
Environmental Samples. USA: CRC Press.
[9] ML, H. (2008 ). Characterisation of esterases as potential biomarkers of pesticide exposure in the lugworm Arenicola
marina (Annelida: Polychaeta). Environmental Pollution , 152, 342350.
[10] Muccio, A.D. (2006). Determination of Organophosphorus Pesticide Residues in Vegetable Oils by Single-Step
Multicartridge Extraction and Cleanup and by Gas Chromatography With Flame Photometric Detector. In A.G. Jos L.
Martnez Vidal (Ed.), Pesticide Protocols. New Jersey , USA: Humana Press Inc.
[11] Mulchandani, A. (2001). Biosensors for direct determination of organophosphate pesticides. Biosensors & Bioelectronics ,
16, 225230.
[12] Mulchandani, A. (2011). Microbial Biosensors for Organophosphate Pesticides. Applied Biochemistry and Biotechnology .
[13] Mulchandani, P. (1999). Biosensor for direct determination of organophosphate nerve agents. 1. Potentiometric enzyme
electrode. Biosensors & Bioelectronics , 14, 7785.
[14] MYERS, D. (1999). ORGANIC COTTON. In D. MYERS (Ed.), ORGANIC COTTON. LONDON: Intermediate
Technology Publications Limited.
[15] Naggar, A. E.-R. (2009). Clinical findings and cholineste rase levels in children of organ ophosphate s and carbamates
poisoning. Eur J Pediatr , 168, 951956.
[16] Philip J. Wakelyn, N.R. (Ed.). (2007). Handbook of Fiber Chemistry. CRC Press Taylor & Francis Group.
[17] S. Gordon, Y.-L. (Ed.). (2007). Cotton: Science and Technology. Cambridge: WoodHead Publishing Ltd.
[18] Turiel, E. (2008). Sample Handling of Pesticides in Food and Environmental Samples. In J.L. Tadeo (Ed.), Analysis of
Pesticides in Food and Environmental Samples. USA: CRC Press.
Studies on Composition of Oil and Fatty Acid

in Bt and Non Bt Cotton (G. hirsutum)
Harijan Nagappa
1
and Khadi B.M.
2

1
Asst. Professor (GPB), ARS, Hanumanamatti, UAS, Dharwad
2
Dean (PG Studies), UAS, Dharwad
AbstractCotton has pride place among cash crops from earliest times. Cotton though mainly grown for fibre is also
ranked as major oilseed crop in the international market. Cotton seed oil can be used for edible purpose after refining.
Cotton seed oil is premium quality oil and it has no cholesterol and is vegetable oil. On an average, oil content of Bt
hybrids (17.43%) were on par with non Bt hybrids (17.4 %) The Bt hybrids viz., JKCH-2245, K-5038, RCH-2,
NCEN-3R, JK-Ishwar and JKCH-266 showed numerically superior over non Bt hybrids. On an average, unsaturated
fatty acid of Bt and non Bt hybrids was on par with each other. The Bt hybrids viz., JKCH-2245, K-5038, RCH-2,
JKCH-22, NCEN-3R and RCH-138 recorded more than 4 per cent of unsaturated fatty acid higher than non Bt
hybrids, while JK-Ishwar, JKCH-1050, JKCH-66 and K-5316 non Bt hybrids had unsaturated fatty acid more than 4
per cent higher than their respective Bt versions. The myristic acid in Bt hybrids was numerically superior over non Bt
hybrids which helps to increase the keeping quality of oil.
The mean of 35 Bt hybrids had monounsaturated oil on par with mean of respective non Bt hybrids. JKCH-2245
Bt, K-5038, RCH-2, JKCK-22, NCEN-3R and RCH-138 were significantly superior over non Bt hybrids. Considering
polyunsaturated fatty acids, JKCH-2245 and JK-Ishwar Bt hybrids were numerically superior over non-Bt hybrids.
From current investigations it can be concluded that there is no significant difference in oil percentage and fatty acid
composition in Bt cotton hybrids when compared to their non Bt versions.
INTRODUCTION
Cotton though mainly grown for fibre is also ranked as major oil seed crop in the international market.
Out of the four major products i.e., meal, hull, oil and lint, oil is the most important. Besides commercial
importance in the leather industry it is also used as a lubricant. Cotton seed oil can also be used for edible
purpose after refining. Cotton seed oil is premium quality as it has no cholesterol, transfree and highly
stable with low flavor reversion vegetable oil. The cotton seed oil has neutral flavor, strong shelf life, low
fryer turnover and extremely versatile. It is a good source of essential fatty acids (70% unsaturated, 26%
saturated) and good source of vitamin E. Cotton seed oil has a fatty acid profile that makes acceptable as
healthful oil and very useful as cooking and frying oil (Boghara etal, 1985). Evaluation of different Bt
and non Bt cotton genotypes for seed oil and both acid profile has been carried out and presented in the
paper.
A gas chromatograph, model GC 2010 (Shimadzu, Kyoto, Japan) was used to separate methyl esters.
The fatty acid methyl ester was identified by a comparison of retention time to standard methyl ester fatty
acid mixtures (Sigma, Aldrich). Concentration of each fatty acid was recorded by normalization of peak
areas as per cent of particular fatty acid.
Phenotyping for fatty acid composition was done using Near Infrared Spectroscopy (NIRS). Eight
fatty acids viz., Palmitic acid (16:0), Stearic acid (18:0), Oleic acid (18:1), Linoleic acid (18:2), Arachidic
acid (20:0), Cyclopropanid (1:1), Behenic acid (22:0), and Myristic acid (14:0) were studied in 35 Bt and
non Bt counterpart cotton hybrids.

