STUDY ON MORPHO-PHYSIOLOGICAL FEATURES AND GROWTH OF MOKARA SP.

ORCHID INFLUENCED BY SPRAY FORMULATION

A Thesis By Examination Roll No. 09Ag.CBot.JD-22 M Semester: July-December, 2010 Registration No. 35893 Session: 2009-2010 Approved as to style and contents by

(Prof. Dr. M. Obaidul Islam) Supervisor

(Prof. Dr. A.K.M. Azad-ud-doula Prodhan) Co-supervisor

(Prof. Dr. Md. Shahidur Rahman)

Chairman Examination Committee and Head, Department of Crop Botany Bangladesh Agricultural University, Mymensingh

November, 2010

STUDY ON MORPHO-PHYSIOLOGICAL FEATURES AND GROWTH OF MOKARA SP. ORCHID INFLUENCED BY SPRAY FORMULATION

A Thesis By Examination Roll No. 09Ag.CBot.JD-22 M Semester: July-December, 2010 Registration No. 35893 Session: 2009-2010

MASTER OF SCIENCE (M.S) IN CROP BOTANY

DEPARTMENT OF CROP BOTANY

BANGLADESH AGRICULTURAL UNIVERSITY MYMENSINGH

NOVEMBER, 2010

ACKNOWLEDGEMENT The author is grateful to the Omnipotent and Almighty Allah, the supreme ruler of the universe, for his immense blessing to keep us alive and successful completion of this research work. The author likes to express profound gratitude, deepest respect and most sincere appreciation to his supervisor Prof. Dr. M. Obaidul Islam, Department of Crop Botany, Bangladesh Agricultural University (BAU), Mymensingh for his scholastic guidance, supervision, constant encouragement and suggestion and intellectual comments throughout the progress of this research work and preparing the manuscript. The author expresses deep sense of gratitude and most sincere appreciation to his honorable co-supervisor Prof. Dr. A.K.M. Azadud-doula Prodhan Department of Crop Botany, Bangladesh Agricultural University (BAU), Mymensingh for his, suggestion, encouragement, constructive criticism and inspiration during the period of research work The author expresses special thanks to Md. Iquebal Kabir, Ripon Kumar Gharmi, S. M. Kamrul Hasan, Harunar-Rashid, Md.Yousuf Ali, for their kind help during the period of the research work. Thankfully acknowledgement is due to United State Department of Agriculture (USDA) for giving partly financial support in present research under the project In vitro regeneration of orchids for commercial production and conservation of endangered species (Project No. 2008/79/USDA). Finally, the author expresses his sincere gratitude to beloved parents, brothers and sisters whose sacrifice and encouragement have been paved the way during study period

The Author

ABSTRACT An experiment was conducted in grill house of the Department of Crop Botany, Bangladesh Agriculture University (BAU), Mymensingh during the period of 30 November, 2009 to 30 September, 2010 to investigate the effect of spray formulation of fertilizer on morpho-physiological feature and growth of two Mokara sp. orchids. The experiment was laid out in Completely Randomized Block Design (RCBD) with two cultivars and four spray formulation where each treatment was replicated three times. Data were collected on plant height, leaf length, leaf width, leaf area, total leaf area, leaf area index, stem diameter, internal structure of stem, number of root, internal structure of root, number of leaf, internal structure of leaf, chlorophyll content, crop growth rate. Results revealed that the morphological and growth parameters of Mokara Diheard Red and Mokara Yellow Anne were significantly varied. Mokara Diheard Red was superior in respect of plant height, leaf wide, leaf area, chlorophyll content and CGR. Mokara Yellow Anne was superior in respect of leaf length, leaf area index, total lead area, stem diameter and number of root. It was observed the

growth of plant height, leaf area, leaf length, leaf wide, leaf area index etc. were the highest with spray formulation-3 (F3) and the lowest were in control (F0). Spray formulation-2 (F2) showed maximum crop growth rate and relative growth rate over control (F0) and the ratio of N-P-K as 10:15: 20, 15:20:25, 10:25:25 and 10:25:30 in spray formulation might be best for growth and development of Mokara sp. orchids.

CONTENTS
CHAPTER TITLE ACKNOWLEDGEMENTS ABSTRACT LIST OF CONTENTS LIST OF TABLES LIST OF FIGURES LIST OF APPENDICES INTRODUCTION REVIEW OF LITERATURE MATERIALS AND METHODS 3.1 Experimental Material 3.2 Experimental site 3.3 Experimental Treatments 3.4 Spray Nutrient Formulation 3.5 Climate 3.6 Water management 3.7 Shading 3.8 Collection of data 3.9 Anatomical investigation 3.10 Experimental Design 3.11 Statistical analysis PAGE I II III VI VIII IX 1 5 12 12 12 12 13 14 14 14 14 18 18 18

1 2 3

CONTENTS (CONTINUED)
CHAPTER TITLE PAGE

RESULTS AND DISCUSSION 4.1 EXTERNAL MORPHOLOGY 4.1.1 Stem 4.1.2 Roots 4.1.3 Leaves 4.1.4 Flowers 4.2 INTERNAL MORPHOLOGY 4.2.1 Roots 4.2.2 Stem 4.2.3 Leaves 4.3 MORPHOLOGICAL FEATURES 4.3.1 Plant Height 4.3.2 Individual Leaf Area 4.3.3 Leaf Length 4.3.4 Leaf Area Index 4.3.5 Leaf Width 4.3.6 Total Leaf Area 4.3.7 Leaf Number 4.3.8 Stem Diameter 4.3.9 Number of Root

19 19 19 19 20 21 21 21 22 23 25 25 26 28 29 33 35 38 40 43

CONTENTS (CONTINUED) CHAPTER TITLE 4.4 Physiological and Growth Parameters 4.4.1 Chlorophyll Content 4.4.2 Crop Growth Rate 4.4.3 Relative Growth Rate 5 SUMMARY AND CONCLUSION REFERENCES APPENDICES

PAGE 44 44 46 47 50 52 58

LIST OF TABLES
TABLE 1 2 3 4 5 6 7 8 9 10 11 12 13 14 TITLE PAGE Composition of spray formulation 13 Main effect of variety on plant height and leaf area 27 Main effect of spray formulation on plant height and 27 leaf area Interaction effect of variety to spray formulation on plant height and leaf area Main effect of variety on leaf area index and leaf length Main effect of spray formulation on leaf area index and leaf length Interaction effect of variety to spray formulation on leaf area index and leaf length Main effect of variety on leaf width Main effect of spray formulation on leaf width Interaction effect of variety to spray formulation on leaf width Main effect of variety on leaf number and total leaf area Main effect of spray formulation on leaf number and total leaf area Interaction effect of variety to spray formulation on leaf number and total leaf area Interaction effect of variety to spray formulation on 28 30 30 31 34 34 35 37 37 38 41

15

leaf number and total leaf area Main effect of spray formulation on number of root and stem diameter

41

LIST OF TABLES (CONTINUED)

TABLE 16 17 18 19 20 21 22 TITLE Interaction effect of variety to spray formulation on number of root and stem diameter Main effect of variety on chlorophyll content Main effect of spray formulation on chlorophyll content Interaction effect of variety to spray formulation on Chlorophyll Content Main effect of variety on CGR and RGR Main effect of spray formulation on CGR and RGR Interaction effects of variety to spray formulation on CGR and RGR PAGE 42 45 45 46 48 48 49

LIST OF FIGURES
FIGURE 1 2 3 4 5 6 7 8 9 10 TITLE External morphology of Mokara Yellow Anne External morphology of Mokara Diheard Red Flower of Mokara sp. Transverse section of leaf of Mokara sp Transverse section of stem of Mokara sp. Transverse section of root of Mokara sp. Showing leaf length of Mokara Yellow Anne Showing leaf length of Mokara Diheard Red Showing leaf breadth of Mokara Yellow Anne Showing leaf breadth of Mokara Diheard Red PAGE 20 20 21 23 23 24 32 32 32 32

LIST OF APPENDICES
APPENDIX TITLE 1 Analysis of variance (Mean square) of plant height, leaf area and leaf number at different 2 growth stage of orchid Analysis of variance (Mean square) of total leaf area, leaf area index and leaf length at different 3 growth stage of orchid Analysis of variance (Mean square) of number of root, stem diameter and leaf width at different 4 growth stage of orchid Analysis of variance (Mean square) of plant height, leaf area and leaf number at different growth stage of orchid 61 60 59 PAGE 58

Chapter I INTRODUCTION

Orchids are one of the important groups of angiosperms (flowering plants) belong to the largest and most diverse family Orchidaceae, consists of about 700-800 genera and more than 25,000 species in the world (Singh and Roy, 2004). They are known for their lovely blooms and are found in diverse habitats. Among the flowering plants, orchids are excellent items for garden and can be grown in beds, pots, baskets, split hallows of bamboo pieces etc. Orchids are the most fascinating, varied and beautiful of all flowers due to its long vase life, attractive structure and excellent color (Singh and Voleti, 1995). Orchids vary in their growth habit, foliage and flower characteristics. Knowledge on these characteristics is basic to the understanding of the different orchid genera and species. According to their growth habit, orchids are divided in to sympodial and monopodial types (Royer, 2003). The monopodial orchids have indeterminate terminal growth. This type of orchids have aerial root and have no pseudo bulb. Whereas sympodial type orchids have pseudo bulb and have no aerial root (Hawkey, 1978). Orchids may be propagated either sexually or asexually. Most of the cultivated orchids are found to be self-sterile. This problem can be overcome by high frequency plant regeneration from somatic embryo, through tissue culture technique. When mass propagation of a new hybrid or a variety is needed within a short time, tissue culture is the only method (Goh et al,. 1992). It has been estimated that it is possible to obtain more than 4 million plants in a year from a single explant (Morel, 1964).This high frequency of propagation may active through tissue culture techniques by using seeds (Stenberg and Kane, 1998), shoot tips (Saiprashed et al., 2002), stem nodes (Pathania et al., 1998),

leaf (Chen et al., 2000), root tips (Chen et al., 2002), lateral buds from young flower stalks (Ichihashi, 1992) etc. Orchids grow over a wide range of climatics condition, ranging from the equator to arctic circle and from low land plains to snow line in mountain areas (Hatch, 1989). Majority of the cultivated orchids were native of tropical countries and found to grow to humid tropical forests of south and central America (Chakrabarti, 1986). They are also distributed through Mexico, India, Myanmar, china, Thailand, Malaysia, Philippines, New Guinea and Australia (Rao, 1977) Orchids having flowers of wonderful beauty and it has very good keeping qualities. They are the most wonderful items for indoor decoration (Patil, 2001). As cut flower and pot plants, orchid is high demandable item. They are also used in different purposes as fragrant, glue, medicine, drinks and flavoring (Goh et al,. 1992). Orchids such as Cymbidium, Dendrobium, Oncidium are marketed globally and the orchid industry has contributed substantially to the economy of many Asian countries (Hew, 1994; Laws, 1995). Thailand is now the most important centre for orchid trade and exports about over 100 million dollars in a year (Singh, 1998). There is a scope of large scale production of orchid in Bangladesh to meet the demand of international market and to earn foreign currency through export (Chowdhury, 1975). Orchids are mainly found in Sylhet, Rangamati, Cox's Bazar, and Madhupur, Tangail, Bandarban, Sundarban, Chittagong and Hilly areas of Bangladesh.

