Volume 3, Issue 1 of Tropical Plant Research

ISSN (E): 2349 1183
ISSN (P): 2349 9265

3(1): 0109, 2016
Research article
Tree species diversity in tropical forests of Barak valley

in Assam, India
Nepolion Borah1*, Debajit Rabha2 and Florida Devi Athokpam2
1
School of Environmental Sciences, Jawaharlal Nehru University, New Delhi, India

Department of Ecology and Environmental Science, Assam University Silchar, Assam, India
*Corresponding Author: nepolionborah@gmail.com
[Accepted: 01 February 2016]
2
Abstract: To enumerate the tree species diversity of tropical forests, 89 belt-transects was laid in
different reserve forests and private forests of the Barak Valley, Assam, Northeast India. A total of
222 tree species were recorded from 152 genera and 65 families. Euphorbiaceae was the most
species rich family with 23 species. Out of 65 families, 30 families were recorded with only one
species while 10 families were recorded with two species. Artocarpus chama was the most
abundant and frequently occurred species. Podocarpus nerifolia was the only gymnosperm tree
recorded in this study while Caryota urena and Pleomele spicata were the monocot tree species.
Five threatened species were recorded from the Valley.
Keywords: Belt-transect - Threatened species - Frequency - Barak valley.
[Cite as: Borah N, Rabha D & Athokpam FD (2016) Tree species diversity in tropical forests of Barak valley in
Assam, India. Tropical Plant Research 3(1): 19]
INTRODUCTION
Assam is part of Indo-Burma the biodiversity hotspot regions, which is situated in the north-eastern corner of
Indian subcontinent and considered one of the richest occurrences of angiosperm plants. The Southern part of
Assam, which is popularly known as Barak Valley, consists of three districts namely Cachar, Karimganj and
Hailakandi. The vegetation of this region is mostly represented by tropical moist evergreen and tropical moist
semi-evergreen forest types (Champion & Seth 1968). The forests of this region are relatively unexplored
harbouring rich plant diversity. The vegetation of this region has been free from anthropogenic disturbances
over centuries. But due to rapid population growth and development activities, some parts of the forests are
under huge anthropogenic pressure such as over exploitation of species for timber, fuel-wood, fodder, bamboo
cutting, settlement etc. (Borah & Garkoti 2011). The floristic composition is one of the major anatomical
characters of the forest community (Dansereau 1960). So it is very important to know the species composition
and its distribution of these forests to take proper management strategies.
A good number of scientific literatures are available on angiosperm flora of Assam (Kanjilal et al. 1934
1940, Hooker 18721887, Rao & Verma 1969, 1976, Choudhury 1982, Dam & Dam 1984). For a modern
floristic assessment, it is important to know the tree wealth of a forest along with their ecological amplitude as
they are the backbone of any forest and provides the microclimate suitable for the survival of other small plants
as well as animals (Bajpai et al. 2015, Dular 2015).When we see the tree diversity exclusively, very few studies
are available from the state (Sarkar & Devi 2014, Rabha 2014).The literature dealing with the tree diversity and
their ecological standings is either very old (Choudhury 1982, Dam & Dam 1984) or focused to a specific area
(Borah & Garkoti 2011, Borah 2012, Borah et al. 2014). Thus, the present study was performed to enumerate
the tree species composition and their ecological status from the tropical forests of this region.
MATERIAL AND METHODS
Geographically Barak Valley of Northeast India (Fig. 1) is surrounded by North Cachar Hills and Jaintia
Hills in the north, in east by Manipur, in the south by Mizoram and in the west by Tripura and Sylhet district of
Bangladesh.
www.tropicalplantresearch.com
Received: 13 October 2015
1
Published online: 29 February 2016
Borah et al. (2016) 3(1): 0109

.
Figure 1. Map of Barak valley, Assam, India.
The soil of the region is sandy clay loam and sandy loam in texture and slightly acidic in nature (pH ranges
from 5.35 to 6.1) with an average bulk density of 1.08 gcm-3 and water holding capacity of 38.75% (Athokpam
et al. 2013). The area has a tropical monsoon climate with high annual precipitation and high temperature.
Climate during AprilOctober is characterized by rainy season with an average rainfall 2330.50 mm. The region
is characterized by moderate temperature with monthly average temperature ranging from 11.932.7 oC (Borah
2012). Climatically, the year may be divided into four seasons. December to February is the winter season,
followed by spring or early summer from March to April/May, then June to September is the South West
Monsoon rainy season or late summer, and October and November constitute the post monsoon or autumn
season (Athokpam & Garkoti 2013).
Different reserve forest of Barak Valley are Innerline Reserve Forest, Barak Reserve Forest, Borail Reserve
Forest, Sonai Reserve Forest, Upper Jiri Reserve Forest, Katakhal Reserve Forest, Longai Reserve Forest,
Badshahi-tilla Reserve Forest, Duhalia Reserve Forest, Patharia Reserve Forest, Tilbhoom Reserve Forest and
Singla Reserve Forest.
Present study was carried out during the years 2010 to 2013 by laying 89 belt-transects of 10 m 500 m
sized in different reserve forests and private forests of Barak Valley. Out of 89 transects 35 were delimited in
Cachar, 29 in Karimganj and 15 in Hailakandi districts. The belt transects were laid in such way that it covers
different microclimates of the studied forest so that different types of vegetation comes within transects. After
lying transect, all the plant species of >10 cm GBH trees sampled and some specimens of each species were
brought to laboratory to prepare herbarium following Jain & Rao (1977). The species were identified with the
help of Flora of Assam (Kanjilal et al. 19341940) and the herbarium of Botanical Survey of India, Shillong.
Borah et al. (2016) 3(1): 0109

.
Species nomenclatures were followed as per Assams Flora (present status of vascular plants) (Chowdhury et
al. 2005). After identifying all the species, physiognomy type, growth form and IUCN status were studied by
available literature. The frequency and abundance were estimated as follows-
RESULTS AND DISCUSSION

Table 1. Number of genus (GN) and species (SN) in different families recorded in Barak valley, Assam, India.
S. No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
Family Name
Euphorbiaceae
Lauraceae
Moraceae
Verbenaceae
Mimosaceae
Rubiaceae
Caesalpiniaceae
Meliaceae
Myrsinaceae
Myrtaceae
Sapindaceae
Anacardiaceae
Rutaceae
Annonaceae
Clusiaceae
Dipterocarpaceae
Magnoliaceae
Papilionaceae
Sterculiaceae
Symplocaceae
Bignoniaceae
Fagaceae
Myristicaceae
Sapotaceae
Theaceae
Apocynaceae
Bombacaceae
Combretaceae
Dilleniaceae
Elaeocarpaceae
Flacourtiaceae
Malvaceae
Memecylaceae
GN
15
7
4
5
4
9
5
6
3
3
5
4
5
4
2
3
2
4
3
1
3
2
2
3
3
2
1
1
1
1
2
2
1
SN
23
17
14
10
9
9
8
8
7
7
7
5
5
4
4
4
4
4
4
4
3
3
3
3
3
2
2
2
2
2
2
2
2
S. No.
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
Family Name
Simaroubaceae
Urticaceae
Agavaceae
Alangiaceae
Araliaceae
Arecaceae
Bixaceae
Boraginaceae
Bromeliaceae
Burseraceae
Cannabaceae
Capparaceae
Datiscaceae
Ebenaceae
Ehretiaceae
Elaegnaceae
Fagaceae
Juglandaceae
Leeaceae
Lythraceae
Moringaceae
Oxalidaceae
Podocarpaceae
Rhamnaceae
Rhizophoraceae
Sabiaceae
Saurauiaceae
Sonneratiaceae
Styraceae
Thymelaeaceae
Tiliaceae
Ulmaceae
GN
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
SN
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
A total of 222 tree species were recorded from present study belonging to 152 genera and 65 families. Out of
65 families, Euphorbiaceae was the most species rich family (15 genus and 23 species) followed by Lauraceae
(7 genus and 17 species), Moraceae (4 genus and 14 species) etc. (Table 1). Among them 30 families contained
only one species while 10 families contained 2 species. Out of 222 species, 146 species were evergreen tree
while 76 species were deciduous tree species. Among the recorded species, 18% were large, 43% were medium
and 39% were small size trees. Artocarpus chama was the most abundant and frequently occurred species
(Table 2). In the present study five species were recorded in the IUCN Red List of Threatened Species. Out of 5
threatened species Dipterocarpus turbinatus was critically endangered, and 2 species were vulnerable namely
Saraca asoca and Aquilaria malaccensis, 1 species was Lower Risk/least concern namely Mangifera sylvatica
and other 1 species was data deficient namely, Hydnocarpus kurzii. Podocarpus nerifolia was the only
Borah et al. (2016) 3(1): 0109

.
gymnosperm tree species recorded in this study while Caryota urena and Pleomele spicata were the monocot
tree species. Some of these species were recorded only in few numbers. A good number of endemic, primitive
angiosperm and threatened species were not recorded in this survey which were recorded from this region in
earlier works of Hooker (19721887), Kanjilal (19341940), Chowdhury et al. (2005) etc. In earlier surveys
some species like Canarium bengalensis, Quercus semiserrata, Garcinia keeniana, Embelia parviflora,
Calliandra umbrosa, Magnolia pterocarpa, Caesalpinia digyna, Ixonanthes khasiana, Acranthera tomentosa
etc. were recorded from this region but in present study these were not recorded. This may be due to that the
population of these species become very less or distribution of these species restricted to only some particular
pockets or these species may be loss from this region. The main cause behind this is destruction and degradation
of forest area by rapid urbanization, human settlement, industrialization (mainly tea industry and paper mills),
shifting cultivation, rubber plantation, reckless and ruthless exploitation of plants of potential economic
importance, fuel wood collection, raising of artificial forests by monoculture of some important species such as
Tectona grandis etc. Socioeconomic condition might be responsible for enhanced utilization of the forest
resources and this may eventually lead to a species-poor state (Murali et al. 2014). It is very important to take
proper management strategies for those less abundant and less frequently occurred species, otherwise these
species will also lost from this region in near future.
Table 2. Physiognomic type (PhT), growth form (GrF), abundance (Ab) and frequency in % (Fr) of encountered species in
Barak valley, Assam, India. (E-evergreen, D-deciduous, L-large tree, M-medium sized tree, S-small tree)
S.No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
Species Name
Acacia auriculiformis A. Cunn. ex Benth.
Acacia sinuata (Lour.) Merr.
Actinodaphne angustifolia Nees
Actinodaphne obovata (Nees) Bl.
Adenanthera pavonina L.
Aegle marmelos (L.) Corr.
Ailanthus integrifolia Lam.
Alangium chinensis (Lour.) Rehder
Albizia chinensis (Osbeck) Merr.
Albizia lebbeck (L.) Benth.
Albizia lucidior (Steud.) Nielson ex Hara
Albizia odoratissima (L. f.) Benth.
Albizia procera (Roxb.) Benth.
Allophylus triphyllus (Burm. f.) Merr.
Alseodaphane owdenii Parker
Alseodaphne andersonii (King ex Hook. f.) Kostel.
Alstonia scholaris (L.) R.Br.
Amoora hiernii Visw. & Ramech.
Ananas sp.
Annona reticulata L.
Anthocephalus chinensis (Lam.) A. Rich. ex Walp.
Aporusaaurea Hook. f.
Aporusa octandra (Buch.-Ham.ex D.Don) Vick.
Aquilaria malaccensis Lam.
Ardisia calorata Roxb.
Artocarpus chamaBuch.-Ham.
Artocarpus heterophyllus Lam.
Artocarpus lacuchaBuch.-Ham.
Averrhoa carambola L.
Baccaurea ramiflora Lour.
Bauhinia purpurea L.
Bauhinia variegata L.
Bischofia javanica Bl.
Bixa orellana L.
Bombax ceiba L.
Bombax insigne Wall.
Bridelia monoica (Lour.) Merr.
Family
Mimosaceae
Mimosaceae
Lauraceae
Lauraceae
Mimosaceae
Rutaceae
Simaroubaceae
Alangiaceae
Mimosaceae
Mimosaceae
Mimosaceae
Mimosaceae
Mimosaceae
Sapindaceae
Lauraceae
Lauraceae
Apocynaceae
Meliaceae
Bromeliaceae
Annonaceae
Rubiaceae
Euphorbiaceae
Euphorbiaceae
Thymelaeaceae
Myrsinaceae
Moraceae
Moraceae
Moraceae
Oxalidaceae
Euphorbiaceae
Caesalpiniaceae
Caesalpiniaceae
Euphorbiaceae
Bixaceae
Bombacaceae
Bombacaceae
Euphorbiaceae
PhT
E
D
E
E
D
D
D
E
D
D
E
D
D
E
E
E
E
E
E
E
E
E
E
D
D
D
E
D
E
E
D
D
E
D
D
D
D
GrF
M
M
M
S
S
M
M
S
M
M
L
M
M
M
M
L
L
L
S
S
L
S
S
M
S
L
M
L
M
S
M
M
M
M
L
L
S
Ab
1.50
2.50
18.00
18.50
1.00
3.00
63.00
1.50
67.00
57.00
19.00
8.00
44.50
11.00
43.50
10.50
46.50
3.00
4.00
2.00
14.00
21.00
21.00
7.50
2.50
396.0
3.00
141.0
1.50
79.50
12.00
76.00
4.50
5.00
67.50
12.00
32.00
Fr
2.25
2.25
12.36
13.48
2.25
3.37
30.34
1.12
37.08
29.21
15.73
6.74
26.97
8.99
31.46
8.99
43.82
3.37
1.12
4.49
17.98
16.85
12.36
11.24
2.25
97.75
3.37
74.16
3.37
41.57
2.25
29.21
5.62
3.37
40.45
12.36
14.61
4
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
Bridelia Montana (Roxb.) Willd.

Bridelia vomentisa Bl.
Butea monosperma (Lam.) Taub.
Callicarpa arborea Roxb.
Callistemon citrinus (Curtis.) Stapf
Camellia sinensis (L.) O. Cuntz
Carallia branchiata (Lour.) Merr.
Caryota urena L.
Cassia fistula L.
Cassia javanica L.
Cassia siamea Lam.
Castanopsis indica (Roxb.) DC.
Castanopsis purpurella (Miq.) Balak.
Cedrela microcarpa C. DC.
Celtis australis L.
Chukrasia tabularis A. Juss.
Cinnamomum cacharensis Parker
Cinnamomumglaucescens (Nees) Hand.-Mazz.
Cinnamomum tamala (Buch.-Ham.) Nees & Eberm.
Cordia dichotoma Forst. f.
Crateva religiosa G. Forst.
Croton joufra Roxb.
Cryptocarpa sp.
Crysophyllum roxburghii G. Don
Cynometra polyandra Roxb.
Derris rubusta (Roxb. ex DC.) Benth.
Desmos longiflorus (Roxb.) Safford.
Dillenia indica L.
Dillenia pentagyna Roxb.
Diospyros toposia Buch.-Ham.
Dipterocarpus turbinatus Gaertn.
Duabanga grandiflora (Roxb. ex DC.) Walp.
Dysoxylum alliaria (Buch.-Ham.) Balak.
Dysoxylum binectariferum (Roxb.) Hook. f.
Dysoxylum gobara (Buch.-Ham.) Merr.
Elaeagnus sp.
Elaeocarpus floribundus Bl.
Elaeocarpus sphaericus (Gaertn.) K. Schum.
Embelia nutans Wall.
Embelia ribes Burm. f.
Embelia tsjeriam-cottam DC.
Endospermum antiquorum L.
Engelhardtia spicata Lech. ex Bl.
Erythrina variegataL.
Eugenia grandis Wight
Euphoria longan (Lour.) Steud.
Eurya acuminata DC.
Evodia meliaefolia Benth.
Excoecaria oppositifolia Griff.
Ficus auriculataLour.
Ficus benghalensis L.
Ficus heterophylla L. f. var. repens Willd.
Ficus hirta Vahl
Ficus hispida Vahl
Ficus lepidosa Wall.
Ficus racemosa L.
Ficus religiosa L.
Ficus semicordata Buch.-Ham. ex J. E. Sm
Flacourtia indica (Burm. f.) Merr.
Borah et al. (2016) 3(1): 0109

.
Euphorbiaceae
D
S
9.00
6.74
Euphorbiaceae
D
S
0.50
1.12
Papilionaceae
E
S
1.00
2.25
Verbenaceae
E
S 91.00 42.70
Myrtaceae
E
S
0.50
1.12
Theaceae
E
S
2.50
5.62
Rhizophoraceae
E
M 76.00 44.94
Arecaceae
E
M
9.50 11.24
Caesalpiniaceae
D
M
8.50
8.99
Caesalpiniaceae
D
M
1.50
2.25
Caesalpiniaceae
D
M 12.50
5.62
Fagaceae
E
M 33.00 19.10
Fagaceae
E
M 140.5 66.29
Meliaceae
D
M
3.50
4.49
Ulmaceae
D
M
9.50 11.24
Meliaceae
D
L
8.50
4.49
Lauraceae
E
S 18.00
8.99
Lauraceae
E
L 31.50 26.97
Lauraceae
E
S
4.00
7.87
Boraginaceae
E
M 46.00 29.21
Capparaceae
E
S
2.00
3.37
Euphorbiaceae
D
M 44.00 29.21
Lauraceae
E
M
9.00
5.62
Sapotaceae
E
L 24.00
8.99
Caesalpiniaceae
E
L 238.0 50.56
Papilionaceae
D
M
1.50
2.25
Annonaceae
E
S 25.00 19.10
Dilleniaceae
E
L 48.00 44.94
Dilleniaceae
E
L 14.00 19.10
Ebenaceae
E
L 28.50 20.22
Dipterocarpaceae
E
L 158.0 15.73
Sonneratiaceae
D
L 111.5 51.69
Meliaceae
E
M 16.50
8.99
Meliaceae
E
M 172.0 59.55
Meliaceae
E
M 133.5 38.20
Elaegnaceae
E
S 74.50 38.20
Elaeocarpaceae
E
M 32.50 23.60
Elaeocarpaceae
E
M
7.50
5.62
Myrsinaceae
E
S
8.00
6.74
Myrsinaceae
D
S 20.50 12.36
Myrsinaceae
E
S 17.00 11.24
Euphorbiaceae
E
M 13.50 15.73
Juglandaceae
E
S
1
1.12
Papilionaceae
D
M 23.50 16.85
Myrtaceae
E
M
7.00
4.49
Sapindaceae
E
S
3.00
4.49
Theaceae
E
S 104.0 50.56
Rutaceae
D
M 10.00 10.11
Euphorbiaceae
E
S 15.50 10.11
Moraceae
E
S 18.00 15.73
Moraceae
E
L
2.50
5.62
Moraceae
E
S
7.50
3.37
Moraceae
E
S 28.00 19.10
Moraceae
E
S 72.00 38.20
Moraceae
E
S 116.5 46.07
Moraceae
D
M 90.50 55.06
Moraceae
D
L 32.50 37.08
Moraceae
E
S
1.50
1.12
Flacourtiaceae
D
S
3.00
4.49
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
126
127
128
129
130
131
132
133
134
135
136
137
138
139
140
141
142
143
144
145
146
147
148
149
150
151
152
153
154
155
Garciniacowa Roxb. ex DC.