91
Studies on Composition of Oil and Fatty Acid in Bt and Non Bt Cotton (G. hirsutum) 569
Cotton seed oil is the most unsaturated oil, among others include safflower, corn, soybean, rapeseed and
sunflower seed oil. Cotton seed oil has a 2:1 ratio of poly unsaturated to saturated fatty acids and
generally consists of 70 per cent unsaturated fatty acid including 18 per cent mono unsaturated (oleic)
and 52 per cent poly unsaturated (linoleic) and 26 per cent saturated (palmatic and stearic ) oils.
The fatty acid composition of cotton seed oil is determined by fatty acids viz., palmatic, oleic and
linoleic acid. The myristic and stearic acid contribute to minor constituent in oil but play major role in
keeping quality of oil and linoleic acid is very important in view of the health of heart.
Comparison between Bt and non Bt cotton hybrids for fatty acid related traits has been presented in
Table.
On an average oil content of Bt was (17.43%) and non Bt was (17.40%). The results revealed that Bt
gene did not affect the seed oil content.
The range of saturated fatty acid varied from 31.89 (K-5038) to 40.98 per cent (K-5316) in Bt
hybrids while in non Bt hybrids it ranged from 33.96 (SBCH-302) to 39.72 per cent (Ankur.651). On an
average Bt hybrids (36.25%) showed slightly less content of saturated fatty acids than non Bt hybrids
(36.90%). Most of the Bt hybrid showed lesser saturated fatty acid than non Bt hybrids except SBCH-
302, JK-Ishwar, JKCH-1050, SBCH-311, KDCHH-9810, JKCH-266, K-5316 and JKCH-99 cotton
hybrids.
The mean of unsaturated fatty acid of Bt hybrids (59.71%), was on par with non Bt hybrids
(59.15%). The unsaturated fatty acid of Bt hybrids ranged between 56.30 (K-5316) and 63.40 per cent
(K-5038), while non Bt hybrids varied from 56.70 (VICH-111) to 63.21 per cent (SBCH-302). The
highest unsaturated fatty acids observed in K-5038 (63.40%) followed by RCH-2 (62.37%) and SBCH-
302 (62.12%) in Bt cotton hybrids. The non Bt hybrid SBCH-302 (63.21%) showed highest unsaturated
fatty acids followed by JKCH-1050 (62.36%) RCH-144 and RCH-118 (61.00%).
On an average around, 13.23 per cent more myristic acid was noticed in Bt hybrids (1.42%) than
their non Bt hybrids (1.28%). However some of the Bt hybrids viz., K-5038, RCH-2, JKCH-1947, RCH-
134, Ankur-651, NCEN-3R, PCH-2171, PCH-2270, RCH-138 and RCH-377 showed 20 per cent higher
myristic acid than their corresponding non Bt hybrids. Conversely non BT hybrids, JKCH-266(29.17%),
JK-Ishwer (21.85%) and K-5316(21.80%) had more than 20 per cent higher myristic acid than their Bt
versions. Brein and Wakelyn (2005) also observed one per cent myristic acid in seed cotton oil.
The mean stearic acid content of Bt and non Bt hybrids was 3.88 and 3.98 per cent, respectively. The
range of stearic acid varied from 3.15(K-5038Bt) to 4.49 per cent (JK-Ishwar Bt) in Bt hybrids, while in
non Bt hybrids, the range was 3.33 (JKCH-266, JKCH-1050) to 4.48 per cent (Ankur-651). The non-Bt
hybrids viz., PCH-2171, NCEN-3R, JKCH-1947, RCH-2, KDCHH-441 and K-5038 exhibited more
stearic acid content than their counter part Bt hybrids. However, Pelin GUNC ERGONUL, Bulent
ERGONUL, (2008) reported 2.89 per cent of stearic acid in cotton seed oil, Brien (2004) and Berrien and
Wokelyn (2005) observed 3 per cent of stearic acid in cotton seed oil.
The range of palmatic acid content was found between 28.74 (K-5038Bt) and 36.46 per cent (K-
5316Bt) per cent. The overall mean of Bt hybrids (32.37 %) and non Bt hybrids (32.92 %) did not differ
much. Sharma et al (2009) observed 19.10 to 29.10 per cent range of palmatic acid Pelin GUNC
ERGONUL, Bulent ERGONUL (2008) reported 23.03 per cent of palmatic acid in cotton seed oil.
Mean of oleic acid (18:1) content in Bt cotton hybrids was 12.99 per cent, while in non Bt hybrids it
was 12.22 per cent. The oleic acid ranged from 10.11 (K-5316) to 16.03 per cent (K-5038) in Bt hybrids
while in non Bt hybrids, it ranged from 10.45 (VICH-5) to15.76 per cent (SBCH-302). Most of Bt
versions showed statistically as well as numerically superior over non Bt versions for oleic acid except
SBCH-302,JK-Ishwar, RCH-144, JKCH-1050, PCH-2270, NCEN-2R, RCH-118, JKCH-266, K-5316,
Jk-Durga and JK-Gowri.
TABLE 1: OIL CONTENT AND FATTY ACID COMPOSITION IN BT AND NON-BT COTTON HYBRIDS
Genotypes Oil Content (%) Bt NBt % over
NBt
Bt NBt % over
NBt
Myristic (14:0) (%) Palmitic (16:0) (%) Stearic (18:0) (%)
Bt NBt % Differ-
ence
Saturated
Fatty
acid
Saturated
fatty
Acid
Unsaurated
Fatty
Acid
Unsaurated
Fatty
Acid
Bt NBt %
Differ-
ence
Bt NBt %
Differ-
ence
Bt NBt %
Differ-
ence
JKCH-2245 19.20 18.15 5.79 34.76 36.26 -4.14 61.90 59.18 4.60 1.69 1.41 19.86 31.30 32.55 -3.84 3.46 3.71 -6.74
SBCH-302 17.85 18.90 -5.56 35.15 33.96 3.50 62.12 63.21 -1.72 1.41 1.66 -15.06 31.51 30.60 2.97 3.64 3.36 8.33
K-5038 19.25 16.65 15.62 31.89 38.03 -16.15 63.40 60.03 5.61 1.74 1.20 45.00 28.74 33.95 -15.35 3.15 4.08 -22.79
KDCHH-441 18.25 17.10 6.73 34.35 37.09 -7.39 61.64 60.13 2.51 1.47 1.46 0.68 30.93 33.03 -6.36 3.42 4.06 -15.76
JK-Indtra 18.85 18.65 1.07 34.21 36.33 -5.84 61.59 59.66 3.23 1.76 1.54 14.29 30.53 32.54 -6.18 3.68 3.79 -2.90
RCH-2 17.75 17.10 3.80 33.46 39.19 -14.62 62.37 59.13 5.48 1.73 1.14 51.75 29.97 34.81 -13.90 3.49 4.38 -20.32
JK-Ishwar 18.00 17.15 4.96 39.53 36.93 7.04 57.60 60.35 -4.56 1.18 1.51 -21.85 35.04 32.91 6.47 4.49 4.02 11.69
JKCH-1947 15.80 17.10 -7.60 35.82 38.28 -6.43 61.24 58.79 4.17 1.66 1.15 44.35 32.12 33.97 -5.45 3.70 4.31 -14.15
RCH-134 16.95 18.05 -6.09 37.52 37.92 -1.05 60.32 59.24 1.82 1.59 1.27 25.20 33.51 33.77 -0.77 4.01 4.15 -3.37
JKCH-22 16.85 19.15 -12.01 36.88 38.44 -4.06 60.95 58.31 4.53 1.60 1.09 46.79 32.85 34.23 -4.03 4.03 4.21 -4.28
Ankur-651 17.95 17.85 0.56 37.56 39.72 -5.44 60.10 58.26 3.16 1.34 0.97 38.14 33.45 35.24 -5.08 4.11 4.48 -8.26
RCH-144 17.55 14.40 21.88 35.65 35.78 -0.36 60.32 61.00 -1.11 1.43 1.58 -9.49 31.86 32.03 -0.53 3.79 3.75 1.07
JKCH-1050 17.00 17.45 -2.58 37.75 34.44 9.61 59.51 62.36 -4.57 1.43 1.66 -13.86 33.76 31.11 8.52 3.99 3.33 19.82