Morpho-physiological features are the primary requisite for better understanding about the potentialities of a variety of orchid. The basic information regarding the morphological character and desirable physiological traits are the most important consideration for orchid cultivation. Foliar nutrient application is very common practice in orchid cultivation as orchid cannot take nutrient significantly from root. Foliar spray nutrients mainly contain nitrogen (N), phosphorus (P), and potassium (K) with different concentration. Application of spray nutrient containing N-P-K with different concentration varied on the basis of growth stage of plants. Urea, TSP and MP are used as a source of nitrogen (N), phosphorus (P), and potassium (K) in spray nutrients. Spray nutrients solution is prepared by mixing of Urea, TSP and MP that plays an important role in growth and development of orchid. In few years ago, orchids were totally unknown to our country but now it is becoming familiar day by day and their production is increased in recent years. To be a successful orchid producer it is pre-requisite to know the morphophysiological feature and behavior of orchid. Literature revealed that there are no research report in Bangladesh on the effect of spray nutrients on growth and development of orchid. By considering above facts and background the present study was undertaken with following objectives.
1. to understand the

morpho-physiological features of orchid (Mokara sp.)

under field condition.

2. to determine the effect of spray formulation on morpho-physiological feature

and growth of orchid (Mokara sp.)

3. to select the suitable concentration of N, P and K in spray formulation for

proper growth of orchid (Mokara sp.)

Chapter II REVIEW OF LITERATURE

Orchids are herbaceous plants posses distinct and special physiological features to adapt themselves in their habitat. Origin of orchids are mountain, hill and deserts where lack of soil, water are prevailed and plants adopted in these habitat have some characters to over come the adverse environmental barrier. Information regarding morpho-physiological features are available in previous literatures related to the present experiment are summarised in this chapter as follows. Razzak (2009) conducted an experiment to study the seasonal effect of micronutrients on the growth and development of orchid (Dendrobium sp). He applied 0.2% spray solution of N-P-K on Dendrobium sp. in summer and winter. Spray nutrient was applied once in a week. After seven months it was found that the rate of growth of orchids affected with nutrient solution application and it was better in summer than winter due to temperature variation. It was concluded that spray nutrient has significant effect concerned to seasonal effect on growth performance of orchids. Kabir (2007) investigated the effect of different liquid fertilizers for growth of orchid. He formulated three liquid fertilizers namely Miracle Grow, Growing orchid food and Flowering food made of Nitrogen, Phosphorus and Potassium at different ratio. These three liquid fertilizers applied on orchids upto blooming and it was found that larger flowers are bloomed in those plants which were treated with flowering food as a source of nutrient.

A pot experiment was conducted by Mohapatra and Saravanan (2006) to determine the suitable growing medium for orchid (Mokara sp.). Different medium namely coconut coir, bhusa, rice husk, tile bits, charcoal, cow dung, brick pieces and groundnut shell single or in combination were used as treatment. They found that maximum plant height and the highest number of new shoots per plant were on the medium as mixture of coconut coir + cow dung + bhusa and Gravel + groundnut shell + cow dung respectively. He also reported the flowering parameters such as early flower bud emergence, number of spikes per plant, spike length and number of florets per spike, were also the greatest with gravel + groundnut shell + cow dung. Ketsa and Kosonmethakul (2006) studied the effect of aluminum sulfate [A12(SO4)3] at 50, 100 and 150 mgL-1 and cobalt chloride (CoC12) at 100, 200 and 300 mgL-1 in the holding solution containing 225 mgL-1 8 mgL-1 hydroxyquinoline sulfate (HQS) and 4% glucose on vase life of Dendrobium 'Sonia Bom Joe' in comparison with the conventional holding solution containing 225 mgL-1 HQS, 30 mgL-1 AgNO3 and 4% glucose was studied at ambient conditions (30.2C and 62% RH) increased vase life and bud opening of orchid flowers effectively. He reported that the holding solution containing 225 mgL-1 HQS, 50 mgL-1, A12(SO4)3 and 4% glucose significantly increased vase life and bud opening of Dendrobium 'Sonia Bom Joe' Ketsa and Kosonmethakul (2004) observed that application of spray nutrients named Physan at the concentration of 100, 200, 300, 400, and 500 ppm weekly interval for 6 months enhanced bud opening and vase life of cut flowers of orchid. Adding 1-7% sucrose to Physan considerably improved water uptake, bud opening and vase life of cut flowers of orchid.

Vaz and Kerbauy (2004) studied the effect of nitrogen, phosphorus and potassium on floral spike induction of Mokara sp. They observed a positive correlation between the total mineral salt concentration and floral spike induction. He concluded that increasing phosphorus with reduction of nitrogen in spray solution have significant effect on floral stimulation. Ghoshal et al. (1989) investigated the effect of liquid nitrogen on floral morphology and behavior of some orchids. He used liquid nitrogen with different concentration and found that those plants are treated with high concentration of liquid nitrogen produced attractive bloom. He also reported that high concentration of liquid nitrogen affect on pollen grain sterility. Wang and Konow (2002) investigated the effect of medium composition on vegetative growth of orchid. They used bark-peat; saw dust and coconut shell as growing medium of orchid. He reported that plant height and number of leaves was maximum on those plants which were grown on saw dust among those medium. Wang and Konow (2004) determined the interaction effect of water-soluble fertilizer and growing medium of Dendrobium sp. They used bark peat and charcoal as growing medium with same concentration of water soluble fertilizer. After six months they found that more leaves greater fresh weights (FW) and larger total leaf areas of Dendrobium sp. on bark peat. Poole and Sheehan (1970) indicated the effect of a low N, high P and K on flower size, flower longevity of orchids. He applied various concentration of N, P and K fertilizer on Vanda sp. upto flowering and found that all plants produces more or less similar size of flowers. It was concluded that Flower size was unaffected by any treatment in this experiment.

Fleischer (1935) studied on the bud opening and vase life of opened buds. He found flower buds near the proximal end of the inflorescence were larger and heavier than those the distal end. He used different chemicals separately or in combination of them and reported that the optimum holding solution was 200 mgL-1 8-hydroxyquinoline sulphate (HQS) + 50mgL-1 vase life . Furukawa et al. (1989) showed that pretreatments of orchid flower with BA significantly suppressed ethylene production and prolonged the vase life of cut florets in Mokara sp. and Dendrobium sp. He found that Combination of BA and spray of silver thiosulfate (STS) had significant effect on the vase life of Cattleya sp. It was concluded that ethylene seems to be involved in senescence of the cut florets but BA suppresses its production which leads to extend vase life. Wong and Lee (2000) evaluated the effect of potting mixes on the growth of Dendrobium sp. and the optimum fertilizer requirement for leaf growth. The different potting media were Cornell Mix (peat moss: vermiculite), Modified Mix (cocopeat: vermiculite), CVS (cocopeat: vermiculite: sand) and cocopeat which were treated with 100 ml of foliar fertilizer (N, P and K) weekly at different concentrations. They reported the average leaf length was higher in Cornell Mix than other medium. Webb and Webb (1993) studied on the maturation of pseudobulb of different orchids as influenced by application of phosphorus (P). They applied different concentration of phosphatic fertilizer on orchid at weekly interval. After eight months they found that high concentration of P leads to attain early maturity of pseudobulb of some orchids. AgNO3 + 8% sucrose solution as this solution increased the percentage of bud opening and prolonged

Matsui et al. (1999) investigated the effect of macronutrients on growth of Dendrobium sp. to identify the optimum concentration for growth of orchid. The growth was promoted at 7.5 mM to 15 mM of N per litre with spray solution weekly whereas growth was suppressed at 0 mM and 30 mM of N per litre spray solution. There was no effect of phosphorous (P) on growth and flowering between 0.5 mM to 2 mM but growth was superior at 10 mM to 20 mM of K per litre spray solution. Therefore it was concluded that N-P-K (7.5- 0.5-20 mM L-1) was suitable for growth of Dendrobium sp. In an investigation Growth, morphology and anatomy of pseudobulb and inflorescence of Oncidium 'Gower Ramsey', studied by Sell and Hermann (1998) in the glasshouse conditions to determine the effect of foliar nutrients on growth and morphological behavior. They observed that mature plants could complete two growth cycles per year. Anatomical observations revealed that a newly elongated vegetative bud contained 1314 nodes in which one node was enlarged to form the pseudobulbs. The pseudobulb contains 5 latent buds with a flower stalk developed from the first nodes (numbered basipetally from the base). They concluded foliar nutrients directly affects on growth.Pseudobulb formation occurred during the unsheathing stage and flower stalks were differentiated when pseudobulbs were elongating. The length of the flower stalk was 4.3 cm when the pseudobulb was fully developed. This young inflorescence contained 23 nodes with floral primardia initiated on the node number of 17 and above. Differentiation and development of inflorescence branches occurred rapidly during the period of flower stalk elongation while differentiation of new floral buds on the main inflorescence apex was slow.

Results suggested that the unsheathing stage was the critical stage for determining flowering. Yin-Tung Wang (1995) conducted an experiment to determined the effect of water-soluble fertilizers on leaf characteristics and applied six water-soluble fertilizers containing N-P-K at different ratio (10-30-20, 15-10-30, 15-20-25, 20-519, 20-10-20, and 20-20-20 in N2-P2O5-K2O respectively) per litre to young seedlings of Phalaenopsis sp. After seven months, he found, leaf size, total leaf area, and shoot and root fresh weight were highly variable among the plants and concluded that the different N-P-K concentration of nutrient solution had a significant effect on leaf number and leaf area. Hlgakl and Imamura (1987) studied on flower production and plant height. He showed that the flower production and plant height increased linearly with increasing of P and N concentration respectively in foliar spray nutrients application. Komori (2002) conducted an experiment during 15 May 1997 to 31 August 1998 to defermine the effect of low light intensity on the growth of Cymbidium orchid cultivars Rose Wine 'Fruity Dror' (RF) and Great Katly 'Little Louransan' (GL). RF and GL plants grown under shaded conditions exhibited lower fresh weight especially root fresh weight than the control treatment. The number of leaves and leaf length were also smaller in RF and Gl than the control. RF and Gl exhibited low development of new leaf than the control. He found that light intensity affect on leaf characteristics significantly.