Garcinia xanthochymus Hook. f.
Garuga pinnata Roxb.
Glochidion khasicum Hook. f.
Glochidion lanceolarium (Roxb.) Voigt
Gmelina arborea Roxb.
Grewia nervosa (Lour.) Panigr.
Glycosmis arborea (Roxb.) Corr.
Gynocardia odorata R. Br.
Haldinia cordifolia (Roxb.) Ridsd.
Heteropanax fragrans Seem.
Hibiscus macrophyllus Roxb.
Holarrhena pubescens (Buch.-Ham.) Wall. ex G. Don
Hydnocarpus kurzii (King) Warb.
Ixora sp.
Knema augustifolia (Roxb.) Warb.
Knema linifolia Roxb.
Kydia calycina Roxb.
Lagerstroemia reginae Roxb.
Lannea coromondelica (Houtt.) Merr.
Leea indica (Burm. f.) Merr.
Litsea cubeba (Lour.) Pers.
Litsea laeta Nees Hook. f.
Litsea monopetala (Roxb.) Pers.
Litsea salicifolia (Roxb. ex Nees) Hook. f.
Litsea sp.
Macaranga denticulata (Bl.) Muell. Arg
Macaranga sp.
Macaranga peltata Roxb.
Maesa indica (Roxb.) A. DC.
Maesa montana A. DC.
Maesa paniculata A. DC.
Mallotus ferrugineus (Roxb.) Muell. Arg
Mallotus philippinensis (Lamk) Muell. Arg
Mallotus roxburghianus Muell. Arg
Mangifera indica L.
Mangifera sylvatica Roxb.
Manihot esculenta Crantz
Melia azedarach L.
Meliosma pinnata (Roxb.) Maxim
Memecylon celastrinum Kurz
Memecylon umbellatum Burm. f.
Mesua ferrea L.
Mesua floribuanda (Wall.) Kostel.
Meyna spinosa Roxb. ex Link
Michelia baillonii (Pierre) Finet & Gagnep.
Michelia champaca L.
Michelia glabra Parment
Micromelum minutum (Forst. f.) Wight & Arn.
Miliusa globosa (DC.) Panigr. & Mishra
Mitragyna rotundifolia (Roxb.) O. Kuntze
Moringa oleifera Lam.
Morus macroura Miq.
Myristica sp.
Neocinnamomum caudatum (Nees) Merr.
Lepisanthes sp.
Oreocnide integrifolia (Gaud.) Miq.
Oroxylum indicum (L.) Vent.
Ostodes paniculata Bl.
Clusiaceae
Clusiaceae
Burseraceae
Euphorbiaceae
Euphorbiaceae
Verbenaceae
Tiliaceae
Rutaceae
Flacourtiaceae
Rubiaceae
Araliaceae
Malvaceae
Apocynaceae
Flacourtiaceae
Rubiaceae
Myristicaceae
Myristicaceae
Malvaceae
Lythraceae
Anacardiaceae
Leeaceae
Lauraceae
Lauraceae
Lauraceae
Lauraceae
Lauraceae
Euphorbiaceae
Euphorbiaceae
Euphorbiaceae
Myrsinaceae
Myrsinaceae
Myrsinaceae
Euphorbiaceae
Euphorbiaceae
Euphorbiaceae
Anacardiaceae
Anacardiaceae
Euphorbiaceae
Meliaceae
Sabiaceae
Memecylaceae
Memecylaceae
Clusiaceae
Clusiaceae
Rubiaceae
Magnoliaceae
Magnoliaceae
Magnoliaceae
Rutaceae
Annonaceae
Rubiaceae
Moringaceae
Moraceae
Myristicaceae
Lauraceae
Sapindaceae
Urticaceae
Bignoniaceae
Euphorbiaceae
Borah et al. (2016) 3(1): 0109

.
E
M 11.50 13.48
E
M 67.00 33.71
D
M 80.50 49.44
E
S 17.50
7.87
E
S 31.50 24.72
D
L 83.00 50.56
D
S 88.50 23.60
D
S
8.50
7.87
E
L 44.00 16.85
D
L
3.00
3.37
E
S
7.50
8.99
D
M 27.50 20.22
D
S 67.00 21.35
E
M 73.00 21.35
E
S
2.00
1.12
E
M 34.00 12.36
E
M 41.00 20.22
D
S 12.00 15.73
D
M 67.00 37.08
D
S 23.50 22.47
E
S 40.00 23.60
E
S
0.50
1.12
E
M 16.00
6.74
E
M 124.0 57.30
E
S 87.50 39.33
E
S 46.00 30.34
E
S 65.00 22.47
E
S
9.00
1.12
E
S 129.5 51.69
D
S
7.00
7.87
D
S
4.00
4.49
D
S
6.50
4.49
E
S 243.0 60.67
E
S
3.00
2.25
E
S 96.50 42.70
E
L 24.00 31.46
E
L
6.50
8.99
E
S
5.50
5.62
D
M
4.00
5.62
E
M
7.50
6.74
E
S 15.50 11.24
E
S
8.50
8.99
E
L 168.5 49.44
E
L 77.00 35.96
D
S
2.00
2.25
E
M
7.00
4.49
E
M 53.00 32.58
E
M
7.00
7.87
E
S 16.00
8.99
D
S 17.00
5.62
D
S 175.0 55.06
D
S
2.50
3.37
E
S 15.00 11.24
E
S
1.00
1.12
E
M
7.00
8.99
E
S
5.00
4.49
E
S
9.00
8.99
D
S 108.0 62.92
E
M 15.50
8.99
156
157
158
159
160
161
162
163
164
165
166
167
168
169
170
171
172
173
174
175
176
177
178
179
180
181
182
183
184
185
186
187
188
189
190
191
192
193
194
195
196
197
198
199
200
201
202
203
204
205
206
207
208
209
210
211
212
Pajanelia longifolia (Willd.) Schum.

Palaquium polyanthum Benth.
Parkia timoriana (DC.) Merr.
Persea bombycina (King ex Hook. f.) Kostel
Phoebe attenuate Nees
Phoebe goalparensis Hutchinson
Phyllanthus emblica L.
Picrasma javanica Bl.
Pithecelobium heterophyllum (Roxb.) Benth.
Pleome lespicata (Roxb.) N.E. Brown
Podocarpus nerifolia D. Don
Polyalthia sp.
Premna benghalensis Cl.
Premna milleflora Cl.
Psychotria monticolaKurz
Pterospermum acerifolium (L.) Willd.
Pterospermum lanceaefolium Roxb.
Pterygota alata (Roxb.) R. Br.
Quercus griffithii Hook. f. & Th.
Randia racemosa (Cav.) f. Vill.
Samanea saman (Jack.) Merr.
Sapindus attenuatus wall.
Sapindus mukorossi Gaertn.
Sapindus sp.
Sapium baccatum Roxb.
Saprosma ternatum Hook. f.
Saraca asoca (Roxb.) de Wilde.
Saurauia roxburghii Wall.
Schima wallichii (DC.) Kuntze
Schleichera trijuga Willd.
Semecarpus anacardium L.
Shorea robusta Gaertn.
Spondias pinnata (L. f.) Kurz
Sterculia urens Roxb.
Sterculia villosa Roxb.
Stereospermum personatum (Hassk.) Chatterjee
Streblus asper Lour.
Styrax serrulatum Roxb.
Symplocos cochinchinensis (Lour.) Moore ssp.
Cochinchinensis Lour.
Symplocos khasiana (Cl.) Brand.
Symplocos sp.
Symplocos cochinchinensis (Lour.) Mooressp. Laurina
(Retz.) Nooteboom
Syzygium cumini (L.) Skeels
Syzygium jambos (L.) Alston
Syzygium kurzii (Duthie) Balak.
Syzygium oblatum (Roxb.) Wall. ex A.M. & Cowan
Syzygium praetermissum (Gage) Balak.
Syzygium syzygioides (Miq.) Merr.
Magnolia hookeri (Cubitt & Smith) Raju&Nayar
Tamarindus indica L.
Tectona grandis L. f.
Terminalia bellirica (Gartn.) Roxb.
Terminalia chebula Retz.
Tetramelesnudiflora R. Br.
Toona ciliata M. Roem.
Trema orientalis (L.) Bl.
Trewia nudiflora L.
Borah et al. (2016) 3(1): 0109

.
Bignoniaceae
E
S 11.00 13.48
Sapotaceae
E
M 197.5 47.19
Fagaceae
D
M
0.50
1.12
Lauraceae
E
M 11.00 11.24
Lauraceae
E
M 13.50
8.99
Lauraceae
E
M 15.50
7.87
Euphorbiaceae
D
M 14.50
8.99
Simaroubaceae
E
S
8.50
5.62
Mimosaceae
D
S
5.00
4.49
Agavaceae
E
S 38.00 12.36
Podocarpaceae
E
M 34.50 17.98
Annonaceae
E
S
0.50
1.12
Verbenaceae
D
M 16.00 13.48
Verbenaceae
D
M 19.50 19.10
Rubiaceae
E
S 16.50 10.11
Sterculiaceae
D
L
7.50
5.67
Sterculiaceae
E
M 79.00 33.71
Sterculiaceae
D
L 97.50 44.94
Fagaceae
E
M 18.00 10.11
Rubiaceae
E
S
7.50
7.87
Papilionaceae
E
L
5.00
4.49
Sapindaceae
D
M
8.50
6.74
Sapindaceae
D
M
0.50
1.12
Sapindaceae
D
M
8.50
6.74
Euphorbiaceae
E
L 136.0 60.67
Rubiaceae
E
S 11.50
8.99
Caesalpiniaceae
E
M 33.50 22.47
Saurauiaceae
E
S 78.50 32.58
Theaceae
E
M 376.5 65.17
Sapindaceae
D
L
7.00
6.74
Anacardiaceae
E
M 187.5 53.93
Dipterocarpaceae
E
L 33.50
3.37
Anacardiaceae
D
M 109.5 53.93
Sterculiaceae
D
M
8.00
7.87
Sterculiaceae
D
M 49.50 35.96
Bignoniaceae
E
L 191.0 62.92
Moraceae
E
M 49.00 28.09
Styraceae
E
S 10.00 10.11
Symplocaceae
E
S
6.00
5.62
Symplocaceae
Symplocaceae
Symplocaceae
E
E
E
S
S
S
10.50
2.50
4.00
10.11
3.37
3.37
Myrtaceae
Myrtaceae
Myrtaceae
Myrtaceae
Myrtaceae
Myrtaceae
Magnoliaceae
Caesalpiniaceae
Verbenaceae
Combretaceae
Combretaceae
Datiscaceae
Meliaceae
Cannabaceae
Euphorbiaceae
E
E
E
E
E
E
E
D
D
D
D
D
D
E
D
M
S
M
M
M
M
S
M
L
L
L
L
M
M
M
93.00
30.00
16.00
1.50
73.50
67.00
18.50
0.50
145.0
114.5
36.50
109.0
161.5
2.00
58.00
49.44
16.85
8.99
3.37
30.34
44.94
16.85
1.12
21.35
48.31
29.21
51.69
66.29
1.12
28.09
213
214
215
216
217
218
219
220
221
222
Vatica lancaefolia (Roxb.) Bl.

Oreocnide sp.
Vitex altissima L. f.
Vitex peduncularis Wall. ex Schauer.
Vitex pinnata L.
Vitex sp.
Wendlandia sp.
Xerospermum glabratum (Kurz) Radlk.
Zanthoxylum rhetsa (Roxb.) DC.
Zizyphus mauritiana Lam.
Borah et al. (2016) 3(1): 0109

.
Dipterocarpaceae
E
M 138.0 50.56
Urticaceae
E
S
2.50
4.49
Verbenaceae
E
M 49.00 25.84
Verbenaceae
E
M 89.00 58.43
Verbenaceae
E
M
5.50
7.87
Verbenaceae
E
M 30.50 24.72
Rubiaceae
E
S
4.50
5.62
Sapotaceae
E
L 62.00 21.35
Rutaceae
E
M 57.50 40.45
Rhamnaceae
D
M 20.50 13.48
CONCLUSION
The Barak Valley of Assam has good number of tree species, the major component of the forests ecosystem.
Depletion of species number and frequency due to the different anthropogenic pressure are the main disquiet.
Utilization of traditional knowledge and legal and full involvement of the local communities in conservation
practices might be very effective to conserve the forests in this region. Despite of rich tree species diversity it
provides various ecosystem services such as habitat to other species, carbon storage, carbon sequestration etc.
and environmental benefits which needs further study.
ACKNOWLEDGEMENTS
Authors thank to Botanical Survey of India, Shillong for species identification. Authors are grateful to the
forest departments of Cachar, Karimganj and Hailakandi districts of Assam for permission and support during
the study.
REFERENCES
Athokpam FD & Garkoti SC (2013) Variation in evergreen and deciduous species leaf phenology in Assam,
India. Trees 27:985997.
Athokpam FD, Garkoti SC & Borah N (2014) Periodicity of leaf growth and leaf dry mass changes in the
evergreen and deciduous species of Southern Assam, India. Ecological Research 29: 153165.
Bajpai O, Kumar A, Srivastava AK, Kushwaha AK, Pandey J & Chaudhary LB (2015) Tree species of the
Himalayan Terai Region of Uttar Pradesh, India: a checklist. Check List 11(4): article 1718.
Barua IC, Choudhury S & Neog B (1988) Primitive Land Plants (Angiosperm) and Their Distribution Pattern in
Assam. Journal of Economic and Taxonomic Botany 12 (1): 8182.
Borah N & Garkoti SC (2011) Tree Species Composition, Diversity, and Regeneration Patterns in Undisturbed
and Disturbed Forests of Barak Valley, South Assam, India. International Journal of Ecology and
Environmental Sciences 37 (3): 131141.
Borah N (2012) Community Structure, Tree Regeneration and Utilization of Forest Resources in Cachar and
Hailakandi Districts of Assam, India. Ph.D. Thesis, Assam University, Silchar.
Borah N, Athokpam FD, Garkoti SC, Das AK & Hore DK (2014) Structural and compositional variations in
undisturbed and disturbed tropical forests of Bhuban hills in south Assam, India. International Journal of
Biodiversity Science, Ecosystem Services & Management 10(1): 919.
Champion HG & Seth SK (1968) A Revised Survey of the Forest Types of India. Govt. of India publications,
New Delhi.
Choudhury S (1982) Cleisostoma spicatum Lindi. in Cachar District, Assam. Indian Forester108 (8): 589592.
Chowdhury S, Nath AK, Bora A, Das PP & Phukan U (2005) Assams Flora. Assam Science Technology and
Environment Council, Guwahati.
Dam DP & Dam N (1984) Plalaenopsis coru-ceri (Breda) Bl. and Rechb. F. -An Orchid Record from Tropical
Rain Forest of Assam India. Bulletin of the Botanical Survey of India 26: 34.
Dansereau P (1960) Biogeography-An Ecological Perspective. The Ronald Press Co. New York
Dular AK (2015) Plantdiversity assessment of Sariska tiger reserve in Aravallis with emphasis on minor forest
products. Tropical Plant Research 2(1): 3035.
Hooker JD (18721887) Flora of British India. Vols. 1-7. London.
Jain SK & Rao RR (1977) A handbook of field and herbarium methods. Today & Tomorrows Printers &
Publishers, New Delhi, 107 p.
Borah et al. (2016) 3(1): 0109

.
Kanjilal UN, Kanjilal PC, Das A & De RN (19341940) Flora of Assam. Vols. 1-4 Govt. Press, Shillong.
Murali KS, Uma Shankar R, Ganeshaiah KN & Bawa KS (1996) Extraction of Non-Timber Forest Products in
the Forests of Biligiri Rangan Hill, India. 2. Impact of NTFP Extraction on Regeneration, Population
Structure, and Species Composition. Economic Botany 50: 252269.
Rabha D (2014) Species composition and structure of Sal (Shorea robusta Gaertn. f.) forests along distribution
gradients of Western Assam, Northeast India. Tropical Plant Research 1(3): 1621.
Rao AS & Verma DM (1969) Materials Towards Monocot Flora of Assam. Bulletin of the Botanical Survey of
India 8: 296303.
Rao AS & Verma DM (1976) Materials Towards Monocot Flora of Assam. Bulletin of the Botanical Survey of
India 18:148.
Sarkar M & Devi A (2014) Assessment of diversity, population structure and regeneration status of tree species
in Hollongapar Gibbon Wildlife Sanctuary, Assam, Northeast India. Tropical Plant Research 1(2): 2636.
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 1017, 2016
Research article
Soil organic carbon stocks in different land uses in Pondicherry

university campus, Puducherry, India
SM. Sundarapandian*, S. Amritha, L. Gowsalya, P. Kayathri, M. Thamizharasi, Javid
Ahmad Dar, K. Srinivas, D. Sanjay Gandhi and K. Subashree
Department of Ecology and Environmental Sciences, Pondicherry University, Pondicherry - 605014
*Corresponding Author: smspandian65@gmail.com
Abstract: Soil Organic Carbon (SOC) stocks (30 cm soil depth) were assessed in different land
uses (teak plantation, eucalyptus plantation, acacia plantation, shrub land and grass land) in
Pondicherry University campus by using Walkley & Blacks method. The soil bulk density was
found to increase significantly (P<0.05) with increasing soil depth in all sites except acacia
plantation and shrub land. Grass land and shrub land showed significantly (P<0.05) greater bulk
density than other study sites. The stock of SOC percent and soil organic matter significantly
(P<0.001) decreased with increasing soil depth. Acacia and eucalyptus plantations showed
significantly (P<0.05) greater SOC percent than other study sites. This may be due to higher litter
inputs and greater biological activity. The stock of total SOC was significantly greater in the grass
land and shrub land than the other study sites. This could be attributed to more bulk density in
these study sites. The present study suggests that maintaining diverse land uses would enrich the
carbon stock of the institution in addition to preservation of biodiversity.
Keywords: Bulk density - Grass land - Plantation - Shrub land - Soil organic matter.
[Cite as: Sundarapandian SM, Amritha S, Gowsalya L, Kayathri P, Thamizharasi M, Dar JA, Srinivas K,
Gandhi DS & Subashree K (2016) Soil organic carbon stocks in different land uses in Pondicherry university
campus, Puducherry, India. Tropical Plant Research 3(1): 1017]
INTRODUCTION
Forests have a vital role in the global carbon cycle and provide various ecosystem services to the society.
India has a vast range of forest types, weather and soil conditions and it is often very difficult to conclude which
forest type is best in carbon sequestration. Long-rotation forests store more carbon in forest biomass and in
associated carbon pools than the short-rotation plantations (Kaul et al. 2011). Carbon sequestration by
plantations serves as a sizeable sink for atmospheric CO2 both in temperate and tropical regions (Houghton et al.
2000, Fang et al. 2001). Carbon stocks have received considerable attention in the recent past as a result of its
commoditization. Plantation programmes are often initiated to create carbon credits that generate significant
income for the developing countries (Niles et al. 2002). Although in the first commitment period of Kyoto
Protocol (20082012), the market for CDM (Clean Development Mechanism) sinks was limited, the importance
of CDM sinks lies largely in reforestation and afforestation activities in the developing countries beyond 2012
(De Koning et al. 2005). In India, several workers have published biomass estimations using allometric
equations for few tree species, which had diameter above 10 cm at breast height (Bargali et al. 1992, Lodhiyal et
al. 1992) that are grown in plantations. However, soil carbon stock assessments in these plantations are very
limited and scattered and the studies are mostly concentrated only on aboveground biomass carbon.
The carbon in the tropical forest soils is roughly equivalent to or less than the aboveground biomass due to
degradation (Ramachandran et al. 2007). Ravindranath et al. (1997) reported that the ratio of soil organic carbon
(SOC) to biomass carbon was 1.25. Tree plantations are known to increase the carbon pool in biomass and soil
(Arora & Chaudhry 2014). Kaul et al. (2010) found that in plantations, the carbon content in the soil was almost
double the biomass carbon but not 2.5 to 3 times the biomass carbon as recorded earlier.
Forest ecosystems adjacent to the cities are more significant as they provide clean environment, have
aesthetic value, act as storehouse of medicinal plants etc. Union territory of Pondicherry is a small piece of land
10
Sundarapandian et al. (2016) 3(1): 1017