SBCH-311 17.10 16.80 1.79 39.49 38.32 3.05 58.52 58.17 0.60 1.25 1.21 3.31 35.06 34.18 2.57 4.43 4.14 7.00
PCH-2270 17.45 19.50 -10.51 37.76 38.46 -1.82 59.14 60.15 -1.68 1.36 1.15 18.26 33.71 34.22 -1.49 4.05 4.24 -4.48
NCEN-3R 16.75 16.40 2.13 35.88 39.31 -8.73 61.53 58.40 5.36 1.69 1.05 60.95 32.11 34.91 -8.02 3.77 4.40 -14.32
KDCHH-9810 18.25 18.60 -1.88 36.18 35.49 1.94 58.27 58.78 -0.87 1.34 1.27 5.51 32.22 31.79 1.35 3.96 3.70 7.03
NCEN-2R 17.25 17.35 -0.58 35.91 35.97 -0.17 58.92 59.53 -1.02 1.27 1.42 -10.56 32.06 32.11 -0.16 3.85 3.86 -0.26
RCH-118 18.10 18.35 -1.36 34.70 35.00 -0.86 60.93 61.00 -0.11 1.52 1.59 -4.40 31.08 31.19 -0.35 3.62 3.81 -4.99
JKCH-266 18.45 15.05 22.59 38.39 34.42 11.53 57.62 60.30 -4.44 1.02 1.44 -29.17 34.36 31.09 10.52 4.03 3.33 21.02
Dhruva 17.95 17.00 5.59 37.45 37.51 -0.16 57.45 57.91 -0.79 1.13 1.14 -0.88 33.09 33.39 -0.90 4.36 4.12 5.83
K-5316 18.05 18.05 0.00 40.98 34.69 18.13 56.30 59.15 -4.82 1.04 1.33 -21.80 36.46 30.57 19.27 4.52 4.12 9.71
KDCHH-9632 17.10 16.60 3.01 35.17 37.64 -6.56 59.96 57.83 3.68 1.32 1.16 13.79 31.42 33.50 -6.21 3.75 4.14 -9.42
JK-Varun 13.75 17.55 -21.65 36.60 36.85 -0.68 58.57 58.75 -0.31 1.18 1.11 6.31 32.68 32.99 -0.94 3.92 3.86 1.55
PCH-2171 17.55 17.65 -0.57 34.66 37.91 -8.57 60.70 58.91 3.04 1.62 1.14 42.11 31.23 34.05 -8.28 3.43 3.86 -11.14
RCH-20 17.25 16.35 5.50 36.21 37.37 -3.10 59.82 58.92 1.53 1.57 1.25 25.60 32.15 33.20 -3.16 4.06 4.17 -2.64
JK-Durga 15.20 15.60 -2.56 37.03 37.67 -1.70 57.94 57.88 0.10 1.13 1.15 -1.74 33.04 33.47 -1.28 3.99 4.20 -5.00
JKCH-99 17.90 17.85 0.28 35.33 34.97 1.03 58.61 59.49 -1.48 1.39 1.33 4.51 31.59 31.26 1.06 3.74 3.71 0.81
RCH-138 17.45 18.15 -3.86 35.21 36.55 -3.67 60.90 58.47 4.16 1.64 1.10 49.09 31.42 32.71 -3.94 3.79 3.84 -1.30
JKCH-1945 16.10 16.70 -3.59 34.52 34.60 -0.23 59.85 59.17 1.15 1.72 1.43 20.28 31.09 30.95 0.45 3.43 3.65 -6.03
JK-Gowri 17.05 17.80 -4.21 37.17 37.33 -0.43 57.83 58.18 -0.60 1.32 1.32 0.00 33.16 33.30 -0.42 4.01 4.03 -0.50
RCH-377 17.85 16.55 7.85 36.91 38.00 -2.87 59.07 57.81 2.18 1.33 1.10 20.91 32.91 33.91 -2.95 4.00 4.09 -2.20
VICH-111 17.40 17.60 -1.14 36.49 37.23 -1.99 58.12 56.70 2.50 1.27 1.14 11.40 32.52 33.20 -2.05 3.97 4.03 -1.49
VICH-5 16.80 17.50 -4.00 35.82 36.08 -0.72 58.54 57.41 1.97 1.23 1.10 11.82 31.76 31.85 -0.28 4.06 4.23 -4.02
VICH-9 18.00 18.30 -1.64 36.28 37.64 -3.61 58.17 57.85 0.55 1.21 1.08 12.04 32.24 33.63 -4.13 4.04 4.01 0.75
Mean 17.43 17.40 0.51 36.25 36.90 -1.59 59.71 59.15 0.95 1.42 1.28 13.23 32.37 32.92 -1.51 3.88 3.98 -2.05
Std. Dev. 1.06 1.68 1.54 0.21 1.41 0.30
TABLE 2
Genotypes Oleic (18:1) (%) Linoleic (18:2) (%) Arachidic (20:0) (%) Behanic (22:0) (%) CPA (1:1) (%)
Bt NBt % Differ-
ence
Bt NBt % Differ-
ence
Bt NBt % Differ-
ence
Bt NBt % Differ-
ence
Bt NBt % Differ-
ence
JKCH-2245 14.86 12.43 19.55 47.04 46.75 0.62 0.92 0.99 -7.07 1.02 1.11 -8.11 1.08 1.14 -5.26
SBCH-302 14.58 15.76 -7.49 47.54 47.45 0.19 0.91 0.87 4.60 1.12 0.98 14.29 1.11 1.07 3.74
K-5038 16.03 12.70 26.22 47.37 47.33 0.08 0.87 1.01 -13.86 1.10 1.14 -3.51 1.03 1.15 -10.43
KDCHH-441 14.55 13.38 8.74 47.09 46.75 0.73 0.90 1.01 -10.89 1.04 1.09 -4.59 1.05 1.13 -7.08
JK-Indtra 14.75 12.85 14.79 46.84 46.81 0.06 0.95 0.99 -4.04 1.02 1.01 0.99 1.04 1.11 -6.31
RCH-2 15.14 11.94 26.80 47.23 47.19 0.08 0.92 1.05 -12.38 1.00 1.05 -4.76 1.04 1.17 -11.11
JK-Ishwar 11.31 13.45 -15.91 46.29 46.90 -1.30 1.08 0.99 9.09 1.09 1.02 6.86 1.17 1.14 2.63
JKCH-1947 14.82 11.57 28.09 46.42 47.22 -1.69 0.96 1.06 -9.43 1.06 1.12 -5.36 1.11 1.15 -3.48
RCH-134 13.73 12.64 8.62 46.59 46.60 -0.02 1.05 1.00 5.00 1.17 1.05 11.43 1.23 1.16 6.03
JKCH-22 14.29 11.35 25.90 46.66 46.96 -0.64 1.03 1.01 1.98 1.27 1.12 13.39 1.22 1.17 4.27
Ankur-651 13.05 10.79 20.95 47.05 47.47 -0.88 0.96 1.01 -4.95 1.07 1.16 -7.76 1.16 1.18 -1.69
RCH-144 13.32 14.32 -6.98 47.00 46.68 0.69 0.96 0.94 2.13 1.09 1.07 1.87 1.05 1.13 -7.08
JKCH-1050 13.19 13.78 -4.28 46.32 48.58 -4.65 0.99 0.94 5.32 1.12 1.00 12.00 1.17 0.99 18.18
SBCH-311 12.44 11.73 6.05 46.08 46.44 -0.78 1.05 0.99 6.06 1.22 1.16 5.17 1.22 1.16 5.17
PCH-2270 12.28 12.73 -3.53 46.86 47.42 -1.18 1.03 0.99 4.04 1.14 1.15 -0.87 1.18 1.14 3.51
NCEN-3R 14.10 11.04 27.72 47.43 47.36 0.15 0.98 1.01 -2.97 1.14 1.18 -3.39 1.07 1.17 -8.55
KDCHH-9810 12.07 12.06 0.08 46.20 46.72 -1.11 1.00 0.93 7.53 1.11 1.04 6.73 1.15 1.12 2.68
NCEN-2R 12.28 12.89 -4.73 46.64 46.64 0.00 0.97 1.00 -3.00 1.13 1.15 -1.74 1.16 1.17 -0.85
RCH-118 13.40 14.34 -6.56 47.53 46.66 1.86 0.93 1.03 -9.71 1.05 1.30 -19.23 1.09 1.19 -8.40
JKCH-266 10.75 13.02 -17.43 46.87 47.28 -0.87 0.97 0.91 6.59 1.14 1.01 12.87 1.16 1.04 11.54
Dhruva 11.01 11.07 -0.54 46.44 46.84 -0.85 1.07 1.06 0.94 1.28 1.22 4.92 1.22 1.21 0.83
K-5316 10.11 12.26 -17.54 46.19 46.89 -1.49 1.11 1.05 5.71 1.23 1.21 1.65 1.20 1.21 -0.83
KDCHH-9632 12.64 11.07 14.18 47.32 46.76 1.20 0.93 1.08 -13.89 1.07 1.16 -7.76 1.12 1.19 -5.88
JK-Varun 12.02 11.11 8.19 46.55 47.64 -2.29 0.98 1.00 -2.00 1.12 1.12 0.00 1.17 1.14 2.63
PCH-2171 13.88 12.02 15.47 46.82 46.89 -0.15 0.96 0.95 1.05 1.08 1.04 3.85 1.10 1.12 -1.79
RCH-20 13.59 12.58 8.03 46.23 46.34 -0.24 1.05 0.98 7.14 1.35 1.13 19.47 1.26 1.17 7.69
JK-Durga 11.35 12.06 -5.89 46.59 45.82 1.68 1.07 1.06 0.94 1.19 1.26 -5.56 1.21 1.23 -1.63
JKCH-99 12.26 12.41 -1.21 46.35 47.08 -1.55 0.97 0.95 2.11 1.10 1.10 0.00 1.19 1.12 6.25
RCH-138 14.40 11.38 26.54 46.50 47.09 -1.25 1.00 0.99 1.01 1.34 1.13 18.58 1.21 1.13 7.08
JKCH-1945 13.29 12.29 8.14 46.56 46.88 -0.68 0.97 0.99 -2.02 1.08 1.13 -4.42 1.14 1.14 0.00
JK-Gowri 11.91 12.02 -0.92 45.92 46.16 -0.52 1.05 1.07 -1.87 1.14 1.13 0.88 1.22 1.22 0.00
RCH-377 12.75 10.91 16.87 46.32 46.90 -1.24 0.97 1.01 -3.96 1.12 1.12 0.00 1.13 1.15 -1.74
VICH-111 11.33 10.59 6.99 46.79 46.11 1.47 1.05 1.07 -1.87 1.13 1.16 -2.59 1.17 1.22 -4.10
VICH-5 11.67 10.45 11.67 46.87 46.96 -0.19 1.02 1.05 -2.86 1.12 1.18 -5.08 1.18 1.20 -1.67
VICH-9 11.59 10.87 6.62 46.58 46.98 -0.85 0.97 1.02 -4.90 1.18 1.14 3.51 1.19 1.19 0.00
Mean 12.99 12.22 6.95 46.72 46.93 -0.45 0.99 1.00 -1.16 1.13 1.12 1.54 1.15 1.15 -0.16
Std. Dev. 1.36 0.47 0.05 0.07 0.05