Patil

(2001)

conducted

an

experiment in

a greenhouse

for the

characterization of flower and observed that six species (Dendi-obium densiflorum, Asocentrum ampullaceum, D. nobile, Phaius tankervilleae, D. pierardii and Ascocentrum ainpullucettin var. Auranticum) and five species (Spcjlhoglolfis speciosa, Aerides mulfiflorum , D. crepidatum, D. Jenkinsii and D. primulinuln) flowered during October-December and JanuaryApril respectively. He reported the largest flowers and the longest spikes (60 cm) were observed in Phaius tankervilleae on April and the smallest flowers were recorded in D. crepidatum and concluded that flowering of orchid is affected by growing season. Lee and Chang (2000) studied the growth of pseudobulb and quality of flower and inflorescence on Oncidium spot determine the effect of temperature on pseudobulb formation and flowering longevity. They observed that high day/night temperature made the pseudobulb thin and long but lower temperature (13-18C) made it short and flat. In lower temperature growth of the pseudobulb was slow and flowering was delayed but the longevity and floret number of the inflorescence was increased. Flower stalks emerged in all temperature treatments except at the extreme temperature. Lower day night temperature favoured inflorescence development and increased branch and floret numbers. In an experiment Prasad et al. (1997) studied on the morphological feature of orchid. They observed that spike length and plant height had the highest heritability and some leaf characteristics are most diversied which was also affected by geographical position.

Sobhana and Rajeevan (1993) conducted an experiment on the performance of' certain epiphytic species in central Kerala as influenced by spray nutrient.They reported flower characteristics such as Inflorescence length and the number, size, colour, fragrance and flowering period are influenced by frequent use of spray nutrients.

Chapter III MATERIALS AND METHOD

3.1 Experimental material Two cultivars of Mokara sp orchid were collected from Dipto orchid nursery Valuka, Mymensingh in the month of November, 2009. The age of clone was about six months. These clones were separated from mother plant and cultured for six months with coconut shell, sawdust and carpenter dust as supporting material. 3.2 Experimental site The experiment was conducted in grill house of Crop Botany Department, Bangladesh Agriculture University, Mymensingh during the period of 30 November, 2009 to 30 September, 2010 to determine the effect of spray nutrients on growth and morph-physiological features of orchid. 3.3 Experimental treatments There are two factors used in this experiment known as variety and formulation of spray nutrients. In the present experiment, two cultivars of Mokara sp were used. As treatment three spray formulations were used along with a control where only tap water was used. Factor A: Two orchid species
a. Mokara Diehard Red

b. Mokara Yellow Anne

Factor B: Three levels of spray nutrient with control

a. Spray Formulation 0 (F 0) b. Spray Formulation 1 (F 1) c. Spray Formulation 2 (F 2) d. Spray Formulation 3 (F 3)

Thus, the total number of treatments was 8 (4x2) and each treatment was replicated three times. The whole experimental beds were divided into three blocks. The size of each block was 2.64 m2. 3.4 Spray nutrient formulation The spray solutions were prepared by mixing of Urea, Tripple Super Phosphate (TSP) and Muriate of Potash (MoP). Formulation was prepared by following the thumb rule of BARI where total amount of fertilizer to be 700 g per 172 litre water was recommended for use as spray. The formulation used in present research contained different ratio of N, P and K. The ratio of N, P and K varied over growth stage of orchid shown in Table 1. Spray formulation used in early stages contained higher N and gradually reduced in later stages. Table 1. Composition of spray formulation. Name of formulation Total urea, TSP and MP 70 gm per 17.2 liter water at N : P : K ratio

F0 (Control) F1 F2 F3 Duration of spray

0.0 15:20:25 10:15:20 5:15:10 December 15 to January 30

0.0 20:20:25 15:20:25 10:15:10 January 31 to March 15

0.0 20:25:30 10:25: 25 10:20:15 March 16 to April 30

0.0 10:30:25 10:25:30 15:20:20 May 01 to June 15

% of N, P and K in Urea, TSP and MP is 46.0, 48.0 and 60.0 respectively. In each times fresh formulation was prepared and sprayed once weekly with a hand sprayer at afternoon. 3.5 Climate The experimental area was located in sub-tropical climatic zone characterize by moderate rainfall and temperature during research period. 3.6 Water management Frequent application of water is essential in orchid cultivation. In November March watering was done once per day and in April - July, watering was done 2 times per day. Watering was done by water cane. 3.7 Shading As an epiphyte, most orchids avoid direct sunlight under natural condition. Orchids prefer dappled shade. For maintaining shade a hand made shade was provided by bamboo. 3.8 Collection of data

Data were collected periodically during the growing period of orchid. The data were recorded on the following morphological parameters. a. Plant height (cm) b. Leaf length (cm) c. Leaf wide (cm)
d. Leaf area (cm2) e. Total Leaf area (cm2)

f. Leaf area Index (LAI) g. Stem diameter (cm) h. Number of root i. Number of leaf j. Chlorophyll content in leaf k. Fresh weight l. Dry weight From collected data Crop Growth Rate (CGR) and Relative Growth Rate (RGR) were calculated. 3.8.1 Plant height The height of plant was measured in cm from ground level to top of the main stem by measuring scale at the interval of 45 days during the study period. 3.8.2 Leaf length The leaf length was measured in cm from leaf base to tip of the leaf by measuring scale at the interval of 45 days during the study period. 3.8.3 Leaf width

The width of leaf was measured by scale along the middle portion of leaf transversely and its average was calculated and expressed in cm at the interval of 45 days during the study period. 3.8.4 Individual leaf area /plant The Individual leaf area was determined by multiplying leaf length with leaf width and expressed in cm 2 . 3.8.5 Total leaf area /plant The total leaf area was determined by summation of all leaf area and expressed in cm 2 .

3.8.6 Leaf Area Index Leaf Area Index is the ratio of leaf area to ground area. Leaf Area Index was determined by dividing Individual leaf area to Individual ground area.
Leaf area Leaf Area Index = Ground area

3.8.7 Stem diameter Stem diameter was measured by using slide calipers at the middle portion of stem. 3.8.8 Number of root Number of Root was estimated by counting all roots in a plant.

3.8.9 Crop Growth Rate Crop Growth Rate (CGR) defined as the increase of plant material per unit of time. It is expressed as g of dry matter produced per day. It can be expressed as following equation. W 2 -W 1 Crop Growth Rate = T 2 -T 1 W1 and W2 are dry weights of plants at times T1 and T2 respectively. 3.8.10 Relative Growth Rate Relative Growth Rate (RGR) defined as the increase of plant material per unit of time per unit area. This parameter indicates a rate of growth per unit of dry matter. So, RGR is the unit of plant material per unit of material present per unit of time. It is expressed as g of dry matter produced by a g of existing dry matter in a day. RGR is the efficiency of dry matter production. It can be expressed as following equation. Log W 2 -Log W 1 Relative Growth Rate = T 2 -T 1 Where, W1 and W2 are dry weights of plants at times T1 and T2 respectively. 3.8.11 Chlorophyll estimation Chlorophyll is the most important photosynthetic apparatus in plant. Chlorophyll content in leaf was estimated at 60 days after transplanting

(DAT) and 90 DAT by following procedure of Arnon (1949). Chlorophyll content in leaf was expressed as mg/g of a sample by using following equation. Chl(a)= 0.0127D663-0.00269D645 Chl(b)= 0.0229 D645-0.00468 D663 Where, D645 =Absorbance at 645 nm wave length D663= Absorbance at 663 nm wave length Total chlorophyll= Chl. a + Chl. b

3.8.11.1 Steps for chlorophyll estimation At first 0.5 mg fresh leaf was taken as sample. Each sample was crushed by mortar and pestle in 10 ml acetone (80%). Then the extract was centrifuged for 10 minutes at 1000 rpm. Extract of each test tube was made a final volume of 10 ml by addition of acetone to fulfill volatilization loss. The reading was recorded at 663 nm and 654 nm in UV- VIS spectrophotometer. 3.9 Anatomical investigation

To investigate the general anatomical structure of root, stem and leaf thin (about 10 M) section were prepared from fresh plant material by sharp blade. The thin sections were stained in 1.0% safranin and mounted on slide. A cover sleep was set on thin section and observed under microscope with high resolution, progressive scan digital camera system (Model Olympus BX 41 and camera DP 20) and photos were taken. 3.10 Experimental design The experiment was laid out in Completely Randomized Block Design (RCBD) with 8 treatments and each treatment was replicated three times. 3.11 Statistical analysis The collected data analyzed by following the analysis of variance (ANOVA) technique and mean differences were adjudged by Duncans Multiple Range Test (DMRT) (Gomez and Gomez, 1984) using a computer operated programme named MSTAT-C (Russel, 1986).

Chapter IV RESULTS AND DISCUSSION

The results obtained from the experiment are described and discussed in this chapter. The morphological and physiological parameters of orchid have been presented and discussed under separate heads and sub-heads with tables and figures. 4.1 External morphological features of Orchid Orchids easily identified by its leaves, stems and roots, flowers, fruits and seeds. They are herbaceous plants having distinct characteristics. The present cultivars namely Mokara Diheard red and Mokara Yellow Anne grew as epiphyte on sawdust, coconut coir and carpenter dust. The general morphological features of the species under study were as follows. 4.1.1 Stem The stem of Mokara sp. was erected, cylindrical having nodes and internodes. Basal parts of the stem were enclosed by leaf sheath. The lower most part of the stem contained 3 types of roots. The color of stem was light green to dark green. 4.1.2 Roots Roots of Mokara sp. under study were thin and cylindrical. There were three types root found in Mokara sp. namely clinging root, absorbing root and aerial root. The color of root was brown to purple. Tip of the root is blunt and smooth in aerial root. Orchid roots arouse from the base of pseudobulb, node.

Fig.1. External morphology of Mokara Yellow Anne

Fig. 2. External morphology of Mokara Diheard Red 4.1.3 Leaves

Leaves were simple, alternate and very thick. Leaves of Mokara sp. orchid have parallel venation. The shapes of orchid leaves were palmate, ovate, obovate.

Fig. 3. Flower of Mokara sp. 4.1.4 Flower Orchid flowers were zygomorphic. The inflorescence of Mokara sp. is racemos. Their segments were arranged in several whorls. The outermost whorl consists of three sepals which together make up the calyx. The second whorl of Mokara sp consists of three petals and it is known as corolla. Calyx and corolla together produced perianth. The anthers of orchids are finger shaped which partially fused with the filaments of stamens and individual pollen grains are monads. 4.2 Anatomical features 4.2.1 Root

The orchid under study had stout roots that were circular in transverse section. The epidermis functions as protective layer and consists of rectangular cells with lignified wall. The cortex remains inside of epidermis which functions as storage and transportation (Fig. 6). An endodermis bounds the cortex on the inside and encloses the stele. The endodermis cell had distinct casperian strips and was rectangular to oblong in shape. The vascular bundles were aerial in which protoxylem distributed throughout the periphery and metaxylem throughout the centre. The velamen tissue consists of 4-6 layers of polygonal radially elongated cells which transport water and nutrient from soil to leaf. 4.2.2 Stem The stem under microscope was showed different components of tissue. Epidermis of Mokara sp orchid was observed as single layered with cuticle. A few stomata were present in the epidermis. Inside the epidermis 7 layers of round, oval, elliptical or irregular parenchyma cells were found known as cortex (Fig. 5). Interior to the epidermis one layer of irregularly shaped parenchymatous cell was observed and called hypodermis. Ground tissue was mostly isodiametric cells. Vascular bundles are distributed throughout the ground tissue. There was no cambium in vascular bundle. The metaxylem present in peripheral region and protoxylem present in centre of stem under microscope. 4.2.3 Leaf The leaf of orchid under study was thick and fleshy. A transverse section of orchid leaf was observed under microscope and different component of tissue

was found. In Mokara sp. a thick cuticle covers the epidermis which consists of small heavy walled cells. A multilayered palisade parenchyma was seen bellow the epidermis. The epidermis was two types known as upper epidermis and lower epidermis. The palisade layer is located bellow the hypodermis and was two or three layers of thick elongated cell. The spongy mesophyll bellow the palisade parenchyma was round. Both the palisade layer and spongy mesophyll are photosynthetic tissue. Vascular bundles were collateral and closed (Fig. 4).