.
with alarming population growth rate. This leads to conversion of agricultural land and areas of aquifers into
real estates, industrialized and institutional areas. Pondicherry University is one of the institutions with a large
land cover of 760 acres. Hence, the university has taken steps to green the entire campus except the built-areas
and paths. Even though the disaster of Thane cyclone in December 2011 uprooted and damaged several large
and very old trees, huge patches of forest still exists. The Flora of the university campus has been documented
by Parthasarathy et al. (2010). Influence of Thane cyclone on tree damage has been assessed by Sundarapandian
et al. (2014a). Biomass and carbon stock assessments of woody vegetation in the University campus have been
done by Sundarapandian et al. (2014b). Recently, many educational institutes in the western world have
assessed their carbon footprints. Recent advances reveal that instead of carbon footprint, ecological footprint
would be more reasonable and applicable. Based on the assessment of ecological footprint, Canada decided to
withdraw from the Kyoto Protocol. At present, Indian institutes also take initiatives to green their campuses and
assess their carbon footprints. Pondicherry University has endeavoured to construct a solar campus in silver
jubilee buildings. Several initiatives are under discussion. At this crucial time, baseline data of carbon stocks of
the campus is one of the important parameters to plan green campus initiatives and estimate ecological footprint.
The present study will be more important in terms of baseline data generation and documentation. This baseline
data will also be helpful to estimate the carbon sequestration potential of forest ecosystems in the campus in the
near future. Therefore, the present study was intended to evaluate the following objectives: (1) to examine the
variations in soil C stock in different land uses and (2) to examine the relationship of soil C stock with various
edaphic factors.
Study area
Figure 1. Aerial view of Pondicherry University Campus. (Source: Google Earth)
Pondicherry University (12o 0.97' N, 79o 51.33' E) is situated 10 km north of Puducherry town, on the
Coromandel coast (Fig. 1) and spans an area of 780 acres, of which the built-areas occupy approximately
1,80,000 m2. The climate is tropical with most rainfall during northeast monsoon (OctoberDecember) and very
less and inconsistent rainfall during southwest monsoon (JuneSeptember). The mean annual rainfall is 1282
mm for the last two decades (19902010).The mean annual maximum and minimum temperatures of
Puducherry are 32.58oC and 24.51oC. The soil is red ferrilitic, sandy and heavily drained. The vegetation of
Pondicherry University is mainly composed of tropical dry evergreen scrub and palm savannas in the west and
south and cashew plantations, rice, sugarcane and groundnut cultivation in the east. For the present study, five
11

.
different land uses in the university viz., teak plantation, eucalyptus plantation, acacia plantation, shrub land and
grass land were selected.
Soil sampling and laboratory analysis
Soil samples were collected at 010, 1020 and 2030 cm depths from each land use using a core sampler
during January and February of the year 2013. Ten sets of samples were collected from each study site and are
mixed together to form a composite soil sample, from which six replicate samples were brought to the
laboratory for further analysis. Before analysis, soil samples were sieved through a 2 mm mesh and then mixed
thoroughly. Soil organic carbon was estimated by using Walkley and Blacks method (Walkley 1947). In this
method, about 6086% of SOC is oxidized and therefore a standard correction factor of 1.32 was used to obtain
the corrected SOC values (De Vos et al. 2007).
For bulk density, in each site, six aggregated undisturbed soil cores were taken by a soil corer with 5 cm
internal diameter. The soil samples were weighed immediately and transported to the laboratory where they
were oven-dried at 105oC for 72 h and re-weighed. In the soils containing coarse rocky fragments, the coarse
fragments were separated by a sieve and weighed. The bulk density of the mineral soil core was calculated with
the help of the formula described by Pearson et al. (2005). Soil carbon stocks were then calculated for each soil
depth based on the thickness of the soil layer, bulk density and carbon concentration. The total carbon content
upto 30 cm depth was finally estimated by summing the carbon concentration of all the layers (Pearson et al.
2005).
Statistical Analysis
The variation in SOC stocks among different land uses and soil depths (010, 1020, 2030 cm) was
examined with analysis of variance (ANOVAs). The relationship between SOC stock and three edaphic factors
(soil moisture, soil pH and bulk density) were examined with correlation analysis followed by linear regression.
RESULTS
The soil moisture (%) in different land uses of Pondicherry University campus ranged from 2.98 (shrub land)
to 5.63 (acacia) up to 30 cm soil depth (Fig. 2). The soil moisture (%) was found to significantly vary (P<0.001)
among the study sites. Eucalyptus plantation and grass land showed almost same soil moisture (%) i.e. 4.0%.
The soil pH in different land uses of Pondicherry University campus ranged from 5.50 (grass land) to 7.34 (teak)
up to 30cm soil depth (Fig. 3). The pH was significantly (P<0.001) greater in teak plantation compared to grass
land and other plantations.
8
Soil moisture (%)
7
6
5
4
3
2
1
0
Teak
Eucalyptus
Acacia
Grassland
Shrubland
Figure 2. Soil moisture in different land uses in Pondicherry University campus, Puducherry, India.
The soil bulk density (BD) in different land uses of Pondicherry University campus ranged from 1.18 (teak)
to 1.49 (grass land) up to 30 cm soil depth (Fig. 4). The soil bulk density increased significantly (P<0.05) with
increasing soil depth in all the sites except acacia plantation and shrub land. Grass land and shrub land showed a
significantly (P<0.05) greater bulk density than the other study sites. The mean range of bulk density in different
depths was 1.16 (shrub land) to 1.26 (acacia) at 010 cm, 1.13 (acacia) to 1.70 (shrub land) at 1020 cm and
1.15 (acacia) to 1.67 (grass land) at 2030 cm. Significant differences (P<0.05) were found to exist in the soil
profile of all the sites except acacia plantation.
12

.
10
Teak
Eucalyptus
Acacia
Grassland
Shrubland
8
Soil pH
6
4
2
0
0-10
10-20.
20-30
Soil depth (cm)
0-30
Figure 3. pH in different land uses in Pondicherry University campus, Puducherry, India.
Bulk density (g/m2)
2.5
Teak
Eucalyptus
Acacia
Grassland
Shrubland
2
1.5
1
0.5
0
0-10
10-20
Soil depth (cm)
20-30
0-30
Figure 4. Soil bulk density in different land uses in Pondicherry University campus, Puducherry, India.
The SOC percent in different land uses of Pondicherry University campus ranged from 1.53 (teak) to 2.1
(acacia) up to 30 cm soil depth (Fig. 5). The SOC stock percent significantly (P<0.001) decreased with
increasing soil depth. Acacia and eucalyptus plantations showed significantly (P<0.05) greater SOC percentage
than the other sites. The mean range of SOC percent in different depths was 0.64 (shrub land) to 1.05 (acacia) at
010 cm, 0.47 (teak) to 0.63 (shrub land) at 1020 cm and 0.27 (teak) to 0.58 (shrub land) at 2030 cm.
2.5
Teak
Eucalyptus
Acacia
Grassland
Shrubland
SOC (%)
2
1.5
1
0.5
0
0-10
10-20.
20-30
0-30
Soil depth (cm)

Figure 5. Soil organic carbon (SOC) in different land uses in Pondicherry University campus, Puducherry, India.
The Soil Organic Matter (SOM %) in different land uses ranged from 4.54 to 6.10 in 030 cm soil depth
(Fig. 6). SOM stocks (%) significantly (P<0.001) decreased with increasing soil depth. Acacia and eucalyptus
plantations showed significantly (P<0.05) greater SOM percentage than the other study sites. Teak plantation
had the least SOM (%). The observed mean range of SOM percentage in different depths was 1.9 (shrub land)
13

.
to 2.96 (acacia) at 010 cm, 1.38 (teak) to 1.86 (shrub land) at 1020 cm and 0.80 (teak) to 1.70 (shrub land) at
2030 cm.
7
Teak
Eucalyptus
Acacia
Grassland
Shrubland
6
SOM (%)
5
4
3
2
1
0
0-10
10-20.
20-30
0-30
Soil depth (cm)

Figure 6. Soil organic matter (SOM) in different land uses in Pondicherry University campus, Puducherry, India.
The total SOC stocks up to 30 cm soil depth in the studied different land uses ranged from 19.47 (teak) to
27.06 (grass land) Mg C ha-1 (Fig. 7). The mean range of total soil carbon in different depths was 7.39 (shrub
land) to 12.56 (acacia) Mg C ha-1 at 010 cm, 6.35 (teak) to 10.74 (shrub land) Mg C ha-1 at 1020 cm and 3.69
(teak) to 8.72 (shrub land) Mg C ha-1 at 2030 cm. The total SOC stocks were significantly greater in grass land
and shrub land than the other study sites.
Total Organic Carbon (t/ha)
35
Teak
Eucalyptus
Acacia
Grassland
Shrubland
30
25
20
15
10
5
0
0-10
10-20.
20-30
Soil depth (cm)
0-30
Figure 7. Total soil organic carbon (t/ha) in different land uses in Pondicherry University campus, Puducherry, India.
Regression analysis indicated that soil pH and soil moisture had a negative relationship with SOC percent,
SOM and total carbon (Mg C ha-1) in Pondicherry University campus (Fig. 8). However, bulk density showed a
positive relationship with total carbon (Mg C ha-1).
DISCUSSION AND CONCLUSION
SOC showed a decreasing trend with increasing soil depth in all the study sites. This may be due the greater
decomposition rate in the upper layer compared to other layers. Similar results have been observed by other
workers as well (Jobbagy & Jackson 2000). The higher percentage of carbon in acacia and eucalyptus
plantations may be due to high litter inputs and more biological activity. In addition, the leaves of acacia and
eucalyptus trees have high lignin content which slows down the decomposition rate, which might have led to the
accumulation of humus throughout the year. This might be one of the reasons for high SOC stocks in these sites.
The range of SOC in tropical dry deciduous and moist forests ranged between 8.9 and 177 Mg C ha -1 in the top
50 cm soil depth (Chhabra et al. 2003) and our results are consistent with the above-stated values. The changes
in SOC stocks might also be due to different vegetation types, litter quality and quantity, soil type and texture,
14

.
soil chemistry, soil moisture, decomposition rates and also landscape position as stated by Beets et al. (2002),
Vesterdal et al. (2008) and Twongyirwe et al. (2013). The change in total SOC stocks (Mg C ha-1) could also be
due to differences in soil bulk density at different soil depths (Jobbagy & Jackson 2000).
0.35
0.25
0.2
0.1
y = -0.1824x +
0.2915
R = 0.0219
0.25
0.2
0.15
0.1
0.25
0.2
0.15
0.1
0.05
0
0
0.1
0.2
Logrithm of Bulk density
(g/cm3)
0.7
0.8
0.78
0.78
0.78
0.74
0.72
0.7
0.68
y = -0.1824x +
0.7636
R = 0.0219
0.66
0.74
0.72
0.7
0.68
0.66
0.64
y = 0.0392x +
0.7153
R = 0.0063
Logarithm of SOM (%)
0.8
0.76
0.74
0.72
0.7
0.68
0.64
0
0.5
1
Logarithm of soil moisture
(%)
0.7
1.42
1.44
1.42
1.36
1.34
1.32
1.3
y = 0.6696x +
1.3044
R = 0.2092
1.28
0
0.1
0.2
(g/cm3)
1.42
1.4
1.38
1.36
1.34
1.32
y = -0.284x + 1.5691
R = 0.2362
Logarithm of TC (t C/ha)
1.44
Logarithm of TC (t C/ha)
1.46
1.38
y = -0.6958x +
1.2832
R = 0.7201
0.76
1.44
1.4
0.9
0.66
0.64
0
0.1
0.2
(g/cm3)
0.8
Logarithm of pH
0.8
0.76
y = -0.6958x +
0.8111
R = 0.7201
0
0.5
1
(%)
Logarithm of SOM(%)
Logarithm of SOM (%)
y = 0.0392x + 0.2432
R = 0.0063
0.05
0.05
0
Logarithm of TC (t C/ ha)
0.3
Logarithm of SOC (%)
0.3
0.15
0.35
0.3
0.35
1.4
1.38
y = -0.7012x +
1.9397
R = 0.519
1.36
1.34
1.32
1.3
1.3
1.28
1.28
0
0.5
1
(%)
0.8
0.9
Logarithm of pH
0.7
0.8
Logarithm of pH
0.9
Figure 8. Regression analysis of soil organic carbon (SOC), soil organic matter (SOM) and total carbon (TC) with bulk
density, soil moisture and soil pH in different land uses in Pondicherry University campus, Puducherry, India.
15

.
SOC (%) showed a negative correlation with soil pH, soil bulk density and soil moisture. SOM (%) also
showed the same trend. To the contrary, the total carbon showed a positive correlation with bulk density, but not
with soil pH and soil moisture. Jobbagy & Jackson (2000) and Li et al. (2010) have also observed a negative
correlation of soil organic carbon with bulk density. The overall SOC was highest in grass land- than the other
sites. Shoji et al. (1993) in Central France have also reported high SOC values in grass land. Gupta & Sharma
(2014) have also concluded that grass lands have the maximum SOC pool based on their assessment of SOC
stocks in different land use systems of Uttarakhand. The present study suggests that maintaining diverse land
uses enriches the soil carbon stocks of the institution in addition to preserving biodiversity.
REFERENCES
Arora P & Chaudhury S (2014) Carbon Sequestration in tree plantations at Kurukshetra in Northern India.
American International Journal of Research in Formal, Applied and Natural Sciences 5(1): 6570.
Bargali SS, Singh SP & Singh RP (1992) Structure and function of an age series of Eucalyptus plantations in
Central Himalaya. I. Dry matter dynamics. Annals of Botany 69: 405411.
Beets PN, Oliver GR & Clinton PW (2002) Soil carbon protection in podocarp/hardwood forest and effects of
conversion to pasture and exotic pine forest. Environmental Pollution 116: 6373.
Chhabra AS, Palria & Dadhwal PK (2003) Soil organic pool in Indian forests. Forest Ecology and Management
173: 187199.
De Koning, Olschewski FR, Veldkamp E, Benitez PM, Lopez-Ulloa MT, Schlichter & DeUrquiza M (2005)
The ecological and economic potential of carbon sequestration in forests-examples from South America.
Ambio 34: 224229
De Vos B, Letten S, Muys B & Deckers JA (2007) Walkley-Black analysis of forest soil organic carbon:
recovery, limitations and uncertainty. Soil Use and Management 23: 221229.
Fang J, Chen A, Peng C, Zhao S & Ci L (2001) Changes in forest biomass carbon storage in China between
1949 and 1998. Science 292: 23202322.
Gupta MK & Sharma SD (2014) Sequestered Organic Carbon Stock in the Soils under Different Land Uses in
Uttarakhand State of India. Journal of Life Sciences Research 1(1): 59.
Houghton RA, Skole DL, Nobre CA, Hackler JL, Lawrence KT & Chomentowski WH (2000) Annual uxes of
carbon from deforestation and regrowth in the Brazilian Amazon. Nature 403: 301304.
Jobbagy EG & Jackson RB (2000) The vertical distribution of soil organic carbon and its relation to climate and
vegetation. Ecological Applications 10: 423436.
Kaul MG, Mohren MJ & Dadhwal VK (2010) Carbon storage and sequestration potential of selected tree
species in India. Mitigation and Adaptation Strategies for Global Change 15: 489510.
Kaul MG, Mohren MJ & Dadhwal VK (2011) Phytomass carbon pool of trees and forest in India. Climate
Change 108: 243259.
Li P, Wang Q, Endo T, Zhao X & Kakubari Y(2010) Soil organic carbon stock is closely related to aboveground
vegetation properties in cold-temperate mountainous forests. Geoderma 154: 407415.
Lodhiyal LS, Singh RP & Rana BS (1992) Biomass and productivity in an age series of short rotation Populus
deltoides plantation. Tropical Ecology 33: 214222.
Niles JO, Brown S, Pretty J, Ball AS & Fay J (2002) Potential carbon mitigation and income in developing
countries from changes in use and management of agricultural and forest lands. Philosophical Transactions
of The Royal Society A Mathematical Physical and Engineering Sciences 360: 16211639.
Parthasarathy N, Arul P, Muthuperumal C & Anbarasan M (2010) Flora of Pondicherry University Campus; A
Pictorial Guide to the Wild and Cultivated Plant Biodiversity. Pondicherry University.
Pearson T, Walker S & Brown S (2005) Source Book for LULUCF Projects. Winrock International, Arlington,
VA, USA.
Ramachandran A, Jayakumar S, Haroon RM, Bhaskaran A & Arockiasamy DI (2007) Carbon sequestration:
estimation of carbon stock in natural forests using geospatial technology in the Eastern Ghats of Tamil Nadu,
India. Current Science 92(3): 323331.
Ravindranath NH, Somasekhar BS & Gadgil M (1997) Carbon flows in Indian forest. Climatic change 35(3):
297320.
16