Studies on Composition of Oil and Fatty Acid in Bt and Non Bt Cotton (G. hirsutum) 571
On an average, linoleic acid (18:2) in Bt and non Bt hybrid was 46.72 and 46.93 per cent,
respectively. It ranged from 46.11 (VCH-111) to 48.58 per cent (JKCH-1050) in non Bt hybrids while Bt
hybrids varied from 46.08 (SBCH-311) to 47.54 per cent (SBCH-302). However, Pelin GUNC
ERGONUL, Bulent ERGONUL, (2008) reported 56.01 per cent of linoleic acid and Brein and Wakelyn
(2005) and Brien (2004)observed 52 per cent in cotton seed oil. On an average arachidic acid content of
Bt and non Bt hybrids was 0.99 and 1 per cent respectively. The mean behanic acid (22:0) in Bt hybrids
was 1.13 per cent and non Bt hybrids was 1.12 per cent. Similarly cycloproponide acid content of Bt
hybrids was 1.15 per cent and non Bt hybrids was 1.15 per cent. So, the content of arachidic acid,
behanic acid and cycloproponide acid did not much get influenced by Bt gene.
As many as 20 Bt hybrids had more than 60 per cent of unsaturated fatty acids. The hybrids viz., K-
5038(63.4%), SBCH-302 (63.2%) and JKCH-1050 (62.36%) non Bt, hybrids showed statistically higher
unsaturated fatty acids than RCH-2 Bt cotton hybrid (62.37%).
CONCLUSION
From the current investigations it can be concluded that there was no significant changes seen for oil
percentage and fatty acid composition in the Bt hybrids when compared to their non Bt versions. It also
appears that the presence of the Bt gene does not affect the oil content and its profile.
REFERENCES
[1] Brien, R.D., 2004, Fats and Oils: Formulating and Processing for Applications, pp. 1567.
[2] Brein, R.D. and Walkelyn, P.J., 2005, Cotton seed oil: An oil for transfer options. Inform. Nov. 2005, 16(11): 667679.
[3] Boghara, D.G., Mehta, N.P. and Pethani, K.V., 1985, Genetic studies on oil content in upland cotton gossypium hirsutum
L., Gujrat Agric. Univ. Res. J., 10(2). 14.
[4] Pelin GUNC ERGONUL, Bulent ERGONUL, 2008. Changes in fatty acid profiles and omega fatty acid contents of
selected vegetable oil during refining process. Electronic Journal of Environmental, Agricultural and Food Chemistry,
7 (13), 2008. [26552660].
[5] Sharma, D.H., Dharminder Pathaki, A.K., Atwal and Sangha, M.K., 2009, Genetic variation for some chemical and
biochemical characteristics in cotton seed oil, Cotton Res. Dev., 23(1): 17.
The Properties of the Naturally-Pigmented Cotton