Cuticle Spongy parenchyma Vascular bundle Palisade parenchyma

Epidermis

Fig. 4. Transverse section of leaf of Mokara sp.

Corte

Vascular Bundle

Epidermis Cuticle

Fig. 5. Transverse section of stem of Mokara sp.

Cuticle Xylem vessels Epidermis Epidermis (Radially Cortrex Endodermis Velamen

Fig. 6. Transverse section of root of Mokara sp.

4.3 Morphological features 4.3.1 Plant height Plant height varied significantly between the two cultivars of orchids. At 45 DAP, plant height of Mokara Diheard Red and Mokara Yellow Anne were 24.88 cm and 19.75 cm respectively. At 90 DAP, it was 27.23 cm and 22.28 cm, respectively. Plant height of Mokara Diheard Red and Mokara Yellow Anne were 29.68 cm and 24.95 cm , respectively at 135 DAP. Maximum plant height of Mokara Diheard red and Mokara Yellow Anne were 32.19 cm and 27.44 cm, respectively at 180 DAP. It was observed that plant height increased progressively with the advancement of age and growth stages. Plant height increased rapidly in Mokara Diheard Red than Mokara Yellow Anne comparatively (Table 2). Different nutrients as spray have significant effects on plant height. At 45 DAP, the highest plant height was 23.15 cm recorded in F1 and lowest was 21.63 cm in F3. At 90 DAP and 135 DAP, maximum plant height 25.91 cm and 28.8 cm respectively were recorded in F1 and minimum plant height was 24.2 cm and 25.51 cm in control (F0). At 180 DAP, maximum plant height was 31.9 cm in F1 and minimum was 26.88 cm in F0 (Table 3). The plant height increased slowly in control (F0) and comparatively rapid in F1 indicated that different formulation was effective in plant height. The interaction effect of variety to formulation revealed that plant height affected significantly. Maximum plant height was 26.8 cm in Mokara Diheard Red at formulation-1 (V1F1) and minimum was 19.03 cm in Mokara Yellow Anne with formulation-2 (V2F2) at 45 DAP. Similar trend in plant height was observed with increasing cultivation period. At 180 DAP, maximum plant

height was 32 cm that was found in V1F1 and minimum was 22.93 cm recorded in V2F0 (Table 4). The present study support the report of Wang and Konow (2004) 4.3.2 Individual leaf area Varietal effect of leaf area was significant. The leaf area ranged from 50.577.1 cm2 and 44.2 - 73.2 cm2 in Mokara Diheard red and Mokara Yellow Anne, respectively over growing period. The Mokara Diheard red (V1) showed maximum leaf area of 60.03 cm2 over leaf area 53.18 cm2 of Mokara Yellow Anne (V2) at 45 DAPS. At 90 DAP, the leaf area of V1 was 60.03 cm2 and of V2 Was 53.18. At 135 DAP, the leaf area of V1 and V2 was 68.82 cm2 and 63.58 cm2, respectively. At 180 DAP, the leaf area of V1 was 77.14 cm2 and of V2 Was 73.20 cm2 (Table 2). From table it was observed that leaf area increment was higher in V2 than V1. Spray formulation was effective in enhancing the leaf area. At 45 DAP, maximum leaf area was 51.81 cm2 with F2 and minimum was 46.89 cm2 was with F1. At 90 DAP, maximum leaf area was 59.52 cm2 in F2 and minimum leaf area of 52.67 cm2 in control (F0). Leaf area gradually increased with increasing age of the plant. At 135 DAP, maximum leaf area was 70.16 cm2 in F3 and minimum leaf area was 58.93 cm2 in control (F0). At 180 DAP, maximum leaf area 80.60 cm2 was found in F3 and minimum 66.85 cm2 in F0 (Table 3). The trend of leaf area increment over vegetative growth was rapid in F3. Leaf area varied significantly due to the interaction effect between variety and treatments. At 45 DAP, the highest leaf area was 51.29 cm2 recorded in V1F1

and the lowest was 41.94 cm2 in V2F3. At 90 DAP, the highest leaf area was 62.77 cm2 and the lowest was 48.54 cm2 recorded in V1F3 and V2F0, respectively. At 135 DAP, the highest leaf area was 74.03 cm2 and the lowest was 54.98 cm2 that recorded in V1F3 and V2F0, respectively. V1F3 and V2F0 also shows the highest 82.84 cm2 and the lowest 63.94 cm2, respectively at 180 DAP (Table 4). The variation in leaf area might occur due to athe vriation in number of leaves and their expansion. The result obtained from the present study is consistent with the results of Sharma and Haloi (2001) who stated at variation in leaf area might be attributed to the difference in number of leaves. Table 2. Main effect of variety on plant height and leaf area.
Variety V1 V2 CV Plant Height (cm) at DAP 45 90 135 180 24.88 27.23 29.68 32.19 19.75 16.91 22.28 14.63 24.95 12.88 27.44 12.09 Leaf area (cm2) at DAP 45 90 135 180 50.54 60.03 68.828 77.1 44.25 10.89 53.18 11.06 63.58 10.59 73.20 10.12

Table 3. Main effect of spray formulation on plant height and leaf area.
Spray formulation F0 F1 F2 F3 CV Plant height (cm) at DAP 45 22.76 b 24.15 a 21.71 b 21.63 b 16.91 90 24.20 25.91 24.45 24.46 14.63 135 25.51 b 28.8 a 27.71 a 27.25 a 12.88 180 26.88 b 31.90 a 30.58 a 29.9 a 12.09 45 46.69 46.89 51.81 47.79 10.89 Leaf area (cm2) DAP 90 52.67 56.94 59.52 57.27 11.06 135 58.93 b 66.81 a 68.90 a 70.16 a 10.59 180 66.85 b 77.17 a 76.08 a 80.60 a 10.12

Legend

V1 = Mokara Diheard Red V2 = Mokara Yellow Anne DAP = Days After Planting

F0 = Control F1 = Spray Formulation-1 F2 = Spray Formulation-2 F3 = Spray Formulation-3

Table 4. Interaction effect of variety to spray formulation on plant height and leaf area.
Variety x spray formulation V1F0 V1F1 V1F2 V1F3 V2F0 V2F1 V2F2 V2F3 CV 45 25.46 a 26.80 a 24.40 a 22.86 ab 20.06 b 19.50 ab 19.03 b 20.40 b 16.91 Plant height (cm) at DAP 90 26.96 a 29.53 a 26.93 a 25.50 a 21.43 b 22.3 b 21.96 b 23.43 ab 14.63 135 28.10 a 32.00 a 30.o a 28.63ab 22.93 b 25.6 ab 25.43ab 25.86ab 12.88 180 29.56 b 35.1 a 32.86 a 31.23 a 24.20 b 28.70 b 28.30 b 28.56 b 12.09 45 49.87ab 51.29 a Leaf area (cm2) at DAP 90 56.81ab 62.41 a 135 62.87 ab 71.18 a 67.22 ab 54.98 b 62.45 ab 66.29 ab 10.59 180 69.76 ab 81.97 a 74.01 ab 82.84 a 63.94 b 72.37 ab

47.76 ab 58.12ab 53.26 ab 44.07 ab 42.89 b 48.11 ab 41.94 b 10.89

62.77 a 74.03 a 48.54 b 51.48ab

60.93 a 70.58 a 78.15 ab 51.78ab 11.06 78.36 ab 10.12

4.3.3 Leaf length Average leaf length recorded at 45, 90 , 135 and 180 DAP was varied significantly between Mokara Diheard Red and Mokara Yellow Anne .The average leaf length of V1 was 22.84 cm and V2 was 22.65 cm at 45 DAP. At 90

DAP, the average leaf length of V1 was 23.63 cm and V2 was 23.15 cm. The average leaf length was 24.07 cm and 23.6 cm at 135 DAP in V1 and V2 respectively. At 180 DAP, the average leaf length of V1 was 24.75 cm and V2 was 24.34 cm (Table 5). From table it was observed that in all days leaf length was more in V1 than V2. So, Mokara Diheard Red (V1) was superior to Mokara Yellow Anne (V2) in respect of leaf length.

At 45 DAP, the highest leaf length was 23.23 cm recorded in F2 and the lowest of 22.44 cm in F3. Similar trend in leaf length was observed with increasing cultivation period. At 180 DAP, maximum leaf length was 25.13 cm that recorded in F2 and minimum 24.03 cm was in F3 (Table 6). Leaf length increased at slow rate with advancement of time and growth stage. The N, P and K concentration gradually changed with culture period and found less little effective on leaf length. The lowest leaf length was observed in F3 which contained the lowest concentration of N, P and K. Results revealed that increasing the concentration of N, P and K in spray formulation leads to increase leaf length. The interaction effect of variety and treatments on leaf length was significant. At 45 DAP, the highest leaf length was 23.9 cm recorded in V1F0 and the lowest 21.78 cm was in V1F3. At 90 DAP, the highest leaf length was 24.78 cm in V1F0 and the lowest 22.2 cm was in V1F3. At 135 DAP, the highest leaf length 25.23 cm was recorded in V1F0 and the lowest 22.66 cm was in V1F3. At 180 DAP, the highest leaf length 26.03 cm was recorded in V 1F0 and the lowest 23.2 cm was in V1F3 (Table 7).