.
Shoji S, Dahlgren RA & Nanzyo M (1993) Genesis of volcanic ash soils. In: Shoji S, Nanzyo M & Dahlgren
RA (eds) Volcanic Ash Soils: Genesis, Properties and Utilization. Elsevier, Amsterdam, pp. 3771.
Sundarapandian SM, Mageswaran K, Gandhi DS & Dar JA( 2014a) Impact of Thane Cyclone on Tree Damage
in Pondicherry University Campus, Puducherry, India. Current World Environment 9(2): 287300.
Sundarapandian SM, Amritha S, Gowsalya L, Kayathri P, Thamizharasi M, Dar JA, K Srinivas, Gandhi DS &
Subashree K (2014b) Biomass and carbon stock assessments of woody vegetation in Pondicherry University
Campus, Puducherry. International Journal of Environmental Biology 4(2): 8799.
Twongyirwe R, Sheil D, Majaliwa JGM, Ebanyat P, Tenywa MM, van Heist M & Kumar L (2013) Variability
of Soil Organic Carbon stocks under different land uses: A study in anafro-montane landscape in
southwestern Uganda. Geoderma 193194: 282289.
Vesterdal L, Schmidt IK, Callesen I, Nilsson LO & Gundersen P (2008) Carbon and nitrogen in forest floor and
mineral soil under six common European tree species. Forest Ecology and Management 255: 3548.
Walkley A (1947) An estimation of methods for determining organic Carbon and Nitrogen in soil. Journal of
Agricultural Science 25: 598609.
17
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 1832, 2016
Review article
Predicting suitability of tree species in various climatic conditions

Sharad Tiwari1 and Rajesh Kumar Mishra2*
1
Institute of Forest Productivity, Lalgutwa, Ranchi, Jharkhand

Tropical Forest Research Institute, P.O. RFRC, Mandla Road, Jabalpur, Madhya Pradesh
*Corresponding Author: rajeshkmishra20@gmail.com
2
Abstract: Climate is a key factor shaping the forest environment; thus changes in the climate are
likely to strongly affect forest ecosystems by altering the physiology, growth, mortality and
reproduction of trees, the interactions between trees and pathogens, and ultimately the disturbance
regimes (winds, wildfires, insect attacks, etc.). The sensitivity to such changes depends on the
level that is considered (landscapes vs. forest, stands vs. single trees) and on the specific site
conditions. These complex influences indicate that a changing climate may lead to non-linear
responses, tipping points, etc., particularly since the longevity of trees implies that many
individuals present today will experience substantial changes of the climate before they will be
replaced by the next generation. Thus, the question arises to what degree current trees and forest
ecosystems are able to cope with a changing climate. In the present work a user-friendly package
PLANTPAK has been developed and tested successfully to evaluate the climatic suitability of
forestry species in central Indian region. The package can be used to store, retrieve and display
information based on simple key strokes. The package provides query on textural as well as map
basis. The package is tested with 15 data records and all the features including data entry,
information retrieval based on species name, location wise, climatic as well as edaphic fields are
working properly. Further the package is also successfully tested for providing map based retrieval
of information of suitable species.
Keywords: Forest ecosystem - Central India - Climatic suitability - PLANTPAK.
[Cite as: Tiwari S & Mishra RK (2016) Predicting suitability of tree species in various climatic conditions.
Tropical Plant Research 3(1): 1832]
INTRODUCTION
The worlds rapidly rising population requires most countries to make the best possible use of their land
resources for agriculture, horticulture, forestry and conservation. Being able to predict where and how well
particular plants are likely to grow in different regions is vital for land use planning. Linking GIS and modules
can help to answer these questions, but decision makers and researchers in developing countries have limited
access to these technologies. Climate has an important influence on tree growth it is particularly useful as a
means to predict where particular tree will grow, as mean climatic condition can now be reliably estimated for
most locations around the world. Being able to identify where particular trees (or plants) will grow is useful, but
many people need to know how well they will grow on particular sites. Generally they do not require highly
precise predictions of yield, but they do need to know whether growth will be good, fair, poor or useless.
Therefore the development of interactive and user friendly decision support system is being proposed, which
will help in predicting suitability of important forestry species of central India in varied climatic and edaphic
conditions.
Dr. Michael Hutchinson (Center for Resource and Environmental Studies, Australian national University)
has developed a package known as ANUSPLIN, which uses Laplacian smoothing splines to interpolate spatially
between data recorded at meteorological stations (Hutchinson 1989, 1992). As part of ACIAR project 9127
mean monthly data were collated from meteorological stations in a single area including China, Thailand,
Vietnam, Laos, Cambodia and Peninsula Malaysia (Zuo et al. 1996). A digital elevation model was prepared by
Zuo et al. (1996) and monthly mean values for all five climatic factors were estimated for a 1/20 th of a degree
Received: 04 November 2015
18
Tiwari & Mishra (2016) 3(1): 1832

.
grid (ca.5 km) of approximately 400 000 points across China and mainland South east Asia. As part of ACIAR
Project 9127 climatic mapping programs were prepared at the CSIRO Division of Forestry for China (100 000
grid points) and Latin America (66000 grid pints interpolated climatic data kindly supplied by Dr. Peter Jones,
CIAT, Colombia).
Most of the programs have been developed for the MS-DOS environment, which is the most common
operating system on PCs used in the developing world. However, a version of the THAI program has recently
also been developed for the Windows environment. Windows allows multitasking which makes it easy to
compare maps produced by different descriptions on the computers screen, as well as providing built-in support
for hundreds of different printers. Significant progress has been made in the development of climatic mapping
software in recent years (Hackett 1988, Booth 1990, Hackett 1991, Booth 1996a, b).
METHODOLOGY
For the present work climatic and edaphic data for entire central region has been collected. Basic database
structure and data retrieval algorithm has been developed. Initially records for 15 selected species with reference
to their climatic and edaphic suitability has been collected. All the forms including Main form, user
management form, data entry and edit form, query shell form has been designed and tested successfully. The
package has been developed and tested successfully for all the operations including data entry, data modification
and retrieval of information.
Requirement analysis
The brief study of the areas involved including the potential of the work, target user group, infrastructure
requirement and other related issues. It was attempted to create an easy to use and user friendly package, which
not only allows maintaining records but also provides for data retrieval, data entry, search for any specific data
among the entire database. It was decided that this database package will be made available to entire institute
and to others interested in deriving information or consultation. The field or parameters of input of information
were discussed at large and at various levels. The database structure was suitably modified to incorporate the
suggestions made to widen the applicability and usefulness of the package. Table 1 containing information
about species was designed for entering species records. Table 2 is also depicted the location details from the
fields.
Table 1. Species information.
Table 2. Location details and climatic conditions.
Species Id
Species Botanical Name
Genus
Vernacular Name
Family
Uses
Combination
Soil- Poorly suitable
Soil- Moderately Suitable
Soil- Most Suitable
Temp Mean Min- Poorly suitable
Temp Mean Min- Moderate suitable
Temp Mean Min- suitable
Temp Mean Max- Poorly suitable
Temp Mean Max- Moderate suitable
Temp Mean Max- suitable
Rainfall- Poorly suitable
Rainfall- Moderate suitable
Rainfall- Most Suitable
Remark
Location ID
Location name
Soil Type
Temp min
Temp max
Rainfall min
Rainfall max,
Suitable Species
Remark
19
Tiwari & Mishra (2016) 3(1): 1832

.
Database structure
The database should be prepared as showed in tables 36.
Table 3. Species master.
Field name
Species_ID
Species_Botanical_Name
Genus
Vernacular_Name
Family
Uses
Combination
Soil_Poorly_suitable
Soil_Moderately_suitable
Soil_Most_Sutable
Temp_Mean_Min_Poorly_l
Temp_Mean_Min_Moderate_l
Temp_Mean_Min_suitable_l
Temp_Mean_Max_Poorly_l
Temp_Mean_Max_Moderate_l
Temp_Mean_Max_suitable_l
Temp_Mean_Min_Poorly_u
Temp_Mean_Min_Moderate_u
Temp_Mean_Min_suitable_u
Temp_Mean_Max_Poorly_u
Temp_Mean_Max_Moderate_u
Temp_Mean_Max_suitable_u
Rainfall_Poorly_l
Rainfall_Moderate_l
Rainfall_Most_Sutable_l
Rainfall_Poorly_u
Rainfall_Moderate_u
Rainfall_Most_Sutable_u
Remark
Preference
Data type & Width

Number (7) Primary key
Varchar2 (50)
Varchar2 (40)
Varchar2 (40)
Varchar2 (40)
Varchar2 (100)
Varchar2 (100)
Varchar2 (100)
Varchar2 (100)
Varchar2 (100)
Number (2)
Number (2)
Number (2)
Number (2)
Number (2)
Number (2)
Number (2)
Number (2)
Number (2)
Number (2)
Number (2)
Number (2)
Number (4)
Number (4)
Number (4)
Number (4)
Number (4)
Number (4)
varchar2 (100)
varchar2 (55)
Table 4. User master.
Field name
USERID
PASSWORD
FNAME
LNAME
DESIGNATION
PREVILLAGE
REMARK
Data type & Width

VARCHAR2 (25) primary key
VARCHAR2 (20)
VARCHAR2 (20)
VARCHAR2 (20)
VARCHAR2 (25)
VARCHAR2 (20)
VARCHAR2 (50)
Table 5. Location master.
Field name
location_ID
location_name
soil_type
temp_min
temp_max
rain_min
rain_max
suitable_species
remark
Data type & Width

Number (6) primary key
varchar2 (35)
varchar2 (40)
varchar2 (5)
varchar2 (5)
varchar2 (9)
varchar2 (9)
varchar2 (35)
varchar2 (50)
20
Tiwari & Mishra (2016) 3(1): 1832

.
Table 6. Error master.
Field name
error_ID
error_date
user_ID
error_details
error_status
solution
remark
Data type & Width

Number (7) primary key
date
varchar2 (25)
varchar2 (100)
varchar2 (15)
varchar2 (100)
varchar2 (100)
Functional identification
The system consists of selections made by the user at different stages in application and the entries that the
user makes. Different selections and entries that the user can make were finalized. It is usually very difficult to
specify system characteristics accurately without actually doing much of the proposed work. Thus, a quick guess
about the systems characteristics was all that was possible at this point. To systematically plan the output of the
proposed system, thorough interaction with some potential target group had been done which resulted in
finalization of procedure of the output display on monitor or printing as reports. The inputs required to produce
the required outputs were listed and the sources of these inputs determined. A tentative, general schedule for
developing the package was decided as follows:
1. User login ID and password.
2. User selects what he wants to do.
3. Data Entry: User enters the data regarding the database.
4. Editing: User can add, edit, species and location details.
5. Information Retrieval.
6. User can report an error.
Data collection
Data were collected from various sources viz. literature, books and journals. The district wise climatic and
edaphic data were collected from NIC site. The state maps were downloaded from state NIC website. Species
data were collected through extensive review of relevant literature.
Feasibility study
Feasibility study was conducted to select the best system meeting performance requirements. Once it was
determined that the project was feasible, project specifications which finalize project requirements were
prepared. Three key considerations were involved in feasibility analysis: Economic, Technical & Operational.
Economic analysis also known as cost / benefit analysis, determined, whether the adoption of the system was
cost justified or not. Technical consideration evaluated existing hardware and software and future requirement.
Operational feasibility specified that whether the proposed system will meet the operating requirements of the
organization.
System design life cycle
In order to transform requirements into a working system both the customer/user and the developer has to be
satisfied. The user/employee has to understand that what the system is suppose do and at the same time the
system developer is to know as to how the system is to work.
Conceptual design
The Conceptual Design tells what the system will do. The System is described in term of its boundary,
entities, attributes and relationship. In this phase it was determined that1. The Data comes from field level survey.
2. The Data is fed to the system.
3. The front end designed, links the user to the database.
4. The user is offered a number of choices like simple viewing of records searching for a particular record,
record entry, deletion of record etc.
5. The format of reports output screen.
21
Tiwari & Mishra (2016) 3(1): 1832

.
Algorithms:
I. Algorithm for adding user
1. START
2. INPUT:
{USERID, PASSWORD, USERNAME, DESIGNATION, REMARK}
3. FORM LEVEL VALIDATION :
i.
IF <data valid> THEN
GO To NEXT STEP
ELSE
User is prompted to correct the entries / data in the field / fields.
4. DATABASE LEVEL VALIDATION:
{As user ID primary key therefore two users IDs cant be identical.}
i.
IF <data valid> THEN
GO To NEXT STEP
ELSE
5. MESSAGE : RECORD ADDED.
6. END.
II. Algorithm for Adding Species
1.
2.
START
INPUT:
{
Species ID
Species Botanical Name
Genus
Vernacular Name
Family
Uses
Combination
Soil- Poorly Suitable
Soil- Moderate Suitable
Soil- Most Suitable
Temp Mean Min- Poorly Suitable
Temp Mean Min- Moderate Suitable
Temp Mean Min- Most suitable
Temp Mean Max- Poorly Suitable
Temp Mean Max- Moderate Suitable
Temp Mean Max- Most Suitable
Rainfall- Poorly Suitable
Rainfall- Moderate Suitable
Rainfall- Most Suitable
Remark
Preference
}
3.
FORM LEVEL VALIDATION :

i. IF <data valid> THEN
GO To NEXT STEP
ELSE
22
Tiwari & Mishra (2016) 3(1): 1832

.
4.
5.
6.
III.
DATABASE LEVEL VALIDATION:

i. IF <Data Valid> THEN
GO To NEXT STEP
ELSE
MESSAGE: RECORD ADDED.
END.
Algorithm for adding location
1. START
2. INPUT :
{LOCATION NAME, RAINFALL, TEMPERATURE, SUITABLE SPECIES, SOIL TYPE, REMARK}
3. FORM LEVEL VALIDATION :
GO To NEXT STEP
ELSE
4. DATABASE LEVEL VALIDATION:
GO To NEXT STEP
ELSE
5.
6.
IV.
MESSAGE: RECORD ADDED.

END.
Algorithm for search / query operations
1. START
2. INPUT:
{BOTANICAL NAME, VARNACULAR NAME, GENUS, RAINFALL, TEMPERATURE, WHATEVER
CARITERIA USER WANTS TO USE.}
3. FORM LEVEL VALIDATION:
GO To NEXT STEP
ELSE
4. FORMING SQL QUERY AND SENDING IT TO RDBMS (ORACLE):
i. IF < Data Found > THEN
GO To NEXT STEP
ELSE
User is prompted no records found try again.
5. DISPLAYING Message.
6. END.
Design approach
Modular is the design approach. The entire system works due to the functionality of its component modules.
Each module was designed to be complete in itself. Its the user action, however that decides the flow of control
in the system since the entire programming is event based. For each action of the user, a particular module is
associated to starts functioning generating the desired results.
23
Tiwari & Mishra (2016) 3(1): 1832

.
START
Manage Accounts
Data Entry
Query shell
Accessories
ABOUT S/W
Logout
Add User
S/W Details
Edit User Privileges

Delete User
Add Species
View User
Add Location
1
Edit Species
Edit Location
By Species
2
By Location
Advanced Search
View Records
Image Gallery
Map
Change Password
Calculator
Paint Brush
Related Site
Error Report
Report Error
View Error
STOP
Figure 1. Flow chart of program.
24
Tiwari & Mishra (2016) 3(1): 1832

.
BOF
Enter the soil Type or Rainfall Rang
or Temperature Range or use
Search in Database table

If
Yes
Records found
Show Results
No
Data not found
Figure 2. Flow chart for advance search.
25
Tiwari & Mishra (2016) 3(1): 1832

.
BOF
Enter the Local Name or soil Type

If
Yes
Records found
Show Results
No
Data not found
Figure 3. Flow chart to search species by local details.
26
Tiwari & Mishra (2016) 3(1): 1832

.
BOF
Enter the Botanical Name or Vernacular

Name or Soil Type

If
Yes
Records found
Show Results
No
Data not found
Figure 4. Flow chart to search by species details.
27
Tiwari & Mishra (2016) 3(1): 1832

.
O LEVEL DFD
SURVEY
Plant-Pak
Figure 5. Flow chart of O level DFD.
1 LEVEL DFD
DATABASE
(Backend)
ADD
Add Species & Location
EDIT
Edit Species & Location Details
SEARCH
Search in Database by Species & Location Criteria
VIEW
View Species & Location Details
Figure 6. Flow chart of 1 level DFD.
28
Tiwari & Mishra (2016) 3(1): 1832

.
2 - LEVEL DFD
ADD
Location
Table
Species
Table
EDIT
SEARCH
VIEW
Figure 7. Flow chart of 2 level DFD.
The database contains important new features that optimize traditional business applications, facilitate
critical advancement for internet-based business, and stimulate the emerging hosted application market. New
database features deliver the performance, scalability, and availability essential to hosted service software made
available to anyone, anywhere. The database offers new transparent, rapid growth clustering capabilities, along
with powerful and cost-effective security measures, zero-data-loss safeguards, and real-time intelligence to
deliver the power needed in today's dynamic marketplace.
Data retrieval
The package provides retrieval of information based on following fields:
By Species Name: User can get access to records by simply selecting the species either by
Local_ Name or by Vernacular_ Name: On selection of this choice, a drop down menu containing all
the available species stored in the database is displayed. User can select the species of his interest from
the drop down menu.
By Family Type: On selection of this option, all the species belongs to a particular family are displayed.
By Location Name: On selection of this option, a window as containing list of all the locations stored in
the database is displayed. User can select the location of his interest from the drop down menu.
By Climatic and Edaphic Condition: On selection of this option, a user friendly window is displayed.
User can enter the Soil type or climate range to retrieve information belonging to those climatic and
edaphic conditions.
Through Map: The package has the option to provide information also through the map. On selection of
this option, the map of selected state is displayed. The user can browse through the map and can retrieve
species suitable to a particular place by simply clicking at that place on the map).
The package has been successfully implemented and tested for 15 species (Acacia catech, Albizia lebbec,
Albizia procer, Ailanthus excels, Boswellia serrata, Gmelina arborea, Holoptelea integrifolia, Lagerstroemia
parviflora, Madhuca latifolia, Moringa oleifera, Pongamia pinnata, Pterocarpus marsupium, Dalbergia
latifolia, Sterculia urens, Emblica officinalis) in Madhya Pradesh and Maharashtra region (Appendix IIV).
The strength of this package is that it is very user friendly more intended to not only function to the best of
user satisfaction but its workability also. It allows the users to not only retrieve data but also manage the
29
Tiwari & Mishra (2016) 3(1): 1832