Cultivated in Nakornsawan Field Crop Research
Center Thailand
Piyanut Jingjit
1
and Parinya Seebunruang
2

1
Faculty of Engineering, Rajamangala University of Technology Thanyaburi, Thailand
2
Nakornsawan Field Crop Research Center, Thailand
E-mail: piyanut.j@en.rmutt.ac.th
AbstractTwo types of naturally pigmented cotton fibers studied in this research were green cotton and brown
cotton. Both fibers were grown, picked, and ginned in the Nakornsawan Field Crop Research Center, Thailand. When
these cotton fibers were observed under optical microscope, the green fibers showed green colour substances scattered
throughout the tissue of the fibers and the brown fibers showed brown color substances concentrated mainly in the
center of the fibers. The fiber colour of the green and brown fibers were not uniform, some fiber are apparently darker
than others. Due to the fineness, length and strength of the green fibers, the fibers were able to be spun, with OE
spinning machine, into fine yarn (no. 24). For the brown fibers, the fibers were found to be coarser, shorter and weaker
and they were only suitable for spun into medium or coarse yarn. The percentage of fiber loss during spinning process
were high, the green fiber 32% and the brown fiber 41.4%, these were partly because of their short fiber index which
were higher than standard. The obtained yarns were used to produce knitted fabrics on a circular knitting machine
using single jersey construction. The color of the obtained fabrics was found to be uniform all through the fabrics. The
fabrics were tested for light fastness, and the light fastness level of the fabric produced from the green fibers was 3-4
and for the fabric produced from brown fiber was 6-7.
INTRODUCTION
The naturally pigmented cotton fibers in this study were selected from limited hues of naturally coloured
cotton cultivated in Thailand. The naturally coloured cotton had been developed and cropped scatteringly
in some areas mostly in the northern and north-eastern regions of the country, in which they still obscure
conserved and maintained theirs provincial handicraft and souvenir markets. These pigmented cottons
had long been struggled to survive, partly as theirs short staple which made them harder to spin and
weave with high speed advanced technology machineries than new developed long stapled white cottons.
Therefore, the naturally coloured cottons cultivated in the country were almost entirely for hand-spun
yarns and traditional woven fabrics as shown in figure 1 and 2. Despite the fact that there were
advantages of the naturally coloured cotton, they never overcome the major advantage of the dyed cotton
as its boundless colour varieties.

Fig. 1: Yarn Spinning by Hand Fig. 2: Weaving Fabric by Traditional Shutter Loom

92
The Properties of the Naturally-Pigmented Cotton Cultivated in Nakornsawan Field Crop Research Center Thailand 573
The fibers selected for this research were naturally coloured cotton in shades of brown and green,
cultivated in 2010 in the Nakornsawan Field Crop Research Center. The fibers were harvested by hand
and separated the fibers from the seeds by small cotton gin at the research center. The fibers then kept in
opaque packaging, as the light might effects shade of the fibers, during transporting to spinning factory
where they would be tested with HVI Spectrum fiber testing instrument. After the fibers had been tested,
they were respectively produced into yarns with open-end spinning machine and then knitted into fabrics
with circular knitting machine as shown in figure 3 and 4.

Fig. 3: Brown Cotton Spinning by OE Machine Fig. 4: Fabric Knitted by Circular Knitting Machine
Considering the test results - the fineness, length and strength, the naturally green cotton fiber was
able to be spun into relatively finer yarn (no. 24) than the brown cotton, which was coarser, shorter and
weaker than the green cotton, so the brown cotton was only applicable for producing medium or coarser
yarns which in this trial the number 20. The yarns were knitted into a green cotton fabric and a brown
cotton fabric with basic single jersey construction. The knitted fabrics were check for the fabric counts
and weights. The average fabric count for the brown fabric were 44.4 course per inch and 29.4 wale per
inch with average fabric weight of 184.64 gram/m
2
and the green fabric were 42.6 course per inch and
31.8 wale per inch with weight of 151.82 gram/m
2
.
THE FIBER PROPERTIES
Microscopic Examination
The fibers were observed and examined under optical microscope for their general and distinctive
appearances - as shown in figure 5. The green fibers showed green colour substances scattered unevenly
but thoroughly the tissue of the fibers. One the other hand, the brown fibers showed brown colour
substances concentrated mainly only in the inner center of the fibers. The both fibers showed marked
distinct colour from the ordinary white cotton. However, the colours of the green and brown fibers were
not consistent - some fibers were apparently darker than others and the colour hues and shades were
different in each fiber. Under the same magnification, it appeared that the brown fibers were coarser and
rounder with higher degree of cell wall thickness than the green ones.

574
Manufactur
The fibers
trades wit
Spinning Co
The avera
favourable
average s
greater tha
Micronaire
The avera
because o
brown fib
different d
micronair
Maturity Ind
The matur
ranges fro
index at 0
which ind
Upper Half M
Fiber leng
of the yar
fiber. The
inch (extr
range of l
range of s
a)
Fig. 5: Phot
ring Quality P
s were measu
th HVI Spect
onsistency Inde
age spinning
e spinning p
spinning con
an the brown

age micronai
of very small
ber had 6.00
degrees of m
re range.
dex
rity index in
om below 0.0
0.80 which in
dicated matur
Mean Length (U
gth affects ya
rn which can
e general com
ra long). The
long fiber. In
short fiber.
World Cot
The Green Cotton
tomicrograph Sho
Parameters
ured influent
trum fiber te
ex (SCI)
g consistenc
otential. On
nsistency ind
n one.
ire of the gre
l perimeter a
0 which con
maturity. Hen
dicates the d
07 (uncommo
ndicated sligh
re state.
(UHML)
arn strength,
n be successf
mmercial len
e upper half
n the contrar
tton Research C

n Fiber
wing the Cross-Se
tial quality p
sting instrum
cy index of
the contrary
dex. Therefo
een fiber wa
and mature o
nsidered as
nce, the green
degree of cell
on) to above
htly immatur
yarn evenne
fully produce
gth range for
mean length
ry the length
Conference on T
ectional and Longi
parameters fo
ment.
the green f
, the brown f
ore, the over
as 2.23 - mea
or medium p
very coarse
n fiber and th
l wall thickn
e 0.95 (very m
re and the br
ess, and the e
ed from give
r cotton was
h of the gree
h of the brow
Technologies fo
b)
tudinal Views of th
or yarn and f
fiber was 14
fiber had onl
rall fiber sp
aning that th
erimeter and
fiber becau
he brown fib
ness within a
mature). The
rown cotton
efficiency of
en fibers is a
between bel
en cotton was
wn cotton wa
or Prosperity
The Brown Cotto
he Naturally Colou
fabric manuf
46 - indicati
ly 42 which c
pinnability o
he fiber was
d immature.
use of its la
ber were in th
cotton samp
e green cotton
had average
f the spinning
also influence
low 0.99 inc
s 1.127 inch
as 0.868 inch