4.3.4 Leaf area index Leaf Area Index (LAI) recorded at 45 DAP, 90 DAP, 135 DAP and 180 DAP was significantly varied between two cultivars of orchids. In V1, the average leaf area index was 0.47, 0.66, 0.94 and 1.21 at 45, 90,135 and 180 DAP respectively. The average leaf area index of V2 was 0.50, 0.74, 1.05 and 1.39 at 45, 90,135 and 180 DAP respectively (Table 5). In comparison between two varieties degree of increasing of leaf area index was more in V2 than V1. Thus varietal effect on leaf area index was significant and V2 may be considered as superior over V1 in this respect. At 45 DAP, the highest leaf area index was 0.50 recorded in F2 and the lowest was 0.45 found in F3. At 90 DAP, the highest leaf area index was 0.76 recorded in F2 and the lowest was 0.63 in F0. At 135 DAP, the highest and lowest leaf area index were 1.05 and 0.89 observed in F2 and F0, respectively. At 180 DAP, the highest leaf area index was 1.37 and the lowest was 1.17 found in F 3 and F0, respectively (Table 6). It was observed that degree of increasing of leaf area index was more in F3. The variation in leaf area index might occur due to the variation in number of leaves and their expansion. The results obtained from the present study was consistent with the result of Sharma and Haloi (2001) who stated that variation in leaf area index could be attributed to the change in leaf number. The results obtained from the present study also supported by Chandra and Das (2000). Table 5. Main effect of variety on leaf area index and leaf length.
Variety Leaf area index at DAP 45 90 135 180 45 Leaf length (cm) at DAP 90 135 180

V1 V2 CV

0.47 0.5 11.62

0.66 0.74 14.25

0.94 1.05 12.61

1.21 1.39 12.77

22.84 22.65 7.65

23.63 23.15 7.52

24.07 23.60 7.49

24.75 24.34 7.13

Table 6. Main effect of spray formulation on leaf area index and leaf length
Spray formulation F0 F1 F2 F3 CV Leaf area index at DAP 45 90 135 180 0.46 a 0.49 a 0.5 a 0.45 a 11.62 0.63 b 0.89 b 0.71 a 0.76 a 0.68 a 14.25 1.02 a 1.05 a 1.02 a 12.61 1.17 b 1.34 a 1.33 a 1.37 a 12.77 Leaf length (cm) at DAP 45 90 135 180 23.00 a 22.33 a 23.23 a 22.44 a 7.65 23.60 a 23.24 a 23.83 a 22.88 a 7.52 24.0 a 23.66 a 24.40 a 23.3 a 7.49 24.65 a 24.36 a 25.13 a 24.03 a 7.13

Legend V1 = Mokara Diheard Red V2 = Mokara Yellow Anne DAP = Days After Planting F0 = Control F1 = Spray Formulation-1 F2 = Spray Formulation-2 F3 = Spray Formulation-3

Table 7. Interaction effect of variety to spray formulation on leaf area index and leaf length.
Variety x Spray formulation V1F0 V1F1 V1F2 V1F3 V2F0 V2F1 V2F2 V2F3 CV 45 0.55 ab 0.43 c 0.44 c 0.46 bc 0.45 bc 0.56 a 0.56 a 0.45 bc 11.62 90 0.68 b 0.64 b 0.62 b 0.69 b 0.59 b 0.78 ab 0.90 a 0.67 b 14.25 135 0.93 bc 180 1.14 b 45 23.9a 22.76a 22.93a 21.78b 22.1 a 21.9 b 23.53 23.1 a 7.65 90 24.78 a 24.18 a 23.36 a 22.2 b 22.43 b 22.3 b 24.3 a 23.56 a 7.52 135 25.23 a 24.56 a 23.83 a 22.66 b 22.76 b 22.76 b 24.96 a 23.93 a 7.49 180 26.03 a 25.36 b 24.4 bc 23.2 c 23.26 c 23.36 c 25.86 a 24.86 bc 7.13 Leaf area index at DAP Leaf length (cm) at DAP

0.92 bc 1.24 ab 0.88 bc 1.02 bc 0.85 c 1.13 ab 1.21 a 1.03 bc 12.61 1.15 b 1.33 ab 1.21 ab 1.44 ab 1.51 a 1.41 ab 12.77

Fig. 7. Showing leaf length of Mokara Yellow Anne

Fig. 8 . Showing leaf length of Mokara Diheard Red

Fig. 9. Showing leaf breadth of Mokara Yellow Anne

Fig. 10. Showing leaf breadth of Mokara Diheard Red

4.3.5 Leaf width

Leaf width ranged from 2.22 - 3.12 cm and 1.95 - 3.02 cm in Mokara Diheard red and Mokara Yellow Anne respectively during the growing period. At 45 DAP, leaf width of V1 was 2.22 cm and 1.95 cm in V2. At 90 DAP, leaf width of V1 was 2.5 cm and 2.29 cm in V2. At 135 DAP, leaf width of V1 was 2.84 cm and 2.68 cm in V2. At 180 DAP, leaf width of V1 was 3.12 cm and 3.02 cm in V2 (Table 8). From table it was observed that rate of increasing of leaf width is more in Mokara Yellow Anne than Mokara Diheard red. Results revealed that Mokara Yellow Anne is superior to Mokara Diheard red in respect of leaf width. Spray formulation significantly affected the leaf width. At 45 DAP, the highest leaf wide was 2.13 cm recorded in F3 and the lowest was 2.05 cm found in control (F0). At 90 DAP, the highest leaf width (2.51 cm) was found in F 3 and the lowest (2.23 cm) in control (F0). Similar trend was observed at 135 DAP. At 180 DAP, the highest leaf width was 3.36 cm was found in F3 and the lowest was 2.71 cm recorded in control F0 (Table 9). Data revealed that leaf width increased at rapidly in F3 than other treatments. Maximum leaf width always recorded in F3 and minimum in control (F0) which revealed that Spray nutrients containing N, P and K have little effect on leaf wide. Leaf width varied significantly by the interaction between variety and treatments. At 45 DAP, the highest leaf width was 2.45 cm recorded in Mokara Diheard red with formulation 1 (V1F3) and the lowest was 1.81 cm found in Mokara Yellow Anne with formulation 3 (V2F3). Similar trend in leaf width was observed with increasing cultivation period (Table 10). At 180 DAP, the highest leaf width (3.56 cm) was recorded in V1F3 and the lowest (2.68 cm) was in V1F0.

Table 8. Main effect of variety on leaf width.

Variety 45 V1 V2 CV 2.22 1.95 7.95 Leaf width (cm) at DAP 90 135 2.5 2.29 7.63 2.84 2.68 7.14 180 3.12 3.02 6.73

Table 9. Main effect of spray formulation on leaf width.

Spray formulation F0 F1 F2 F3 CV 45 2.05 a 2.1 a 2.05 a 2.13 a 7.95 Leaf width (cm) at DAP 90 2.23 b 2.44 a 2.4 a 2.51 a 7.63 135 2.45 c 2.81 ab 2.75 b 3.01 a 7.14 180 2.71 c 3.15 ab 3.07 b 3.36 a 6.73

Legend V1 = Mokara Diheard red V2 = Mokara Yellow Anne DAP = Days After Planting F0 = Control F1 = Spray Formulation-1 F2 = Spray Formulation-2 F3 = Spray Formulation-3

Table 10. Interaction effect of variety to spray formulation on leaf width.

Variety x spray formulation V1F0 V1F1 V1F2 V1F3 V2F0 V2F1 V2F2 V2F3 CV 45 2.10 bc 2.25 ab 2.08 bc 2.45 a 2.0 bc 1.95 bc 2.03 bc 1.81 c 7.95 Leaf width (cm) at DAP 90 2.30 bc 2.58 ab 2.31 bc 2.83 a 2.16 c 2.3 bc 2.5 ab 2.2 c 7.63 135 2.50 cd 2.90 b 2.70 bcd 3.26 a 2.41 d 2.73 bcd 2.81 bc 2.76 bcd 7.14 180 2.68 d 3.23 ab 3.03 bcd 3.56 a 2.75 cd 3.06 bcd 3.11 bc 3.16 b 6.73

4.3.6 Total leaf area The development of total leaf area over time in orchid was significantly affected by varieties. The total leaf area ranged from 523.55-1346.96 cm2 and 565.16 - 1540.38 cm2 in Mokara Diheard red and Mokara Yellow Anne, respectively over growing period. At 45 DAP, the total leaf area of V1 was 523.55 cm2 and of of V2 was 565.16 cm2. At 90 DAP, the average total leaf area of V1 was 735.01 cm2 and of V2 was 809.60 cm2. At 135 DAP, average total leaf area of V1 was 1029.82 cm2 and of V2 was 1169.59 cm2. At 180 DAP average total leaf area of V1 was 1346.96 cm2 and of V2 Was 1540.38 cm2. (Table 11). From table it was observed degree of leaf area increment was higher in Mokara Yellow Anne than Mokara Diheard red affected leaf width. and spray formulation

Spray formulation significantly affected the leaf width. At 45 DAP, maximum average total leaf area was 562.3 cm2 was found in control (F0) and minimum average total leaf area was 508.09 cm2 in F3. At 90 DAP, maximum total leaf area 844.59 cm2 was found in F2 and minimum 691 cm2 in control (F0). At 135 DAP, maximum average total leaf area 1160.46 cm2 was found in F2 and minimum was 965.90 cm2 found in control (F0). At 180 DAP, maximum average total leaf area was 1524.03 cm2 found in F3 and minimum was 1298.03 cm2 observed in F0 (Table 12). From table it was observed that trend of leaf area increased over vegetative growth was relatively higher in F3. In all treatments, average total leaf area increased in later growth stage during which phosphorus (P) and potassium (K) concentration was increased in spray solution which indicated that P, and K in spray nutrients increases leaf area. Total leaf area varied significantly by the interaction between variety and formulation. At 45 DAP, the highest total leaf area was 627.50 cm2 recorded in V2F2 and the lowest was 478.0 cm2 observed in V1F1. At 90 DAP, the highest and lowest leaf area were 1000.54 cm2 and 625.8 cm2 recorded in V2F2 and V2F0 respectively. At 135 DAP, the highest and lowest total leaf area were 1341.67 cm2 and 947.66 cm2 in V2F2 and V2F0, respectively. V2F3 and V1F0 also showed the highest (1665.61 cm2) and lowest (1256.78 cm2) total leaf area respectively at 180 DAP (Table 13). The variation in leaf area might occur due to the variation in number of leaves and their expansion. The result obtained from the present study is consistent with the results of Sharma and Haloi (2001) who stated that variation in leaf area might be attributed to the difference in number of leaves.