.
database by updating to whenever necessary. The user is provided with all the relevant option of adding,
deleting, making changes to and also updating the records. The point of vulnerability of this application is that
care has to be taken while feeding data to the database. The database incorporates two table and care should be
taken that while one table is being fed the relevant filed of the second table also be fed at the same time
otherwise it may cause inconvenience and irrelevant data retrieval.
This application on integration with GIS application can be enhanced to work like a Specialist Decision
Support System. The structure of the database can be planned in a better and efficient way; there still is a scope
to redefine it. This application was tested to work successfully with a fifteen tree species. The workability of the
system has been checked from each and every aspect and was found to work fine generate the desired output.
The application of this model shows good results in the research resulted in satisfactory results for most of
the species, but not for others. Hence, the need and justification that if further developed it can be proved as
quite robust and applicable at any level and with high percentage of accuracy. The evaluation of the results can
still be debated, and various experts will have a say regarding their respective field of expertise. However, this
Decision Support System, with further improvement, i.e. providing more detailed information, will cover the
gaps which can be seen here in some of the suitability maps for the species. The lack of proved mathematical
formula is also the issue in this DSS; a formula was created which would grow more complex and accurate as
further input is provided. This way of producing suitability maps for tree species purposes has not been carried
out in the past in our country. The method needs a lot more input data in order for it to be deemed as absolutely
reliable (or close to that). Such input would require implementing detailed species characteristics, detailed
climate-vegetation data, habitat specifications, slope and aspect data, various constraint data (agricultural land,
urban areas, industrial areas, and protected areas), etc. However, this research shows that this is a very good
approach for creating suitability of tree species in different climatic conditions which provide satisfactory
accuracy. That was the case in our research, where some of the suitability resulted in very good spatial
distribution and accuracy, while other for some of the species deemed areas as suitable when in fact they arent
such. The tree species used in this paper has justified their choice, in terms of offering various possibilities for
afforestation throughout the country. Finally, the approach used in this research resulted in the creation of a
model which was carried out as such for the first time in our country, and with its development will prove to be
robust enough to be used during the afforestation of any localities. In the end it can be concluded that the quality
of the model is dependent on the quality of the input. Therefore, more research should be done in improving the
theoretical background of the modelling process and the input parameters.
Conclusion
The decision support systems make use of variety of technology and new technologies playing important
role in decision making. PLANTPAK as this new decision support system has been named will be immensely
useful for farmers, tree planters and entrepreneurs in the arena of plantation forestry to decide the suitability of
tree species according to locality, climate and edaphic characteristics in central India. However, the package
could easily be enriched by incorporation of other area and species. In the recent times, the need of site specific
information along with geographical details has grown up immensely. The textual data coupled with geographic
detail can provide a very clear picture of the object under consideration. Many areas of DSS application are
concerned with geographic details. GIS based DSS can make use of spatial data processing. Clearly SDSS will
be an important subset of DSS in future. The trend in the future is likely to be growth in the use of SDSS by
users without any special skill in GIS applications. SDSS don't require in-depth commands to operate, yet allow
users to negotiate very sophisticated geographic analysis. In the future, the use of SDSS will be extended to
applications where the spatial information is only an interim stage. Users dealing with this broader set of
applications need to be given control over the important variables in the decision, while other processing is
performed without the need for extensive user interaction. With the development of such systems, new classes
of decision and new class of users will be supported effectively.
REFERENCES
Anonymous (1993) Plant Resources of South-East Asia. No. 5 (1&2). Pudoc Sc. Publication, Wegeningen.
Booth TH (1990) Mapping regions climatically suitable for particular tree species at the global scale. Forest
Ecology & Management 36: 4760.
30
Tiwari & Mishra (2016) 3(1): 1832

.
Booth TH (1996a) The development of climatic mapping programs and climatic mapping in Australia. In: Booth
TH (ed) Matching Trees and Sites. Proceedings of an International Workshop held in Bangkok, Thailand,
2730 March 1995. ACIAR Proceedings No. 63, pp. 3842.
Booth TH (1996b) The development of climatic mapping programs and climatic mapping in Australia. In:
Booth TH (ed) Matching Trees and Sites. ACLAR Proceedings No. 63.
Brandis D (1971) Indian Trees. Bishen Singh Mahendra Pal Singh, Dehradun.
Burley J & Styles BT (1976) Tropical Trees: Variation, Breeding and Coservation. Academic Press, London.
Gamble JS (1984) A Manual of Indian Timbers. Bishen Singh Mahendra Pal Singh, Dehradun.
Hackett C (1988) Matching Plants and Land: Development of a broad scale system from a crop project for
Papua New Guinea. CSIRO Division of Water and Land Resources. Natural Resources Series no.11,
Melbourne, 82 p.
Hackett C (1991) Plantgro: a software package for the prediction of plant growth. CSIRO, Melbourne.
Hutchinson MF (1989) A new objective method for spatial interpolation of meteorological variables from
irregular network applied to the estimation of monthly mean solar radiation, temperature, precipitation and
wind run. CSIRO Division of Water and Land Resources, Tech.Memo.89/5, CSIRO, Canberra 10 p.
Hutchinson MF (1992) Documentation for SPLINA and SPLINB- two programming the ANUSPLIN software
package. CRES, Australian, National University Canberra.
Luna RK (1996) Plantation Trees. International Book Distributors, Dehradun.
Maslekar AR (1996) Foresters Companion. Surya International Publications, Dehradun.
McCann C (1985) Trees of India. Periodical Expert Book Agency, New Delhi.
Prakash R & Hocking D (1986) Some Favourite Trees For Fuel And Fodder. Society For Promotion of
Wasteland Development, New Delhi.
PurKayastha SK (1996) A Manual of Indian Timbers. Sribhumi Publication Company, Kolkata.
Seth SK, Raizada MP & Waheed Khan MA (1962) Trees for Van Mahotsava. FRI and Colleges, Dehradun.
Singh RV (1982) Fodder Trees of India. Oxford and IBH Publication Company, New Delhi.
Singhal RM & Khanna P (1991) Multipurpose Trees and Shrubs. ICFRE, Dehradun.
Troup RS (1986) The Silviculture of Indian trees. International Book Distributors, Dehradun.
Zuo H, Hutchinson MF, McMahon JP & Nix HA (1996) Developing a mean monthly climatic database for
China and Southeast Asia. In: Booth TH (ed) Matching Trees and Sites. ACIAR Proceedings No. 63.
31
Tiwari & Mishra (2016) 3(1): 1832

.
Appendix I: Classification of different districts of Madhya Pradesh according to rainfall (mm).
S. No. Rain fall range
Districts
1.
8001000
Gwalior, Bhind, Shivpuri, Morena, Sheopurkala,
Khandwa, Burhanpur, Khargone, Jhabua
2.
8001200
Mandsaur, Ratlam, Ujjain, Dewas, Indore, Sajapur,
Rajgarh, Dhar
3.
8001400
Chhattarpur, Datia, Tikamgarh,
4.
10001200
Betul, Chhindwara
5.
10001400
Rewa, Satna, Panna, Seoni, Katni
6.
12001400
Bhopal, Sehore, Raisen, Vidisha, Guna, Ashoknagar,
Sagar, Damoh
7.
12001600
Balaghat, Shahdol, Anuppur, Sidhi, Mandla, Jabalpur,
Narsinghpur, Hoshangabad, Harda
Appendix II: Soil types of different districts of Madhya Pradesh.
S. No. Soil Type
Districts
Shallow & medium black
Betul, Chhindwara, Seoni
1.
Deep medium black
Narsinghpur, Hoshangabad, Harda, Shahdol, Umaria, Jabalpur, Katni,
2.
Sagar, Damoh, Vidisha, Raisen, Bhopal, Sehore, Rajgarh, Ujjain,
Dewas, Shajapur, Mandsaur, Neemach, Ratlam, Jhabua, Dhar, Indore,
Khargone, Barwani, Khandwa
Gwalior, Morena, Sheopurkala, Bhind
Alluvial soil
3.
Mandla, Dindori, Balaghat, Rewa, Satna, Panna, Chhatarpur,
Mixed red & black
4.
Tikamgarh, Shivpuri, Guna, Datia, Sidhi
Appendix III: Classification of different districts of Maharashtra according to rainfall (mm).
S. No. Rain fall range
Districts
1.
500800
Ahmednagar, Aurangabad, Akola, Beed, Buldhana,
Dhule, Jalgaon, Jalna, Osmanabad, Pune, Sangli, Solapur
2.
8001200
Amravati, Chandrapur, Hingoli, Latur, Nashik, Nanded,
Parbhani, Washim, Wardha, Yavatmal
3.
12001500
Bhandara, Gondia, Gadchiroli, Kolhapur, Nagpur, Satara
4.
25003000
Sindhudurg, Thane
5.
30003500
Raigad, Ratnagiri
Appendix IV: Soil types of different districts of Maharashtra.
Districts
S. No. Soil Type
Akola, Amravati, Beed, Buldhana, Jalgaon,
1.
Black to red
Latur, Osmanabad, Solapur
2.
Light laterite, Reddish brown, Greyish black Ahmednagar, Satara
3.
Brown to red
Bhandara, Gadchiroli
4.
Reddish brown to black, Greyish black
Kolhapur, Nashik, Pune, Sangli
5.
Coarse & shallow
Raigad, Thane
Aurangabad, Jalna, Nanded, Parbhani,
6.
Black soils, Black to red
Yavatmal
Ratnagiri, Sindhudurg
7.
Laterite, light laterite & reddish brown
Chandrapur, Nagpur, Wardha
8.
Black Soils, Brown to red
9.
Black to red, Greyish black
Dhule
32
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 3339, 2016
Research article
Eight new records of fresh water filamentous algae

(Oedogonium Link) from India
Priya Jitendra* and V. K. Anand
Department of Botany, University of Jammu, Jammu & Kashmir, India
*Corresponding Author: krishnas39@gmail.com
Abstract: The present paper deals with the eight species of Oedogonium which are being reported
for the first time from India. All the taxa are arranged in broad groups according to their
morphological peculiarities and sexual reproduction i.e. Macrandrous (Homothallic and
Heterothallic). During the present investigation 5 macrandrous homothallic {Oedogonium
subvaucherii Claass., O. pseudofragile Claass., O.upsaliense (Wittr.) Hirn., O.visayense
Britt., O.amplius (Tayl.) Tiff.} and 3 macrandrous heterothallic forms {O. cf capillare (L)
Kutz., O. angustistomum Hoff., O. magnusii Wittr. var. major Bock and Bock} have been
collected for the first time from India.
Keywords: Filamentous algae - Oedogonium - Homothallic - Heterothallic - New record.
[Cite as: Jitendra P & Anand VK (2016) Eight new records of fresh water filamentous algae (Oedogonium Link)
from India. Tropical Plant Research 3(1): 3339]
INTRODUCTION
Oedogoniales, an order of filamentous freshwater green algae is well defined with several unique features,
including asexual reproduction through the production of zoospores that possess a subapical ring of many short
flagella called as stephanokont. Oedogoniales exhibited a specialized type of oogamy and an elaborate method
of cell division which results in the accumulation of apical caps. The order is comprised of one family
Oedogoniaceae which includes three genera namely Oedogonium Link, Bulbochaete Agardh and Oedocladium
Stahl (Silva & Moe 2003). These three genera are different from each other morphologically, but also share the
several characteristics that distinguish Oedogoniales from rest of the green algae. On the basis of their peculiar
characteristics, members of the Oedogoniales are important not only from academic point of view but also are of
great ecological significances especially in the field of limnology since they occupy specific niches, food for a
number of aquatic organisms (Olojo et al. 2003, Kone & Teugels 2003, Awasthi et al. 2006), used for the
removal of heavy metals, production of antibiotics (Redondo et al. 2006) and being used as indicator of water
quality (Bajpai et al. 2013, Srivastava et al. 2014). This large, economically important order with its unique
features attracted the phycologists of all over the world including India, but unfortunately this order remained
unexplored in India. Many stray reports on the Oedogoniales have appeared from various parts of India
(Randhawa 1940, Kamat 1967, Kamat & Patel 1973, Shukla 1971, Bharati & Pai 1972, Shukla et al. 1988, Saha
& Pandit 1987, Mahato et al. 1998, Prasad & Misra 1992, Misra et al. 2002) but no consolidated work has ever
been done on Oedogoniales and Oedogonium in particular. Keeping in mind, the paucity of work done on
Oedogonium, the present research problem has been undertaken to work out the Oedogonium species of Jammu
region. Hence, a survey has been made of six districts of Jammu.
Jammu, the winter capital of Jammu & Kashmir State, is situated at a longitude 7476 15 E and latitude
32 15 to 30 30 N, and altitude 304.8 to 3658.5 metres above the mean sea level, It represents subtropical to
temperate climate with plains to mountainous terrain, which exhibiting remarkable longitudinal variations and
vegetation types. Due to the prevalent of varied climatic conditions, there exist number of natural and manmade
water bodies like ponds, rivers, ditches, pools, streams, nallahas, lakes and temporary water bodies. These water
bodies inhabit a great deal of algal diversity, of which Chlorophyceae is the most dominating.
33
Jitendra & Anand (2016) 3(1): 3339

.
Algal samples were collected from different localities of Jammu. Algae were picked up simply by hand from
shallow edges of the ponds or occasionally by plankton net and then preserved in 4% formaldehyde solution.
Preliminary screening of Oedogonium species from the samples was made in the Aquatic biology lab,
Department of Botany, University of Jammu, Jammu. Microphotographs of vegetative and reproductive
structures of different species of Oedogonium were taken using a microphotographic camera PM6 type,
Olympus make and Nikon FM3A (E). The film roll used was Nova Silver Plus, 125 ASA black and white.
Laboratory culture
In the laboratory, samples were first observed under a compound microscope for the presence of
reproductive structures. The vegetative filaments were transferred to petri dishes or flasks, rich in CO2
atmosphere for the formation of reproductive structures (Kumar & Singh 1984). The cultures were kept in the
culture room, illuminated with a fluorescent, white, cool 15w tube light of 464 lux with a temperature of 15C
25C and a photo period of 16:8 L/D regime. After 1020 days, the crude cultures were examined
microscopically. The incubated cultures were observed by pulling some filaments from several locations from
the same sample and made a whole mount on the glass slide. Samples were checked regularly for the next
following few weeks particularly for the late bloomers since each sample may contain many species and also all
species may not fruit at the same time (different species of Oedogoniales undergo fertility at different interval).
RESULTS
Enumeration of species
1. Oedogonium subvaucherii Claass (Gonzalves 1981, p. 183185, f. 951)
Fig. 1A
Accession No.: JUH.-Bot.-11242; Sample No.: J 14 , U 2

Habitat: Seasonal pond formed opposite petrol pump at Kuliyan, paddy field at Mansar .
Habit: Free Floating.
Vegetative filament: Vegetative cells cylindrical, 40 56 2022. Antheridia: Seriate 24 in
number, 26 1820. Oogonium: Single, globose, 3032 4244, poriferous, pore superior.
Oospore: Globose, smooth walled, 2022 2022.
Distribution: S. Africa (Transvaal Province), India {Jammu & Kashmir - Kuliyan (Jammu), Mansar
(Udhampur)}.
Variations: Present specimen resembled the type specimen described by Gonzalves (1981) in all
characters. It also seems to be close to O.vaucherii but differs from the same in the shape of
oogonia. In O. subvaucherii, oogonia is globose whereas in O. vaucherii oogonia is obovoid to
subobvoid globose.
2. O. pseudofragile Claass. (Gonzalves 1981, p. 180, fig. 9.44)
Fig. 1B
Accession No.: JUH.-Bot.-11234; Sample No.: R2

Habitat: Small pools formed along the side of river at Thanda Pani.
Habit: Free floating.
Vegetative filament: Cylindrical, slightly capitellate, 2244 1016. Antheridia: Hypogynous, seriate, 45
in number, 46 2030. Oogonium: Globose to subglobose, 2636 3042, poriferous, pore
supramedian to superior. Oospore: Globose to subglobose, 2024 2030.
Distribution: Africa (Transvaal Province), India {Jammu & Kashmir- Thanda Pani (Rajouri)}.
Variations: Present specimen coincides well with the description of type specimen given by Gonzalves (1981)
i.e. in having globose, poriferous oogonia, macrandrous habit. There were slight little variations in the size of
vegetative and reproductive parts.
3. O. upsaliense (Wittr.) Hirn, 1900, p.115, pl.XII, fig.60; Tiffany 1930, p.77, pl.XVI, fig.157 ;
Gemeinhardt 1939, p.115, fig.101; Tiffany & Britton 1952,p.66, pl.18, fig.144; Gauthier - Lievre
1963, p.293, pl.39, fig. 63h; Gonzalves 1981, p.184185, fig. 9.53.
Fig. 1C
Accession No.: JUH.-Bot.-11239; Sample No.: P 2
Habitat: Paddy Field at Sheesh Mahal, Poonch.
34
Jitendra & Anand (2016) 3(1): 3339

.
Vegetative filament: Vegetative cells cylindrical, 44 96 1224; Antheridia: Subepigynous,
rarely seriate, 2024 1820. Oogonium: Subovoid to suboblong- ellipsoid, 4068 2436,
poriferous, pore superior. Oospore: Suboblong-ellipsoid, smooth walled, 4452 2434.
Distribution: Algeria, Green Land, Ohio, Illinois, Michigan, New Hamphire, Iberia,
Czechoslovakia, Denmark, Finland, France, Germany, Poland, Sweden, India {Jammu & Kashmir
- SheeshMahal (Poonch).
Variations: Ellipsoid, poriferous oogonia and smooth walled oospore enabled present specimen to
resembled O. upsalisnse, O. oviforme, O. subellipsoideum and O. sodiroanum. On the basis of
dimensions of vegetative and reproductive parts it seems to be more close to O. upsaliense as
compared to O. oviforme and O. subellipsoideum species. The present specimen resembled the type
specimen described by Gonzalves (1981) in morphology as well as dimensions of vegetative and reproductive
parts.
4. O. visayense Britton, 1948, p.716, fig.2-5; Gonzalves 1981, P. 189, Fig.- 9.57.
Fig. 1D
Accession No.: JUH.-Bot.-11244; Sample No.: J16, J17, K3, U3