n Fiber
ured Cottons
facturing pro
ing that the
considered a
of the green
very fine wh
On the other
arge in perim
he opposite e
ple. The matu
n had averag
maturity ind
g process. Th
ed by the len
h (short) to a
h which was
h which was
ocesses and
fiber had
as very low
fiber was
hich either
r hand, the
meter with
ends of the
urity index
ge maturity
dex at 0.93
he fineness
ngth of the
above 1.26
within the
within the
The Properties of the Naturally-Pigmented Cotton Cultivated in Nakornsawan Field Crop Research Center Thailand 575
Uniformity Index
The uniformity index means the distribution of length of fibers in the test sample expressing in
percentage. The degree of uniformity index of the green fiber was 82.9 implying that the fiber was
within the range of medium. In the contrary, the brown cotton was in the range of very low with index at
76.1. Therefore, the green cotton was likely to produce higher quality yarn than the yarn produced from
the brown cotton.
Short Fiber Index
The short fiber index indicates the amount of fibers (by weight) that are less than 0.5 inch in length,
expressing in percentage. The short fiber index of the tested fibers should not exceed 8.0% but both of
them - the green cotton and the brown cotton - were 9.8% and 13.6% respectively, denoted that the
quality of the both fibers were likely to be depreciated and to have high percentage of fiber lose during
yarn production, which conformed to the calculation of the percentage of fiber loss during the actual
spinning processes - the green fiber 32% and the brown fiber 41.4.
Strength
The fiber strength, expresses as breaking tenacity, considered to be one of the most crucial factor in
determining potential strength of yarn, reports in grams-force per tex, and range from 33 and above (very
strong) to 20 and below (very weak). The strengths of the green and brown cottons were 30.7 gram/tex
(strong) and 24.1 gram/tex (weak), respectively.
Elongation
The elongation of the fiber is the distance of the fiber extended before break, expressing in percentage.
The percentage of fiber elongation ranges from 5 and below (very low) to 7.6 and above (very high). The
average percentage of fiber elongations of the green and brown cottons were 7.6% (very high) and 7.1%
(high) respectively.
Moisture
There is no commercial moisture regain assessment value for raw cotton required in general cotton fiber
trade, as the wax layer intact to the fiber barricades liquid water to enter the fiber make it difficult to
achieve standard moisture content value of 8.5% which, otherwise, easier to achieve by processed cotton
fibers. The values of moisture in the test sample of the raw green and brown cottons were 5.8% and 5.9%
respectively.
Trash Count, Trash Area, and Trash Grade
The contamination in the raw cotton fiber is all waste and its removal aggravates cost. The green cotton
had average trash count of 52, trash area of 0.77%, and trash grade 6, while the brown cotton had average
of trash count of 38, trash area of 0.45%, and trash grade 4.
THE FABRIC PROPERTIES
Microscopic Examination
The greige fabrics knitted from the green and brown cotton fibers were observed under stereo microscope
for their general and distinctive appearances - as shown in figure 6. Considered with similar fabric
construction, the greige green and brown fabrics showed distinctive colours from the ordinary greige
white cotton fabric.
a) b)
c)
Fig. 6: Photomicrograph Showing Greige Cotton Fabrics a) White Cotton b) Naturally Green Colour Cotton, and c) Naturally Brown Colour Cotton
Colour Fastness to Light
The greige fabrics were tested to determine the resistance of the colour to the action of an artificial light
source representative of natural daylight (D65), standard test method ISO 105-B02: Colour fastness to
artificial light: Xenon arc fading lamp test. The light fastness level of the fabric produced from the green
fibers was 3-4 and for the fabric produced from brown fiber was 6-7, indicated that the green fabric fade
much quicker than the brown fabric when exposed to light.
CONCLUSION
The two types of naturally pigmented cotton fibers studied in this research had distinctive appearance,
characteristic, and properties form the common white cotton fibers and also different from each other.
The green colour cotton fiber was likely to be produced into the better quality yarn than the brown colour
cotton fiber considering the spinning consistency index, fineness, length, strength, and elongation.
However, the most disadvantage of the green cotton fiber was its light fastness which was slightly
inadequate for textile products to be exposed to sunlight and possibly fading quickly if it was hung dry
outdoor. Therefore, the application for the green fiber was limited. On the other hand, the yarn
production parameters of the brown fiber seems to be deficient but its magnificent light fastness
properties made it be applicable to wider range of products than the green one.
ACKNOWLEDGEMENT
This work was supported by the Thailand Textile Institute, and any attempt at any level cannot be
satisfactorily completed without the contribution and collaboration of the Nakornsawan Field Crop
Research Center, KongKiat Textile Co., Ltd., and Thanuluk Public Company Ltd.
REFERENCES
[1] Morton, W.E. and Hearle, J.W.S. (2008) Physical properties of Textile Fibres. Fourth edition, Cambridge, the Textile
Institute.
[2] Myer, Dorothy. And Stolton, Sue. (1999) Organic Cotton: From Field to Final Product. London, Intermediate Technology
Publications Ltd.
[3] Saville, B.P. (1999) Physical Testing of Textiles. Cambridge, the Textile Institute.
[4] Fan, Qinguo. (2005) Chemical Testing of Textiles. Cambridge, the Textile Institute.
[5] Smith, C. Wayne. And Cothren, J. Tom. (1999) Cotton: Origin, History, Technology, and Production. John Wiley & Son
Inc.
[6] Lewin, Menachem. And Pearce, Eli M. (1998) Handbook of Fiber Chemistry. New York, Marcel Dekker Inc.
[7] Wakelyn, P.J. and Gordon, M.B. (1995) Cotton, naturally, Textile Horizons, Volume 15, no 1, pp 36-38.
Author Index
A
Abdelatif, A., 265
Aboe, M., 500
Adegnika, M.A., 244
Afzal, M.N., 392
Akantetou, K.P., 244
Amthor, J.S., 416
Amudha, J., 129, 136
Amutha, M., 210
Anandalakshmi, Radhamani, 304
Andrysiak, Jerzy, 487
Anuradha, N., 454
Arajo, Alderi Emdio De, 290
Areerak, S., 92
Ariela, N., 258
Arude, V.G., 537
Ashtaputre, A.S., 287
Ashtaputre, Sudheendra, 8
Ayeva, B., 244
B
Babiker, E., 265
Baktavachalam, Gajendra, 184
Balachandran, Y.L., 43
Balasubramani, G., 129, 136
Balasubramanya, R.H., 493, 510
Balu, Amala P., 153
Bange, M.P., 416