Table 11. Main effect of variety on leaf number and total leaf area.
Variety V1 V2 CV 45 10.25 12.66 14.69 Leaf number at DAP 90 135 180 12.41 15.00 17.58 15.33 11.77 18.41 10.37 21.25 8.48 Total leaf area (cm2) at DAP 45 90 135 180 523.55 735.01 1029.82 1346.96 565.16 11.62 809.60 14.41 1169.59 12.91 1540.38 12.08

Table12. Main effect spray formulation leaf number and total leaf area.
Spray formulation F0 F1 F2 F3 CV Leaf number at DAP 45 90 135 180 11.83 a 13.50 16.50 19.50 12.83 a 11.66 a 10.50 b 14.69 14.16 14.50 13.33 11.77 a 17.86 a 17.16 19.33 19.83 Total leaf area (cm2) at DAP 45 90 135 180 562.3 a 691 b 965.9 b 1298.03 549.01 a 558.02 a 508.09 b 11.62 789.06 ab 844.59 a 764.57 ab 14.41 1135.1 ab 1160.46 a 1137.37 ab 12.91 c 1482.42 a 1470.2 a 1524.03 a 12.08

a 15.oo b 19.00 10.37 8.48

Legend V1 = Mokara Diheard Red V2 = Mokara Yellow Anne F0 = Control F1 = Spray Formulation-1

DAP = Days After Planting

F2 = Spray Formulation-2 F3 = Spray Formulation-3

Table 13. Interaction effect of variety to spray formulation on leaf number and total leaf area.
Leaf number at DAP 45 12.0 abc 9.0 c 10.33 90 13.33 bc 11.33 c 12.66 135 15.66 b 14.33 b 15.33 b 14.66 b 17.33 ab 20.0 a 19.0 a 17.33 ab 10.37 180 18.0 cd 16.66 d 17.66 cd 18.0 bcd 21.0 ab 22.0 a 22.0 a 20.0 abc 8.48 45 613.7 ab 478.0 c 488.54 c 513.96 abc 510.9 abc 620.02 ab 627.50 a 502.22 bc 11.62 Total leaf area (cm2) at DAP 90 756.19 bc 708.82 bc 688.64 bc 786.39 bc 625.80 c 869.31 ab 1000.54 a 742.75 bc 14.41 135 984.15 bc 1022.99 bc 979.26 bc 1132.9 180 1256.78 b 1369.14 ab 1274.78 b 1487.13

Variety x spray formulation V1F0 V1F1 V1F2 V1F3 V2F0 V2F1 V2F2 V2F3 CV

bc c 9.66 c 12.33 11.66 abc 14.66 a 13.0 ab 11.33 bc 14.69 c 13.66 bc 17.0 a 16.33 ab 14.33 abc 11.77

abc ab 947.66 c 1339.29 1247.20 ab 1341.67 a 1141.85 abc 12.91 ab 1595.7 ab 1665.61 a 1560.93 ab 12.08

4.3.7 Leaf number The number of leaves per plant was statically significant at 45, 90,135,180 DAP. The leaf number ranged from 10.25-17.58 and 12.66-21.25 in Mokara

Diheard red and Mokara Yellow Anne respectively over growing period. At 45 DAP, the average leaf number of Mokara Diheard red was 10.25 and of Mokara Yellow Anne was 12.66. In Mokara Diheard red, the average number of leaves per plant was 12.41, 15 and 17.58 at 90,135 and 180 DAP respectively. In Mokara Yellow Anne, the average number of leaves per plant was 15.53, 18.41 and 21.25 at 90,135 and 180 DAP respectively (Table 11). Though both varieties were treated with same spray nutrients but production of leaf was higher in Mokara Yellow Anne than Mokara Diheard Red. Results revealed that Mokara Yellow Anne might be superior over Mokara Diheard Red in respect of leaf production. At 45 DAP, the averave highest leaf number was 12.83 recorded in F 1 and the lowest was 10.5 found in F3. At 90 DAP, the highest leaf number was 14.5 in F2 and the lowest was 13.33 in F3. At 135 DAP, the average highest leaf number was 17.86 was in F1 and the lowest was 15 found in F3. At 180 DAP, the highest (19.83) and the lowest (19) leaf number was also in F2 and F3, respectively (Table 12). Since level of nutrient concentrations in spray solution was altered over growth stage and number of leaves also increased with alteration of nutrient concentrations in spray formulation. It might be concluded that spray nutrients was t effective on leaf production. The present result supports the report of Singh and Kohli (1999). Leaf number varied significantly due to the interaction effect between variety and formulation. At 45 DAP, the highest leaf number was 14.66 and the lowest leaf number was 9 in V2F1 and V1F1, respectively. The highest leaf number at 90,135, 180 DAP were 17, 20 and 22, respectively and recorded in of V2F1. The lowest leaf number at 90,135, 180 DAP were 11.33, 14.33, 16.66, respectively

and in V1F1 (Table 13). The variation in total number of leaves might be concerned with genetically variation, physiological function and growth characters of plant.

4.3.8 Stem diameter Varietal effect of stem diameter was significantly varied. In Mokara Diheard Red, the average stem diameter per plant was 0.81, 0.97, 1.1, 1.25 at 45, 90, 135 and 180 DAP, respectively. In Mokara Yellow Anne, the average stem diameter per plant was 0.96, 1.11, 1.27 and 1.43 at 45, 90,135 and 180 DAP, respectively (Table 14). In comparison between two varieties degree of increasing of stem diameter was more in Mokara Yellow Anne than Mokara Diheard Red. At 45 DAP, the highest stem diameter was 0.96 cm recorded at spray formulation 1 (F1) and the lowest was 0.81 cm in control (F0). Spray formulation-3 (F3) showed maximum stem diameter (1.11 cm) and F2 showed minimum stem diameter (0.97 cm) at 90 DAP. At 180 DAP, maximum (1.41` cm) and minimum (1.27 cm) stem diameter was observed in F3 and F0, respectively (Table 15). At 90 and 135 DAP, stem diameter in control plants was more than F1 and F2 which indicates that spray nutrients containing N, P, and K have little effect on stem diameter. The interaction between variety and formulation on stem diameter was varied significantly. At 45 DAP, the highest stem diameter was 1.05 cm and the

lowest was 0.75 cm recorded in V2F1 and V1F0, respectively. At 90 DAP, the highest and the lowest stem diameter was 1.2 cm and 0.91 cm also recorded in V2F3 and V1F2, respectively. At 135 DAP, the highest stem diameter was 1.35 cm and the lowest was 1.08 cm recorded in V2F3 and V1F1, respectively. At 180 DAP, the highest stem diameter was 1.5 cm and the lowest was 1.2 cm observed in V2F3 and V1F0, respectively (Table 16).

Table 14. Main effect of variety on number of root and Stem diameter.
Variety V1 V2 CV Number of root at DAP 45 90 135 180 1.08 3.08 5.25 7.58 2.08 14.66 3.91 12.47 5.75 20.99 8.66 19.62 Stem diameter (cm) at DAP 45 90 135 180 0.81 0.97 1.10 1.25 0.96 13.56 1.11 11.23 1.27 8.37 1.43 8.12

Table 15.Main effect of spray formulation on number of root and stem diameter.
Spray formulati on F0 F1 F2 45 1.33 b 1.5 ab 2.33 a Number of root at DAP 90 135 180 3.0 3.50 4.16 5.33 5.50 5.66 8.0 8.33 8.33 Stem diameter (cm) at DAP 45 90 135 180 0.81 0.96 0.83 1.0 1.08 0.97 1.16 1.21 1.13 1.27 1.37 1.3

F3 CV

1.16 b 14.66

3.33 12.47

5.50 20.99

7.83 19.62

0.93 13.56

1.11 11.23

1.25 8.37

1.41 8.12

Legend V1 = Mokara Diheard Red V2 = Mokara Yellow Anne DAP = Days After Planting F0 = Control F1 = Spray Formulation-1 F2 = Spray Formulation-2 F3 = Spray Formulation-3

Table16. Interaction effect of variety to spray formulation on number of root and Stem diameter.

Variety x spray formulatio n V1F0 V1F1 V1F2 V1F3 V2F0 V2F1 V2F2 V2F3 CV

Number of root at DAP 45 90 135 180

Stem diameter (cm) at DAP 45 90 135 180

1.33 b 1.0 b 1.33 b 0.66 b 1.33 b 2.0 b 3.33 a 1.66 b 14.66

3.0 ab 3.33 ab 3.33 ab 2.66 b 3.0 ab 3.66 ab 5.0 a 4.0 ab 12.47

5.66 5.0 5.33 5.0 5. 0 6.0 6.0 6. 0 20.99

8.66 7.0 7.33 7.33 7.33 9.66 9.33 8.33 19.62

0.75 b 0.93 b 0.88 1.0 ab

1.11 b 1.08 b 1.08 b 1.15 b 1.21 ab 1.35 a 1.17 ab 1.35 a 8.37

1.2 b 1.26 b 1.2 b 1.33 ab 1.35 ab 1.48 a 1.4 ab 1.5 a 8.12

ab 0.76 b 0.91 b 0.85 ab 0.88 1.03 ab 1.06

ab ab 1.05 a 1.16 a 0.90 ab 1.01 a 13.56 1.03 ab 1.2 a 11.23

4.3.9 Number of root

The number of root varied from 1.08 - 7.58 and 2.08 - 8.66 during growth stage in Mokara Diheard red and Mokara Yellow Anne, respectively. The average number of root of Mokara Diheard red was 1.08, 3.08, 5.25 and 7.58 at 45, 90,135 and 180 DAP, respectively. In Mokara Yellow Anne average number of root were 2.08, 3.91, 5.75 and 8.66 at 45, 90, 135 and 180 DAP, respectively (Table 14). Though both varieties were treated with same treatments but from table it was observed that degree of root production was higher in Mokara Yellow Anne than Mokara Diheard red. So, varietal effect on number of root was significant. Different spray formulations have significant effects on number of roots. At 45 DAP, maximum average number of roots was 2.33 recorded in F2 and minimum was 1.16 in F3. At 90 DAP, the highest number of roots was 4.16 observed in F2 and the lowest was 3.0 found in control (F0). At 135 DAP, the highest (5.66) number of roots showed in F2 and the lowest (5.33) was found in F0. At 180 DAP, the highest number of roots was 8.33 showed in F1 and F2 and the lowest was 7.83 found in F3 (Table 15). From table it was observed that root development increased rapidly at later growth stage than early growth stage. At 45 DAP, the highest and the lowest number of roots was 3.33 and 0.66 in V2F2 and V1F3, respectively. Similar trend in number of root was observed with increasing cultivation period. At 180 DAP, the highest number of roots was 9.66 recorded in V2F1 and the lowest number of roots was 7 investigated in V1F1 (Table 16). The interaction effect between variety and formulation on root development was varied significantly. The similar results also reported by Nandi (1999).

4.4 Physiological and growth parameters

4.4.1 Chlorophyll content Chlorophyll content in leaf of Mokara Diheard Red and Mokara Yellow Anne was varied significantly. At 45 DAP, average Chlorophyll a, Chlorophyll b and total Chlorophyll content of Mokara Diheard Red were 11.47, 5.25 and 16.72 (mg/g), respectively. At 45 DAP, average Chlorophyll a, Chlorophyll b and total Chlorophyll content of Mokara Yellow Anne were 8.12, 5.20 and 13.32 (mg/g), respectively. The average Chlorophyll a, Chlorophyll b and total Chlorophyll content of Mokara Diheard Red were 13.09, 6.02 and 19.12 (mg/g), respectively at 90 DAP. At 90 DAP, average Chlorophyll a, Chlorophyll b and total Chlorophyll content of Mokara Yellow Anne were 10.94 , 6.65 and 17.9 (mg/g ), respectively (Table 18). The total chlorophyll content of Mokara Diheard Red was relatively higher than Mokara Yellow Anne which indicated varietal effect on chlorophyll content was significant. At 45 DAP, the average highest chlorophyll a and chlorophyll b were 13.21 (mg/g) and 6.01(mg/g) were observed in F3. The average lowest chlorophyll a and chlorophyll b were 5.56 (mg/g) and 3.46 (mg/g) obtained in control (F0) at 45 DAP. At 45 DAP, the highest (19.22 mg/g) and the lowest (9.023 mg/g) total chlorophyll content were also found in F3 and control (F0), respectively. At 90 DAP, the average highest chlorophyll a and chlorophyll b were 15.66 (mg/g) and 8.0 (mg/g) observed in F3 and F1, respectively. At 90 DAP, the average lowest chlorophyll a and chlorophyll b were 7.69 (mg/g) and 4.28 (mg/g), observed in control (F0). At 90 DAP, the total chlorophyll content (22.73 mg/g)

was highest in F3 and the lowest was 11.96 mg/g in control (F0), respectively (Table 17). In all treatments chlorophyll content was lowest in control plants indicated that N, P and K in spray formulation was effective on chlorophyll content. The interaction between variety and spray formulation on total chlorophyll content was varied significantly. At 45 DAP, the highest total chlorophyll content was 25.3 mg/g found in V1F3 and the lowest was 8.68 mg/g recorded in V1F0. At 90 DAP, the highest total chlorophyll content was 26.31 mg/g and the lowest chlorophyll content was 9.88 mg/g found in V1F3 and V1F0, respectively (Table 19). Table 17. Main effect of variety on Chlorophyll content.
Spray formulation F0 F1 F2 F3 CV Chl a 5.56 d 11.30 b 9.11 c 13.21 a 0.36 45 DAP Chl b 3.46 d 5.80 b 5.64 c 6.01 a 0.81 Chlorophyll content (mg/g) Total Chlorophyll 9.02 d 17.11 b 14.75 c 19.22 a 0.44 Chl a 7.69 d 13.04 b 11.68 c 15.66 a 2.45 90 DAP Chl b 4.28 d 8.0 a 6.0 c 7.06 b 0.67 Total Chlorophyll 11.96 d 21.05 b 17.80 c 22.73 a 2.03

Table 18. Main effect of spray formulation on Chlorophyll content.