Habitat: Seasonal pond along the road side opposite petrol pump at Kuliyan, bouley at Kathua, ditches at
Samba, seasonal pond at Majalta.
Vegetative filament: Vegetative cells cylindrical, cell wall smooth, 4274 1014; basal cell elongate;
suffultary cells often inflated; Antheridia: Sub-epigynous to sub-hypogynous or scattered, single or multi seriate,
46 1618. Oogonium: Sub-depressed or sub-pyriform, smooth walled, 4042 3036, poriferous,
pore superior. Oospore: Globose to sub-globose, 1830 2030; three layered, outer layer & middle layer
smooth, inner layer slightly wavy.
Distribution: Philippines Island, India {Jammu & Kashmir- Kuliyan, Samba (Jammu); Majalta (Udhampur);
Kathua}.
Variations: O. visayense is characterized by having inflated suffultary cells and depressed globose oospore
(Gonzalves 1981). It closely resembled with the O. obsoletum, O. plusiosporum, O. tyrolicum, O. urbium and O.
varians in all characters except for inflated suffultary cells. The collected specimen has the dimensions within
the prescribed limit by Gonzalves (1981) with the narrow variations in the size of vegetative cells being slightly
longer and broader than the type specimen.
5. O. amplius (Tayl.) Tiff. Gonzalves 1981, p. 204205, f. 977.
Fig. 1E
Accession No.: JUH.-Bot.-11249; Sample No.: J 23

Habitat: Seasonal Pond formed after monsoon at Kuliyan.
Habit: Free Floating.
Vegetative filament: Vegetative cells cylindrical, 28.6 58.5 6.514.3; suffultory cells not
inflated, 48 14.3; Antheridia: Seriate, subepigynous, 48 1011. Oogonium: Depressed
globose to subpyriforme; 30.5351 3440.3, operculate, division superior. Oospore: Globose,
not filling the oogonium, 28.530.5 30.531.5.
Distribution: Canada, India {Jammu & Kashmir- Kuliyan (Jammu)}.
Variations: Characters of this specimen resembled to that of O. pyrulum, O. amplius and O.
pyriform but it seems to be more close to O. amplius than the other two especially in shape of
oogonia, dimensions of vegetative and reproductive org ans.The present specimen is slightly smaller
in size than the type specimen discussed by Gonzalves (1981).
6. O. cf capillare (L) Kutz. Novis 2003, p.344-345, fig. 5Q-V, 6A-C.
Fig. 1F G
Accession No.: JUH.-Bot.-11255; Sample No.: J30, U7

Habitat: Lake at Surinsar, pools formed along the side of slow moving stream at Ram Nagar.
35
Jitendra & Anand (2016) 3(1): 3339

.
Vegetative filaments: Vegetative cells cylindrical slightly capitellate, chloroplast starts degenerating, female
filament 2060 1224. Oogonium: Globose sub cylindrical, 5060 5060, poriferous, pore
superamedian. Oospore: Depressed globose to globose, not filling the oogonium, 3040 3040 .
Distribution: New Zealand, India {Jammu & Kashmir- Surinsar (Jammu)}.
Variations: The O. capillare complex consists of large forms. Oogonia slightly swollen than the vegetative cells.
In this case, although the males were abundant but oospores were rarely seen. It is possible that the oospores
observed were actually deformed or the infected vegetative cells. More female material required to identifiy the
material.
Figure 1. A, Oedogonium.subvaucherii Claass.; B, O. pseudofragile Claass.; C, O. upsaliense (Wittr.) Hirn; D, O.

visayense Britt.; E, O. amplius (Tayl.) Tiff.; FG, O. cf capillare (L) Kutz.
36
7. O. angustistomum Hoff Gonzalves 1981, p. 562, fig. 9507.
Jitendra & Anand (2016) 3(1): 3339

.
Fig. 2A B
Accession No.: JUH.-Bot.-11262; Sample No.: J 37

Habitat: Along the road side ditches at Pragwal wetland .
Habit: Epiphytic.
Vegetative filament: Vegetative cells cylindrical slightly capitellate, apically obtuse terminal cell,
elongate basal cell with bulbous bare, smooth walled, those of female filaments 70 -150 20-40;
those of male filament 60150 2026. Antheridia: Antherozoids seriate 1025, 48 2026.
Oogonium: Subglobose, smooth walled, poriferous, pore superior 60 90 5070. Oospore:
Globose to subglobose, smooth walled, 5080 4464 .
Distribution: Tennessee, India {Jammu & Kashmir- Pragwal (Jammu)}.
Variations: Characteristic apically obtuse terminal cell and long chain of antheridia up to 30 no.
distinguished this species. This species resembled O. angustistomum the type specimen described
earlier by Gonzalves (1981) in the morphology as well as in the dimensions of vegetative and
reproductive parts with the little variations.
8. O. magnusii Wittr. var. major Bock and Bock, Tiffany 1930, p.68, pl.XII, fig.115; Gemeinhardt 1939, p.95,
fig. 65; Tiffany and Britton 1952, p.61, fig.116; Gautheir- Lievre 1963, p.343, pl.55 fig. 89a; Gonzalves 1981,
325-327, fig. 206b.
Fig. 2C D
Accession No.: JUH.-Bot.-11265; Sample No.: R3, R4, R5, K6
Habitat: Small puddles formed after moonson at Jawahar Nagar , Small boli at Kathua, Small pools formed
along the side of slow moving stream at Thandapani.
Vegetative filament: Cylindrical and sometimes slightly capitellate, smooth walled; female vegetative filament
2850 1418; male vegetative filament 2040 68; suffultary cells inflated, suffultary cells 22
20u. Antheridia: Solitary sometimes seriate, 48 1012. Oogonium: Depressed globose, 2830 34
38, poriferous, pore supramedian. Oospore: Globose, 2628 2030, three layered, smooth walled.
Distribution: Morocco, Canada, Illinois, Massachusetts, Michigan, Finland, France, Germany, Poland, Sweden,
Yugoslavia, India {Jammu & Kashmir- Thandapani, Jawahar Nagar (Rajouri), Kathua}.
Variations: Dimensions of present specimen falls within the prescribed range for the type species described
earlier by Gonzalves (1981) & Tiffany and Britton (1952).
Figure 2. AB, Oedogonium. angustistomum Hoff.; CD, O. magnusii Wittr. var. major Bock and Bock.
37
Jitendra & Anand (2016) 3(1): 3339

.
AKNOWLEDGEMENTS
Thanks are due to Head, Department of Botany for providing the necessary facilities and valuable guidance
and one of us (PJ) is thankful to University of Jammu for financial assistance.
REFERENCES
Awasthi M, Dar DN & Singh RK (2006) Qualitative algal analysis the fish-gut: Tested in the rice fish cropping
system. International Journal of Environmental Science and Technology 3(1): 8994.
Bajpai O, Mishra S, Mohan N, Mohan J & Gupta RK (2013) Phyco chemical characteristics of Lakhna Devi
temple water tank, Lakhna, Bakewar, Etawah, U.P. with reference to Cynobacterial Diversity. International
Journal of Environment 1(1): 2028.
Bharti SG & Pai KM (1972) On the occurrence of some Oedogoniaceae in certain soils of Mysore State.
Journal of Karnataka University Science 17: 132139.
Britton ME (1948) New Species of Oedogonium from Leyte. The Philippine Chicago. Chicago
Gauthier-Lievre L (1963) Oedogoniaceae africianes. Nova Hedwigia 6: 1104.
Gauthier-Lievre L (1964) Oedogoniaceae africianes. Nova Hedwigia 7: 153272, 273481, 545588.
Gemeinhardt K (1939) Oedogoniales In: RABENHURSTs Kryptogamenflora Von Deutschland under
Schweiz. Akad. Verlags. M.B.H. Leipzig. 453 pp. Geitler, L. 1923. DerZellbau on Glavco-cystis
nostochinearum and Gloeochaete Wittrockiana etc. Arch. Protistenk. 47: 124.
Gonzalves EA (1981) Oedogoniales. ICAR, New Delhi.
Hirn KE (1900) Monographic and Iconographie der Oedogoniacen. Acta Societatis Scientiarum Fennicae 27: 1
395.
Kamat ND & Patel MZ (1973) Soil algae of a rice field at different depths. Botanique 4: 10106.
Kamat ND (1967) The algae of Mount Abu. Proceedings of Rajashthan Academy of Science 11: 4954.
Kone T & Teugels GG (2003) Food habits of brackish water tilapia Sarotherodon melanotheron in riverine and
lacustrine environments of a East African Coastal basin. Hydrobiolgia 490: 7585.
Kumar HD & Singh HN (1984) A Text book on Algae, Third Ed.: 242-243. East-West Press Pvt. Ltd., New
Delhi.
Mahato P, Das RN & Mahato AK (1998) New records of macrandous and monoecious species of Oedogonium
from Bihar, India. Phykos 37 (1&2): 115123.
Misra PK, Srivastava AK, Mehrotra RK & Singh SK (2002) Genus Oedogonium Link from North Eastern, Uttar
Pradesh. Geophytology 30 (1&2): 103109.
Novis P (2003) A taxonomic survey of Oedogonium (Oedgoniales, chlorophyta) in the South Island and
Chatham Islands, New Zealand. New Zealand Journal of Botany 41: 335358.
Olojo EAA, Olwin KB & Osikoya OJ (2003) Food and feeding habits of Synodontis nigrita from the Osum
River, SW Nigeria. NAGA, World fish centre Quarterly, 26(4): 2124.
Prasad BN & Misra PK (1992) Fresh water algal flora of Andaman and Nicobar Islands vol. II. Bishan Singh
and Mahendra Pal Singh, Dehradun, India.
Randhawa MS (1940) Perennation in Oedocladium operculatum Tiffany. Current Science 9: 326328.
Redondo PN, Figueroa G, Jarero, JR & Simeon RL (2006) In vitro analysis of the antibacterial activity of O.
capillare against Pathogenic bacteria in fish. Veterinaria Mxico 37(2): 209221.
Saha LC & Pandit B (1987) Pond and Riverine algae of Bhagalpur. Phykos 26: 152158.
38
Jitendra & Anand (2016) 3(1): 3339

.
Shukla AC (1971) Systematic description of algae from Panki rice fields, India. Revue Algologique 10(3): 257
270.
Shukla HM, Tiwari GL, Pandey UC & Habib I (1988) Oedogoniales of Mauri Lake, Pratapgarh (U.P.) India.
Proceedings of Indian Academy of Science (Plant Science) 98(6): 465470.
Silva P & Moe RL (2003) Oedogoniales. Mc Graw Hill Encyclopedia of Science & Technology online.
Srivastava N, Suseela MR & Toppo K (2014) Fresh water cyanobacteria of Sai River near Lucknow, Uttar
Pradesh. Tropical Plant Research 1(2): 1116.
Tiffany LH & Britton B (1952) Algae of Illinois. The University press of Chicago, Chicago.
Tiffany LH (1930) The Oedogoniaceae, a monograph including all the known species of the gener a
Bulbochaete. Oedocladium and Oedogonium. 253 pp. Columbus, Ohio.
39
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 4047, 2016
Research article
Effect of different growth stages on rice crop on soil

microbial and enzyme activities
N. F. Islam1* and Borthakur2
1
Department of Botany, N.N. Saikia College, Titabar-785630, Jorhat, Assam, India

2
Department of Botany, Gauhati University, Guwahati, Assam, India
*Corresponding Author: nazimforidislam@yahoo.co.in
Abstract: The influence of growth stages of rice on soil microbial biomass carbon (MBC), soil
microbial biomass nitrogen (MBN) and enzyme activities (amylase, dehydrogenase, alkaline and
acid phosphatase) in a sub-tropical rice field has been screened. The above soil parameters were
investigated at two soil depth (0-10 and 10-20 cm) and in five growth stages of rice crop, i.e. 30,
60, 90, 120 and 150 days after transplantation (DAT) of rice seedlings. Results shows that,
contents of soil organic carbon, total nitrogen, MBC and MBN were highly influenced by the
flowering stage (90 DAT) of the rice crop, at both 0-10 cm and 10-20 cm soil depths, but decline
gradually when the crop reaches maturity (120 DAT) and late maturity stages (150 DAT). The soil
organic carbon and total nitrogen were positively correlated with MBC and MBN. At both the
sampling depths soil amylase activity showed a peak value at seedling and flowering stages (30
and 90 DAT) whereas dehydrogenase and phosphatase showed at flowering stage (90 DAT). Both
dehydrogenase and phosphatase showed significant correlation with MBC and MBN.
Keywords: Oryza sativa - Soil microbial biomass - Soil enzyme activities.
[Cite as: Islam NF & Borthakur (2016) Effect of different growth stages on rice crop on soil
microbial and enzyme activities. Tropical Plant Research 3(1): 4047]
INTRODUCTION
Soil microbial biomass is the living component of soil organic matter. As organic matters are the preferred
energy source for the microorganisms, ecosystems with high organic substances tend to have higher microbial
biomass contents as well as its activities. Usually the highest microbial activity in soil takes place in the surface
horizon compared to deeper horizons (Januszek 2011). Most of the enzymatic transformations in soil are
accomplished by microbial biomass due to which a part of the organic materials are stabilized as humus and the
remaining carbon and other nutrients are utilized by microorganisms for their own growth (Anderson & Domsch
1980). Seasonal changes in soil moisture, temperature and available residue have a strong influence on soil
microbial biomass and its activity (Diaz-Ravina et al. 1995). Moreover, reduction in microbial biomass and
enzyme activities due to excessive cultivation practices have been reported earlier (Gupta & Germida 1988).
There is insufficient information on the dynamics of soil microbial biomass in sub-tropical rice field. On the
other hand estimation of soil microbial biomass alone couldnt indicate microbial activity in soil, hence study of
soil enzyme like dehydrogenase and phosphatase is essential to get a clear picture of soil microbial activities.
Soil enzymes are known to play an important role in the biochemical functioning of soils (Wang et al. 2011),
including organic residue decomposition (Lei 2011), cycling of nutrients (Makoi & Ndakidemi 2008), and
maintenance of soil structure (Dick et al. 1994, Balota et al. 2004). Their activity is controlled by many factors
such as soil physico-chemical properties (Amador et al. 1997), soil microbial community (Kourtev et al. 2002),
type of vegetation (Sinsabaugh et al. 2002) and ecological disturbances (Boerner et al. 2000). Soil enzymes may
originate from plants, animals or microbes and can exist in bound or free form within the soil. Soil
microorganisms produce quite a number of extra cellular enzymes to decompose the complex organic matter
before it is utilized as a source of energy (Lalitha & Santhaguru 2012). Soil enzymes are specific with respect to
the types of reactions they participate. For instance, amylase is a starch hydrolyzing enzyme which hydrolyzes
1-4D glucosidic linkage of amylase and amylopectin and consists of -amylase and -amylase. Although www.tropicalplantresearch.com
40
Islam & Borthakur (2016) 3(1): 4047

.
amylase is mainly synthesized by plants, studies indicate that -amylase is synthesized by plants, animals and
microorganisms (Pazur 1965). The microbial activity of soil, to a large extent, is dependent on the quantity of
available carbon, and is reflected by dehydrogenase activity (Januszek 2011). Dehydrogenase is involved in
biological oxidation of soil organic matter, and is responsible for dehydrogenation of organic matter by
transferring hydrogen and electrons from substrates to acceptors (Maurya et al. 2011). Phosphatase, originating
from microorganisms and root exudates, cleaves the phosphate from organic substrates and is involved in the P
cycle in soil (Huang et al. 2011). Thus, microbial biomass and soil enzyme activities can potentially provide an
integrated biological assessment of soil quality (Chhotaray et al. 2011). Evidence suggests that plant species
have significant impacts not only on soil physicochemical properties but also more directly on the composition
of soil microbial community and their activities (Ushio et al. 2008, Ushio et al. 2010). Moreover, rhizosphere
zone of plants have profound effect on microbial population and activities (Viyas & Gupta 2014). Unlike other
terrestrial ecosystems, relatively less effort has been made to elucidate possible influence of growth stages of
rice crop on belowground processes in a sub-tropical rice ecosystem. Moreover, limited information is available
on the depth wise changes in microbial biomass and soil enzyme activity of agro-ecosystems especially in the
widely prevalent rice based cropping system of sub-tropical region.
So, the objectives of the present investigation were to determine the influence of different growth stages of
rice on soil microbial biomass and enzyme activities from different soil depth in a subtropical rice field
conditions, as well as to establish the relation between soil chemical parameters with microbial biomass and
enzyme activities.
Site description
The study was carried out in Jalukbari experimental field, Guwahati, Assam. The area is situated in south of
the river Brahmaputra (2612 N, 9150 E) with an average altitude of 54 m (a.s.l). The soils of the
experimental site are of old alluvial type and sandy loam in texture (Table 1). The climate of the area is
monsoonal, characterized by long rainy season (MaySeptember) and dry and cold winter (November
February). The average annual rainfall was about 1782 mm during the study period, with highest rainfall
(315.5mm) in the month of September (Fig. 1). The hottest month of the year was August with a mean
maximum temperature of 33.0C and the coldest month was January with a mean minimum temperature of
9.9C. Maximum relative humidity recorded during the study period was 85% in the month of July.
Table 1. Physical characteristics of soil.
Characteristics
Soil test values
Coarse sand (%)

5.49
Fine sand (%)

61.00
Silt (%)
21.10
Clay (%)
14.98
Textural class
Sandy loam
Figure 1. Mean monthly rainfall (mm), temperature (C) and relative humidity (%) during the
cropping season in the study area.
Soil sampling and physico-chemical analysis

Soil samples were collected from the experimental field at 30, 60, 90, 120, 150 days after transplanting
(DAT) of rice seedlings during the cropping season. At each sampling date, five soil samples were randomly
41