Bange, Michael P., 72
Bansal, K.C., 136
Barnes, E., 442
Barroso, 298
Berthold, F., 543
Bhanare, Kanchan, 293
Bharimalla, A.K., 471
Bhat, S.R., 62
Bheemanna, M., 193, 216
Bhuvaneshwari, M., 17
Bihariya, Priyanka, 62
Bindu, V., 477
Bonacic, Ivan, 13
Bonni, G., 244
Breslauer, H., 232
Brevault, T., 244
Bulone, V., 543
Bunker, G.K., 165
C
Carneiro, 298
Carroll, S.C., 220
Chakraborty, Pranjib, 120
Chandran, S.K., 149
Chaporkar, Chandrashekhar, 3
Char, Bharat, 3
Chattannavar, N.S., 287
Chattopadhyay, S.K., 477
Chaudhary, Babita, 111
Chawda, Chetan, 184
Chiliveri, Prashanth, 293
Chinnusamy, C., 399
Chowdary, R., 216
Chuekittisak, R., 92
Cleber, Furlanetto, 298
Collins, G.D., 410
Conaty, W.C., 369
Cothren, J. Tom, 72
Cottee, Nicola S., 72
Cracogna, Mariano, 13
D
Dai, Jianlong, 420
Dargush, P., 323
Das, Tarun Kumar, 95
Dayakar, S., 261
Deshmukh, N.D., 69
Deshmukh, Rachana, 341
Dhamayanthi, K.P.M., 51
Dharajothi, 281
Dong, Hezhong, 420, 432
Dupont, P.L., 392
E
Edmisten, K.L., 410
Esdras, H., 298
F
Fabiane, Mota C., 298
Farooq, Jehanzeb, 35
Ferreira, Alexandre Cunha De Barcellos, 290
Fok, Michel, 435
Frydrych, Iwona, 487
G
Gautier, C., 244
Ghanim, M., 232
Gholamhossein, Hosseini, 85
Gill, J.S., 56
Gokte-Narkhedkar, Nandini, 293
Gopalakrishnan, N., 43, 272, 350
Gotmare, V.D., 477
Goulas, C., 104
Gourlot, 500
Goz, J.P., 500
Grace, G.A.D., 174
578 Author Index
Gujar, G.T., 165
Gumber, R.K., 56, 338
Gurjar, R.M., 510
H
Hadge, G.B., 471
Halakude, I.S., 69
Hallad, A., 203
Hallikeri, S.S., 347
Hanchinal, S., 216
Hanchinal, S.G., 193
Hassan, Syed Zameer Ul, 561
Hma, O., 244
Horowitz, A.R., 232
Hosagoudar, G.N., 287
Hosamani, A.C., 193
Hosamani, A.K., 216
Houndete, T.A., 244
Hubl, E., 500
I
Ibalo, Silvia, 13
Ishaaya, I., 232
Iyer, K.R.K., 524
J
Jadhav, S.B., 524
Janaki, P., 364
Jaybhaye, Ashok, 3
Jeengar, A.K., 477
Jeyaraman, S., 399
Jingjit, Piyanut, 572
Joseph, Achaleke, 255
Jothi, B. Dhara, 210
Juttupornpong, S.A., 92
K
Kadam, V.V., 477
Kalbande, Bipinchandra, 120
Kalia, V., 165
Kalroo, A.M., 268
Kannan, Nallathambi, 17
Karande, Vilas, 471
Karpe, Amol A., 80
Kasivivat, A., 92
Katageri, Ishwarappa, 8
Katsiotis, A., 104
Katturi, Prashanth, 304
Kavithamani, D., 153
Khadi, B.M., 62, 203, 568
Khadi, Basavaraj, 8
Khockakang, K., 92
Khuhro, S.N., 268
Khumla, N.T., 92
Kontsedalov, S., 232
Kranthi, K.R., 129, 136, 198, 203, 272, 281,
293, 341
Kranthi, S., 203, 281
Krishnamoorthy, R., 17
Kudzayi, Mandiveyi Jeremiah, 115
Kuligoud, V.B., 359
Kulkarni, Pandurang, 3
Kulkarni, Vikas V., 100
Kumar, Bharath, 8
Kumar, Rishi, 198, 210, 281
Kumar, T.S. Manoj, 537
Kumar, V., 149, 376, 396
Kumseub, B., 92
Kurulekar, Meera, 304
Kankanallu, Ravi S., 304
L
Lalage, S.B., 69
Lapbunjob, S., 92
Laxman, S., 62
Leva, R.L., 396
Levi, Avishag, 89
Liora, S. Harpaz, 258
Lukonge, E., 500
M
Madhura, C., 100
Mahalingam, Gunasekaran, 28
Mahan, J.R., 369
Mahangade, R.R., 493
Mahmood, Abid, 35
Mahmood, R., 268
Manickam, S., 95
Maralappanavar, Manjula S., 100
Marc, Giband, 298
Marina, D.G., 298
Mario, R., 258
Mauricio, A. Tcach, 13
Mayee, C.D., 272
Mcspadden, W.O., 220
Meena, S., 364
Meshram, Mithila, 120
Mhaske, S.T., 471
Michalakopoulos, P., 104
Michel, Nicole, 298
Mikkilineni, Venugopal, 3
Militky, Jiri, 561
Mohan, Punit, 95
Mohan, S., 229
Monga, D., 272, 281
Morello, Camilo De Lelis, 290
Moudgal, R.K., 184
Muhammad, Dil Baugh, 392
Muralimohan, K., 161
Muthukrishnan, P., 399
Author Index 579
N
Nachane, Nayana D., 493
Nachane, R.P., 531, 353
Nagappa, Harijan, 568
Nagrare, V.N., 281
Nagrare, V.S., 210
Naik, Amruta, 304
Naik, C.B., 281
Nakapan, P., 92, 350, 386
Nandeshwar, Sukhadeo, 120
Nandini, S., 229
Naphade, Prafulla, 3
Narala, Anuradha, 449
Narayanan, K., 100
Nawkarkar, Prachi, 293
Neilsen, J.E., 369
Nithya, C., 399
Noor-Ul-Islam, 35
O
Ochou, G., 244
Ojeda, Alex Montenegro Daniel, 13
O'Leary, P., 442
Ormiston, S., 416
P
Pabich, Anna, 487
Pal, Vijender, 198
Palve, S.M., 62
Panlai, N., 92
Parajulee, M.N., 220
Parmar, R.R., 396
Patel, D.H., 149
Patel, D.U., 149
Patel, J.G., 376, 396
Patel, K.G., 149
Patel, N.N., 149
Paterson, Andrew H., 89
Pathak, D., 95
Patil, B.V., 193
Patil, Bhuvaneshwaragouda, 8
Patil, R.G., 376, 510
Patil, Rajesh S., 8, 62
Patil, S.B., 203, 287
Patil, S.S., 100
Patil, Shreekanth, 8
Paulo, A.V., 298
Pawar, Kasu, 8
Pawar, N.K., 287
Paytas, Marcelo, 309
Pejman, Behdarvand, 85
Petkar, R.B., 454
Poisson, A.F., 13, 43, 80, 136, 341
Prabhuraj, A., 161
Praharaj, C.S., 350
Prasad, N.V.V.S.D., 174, 239, 261
Prudent, P., 244
Pulcha, P., 92
R
Raj, Sheela, 493
Raj, V.C., 396
Raja, D., 17
Rajagopalan, Prem A., 304
Rajan, Sundara, 184
Rajan, T. Sonai, 210
Rajarathinam, S., 153
Rajesh, 287
Rajput, J.C., 69
Ramamurthy, V.V., 281
Ramasami, M., 17
Ramekar, Rahul, 3
Rami, H.A., 258
Rangasamy, S., 104
Rani, S. Usha, 461
Rao, A.S., 406
Rao, G.M.V. Prasad, 239, 174, 281
Rathore, Pankaj, 56, 338
Raza, I., 392
Reddy, A.R., 449, 254
Reddy, V. Chenga, 174, 239
Reed, J., 442
Regina, M.D.G., 298
Renou, A., 244
Riar, R., 410
Rippa, M., 232
Roee, S., 258
S
Sahare, S., 129
Samuel, Nibouche, 255
Sanapapamma, K.J., 100
Sangsoda, P., 92
Sankaranarayanan, K., 350, 386
Santhy, V., 62
Saranga, Yehoshua, 89
Seburuang, P., 92
Seebunruang, Parinya, 572
Selvakumar, P., 17
Shaikh, A.J., 510
Sharma, Mukesh Kumar, 111
Sharma, S., 261
Shekhar, L., 8
Shrestha, R.B., 220
Shukla, S.K., 537
Shweta, C., 136
Silva, 298
580 Author Index
Silvie, P.J., 244
Singh, A.S., 95
Singh, B.P., 165
Singh, Harmandeep, 338
Singh, Jagmail, 111
Singh, Kanchan, 120
Singh, Kulvir, 338
Singh, Suman B., 80
Singh, T.V.K., 261
Singh, V., 281
Sirichumpan, W., 92
Sluijs, Der, 543
Smith, C., 323
Somshekhar, 100
Srinivasa, Y.B., 161
Srivastva, Preeti, 111
Subramaniam, A., 508
Subramanian, V., 17
Sujatha, T., 174
Surulivelu, T., 210
Sutaria, C.M., 396
Sutton, B.G., 369
Suveetha, M., 386
Swami, Dinesh, 198
T
Tan, D.K.Y., 369, 416
Tan, Daniel K.Y., 72
Tandulkar, N.R., 341
Tarrago, Jose, 309
Thaitad, S., 92
Thierry, Brvault, 255
Thiyagu, Krishnasamy, 28
Nadarajan, Nagasamy, 28
Thompson, Gary D., 184
Tiwari, Sapna, 111
Togola, M., 244
Traisiri, A., 92
U
Udikeri, S.S., 203, 281
Upadhye, D.L., 477
Upperi, S.N., 359
Usadadiya, V.P., 396
V
Van, M.H.J., 543
Vandal, N., 203
Varadarajan, P.V., 493
Vastrad, Jyoti V., 100
Velmourougane, K., 386
Venkatakrishnan, S., 553
Venugopalan, M.V., 341
Verma, S.K., 95
Vigneshwaran, N., 471
Virkhede, Sonali, 62
Visser, F., 323
W
Wang, Guiyan, 435
Wasnik, S.M., 461
Weijiang, L.I., 432
Weintraub, P., 232
Wells, R., 410
Y
Yadav, A., 477
Yadav, M.S., 341
Yeates, S.J., 312
Yelekar, S.M., 454
Yves, Carrire, 255
Z
Zanwar, P.R., 281
Zhang, Huijun, 420
Zhenhuai, L.I., 432