Variety Chl a 11.47 8.12 0.36 Chlorophyll content (mg/g) 45 DAP Chl b Total Chl. 5.25 16.72 5.20 13.32 0.81 0.44 Chl a 13.09 10.94 2.45 Chl b 6.02 6.65 0.67 90 DAP Total Chl. 19.11 17.9 2.03

V1 V2 CV

Legend

V1 = Mokara Diheard red V2 = Mokara Yellow Anne DAP = Days After Planting Chl. = Chlorophyll content of leaf.
Variety x Chl. a formulation V1F0 5.92 f V1F1 V1F2 V1F3 V2F0 V2F1 V2F2 V2F3 CV 12.07 b 10.58 c 17.31 a 5.19 g 10.54 c 7.65 e 9.10 d 0.36 Spray

F0 = Control F1 = Spray Formulation-1 F2 = Spray Formulation-2 F3 = Spray Formulation-3

Table 19. Interaction effect of variety to spray formulation on Chlorophyll

Chlorophyll content (mg/g) 45 DAP Chl. b Total Chl. 2.76 h 5.63 d 4.63 e 7.99a 4.17 f 5.97 c 6.65 b 4.03 g 0.81 8.68 h 17.7 b 15.21 d 25.3 a 9.36 g 16.51 c 14.30 e 13.13 f 0.44 Chl. a 6.77 e 13.12 c 14.58 b 17.89 a 8.61 d 12.96 c 8.78 d 13.44 c 2.45 90 DAP Chl. b 3.11 h 7.39 c 5.19 g 8.42 b 5.45 f 8.62 a 6.82 d 5.71 e 0.67 Total Chl. 9.88 g 20.52 c 19.79 d 26.31 a 14.05 f 21.58 b 15.82 e 19.15 d 2.03

4.4.2 Crop growth rate (CGR) The crop growth rate (CGR) derived in Mokara Diheard red and Mokara Yellow Anne assessed from 45 to 180 DAP and it was observed that growth rate of orchid was very low. The crop growth rate of Mokara Diheard red was 0.29 (g plant-1/day) and of Mokara Yellow Anne was 0.17 (g plant-1/day) during research period. Mokara Diheard red might be considered superior over Mokara Yellow Anne in respect of growth rate. Varietal effect of crop growth rate was varied significantly (Table 20).

Different spray formulations have significant effect on growth rate. The crop growth rate among different treatments ranged from 0.14-0.29 g plant-1/day. Maximum crop growth rate was 0.29 g plant-1/day found in F2 and minimum crop growth rate was 0.14 g plant-1/day was in control (F0) (Table 21) which indicated that spray nutrients formulation containing N, P and K directly affected on crop growth rate. The interaction between variety and treatments on crop growth rate was varied significantly. Maximum crop growth rate was 0.37 g plant-1/day recorded in V1F2 and minimum crop growth rate was 0.12 g plant -1/day recorded in V2F0 (Table 22). 4.4.3 Relative growth rate (RGR) The relative growth rate (RGR) of Mokara Diheard red and Mokara Yellow Anne assessed from 45 to 180 DAP have been presented in Table 20. The relative growth rate of Mokara Diheard red was 6.88 (mg g-1/day) and of Mokara Yellow Anne was 5.89 (mg g-1/day). Different spray formulations have significant effect on relative growth rate. The relative growth rate among different treatments ranged from 6.04-6.76 mg g-1/day. Maximum relative growth rate was 6.76 mg g-1/day was found in F2 and minimum crop growth rate was 6.04 mg g-1/day recorded in control (Table 21) which indicated that spray nutrients formulation containing N, P and K directly affect on relative growth rate.

The interaction between varieties to spray formulation on relative growth rate was varied significantly. Maximum relative growth rate was 7.14 mg g-1/day found in V2F3 and minimum growth rate was 4.96 mg g-1/day recorded in control V1F0 (Table 22).

Table 20. Main effect of variety on CGR and RGR.

Variety V1 V2 CV CGR (g plant-1/day) 0.29 0.17 17.82 RGR (mg g-1/day) 6.88 5.89 8.50

Table 21. Main effect of spray formulation on CGR and RGR.

Spray formulation F0 F1 F2 F3 CV CGR (g plant-1/day) 0.14 c 0.21 b 0.29 a 0.28 a 13.2 RGR (mg g-1/day) 6.04 d 6.12 c 6.76 a 6.62 b 6.50

Legend V1 = Mokara Diheard red V2 = Mokara Yellow Anne DAP = Days After Planting F0 = Control F1 = Spray Formulation-1 F2 = Spray Formulation-2

F3 = Spray Formulation-3

Table 22. Interaction effect of variety to spray formulation on CGR and RGR.
Variety x spray formulation V1F0 V1F1 V1F2 V1F3 V2F0 V2F1 V2F2 V2F3 CV CGR (g plant-1/day) 0.17 de 0.28 bc 0.37 a 0.35 ab 0.12 e 0.15 de 0.22 cd 0.21 cd 17.82 RGR (mg g-1/day) 4.96 g 6.03 f 6.48 c 6.1 e 7.13 a 6.21 d 7.05 b 7.14 a 5.50

Chapter V SUMMARY AND CONCLUSION

An experiment was conducted in grill house of the Department of Crop Botany, BAU, during the period of 30 November, 2009 to 30 September, 2010 to determine the effect of spray nutrients on morpho-physiological features and growth of orchid. The experiment comprised of two orchid

cultivars and four levels of spray nutrients with control. The experiment was laid out in Completely Randomized Block Design (RCBD) with 8 treatments and each treatment was replicated three times. Data were taken on some selected parameters and statistical analyses were performed by using MSTAT-C program. The results of the experiment were summarized below. All the studied parameters were significantly influenced by spray formulation consists of N, P and K. Morpho-physiological features like plant height, leaf length, leaf wide, leaf area, leaf number, leaf area index, stem diameter, number of root and growth parameters viz. crop growth rate, relative growth rate, chlorophyll content in leaf were affected by spray formulation. The effect of cultivars on those morpho-physiological and growth features also significant. Increasing plant height, leaf wide, leaf area, leaf number, leaf area index, stem diameter, number of root and RGR was higher in Mokara Yellow Anne. CGR, leaf length, total chlorophyll content were higher in Mokara Diheard red. Different spray formulation with different ratio of N, P and K also significantly affected morpho-physiological features and growth parameters. Leaf length was slightly affected by spray formulation. Degrees of increasing leaf length, number of root were higher in spray formulation-1 (F1). Rate of increasing of plant height, crop growth rate, relative growth rate were more in spray formulation-2 (F2). Rate of increasing of leaf width, total leaf area, leaf number, leaf area index, stem diameter and total chlorophyll content were more in spray formulation-3 (F3).

Plant height, stem diameter, total leaf area and CGR were best in V 2F2 among all treatments. Leaf area, leaf area index and RGR were best in V2F3 among all treatments. Leaf length and leaf width were best in V2F1 and V2F3, respectively. Chlorophyll content was superior in V1F3 among all treatments. Based on experimental results it may be concluded that i) the morphological and physiological attributes was significantly affected by spray formulation.
ii)

spray formulation-2 (F2) was the best for growth of orchid.

iii) the ratio of N, P and K at 10:15: 20, 15:20:25, 10:25:25, and 10:25:30 for spray formulation might be the best for growth of orchid.

REFERENCES
Anonymous. 2007. Orchid care techniques. Pp. 9 -11. Arnon, D.I. 1949. Copper enzyme in isolated chloroplast polyphenol oxidase in Beta vulgaris, Plant Physiol. 24:1- 5.

Chakraborti, S. 1986. Rokomary phool chash, Nirmol book agency, 89, Mohatma Gandhi road, Kolkata. Pp. 12 14. Chandra, S. and Das, A. K. 2000. Correlation and interrelation of physiological parameters in rice under rainfed transplanted condition. J. Crop Res. Assam Agric.Univ. 19(2): 252 - 254. Chen, J.T., Chang, C. and Chang, W. C. 2000. Efficient plant regeneration through somatic embryogenesis from callus cultures of Oncidium (Orchidaceae). Plant Sci. 160 (1): 87- 93. Chen, J.T., Chang, C. and Chang, W. C. 2002. Efficient plant regeneration through embryogenesis from callus culture of Oncidium (Orchidaceae). Plant sci. 160 (1): 87- 93. Chowdhury, M. 1975. "Baldah Garden", Dhaka, Bangladesh. pp. 12-15. Fleischer, W.E.1935. The relation between vase life and different culture solution. Gen. Physiology. 18: 573-597. Furukawa, K., Ishiko, E. and Furuya, S., 1989. Culturing of Cymbidium to use Coating Fertilizer. Mukoyama Orchids Ltd., Japan.pp. 27. Ghoshal, K.K. and Das, A.1989. Floral morphology and behavior of some orchids in 18 Plains. Environ. Eco. 7(1): 140-144. Goh, CJ., Sim, A.A. and G. Lim. 1992. Mycorrhizal associations in some tropical orchids. Lindleyana, 7(1):13-17. Hatch, E. D. 1953. Orquideas Subterranean. 15 (1): 2-11.