.
collected from 010 and 020 cm depths using a soil corer. Collected samples from each depth were then put
separately into sterilized plastic bags, tied to prevent moisture loss and transferred to laboratory as soon as
possible for estimation of microbial biomass and enzyme activities or kept at 4C until analysis for one week.
The soil samples were mixed thoroughly to obtain a homogeneous sample, and sieved through a 2 mm sieve.
Mechanical fractions were determined according to the standard procedure (Bouyoucos 1962). The pH of the
soil samples was recorded on a digital pH meter in 1:5 (w/v) soil-water suspensions. Soil organic carbon was
analysed using rapid titration method of Walkley & Black (1934), as described by Jackson (1973). Total
nitrogen was estimated by Kjeldahl method (Jackson, 1973).
Estimation of microbial biomass and soil enzyme activities
Microbial biomass carbon (MBC) and microbial biomass nitrogen (MBN) were determined by fumigationextraction method (Brookes et al. 1985, Vance et al. 1987). Enzyme activities like amylase is assayed by as per
method of Kelly & Rodriquez (1975) and estimation of the reducing sugars released was estimated using the
DNS method (Gascoigne & Gascoigne 1960). Dehydrogenase activity was assayed under standard conditions by
the method of reduction of 2,3,5-triphenyltetrazolium chloride (TTC) to triphenylformazan (TPF) (Casida 1977)
and acid phosphatase and alkaline phosphatase were assayed as per the method of Tabatabai & Bremner (1969).
Statistical analysis
Statistical analysis was performed with SPSS program Version 17. Regression analyses and Pearson
correlations were used to determine the interrelationships among the measured soil properties.
RESULTS
Figure 2. (AD): Chemical characteristics of the soil samples collected from 010 and 1020 cm depths at 30, 60, 90, 120
and 150 days after transplanting rice seedlings: A, pH; B, total organic carbon (Corg); C, total nitrogen (Ntot); D, C/N ratio;
(EH): Changes in the values of soil microbial biomass: E, Biomass-C; F, Bomass-N; G, Biomass-C/Corg (%); H, BiomassN/ Ntot (%).
42

.
The basic chemical properties of the soils showed a range of soil conditions in the experimental site (Fig.
2AD). Soil pH ranged from 5.1 to 5.7 and 5.3 to 6.0 in 010 and 1020 cm, respectively. The concentration of
soil organic carbon and total nitrogen in 010 cm layer was higher than that in 1020 cm layer. At 90 DAT
when the rice crops attain flowering stage, high organic carbon (0.90% and 0.60%) and total nitrogen (0.10%
and 0.07%) levels were recorded in both the depth as compared to the preceding stages. Soil organic carbon
showed high correlations with microbial biomass carbon, microbial biomass nitrogen, dehydrogenase and
phosphatase activity (Table 2.). However, no such correlations were observed in case of total nitrogen. The C/N
ratio was high at 30 DAT (14.8 and 14.6) i.e. when the rice crop was at seedling stage.
Table 2. Pearsons linear correlation coefficients (r) between soil chemical properties, microbial biomass, and soil enzymes.
Variables
A
B
C
D
E
F
G
1.000
0.650
0.307
0.384
0.698
0.451
0.688
pH (A)
1.000
0.863
0.939* 0.951* 0.348
0.934*
Organic carbon (B)
1.000
0.920* 0.818
0.174
0.748
Total nitrogen (C)
1.000
0.910
0.384
0.900
Microbial biomass carbon (D)
1.000
0.603
0.988**
Microbial biomass nitrogen (E)
1.000
0.658
Amylase (F)
1.000
Dehydrogenase (G)
Acid Phosphatase (H)
Alkaline phosphatase (I)
Note: ** & * Correlation is significant at the 0.01 & 0.05 level respectively (2-tailed), n = 5.
H
0.420
0.947*
0.834
0.951*
0.840
0.209
0.847
1.000
I
0.540
0.981**
0.851
0.939*
0.881*
0.216
0.873
0.987**
1.000
Figure 3. Relationships between: A, organic-C and biomass-C; B, organic-C and biomass-N; C, total-N and biomassC; D, total-N and biomass-N.
Soil MBC and MBN contents were shown in figure 2EH and the correlations with soil parameters are
shown in table 2. MBC content varied from 249 to 317 mg kg-1dry soil in 010 cm and 193 to 298 mg kg-1dry
soil in 1020 cm depth, highest being at the flowering stage (90 DAT). Similarly, MBN varied from 32 to 42
mg kg-1dry soil and 29 to 38 mg kg-1dry soil in 010 and 1020 cm soil depth respectively, with highest value at
the flowering stage. Thus the soil organic carbon, total nitrogen MBC and MBN content were higher in surface
soil layer (010cm) compared to sub-surface soil layer. According to earlier findings (Wang et al., 2011) the
43

.
value of MBC and Corg decrease with soil depth. These results are in consistent with our present findings.
Moreover, their concentrations were highest at 90 DAT (flowering stage) and lowest in 150 DAT (late maturity
stage). This may be attributed to increase in the root exudates in the flowering stage leading to more intense
microbial activity which gradually decline when the crop attained maturity to late maturity stages. High soil
microbial biomass during flowering stage may also be attributed to sufficient moisture availability due to rainy
summer period. The result is in consistence with earlier researcher (Singh et al., 1989). There was a positive
correlation of soil organic carbon and MBC (R2 = 0.66, P< 0.05) or MBN (R2 = 0.73, P< 0.05, and of total
nitrogen and MBC (R2 = 0.88, P< 0.05) or MBN (R2 = 0.79, P< 0.05) in both the soil depth (Fig. 3). These
relationships were in consistent with many studies, where soils with high organic matter content supported high
levels of microbial biomass (Brookes et al. 1985, Wardle 1992, Bauhus & Khanna 1999, Friedel et al. 2006).
Microbial C/N ratio ranged from 7.2 to 7.9 in 010 cm layer and 6.4 to 8.8 in 1020 cm layer. Witt et al. (2000)
reported microbial C/N ratio of 3.3 to 20.7 in Philippines rice soil. Inubushi et al. (1991) obtained unusually
wide microbial C/N ratios of 912 in three waterlogged Japanese rice soils. Microbial quotient (MBC/ total
organic-C and MBN/total N) has been reported as a useful indicator of changing soil processes and soil quality
than microbial biomass or total organic matter alone (Anderson & Domsch 1989, Sparling 1992). In our study
MBC/ organic-C varied from 3.3 to 3.8% and 3.6 to 4.9%, and MBN/total-N varied from 4.2 to 4.6 % and 5.2 to
9.6% in 010 and 1020 cm depth respectively. Sparling (1997) suggested that if a soil is being used
exploitively, microbial C pools will generally decline at a faster rate than total organic matter, and microbial
quotient will decrease accordingly. Depending on the nutrient status and soil management, metabolic quotient
values below 2.0% are regarded as critical (Anderson 2003). Soil pH is generally considered as one of the
important regulator of microbial activity. In the present study total microbial biomass was lower under acidic
soil conditions and with the increase in soil pH the microbial quotient was found to increase. The result is in
agreement with the findings of Anderson (2003). Thus, seasonal variations, plant developmental stage, organic
matter content and soil depth have been found to have a great influence on soil microbial biomass (Yang et al.
2010).
Figure 4. Changes in the enzyme activities of the soil samples collected from 010 and 1020 cm depths at 30, 60, 90, 120
and 150 days after transplanting rice seedlings: A, amylase activity; B, dehydrogenase activity; C, alkaline phosphatase
activity; D, acid phosphatase activity.
44

.
Figure 4 reveals soil enzyme activities at different stages of plant growth and in two different soil depths.
The activity was more in 010 cm than 1020 cm depth. The progressive decline in amylase activity with
increasing soil depth in rice agroecosystem has been reported earlier (Chhotaray et al. 2011). The amylase
activity increased with the increase in the rice growth and declined at late maturity stage. The activity was more
in the soils collected at 30 DAT (4280.21 and 2543.67 mg glucose kg-1 soil d-1) and 90 DAT (4390.25 and
2130.63 mg glucose kg-1 soil d-1). At 150 DAT the level of activity declined (3024.43 and 1121.45 mg glucose
kg-1 soil d-1) in the two soil depth. The dehydrogenase activity of the soil ranged from 315.10 mg TPF kg -1 soil
d-1 to 572.95 mg TPF kg-1 soil d-1 in 0-10 cm depth and from 124.25 mg TPF kg-1 soil d-1 to 332.56 mg TPF kg-1
soil d-1 in 1020 cm depth. There was progressive decrease in the dehydrogenase activity from 90 DAT to 150
DAT (flowering to late maturity stage of the rice). The higher dehydrogenase activity at 90 DAT was likely due
to high C input in the soil in the form of root mass that enhanced microbial activity. The relation between
dehydrogenase activity and C inputs has been well established (Maurya et al. 2011). Phosphatase activity
(alkaline and acid phosphatase) was maximum during flowering stage in both 010 cm (260.64 and 314.49 mg
PNP kg-1 soil d-1) and 1020 cm (183.45 and 269.63 mg PNP kg-1 soil d-1) depth. High phosphatase activity
during flowering stages may be due to faster growth rate of plants and microbes owing to wet rainy period. As
because, heavy rain in wet season often leads to nutrient loss and low available P detected in the wet season
intensifies the phosphatase activity to meet the increasing P demand by plant and microbes growth (Huang et al.
2011). Both dehydrogenase and phosphatase showed significant correlation with MBC and MBN but no such
correlation was showed by amylase. In our study no significant correlation between enzyme activities and soil
pH were found. This may be due to the narrow range of pH values, 5.16.0 (Balota et al. 2004).
It is evident from the present study that soils collected at 90 DAT showed significantly higher enzymatic
activities compared to those collected from other growth stages. At 90 DAT rice roots might have secreted more
organic acid and carbohydrate, which stimulated higher soil enzymatic activities. The results corroborate with
the findings of Zeng et al. (2005), Wang et al. (2011). Moreover, microbial biomass and enzyme activities were
greater in the surface layer (010 cm). This is due greater nutrient availability in the surface layer which
enhanced microbial activity (Ralte et al. 2005).
CONCLUSIONS
Increase in microbial biomass and enzyme activities indicates high rate of release of nutrients by rice crops
which facilitate microbial activities. In addition, varying soil depth to certain extent is expected to influence
increase in bioactivity. However, these findings are not sufficient to get a clear picture of the whole microbial
activities in sub-tropical rice ecosystem, because present results were obtained from a short term study. Detail
field level studies are necessary to expand the knowledge in this aspect.
ACKNOWLEDGEMENTS
We are thankful to Head of the Botany Department, Gauhati University, Guwahati, Assam, for providing us
all the necessary facilities required for completion of present work.
REFERENCES
Amador JA, Glucksman AM, Lyons JB & Gorres JH (1997) Spatial distribution of soil phosphatase activity
within a riparian forest. Soil Science 162: 808825.
Anderson TH & Domsch KH (1989) Ratios of microbial biomass carbon to total organic-C in arable soils. Soil
Biology and Biochemistry 21: 471479.
Anderson TH (2003) Microbial eco-physiological indicators to assess soil quality. Agriculture Ecosystem and
Environment 98: 295293.
Balota EL, Kanashiro M, Filho AC, Andrade DS & Dick RP (2004) Soil enzyme activities under long term
tillage and crop rotation systems in subtropical agro-ecosystems. Brazilian Journal of Microbiology 35:
300306.
Bauhus J & Khanna PK (1999) The significance of microbial biomass in forest soils. In: Rastin N & Bauhus J
(ed) Going underground: ecological studies in forest soils. Research Signpost, Trivandrum, pp. 77110.
Boerner REJ, Decker KLM & Sutherland EK (2000) Prescribed burning effects on soil enzyme activity in a
southern Ohio hardwood forest: a landscape-scale analysis. Soil Biology and Biochemistry 32: 899908.
45

.
Bouyoucos GJ (1962) Hydrometer method improved for making particle size analysis of soils. Agronomy
Journal 54: 464465.
Brookes PC, Landman A, Pruden G & Jenkinson DS (1985) Chloroform fumigation and release of soil N: a
rapid direct extraction method to measure microbial biomass N in soil. Soil Biology and Biochemistry 17:
837842.
Casida LE (1997) Microbial metabolic activity in soil as measured by dehydrogenase determination. Applied
and Environmental Microbiology 34: 630636.
Chhotaray D, Mohapatra PK & Mishra CS (2011) Farm management to control of soil microbial density and
metabolic activities in rice-rice agroecosystem. International Journal of Microbial Research 2: 8692.
Diaz-ravina M, Acea MJ & Carballas T (1995) Seasonal changes in microbial biomass and nutrient flush in
forest soils. Biology and Fertility of Soils 19: 220226.
Dick RP, Sandor JA & Eash NS (1994) Soil enzyme activities after 1500 years of terrace agriculture in the
Colca Valley, Peru. Agricultural Ecosystem and Environment 50: 123131.
Friedel JK, Ehrmann O & Preffer M (2006) Soil microbial biomass and activity: the effect of site characteristics
in humid temperate forest ecosystems. Journal of Plant Nutrition and Soil Science 169: 175184.
Gascoigne JA & Gascoigne MM (1960) Biological degradation of cellulose. Butterworths Publication Co.
London.
Gupta VVSR & Germida JJ. (1988) Distribution of microbial biomass and its activity in different soil aggregate
size classes as affected by cultivation. Soil Biology and Biochemistry 20: 777786.
Huang W, Liu J, Zhou G, Zhang D & Deng Q (2011) Effects of precipitation on soil acid phosphatase activity in
three succession forests in Southern China. Biogeosciences 8: 19011910.
Inubushi K, Brookes PC & Jenkinson DS (1991) Soil microbial biomass C, N and ninhydrin-N in aerobic and
anaerobic soils measured by the fumigation-extraction method. Soil Biology and Biochemistry 23: 737741.
Jackson ML (1973) Soil Chemical Analyses. Prantice Hall of India, New Delhi, India, 485 p.
Januszek K (2011) The enzyme activity of the forest soils of Southern Poland as a measure of soil quality.
Electronic
Journal
of
Polish
Agricultural
Universities.
Available
from:
http://www.ejpau.media.pl/volume14/issue2/art-01.html (accessed: 20 Jan. 2014).
Kelley WD & Rodriguez-khabana R (1975) Effects of potassium azide on soil microbial population and soil
enzymatic activities. Canadian Journal of Microbiology 21: 565570.
Kourtev PS, Ehrenfeld JG & Haggblom M (2002) Exotic plant species alter the microbial community structure
and function in the soil. Ecology 83: 31523166.
Lalitha S & Santhaguru K (2012) Improving soil physical properties and effect on tree legume seedlings growth
under barren soil. Agricultural Science Research Journal 2: 126130.
Lei T (2011) Understanding soil organic matter mineralization in agro ecosystems:Soil enzyme perspectives,
Ph.D Thesis. North Carolina State University, United States.
Makoi JHJR & Ndakidemi PA (2008) Soil enzymes: Examples of their potential roles in the ecosystem. African
Journal of Biotechnology 7: 181191.
Maurya BR, Singh V & Dhyani PP (2011) Enzymatic activities and microbial population in agric-soils of
Almora District of Central Himalaya as influenced by altitudes. International Journal of Soil Science 6:
238248.
Pazur JH (1965) Enzymes in the synthesis and hydrolysis of starch. In: Whistler R & Paschall EF (eds) StarchChemistry and Technology, Fundamental aspects. Academic Press, New York, U.S.A., pp. 133175.
Ralte V, Pandey HN, Barik SK, Tripathi RS & Prabhu SD (2005) Changes in microbial biomass and activity in
relation to shifting cultivation and horticultural practices in subtropical evergreen forest ecosystem of northeast India. Acta Oecologica 28: 163172.
Singh JS, Raghubanshi AS, Singh RS & Srivastava SC (1989) Microbial biomass acts as a source of plant
nutrients in dry tropical forest and savanna. Nature 338: 499500.
Sinsabaugh RL, Carreiro MM & Repert DA (2002) Allocation of extracellular enzymatic activity in relation to
litter composition, N deposition and mass loss. Biogeochemistry 60: 124.
Sparling GP (1992) Ratios of microbial biomass carbon to soil organic carbon as an indicator of changes in soil
organic matter. Australian Journal of Soil Research 30: 195207.
46

.
Sparling GP (1997) Soil microbial biomass, activity and nutrient cycling as indicators of soil health. In:
Pankhurst CE, Doube BM & Gupta V (eds) Biological indicators of soil health. CAB International,
Wallingford, U.S.A., pp. 97119.
Tabatabai MA & Bremner JM (1969) Use of p-nitrophenyl phosphate for assay of soil phosphatase activity. Soil
Biology and Biochemistry 1: 301307.
Ushio M, Kitayama K & Balser TC (2010) Tree species effects on soil enzyme activities through effects on soil
physicochemical and microbial properties in a tropical montane forest on Mt. Kinabalu, Borneo.
Pedobiologica 53: 227233.
Ushio M, Wagai R, Balser TC & Kitayama K (2008) Variations in the soil microbial community composition of
a tropical montane forest ecosystem: does tree species matter? Soil Biology and Biochemistry 40: 2699
2702.
Vance ED, Brookes PC & Jenkinson DS (1987) An extraction method for measuring soil microbial biomass C.
Soil Biology and Biochemistry 19: 697702.
Vyas D & Gupta RK (2014) Effect of edaphic factors on the diversity of VAM fungi. Tropical Plant Research
1(1):1425.
Walkley A & Black IA (1934) An examination of Degtjareff method for determining soil organic matter and a
proposed modification of the chromic acid titration method. Soil Science 37: 2937.
Wang C, Wang G, Liu W & Wu P (2011) The effects of plant-soil-enzyme interactions on plant composition,
biomass and diversity of Alpine meadow in the Qinghai-Tibetan Plateau. International Journal of Ecology
2011: 110.
Wardle DA (1992) A comparative assessment of factors which influence microbial biomass carbon and nitrogen
levels in soil. Biological Review 67: 321358.
Witt C, Gaunt JL, Galicia CC, Ottow JCG & Neue HU (2000) A rapid chloroform-fumigation extraction method
for measuring soil microbial biomass carbon and nitrogen in flooded rice soils. Biology and Fertility of Soils
30: 510519.
Yang K, Jiaojun Z, Min Z, Qiaoling Y & Osbert JS (2011) Soil microbial biomass carbon and nitrogen in forest
ecosystems of Northeast China: a comparison between natural secondary forest and larch plantation. Journal
of Plant Ecology 3: 175182.
Zeng L, Liao M, Chen C & Huang C (2005) Variation of soil microbial biomass and enzyme activities at
different growth stages of rice (Oryza sativa). Rice Science 12: 283288.
47
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 4851, 2016
Research article
Phytochemical screening and antifungal activities of