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WORLD COTTON RESEARCH CONFERENCE-5

Renaissance Convent ion Cent re, Mumbai 7-11 November 2011

Genetic Diversity Analysis in Cotton Germplasm

Creating Novel Diversity and using Comprehensive

New Cotton Germplasm as an Intermediate Cycle

Introgression of High Fibre Strength Trait

HVI Spectrum in ICC mode.

Estimation of Genetic Parameters for Yield

26.90 0.50 25.50 0.43 24.70 0.56 23.70 0.56

20.70 0.37 27.80 0.42 16.20 0.53 28.70 0.54

41.70 0.54 45.70 0.73 28.90 0.64 40.80 0.84

23.64 0.31 22.41 0.32 19.69 0.29 26.75 0.38

24.63 0.44 27.00 0.51 20.58 0.39 27.34 0.56

22.77 0.50 25.07 0.50 20.33 0.40 29.42 0.58

4.82 0.09 4.74 0.06 4.68 0.08 4.84 0.05

4.13 0.06 4.01 0.05 4.09 0.04 4.07 0.04

3.73 0.03 3.71 0.05 4.57 0.05 4.55 0.05

2.79 0.04 3.08 0.03 2.77 0.04 3.66 0.04

3.02 0.06 3.44 0.05 3.37 0.05 3.73 0.05

2.74 0.06 2.86 0.04 2.61 0.04 3.51 0.05

104.65 3.32 100.76 2.34 100.37 2.87 95.14 2.21

61.28 2.41 96.53 2.06 56.28 2.28 96.99 1.17

134.97 2.64 149.38 2.92 114.16 2.80 169.78 3.45

50.31 1.19 60.36 1.44 45.49 1.21 81.74 1.92

60.99 1.93 83.60 2.29 54.12 1.60 90.36 2.56

47.38 1.87 62.75 2.06 42.69 1.47 87.73 2.76

36.97 0.40 36.31 0.33 34.87 0.47 34.06 0.62

33.77 0.47 32.87 0.32 30.56 0.32 33.99 0.45

33.29 0.35 32.00 0.37 34.07 0.57 33.22 0.38

30.22 0.20 31.54 0.19 30.44 0.17 30.78 0.17

32.93 0.33 33.94 0.30 31.25 0.22 32.75 0.23

29.68 0.29 31.55 0.28 31.67 0.23 30.80 0.26

25.46 0.36 25.36 0.31 30.00 0.42 30.06 0.49

37.28 0.32 34.56 0.29 37.08 0.40 34.54 0.28

34.20 0.24 33.56 0.25 36.26 0.42 35.10 0.37

30.34 0.26 29.49 0.23 31.25 0.35 31.45 0.24

28.47 0.45 28.18 0.38 31.19 0.53 31.28 0.40

34.94 0.16 31.96 0.38 33.60 0.42 32.16 0.38

18.36 0.21 18.22 0.10 21.18 0.33 21.18 0.33

27.12 0.15 26.94 0.29 27.20 0.15 26.86 0.27

24.26 0.26 25.00 0.35 24.36 0.41 24.70 0.41

22.49 0.26 21.90 0.20 22.91 0.51 23.44 0.23

20.83 0.39 21.06 0.32 21.19 0.66 22.23 0.30

25.48 0.39 25.39 0.33 25.18 0.54 25.50 0.41

Introgression of Desirable Characters

40 World Cotton Research Conference on Technologies for Prosperity

Temporal Changes in Metabolically Important

Study of Interspecific Hybrids (Gossypium hirsutum x

Thermosensitive Genetic Male Sterility System

Heterosis for Yield and Yield Attributing Traits

Multi-Level Determination for Heat Tolerance

Genetic Variability in Single Plant Selections

Genetic Parameters of Physiological Traits for Salinity

Tak FA84-4: New Jassid Tolerant Cotton Variety

High Boll Weight and High Ginning Outturn

Development of Naturally Coloured Gossypium

Divergent Selection for Yield and Earliness

Development of Recombinant Inbred Lines

Elite Cotton Varieties in the Zimbawean Private

Development of Biotic Stress Resistance Transgenic

Cloning and Characterization of Cellulose Synthase

Cotton Transgenic with DRE-binding Transcription

Study of Heterosis in Inter Varietal Crosses

Assessment of Genetic Diversity for Improved Fibre