Gomez, A. K. and Gomez, A. A. 1984. Statistical Procedures for Agricultural Research, 2nd Ed., John Wiley and Sons, Inc., NY. pp. 8-20. Hagakl, Y. and Imamura, J.S. 1987. NPK requirements of vanda miss Joaquin orchid plants. Department of Horticulture. Hawaii Institute of Tropical Agriculture and Human Resources. University of Hawaii. Hilo.pp. 34 36. Hatch, E. D. 1989. Orquideas subtermeas. Orquidea. 15(1): 2-11. Hawkey, A.D. 1978. Encyclopedia of cultivated orchids. In: Commercial

Flowers. 34(2) 65 - 68. Hew, C.S. 1994. Orchid cut flower production in ASEAN Countries. In Orchid Biology Review and Perspectives. J. Arditti John, (ed). Wiley and Son Inc, New York. 6: 363 - 401. Ichihashi, S. 1992. Micropropagation of Phalaenopsis through the culture of lateral buds from young flower stalks. Lindleyana. 7(4): 208 - 215. Kabir, T. 2007. Orchid . 2nd edition, Shilpa Taru prokashani, 27110, Taimahal road, block-c, Muhammadpur, Dhaka. 1207. Pp. 32 34. Ketsa, S. and Kosonmethakul, N. 2004. Home orchid Revised Ed.) New York: Prentice Hall, pp. 32 34. Ketsa, S. and Kosonmethakul, N. 2006. Home orchid growing (Fifth Revised Ed.) New York: Prentice Hall. pp. 33 35.
Komori, K. 2002. Effect of low light intensity in summer on the growth of mericloned Cymbidium young plantlets. Environ and Bio. 40(1): 107- 110.

growing . (Fourth

Laws, N. 1995. Cut flower as Orchid in the World market. Flora Culture International. 5(12): 12 - 15. Lee, N. and Chang Y.C. 2000. Effect of temperature on growth of pseudobulb and inflorescences development of Oncidium 'Gower Ramsey'. J. Chinese. Soc. Hort. Sci. 46(2): 221 - 230. Matsui, N.,Yoneda, K. and Kubota, S. 1999. Effect of N, P and K concentration in culture solution on growth and flowering of Dendrobium sp. Bioresearch Science, Nihon University, Fujisawa, Japan.p. 31. Mohapatra, S. and Saravanan, S. 2006. Effect of different potting media on growth and flowering of Dendrobium orchid. Department of Horticulture, Allahabad Agricultural Institute, Deemed University, Allahabad, India. Orissa-Journal-of-Horticulture. 34(2). 65 - 68. Morel, G.1964. Tissue culture: A new means of clonal propagation of orchids. Amer. Orchid Soc. Bull., 33: 473-478. Nandi, M.K. 1999. Effect of different levels of nitrogen, phosphorus and potassium on plant growth and seed yield of tomato (Lycopersicon esculentum) M.S. Thesis, Dept. of Hort., Bangladesh Agric. Univ. Mymensingh. Pathania, N. S., Sehgal , O.P., P.Debojit, B.S. Dilta and Paul, D. 1998. Studies on micropropagation in Dendrobium cv. Sonia. J. Orchid Soc. India, 12 (1-2): 35 38.

Patil, P.V. 2001. Orchids of India in Commercial Flowers 2nd Ed. Calcutta, India. Pp 193. Poole, H. A. and Sheehan T. J.1970. Effects of levels of phosphorus and potassium on growth, composition and incidence of leaf-tip die-back in cattleya orchids. University of Florida Gainesville. Prasad, A., Arya, S., and Katiyar, J.N. 1997. Studies on genetic component and genotype correlation in orchids. Orchid Soc. India. 11(4-5): 82-83. Rao, A.N. 1977. Orchids. In Plant Cell Tissue and Organ Culture. (Ed.) Chandha and Singh, NewYork. p 126. Razzak, A. 2009. Personal communication, General Manager of BRAC Agricultural Research and Development Center, Joydebpur, Gazipur, Bangladesh. Royer, S.J (ed.). 2003. An Orchid Handbook, Michiana Orchid Society, p. 34 Russel,D.F. 1986. MSTAT-C. MSTAT Director. Crop and Soil Science. Department of Michigan State University, USA. Saiprasad, G.V.S., Raghuveer, P. and Polisetty. R. 2002. Effect of various nutrient media on the production of protocomb Like Body (PLB) and multiple shoots in Orchids.J. Ornamental Hort. 5(2): 72-73. Sell, Y. and Hermann , S. 1998. Some parameters for the characterization of the floral axes of Plalaenopsis (Orchidaceae) with a view to a study of their morphological potential. Indian Agricultural Research Institute, New Delhi. P. 75.

Sharma, S.K. and Haloi, B. 2001. Characterization of crop growth variable in some selected rice cultivars of Assam. Indian J. Plant Physiol. 6: 166 -171. Singh, B. and Voleti, S. R. 1995. Research and achievements. Division of Floriculture and Landscaping. Indian Agricultural Research Institute, New Delhi. p. 15. Singh, F. 1998.Orchids Ornamental Horticulture in India. P. 37. Singh, P. and Roy, S. K., 2004, Regeneration of an indigenous orchid of Vanda Plant Tissue Cult. 14(1): 55-61. Singh, R. and Kohli, A. U.K. 1999. Effect of NPK regimes on growth and development of orchid .J. Hill Res. 12(1): 63 - 66. Sobhana, A. and P. K. Rajeevan. 1993. Performance of certain epiptytic species in central Kerala. J. Orchid Soc. India. 7(1-2): 31- 35. Stenberg, M.L. and M. E. Kane. 1998. In vitro seed germination and greenhouse cultivation of Encyclia boothiana var. erythronioides, an endangered Florida orchid. Lindleyana. 13 (2): 101- 112. Vaz, A. P. A. and Kerbauy, G.B. 2004. Senescence and post-harvest physiology of cut flowers. Horticultural Review, pp. 59 - 65. Wang, Y . T. and Konow, E. A. 2002. Fertilizer source and medium

composition affect vegetative growth and mineral nutrition of a hybrid moth orchid. J. Ame. Soc. Hort. Sci. 127(3): 442- 447.

Wang, Y. T. and Konow, E. A .2004. Impact of a high phosphorus fertilizer and timing of termination of fertilization on flowering of a hybrid moth orchid. J. Ame. Soc. Hort. Sci. 130(2): 52- 60. Webb, M. and Webb, A. 1993. Dendrobium speciosum complex. Amer.

Orchid Soc. Bull., 52 (5): 115-122. Wong, L.S. and Lee, S.M. 2000. Effects of nutrients and potting media on cultivation of Dendrobium sp. Singapore-Journal-of-Primary-Industries. 28: 1. Yin-Tung, Wang. 1995. Effects of six fertilizers on vegetative growth and flowering of phalaenopsis orchids. J. Ame. Soc. Hort. Sci. 54 (6): 119132.

Appendix 1. Analysis of variance (Mean square) of plant height, leaf area and leaf number at different growth stage of orchid
Analysis of variance Factor A (Variety) 1 Factor B (Spray formulation ) 158.1 ** 147.01* 133.95 ** 135.37** 237.57* 281.12** 165.27 * 93.18 * 35.04** 51.04 ** 70.04 ** 80.66 ** Degree of freedom Plant height 45 DAP 90 DAP 135 DAP 180 DAP 45 DAP 90 DAP Leaf area 135 DAP 180 DAP 45 DAP Leaf number 90 DAP 135 DAP 180 DAP

3.44 *

3.66 ns

11.2 *

27.07 **

1.22 ns

49.12 ns

152.45 *

207.08 *

2.486*

1.81 ns

1.93 *

0.72 ns

Interaction (AxB)

5.96 **

6.93 *

3.43 *

3.73 *

37.02 **

64.46 *

49.76 **

50.56 **

9.37*

7.81 *

4.37 **

3.2 *

Error 16 14.24 13.12 12.38

12.98

26.67

39.21

49.19

57.9

2.83

2.66

3.0

2.7

Legend * Indicates significant at 5 % level * * Indicates significant at 1 % level

Ns Indicates

non -significant

58

Appendix 2. Analysis of variance (Mean square) of total leaf area, leaf area index and leaf length at different growth stage of orchid
Analysis of variance Factor A (Variety) Degree of freedom 1 Factor B (Spray formulation) 3 Interaction (AxB) 3 21629.82 ** 59844.41 ** 52437.71 * 33338.43 * 0.017** 0.043* 0.051 * 0.02 ** 2.97 * 5.43 * 5.67 * 7.95 * 45 DAP 10388.35 ** Total leaf area 90 DAP 135 DAP 33383.50 * 180 DAP 224479.37 ** 45 DAP Leaf area index 90 DAP 135 DAP 180 DAP 45 DAP Leaf length 90 DAP 135 DAP 180 DAP

117214.88 *

0.007 **

0.038 *

0.082*

0.18 **

0.21 ns

1.4 ns

1.3 ns

1.0 ns

3690.55 *

24353.24 **

48532.35 *

59745.47 *

0.003 ns

0.016 *

0.03 *

0.04 **

1.13 ns

1.04 ns

1.32 ns

1.3 ns

Error 16 4001.67 12392.89 20144.04 30407.56 0.003 0.01 0.016 0.026 3.03 3.09 3.18 3.06

Legend * Indicates significant at 5 % level * * Indicates significant at 1 % level Ns Indicates non -significant

59

Appendix 3. Analysis of variance (Mean square) of number of root, stem diameter and leaf width at different growth stage of orchid

Analysis of variance Factor A (Variety)

Degree of freedom 1

Number of root 45 DAP 6.0 ** 90 DAP .16 ns 135 DAP 1.5 ns 180 DAP 7.04 * 45 DAP 0.13 **

Stem diameter 90 DAP 0.12 ** 135 DAP 0.16 ** 180 DAP 0.20 ** 45 DAP 0.44 **

Leaf width 90 DAP 0.28 ** 135 DAP 0.15 ns 180 DAP 0.06 ns

Factor B (Spray formulation)

1.61 *

1.44 ns

0.11 ns

0.375 ns

0.033 ns

0.027 ns

0.017 ns

0.026 ns

0.009 ns

0.08 *

0.32 **

0.43 **

Interaction (AxB) Error

1.0 ns

0.94 ns

0.94 ns

4.59 ns

0.001 *

0.001 **

0.01 **

0.001 **

0.1 *

0.17 **

0.09 **

0.07 **

16

0.50

1.29

1.33

2.54

0.014

0.014

0.01

0.012

0.02

0.03

0.03

0.04

Legend * Indicates significant at 5 % level * * Indicates significant at 1 % level Ns Indicates non -significant

60

Appendix 4. Analysis of variance (Mean square) of plant height, leaf area and leaf number at different growth stage of orchid

Analysis of variance Factor A (Variety) Factor B (Spray formulation )

Degree of freedom 1

Chlorophyll content (45DAP) Total Chl(a) Chl(b) chlorophyll 67.19 ** 0.014** 69.21 **

Chlorophyll content(90DAP) Total Chl(a) Chl(b) chlorophyll 27.57** 2.319** 13.061**

CGR

RGR

0.084**

5.9 **

64.73 **

8.43 **

116.06 **

66.37 **

15.29 **

134.98 **

0.028**

0.76 **

Interaction (AxB) Error

17.005 ** 0.001

10.92 ** 0.002

52.32 ** 0.004

19.2 ** 0.087

7.71 ** 0.002

38.34 ** 0.139

0.003 ns 0.002

1.1 ** 0.001

16

Legend * Indicates significant at 5 % level * * Indicates significant at 1 % level Ns Indicates non -significant

61