five plant species
Edward N. Okey* and Idoroenyin E. Asuqwo
Department of Biological Sciences, Akwa Ibom State University, Ikot Akpaden, Mkpat Enin LGA, Nigeria
*Corresponding Author: nsofang2008@yahoo.com
Abstract: Aspergillus niger and Rhizopus oryzae are two common pathogens of post-harvest fruits
diseases in Mkpat Enin area of Akwa Ibom State in Nigeria. Traditional control measures of these
pathogens have emphasized the use of agro-chemicals with high environmental toxicity. The
search for alternative control methods using local plant products which are environmentally
friendly is therefore, necessary. Antifungal properties of leave extracts of five plant species;
Mimosa pudica, Phyllanthus amarus, Emilia sonchifolia, Bryophyllum pinnatum and Amaranthus
lhybridus leave extracts were investigated using disc diffusion method while screening for
bioactive compounds was carried out using standard phytochemical procedures. Antifungal
activities were tested against the two pathogens at 2%, 4%, 6%, 8% and 10% concentrations. All
extracts indicated significant growth inhibition on the two pathogens with 10% recording the
highest. Phytochemical screening showed the presence of tannin in all the plant extracts tested
while saponin was found in all except Bryophyllum pinnatum extract. Flavonoids were present in
all excluding Bryophyllum pinnatum extracts. Leave extracts of the tested plants can be explored
as potential crop disease control measures being natural products which are less hazardous.
Keywords: Plant extracts - In-vitro analysis - Diseases.
[Cite as: Okey EN & Asuqwo IE (2016) Phytochemical screening and antifungal activities of five plant species.
INTRODUCTION
Although the screening of plants for bioactive agents is one of the most intensive areas of natural product
research currently, the field is far from being exhausted. Only around ten percent of plants have been
investigated (Sandberg & Bruhn 1979). Also, the WHO (2010) report noted that although, traditional medicines
represent 60% of the world primary health care system, the plant species with possible biological activities
remain largely unexplored. Natural products play a significant role in the drug discovery and development
process (Newman & Cragg 2012, Mehra et al. 2014, Truven 2015, Bajpai et al. 2015). There are no doubts that
plants represent an unlimited source of novel chemical entities with potential as drug leads.
While the extraction of natural products from plants may appear simple, Sticher (2008) reported that, the
presence of numerous inactive components makes the isolation and screening of target compounds extremely
cumbersome. It is therefore, necessary to modify existing protocols and develop new ones for effective
screening. Consequently, choosing plants for scientific evaluation of biological activities will depend on a
number of criteria such as ethno-pharmacological usage by local population, species availability, mode of
preparation and administration (Van der Watt & Pretorious 2001). A number of secondary products such as
tannins, phenolics, flavonoids, alkaloids, essential oils, steroids, saponins have been reported as possessing antimicrobial activities (Liu 2003, Akindele & Adeyemi 2007, Okey 2014)
The objective of this study is therefore, to evaluate five indigenous plant species (Emilia sonchifolia,
Mimosa pudica, Phyllanthus amarus, Bryophyllum pinnatum and Amaranthus hybridus) for their bioactive
properties through phytochemical screening and invitro assays on fungal pathogens. These plant species are
selected for their availability and associated traditional healing properties.
Received: 23 December 2015
48
Okey & Asuqwo (2016) 3(1): 4851

.
Collection and identification of plant materials
Five plant species were collected from the Botanic garden of the Department of Biological Sciences, Akwa
Ibom State University and identified by taxonomic characters as Emilia sonchifolia (Red tassel flower), Mimosa
pudica (Sensitive plant), Phyllanthus amarus (Stone-breaker), Bryophyllum pinnatum (Life plant) and
Amaranthus hybridus (Female finger). These plant species were selected based on availability and historic
medicinal applications.
Preparation of Plant Extracts
Fresh leaves were collected from each species, washed thoroughly and air-dried at room temperature for one
week. These were separately triturated into fine powder in a mortar and pestle. The powders were stored
separately in sterile containers, kept away from light and moisture for further analysis. 5g of each powder leaves
were then treated with 500 ml of hexane and allowed to stand for 14 days at room temperature with frequent
agitation. The extracts were filtered using Whatman filter paper No 1 and were then concentrated in a water bath
at 50oC. These were subsequently brought to dryness at 60oC in an oven. The crude extracts were then
phytochemically screened for the presence of flavonoids, tannins and saponins, using modified protocols of
Edeoga et al. (2005).
Bioassay
The effects of leaf extracts on two fungal pathogens were assessed based on zone of inhibition. In vitro
antifungal activity was conducted on Potato Dextrose Agar (PDA) plates. Plates of Potato Dextrose Agar
(PDA) were prepared following standard procedures. 3-day old fungal cultures were used in this experiment.
0.5 ml of each extract at 2%, 4%, 6%, 8% and 10% concentrations were separately added to sterile filter paper
disc measuring 5mmin diameter. The plates were incubated at room temperature for 72 hours and the activities
of the extracts were determined by measuring the diameter of zone of inhibition. For each plate standard
antibiotic ketoconazole (10g ml-1) was used as control and each experiment replicated three times.
Statistical Analysis
Data was subjected to ANOVA (Analysis of Variance) to determine the significance of treatment effect.
Phytochemical screening
Phytochemical screening of plant extracts revealed the presence of favonoids, saponins and tannins (Table
1). Tannin was present in all the plant extracts tested while saponin was found in all except A. hybridus extract.
Flavonoids were present in all except P. amarus and B. pinnatum extracts. Results also showed that E.
sonchifolia and M. pudica contained all three constituents tested. These constituents have been reported in
several plant species including those presently tested in different parts of the world (Vander & Pretorious 2001,
Okwu & Josiah 2006).
Table 1. Phytochemical constituents in five plant species leaf extracts.
Plant Species
Phyllantus amarus
Emilia sonchifolia
Mimusa pudica
Bryophyllum pinnatum
Amaranthus hybridus
Note: + = Present; - = Absent.
Saponin
+
+
+
+
-
Constituents
Tannin
+
+
+
+
+
Flavonoids
+
+
+
These compounds have also been implicated in the control of both animals and plant diseases such as;
anticonvulsant (Asije et al. 2006), anti-inflammation (Akindele & Adeyenmi 2007), hepatoxicity (Adeneye et
al. 2006). Thus, the antifungal activities of the extracts reported in this study could be associated with the
presence of favonoids, tannins, and saponins. It is therefore, not surprising why these plants have been used by
both traditional healers and contemporary medicine (Liu 2003). The fact that all plant species tested did not
contain the same constituents implies different levels of antifungal activities. Hence, the need for screening to
determine most efficient plant species.
49
Okey & Asuqwo (2016) 3(1): 4851

.
Effect of plant extracts of Pathogenic fungi
Results of the effect of five plant extracts indicate growth inhibition of A. niger at different concentrations
(Table 2). The level of inhibition significantly increased with increase in concentration, 10% being the highest
in all the extracts tested. Inhibitory effects of E. sonchifolia and M. pudica extracts were the highest at 10%
concentration and were not significantly different from those of the control (ketoconazole). Extracts of P.
amarus recorded the least inhibition at 10% concentration. The significant high inhibitory effects of E.
sonchifolia and M. pudica could be associated with the fact that these extracts contained all the constituents
identified compared to the other extracts tested (Table 1). These results imply that all extracts tested can be
employed as biocontrol agents against A. niger, although, E. sonchifolia and M. pudica more recommended.
Table 2. Antifungal activities of extracts on Aspergillus niger.
Zone of inhibition at different concentrations (cm)

2%
4%
6%
8%
Phyllantus amarus
5.0a0.00
6.0a0.00
8.0b0.00
10.0c0.00
Emilia sonchifolia
6.5a0.50
12.0d1.00
15.0e1.00
17.0f0.70
a
d
e
Mimusa pudica
6.8 0.70
11.5 0.80
14.6 0.00
16.9f0.60
a
b
b
Bryophyllum pinnatum
5.6 0.10
9.0 0.00
9.2 0.60
11.2c0.30
Amaranthus hybridus
6.0a0.10
9.0b0.10
10.5c1.00
13.5d0.50
a
d
e
Control
6.7 0.00
12.8 0.00
15.5 0.60
17.2f0.00
Note: Different letters are significantly different (p<0.05).
Plant Extracts
10%
14.6e0.00
18.6g5.10
16.9f0.60
17.5f0.00
17.4f0.00
19.9g0.00
Table 3. Antifungal activities of extracts on Rhizopus oryzae.
Zone of inhibition at different concentrations (cm)

2%
4%
6%
8%
Phyllantus amarus
5.0a0.00
6.5b1.00
8.6 c 1.00
10.0 d0.00
c
e
f
Emilia sonchifolia
7.9 0.00
12.8 0.70
16.0 0.00
18.0 g0.40
c
e
f
Mimusa pudica
8.1 0.00
13.0 0.00
16.2 0.40
17.6 g0.20
Bryophyllum pinnatum 5.0 a0.00
6.0 b0.00
8.5 c 0.60
10.0 d0.00
a
b
d
Amaranthus hybridus
5.0 0.00
7.0 1.00
10.0 0.00
16.0 f0.50
c
e
f
Control
8.0 0.00
13.5 0.40
16.3 0.20
18.2 g0.50
Note: Different letters are significantly different (p<0.05).
Plant Extracts
10%
17.0 f0.50
19.6h 0.30
19.7 h0.70
18.0 g0.00
18.4 g0.30
20.0h0.00
Results of growth inhibition effects of the extracts on R. oryzae are shown in table 3. The trends were
generally similar to those recorded on A. niger with 10% concentration having the highest inhibition in all the
extracts tested. Extracts of E. sonchifolia, M. pudica, were significantly higher than the others tested. These
results support previous reports by (Asije et al. 2006, Akindele & Adeyenmi 2007, Adeneye et al. 2006,
Agrawal et al. 2004). There is collaborative evidence that the antifungal activities of the tested extracts are
associated with the presence of saponin, tannin and flavonoids. Formulations of these plants can therefore, be
recommended for both plant and animal disease control. The advantages of these plants as biocontrol measures
are enormous. Apart from being environmentally friendly, these plants are common weeds that can easily be
obtained and hence reducing the cost burden.
CONCLUSIONS
Amaranthus hybridus and Mimosa pudica, are the most recommended plant species to develop
pharmaceutical products against plant and animal diseases, while Phyllanthus amarus, Bryophyllum pinnatum
and Emilia sonchifolia can be further processed in this regard.
ACKNOWLEDGEMENTS
Authors are thankful to Head, Department of Biological Sciences, Akwa Ibom State University, Ikot
Akpaden, Mkpat Enin LGA, Nigeria for providing the necessary facilities required to conduct the present study.
REFERENCES
Adeneye AA, Benebo AS & Agbaje EO (2006) Protective effect of Aqueous leaf and seed extracts of
Phyllantus amarus on Alcohol-induced hepatoxicity in rats. West African Journal of Pharmacology and
Drug Research 22: 4250.
50
Okey & Asuqwo (2016) 3(1): 4851

.
Agraral A, Umarani D & Cimanga RK (2004) Evaluation of the inhibitory effect of the plant Phyllantus amarus
against deematophyphytic fungi Microsporum gypseum. Environmental Science 17: 359365.
Akindele AJ & Adeyemi OO (2007) Anti inflammatory activity of the aqueous leaf extracts of Brysocarpus
coccineus. Filoterapia 78: 2528.
Asije O, Adelusi SA & Usifofoh CO (2006) Anticonvulsant activity of Emilia sonchifolia in mice and chicks.
Pakistan Journal of Scientific and Industrial Research 49 (4): 269275.
Bajpai O, Pandey J & Chaudhary LB (2015) Ethnomedical uses of tree species by Tharu tribes in the Himalayan
Terai region of India. Research Journal of Medicinal Plant 10 (1): 1941.
Edeoga HO, Okwu DE & Mbaebie BO (2005) Phytochemical constituents of some Nigerian medicinal plants.
African Journal of Biotechnology 4(7): 685688.
Liu RH (2003) Health benefits of fruits and vegetables are from additive and synergistic combination of
phytochemicals. American Journal of Clinical Nutrition 78: 517S520S.
Mehra A, Bajpai O & Joshi H (2014). Diversity, utilizationand sacred values of ethno-medicinal in Kumaun
Himalaya. Tropical Plant Research 1(3): 8086.
Newman DJ & Cragg GM (2012) Natural products as sources of new drugs over the 30 years from 1981-2010.
Journal of Natural products 75: 311355.
Okwu DE & Josiah C (2006) Evaluation of the chemical composition of two Nigerian medicinal plants. African
Journal of Biotechnology 5(4): 357361.
Sandberg F & Bruhn JG (1979) Screening of plants for biologically active substances. In: Soforowo EA (ed)
African medicinal plants. University of Ife Press, Lagos, pp. 119.
Sticher O (2008) Natural product isolation. Natural Product Report 25: 517554.
Truyen DM, Mausor M & Ruddin AS (2015) A note on Aroids ethnobotany in the Hau River, Vietnam.
Tropical Plant Research 2(1): 5863.
Vander Watt E & Pretorious JC (2001) Purification and identification of active antibacterial components in
Carpobrotus edulis L. Journal of Ethnopharmacology 76: 8791.
WHO (2010) AFRO, African Traditional Medicine Day, The African Health Monitor. Special Series.
51
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 5266, 2016
Research article
Leaf traits and foliar herbivory in tropical dry evergreen

forest of India
K. Anil and N. Parthasarathy*
Department of Ecology and Environmental Sciences, Pondicherry University, Puducherry 605014, India
*Corresponding Author: parthapu@yahoo.com
Abstract: We investigated leaf traits of 110 plant species and the seasonal patterns of leaf damage
by diverse foliar herbivores in tropical dry evergreen forest (TDEF) on the Coromandel Coast of
India. The leaves of 110 plant species of TDEF are consumed by fifty-four species of foliar
herbivores that includes beetles, larvae and grasshoppers. Mean leaf damage ranged from 1.8% to
21% during the study period (201214). The mean leaf damage varied by season with a high value
of 16.1910.44% in monsoon, 9.666.66 % in pre-monsoon, 5.244.10 % in post-monsoon and
1.971.52 % in summer. Among tree species, Memecylon umbellatum showed maximum leaf
damage and among liana species, Combretum albidum suffered maximum leaf damage.
Information on different leaf resource users and their food-plants provide an insight into the
complex web of forest biotic interactions and such data will be valuable for biological
conservation.
Keywords: Foliar herbivores - Leaf damage - Leaf traits - Seasonal variation - Tropical dry
evergreen forest.
[Cite as: Anil & Parthasarathy (2016) Leaf traits and foliar herbivory in tropical dry evergreen forest of India.
INTRODUCTION
Herbivory among plant species can greatly affect the functioning of forest ecosystems (Schuldt et al. 2012)
and might play an important role in structuring tropical forests (Wright 2007, Viola et al. 2010). Different
studies found that herbivory differed among species (Barone 1998, Marquis et al. 2001, Xiang & Chen 2004,
Unsicker & Mody 2005) and was associated to various leaf traits (Marquis et al. 2001, Forkner et al. 2004,
Xiang & Chen 2004). Plants have the capacity to resist or tolerate the foliar herbivory and they can show large
differences in the rate of damage by differing their leaf traits (Loranger et al. 2012). According to Belsky (1986)
herbivory benefits certain plant species by increasing their net primary productivity and seed production by
removing senescent tissue and these changes increase plant fitness and supports the herbivore-plant mutualism.
Leaf herbivory induces changes in leaf defensive compounds (Karban & Baldwin 1997) and that can affect
pollination by inducing nectar defensive compounds (Adler et al. 2006). Recent analysis indicates that
secondary metabolites are of less importance as a defence against herbivory than morphological and life-history
traits (Carmona et al. 2011). A positive relationship between the rate of insect herbivory and plant diversity was
also documented in subtropical forests of south-east China (Schuldt et al. 2010). To understand the effect of
foliar herbivores on plant species, it is essential to quantify the variation in the rate of leaf damage and the
preference of insect herbivores that affect the foliage of those plants (Marquis 1991, Cuevas-Ryes et al. 2006).
The level of insect herbivory is generally lower than most vertebrate herbivory, because structural barriers
that are insignificant to vertebrates may be significant to insects (Peeters 2002). Therefore leaf traits including
surface, textures, anatomy and morphology have the capacity to block insect feeding. Hoffman & McEvoy
(1985) held that trichomes in leaf surface prevent the feeding of leaves by some sucking insects, but not for
other herbivores. Leaves containing waxes and thick cuticle on their surface also protect them from insect
herbivory (Edwards 1982, Potter & Kimmerer 1988). Leaf texture is an important trait of plant species that plays
a major role in plant-herbivore interaction (Peeters 2002).
52
Anil & Parthasarathy (2016) 3(1): 5266

.
The relationship between local host plant and their foliar herbivores have been studied in different tropical
forests (Janzen 1988, Marquis 1991, Basset 1991, Basset 1992, Barone 1998). Host specificity of herbivores is
frequently characterised by the distribution of herbivore densities throughout a set of local plant species
(Novotny et al. 2003). Host-plant interaction is an important ecological characteristic of herbivore species as it
determines their resource base and acts as a key factor in regulating their density and their relation with other
herbivores (Novotny 2002).
The impact and patterns of leaf damage of foliar herbivores have been documented in a few tropical forests
(Coley 1980, Janzen 1981, Cruz & Dirzo 1987, Coley & Baron 1996) and poorly documented in tropical dry
forest (Janzen 1981, Filip et al. 1995). Recent studies suggest that the top-down effects of plant resource users
are integral to the structure and functioning of ecosystem (Duffy 2002). No information is available on foliar
herbivory in tropical dry evergreen forest vegetation and hence the present study was undertaken with an
objective of investigating leaf traits and foliar herbivory and their relationship with respect to season. Here, we
assessed leaf traits, and diversity of foliar herbivores, the extent of leaf damage across various seasons and the
relationship between them in tropical dry evergreen forest (TDEF) on the Coromandel Coast of peninsular India.
In this study we addressed the following questions (1) Does the foliar herbivory in tropical dry evergreen forest
differ with season? (2) How variable are the leaf traits of TDEF plant species and (3) to check whether the leaf
traits had significant influence on the extent of leaf damage in the studied forest?
Study area
This study was conducted in tropical dry evergreen forest located in Villupuram (1156 N and 7953 E)
and Cuddalore (1143 N and 7949 E) districts of Tamil Nadu on the Coromandel Coast of peninsular India.
For the present study, we selected a total of nine tropical dry evergreen forest sites, as to cover all plant species
of TDEF. Site Puthupet (PP- 1203 N and 7952 E), Oorani (OR- 1209 N and 7952 E) and Vada Agaram
(VA- 7210 N and 7955 E) are located respectively 15, 28 and 32 km north of Puducherry town (1156 N
and 7953 E) and six other sites Kuzhandhaikuppam (KK- 1143 N and 7938 E), Thirumanikkuzhi (TM1143 N and 79 41 E), Suriyanpet (SR- 1144 N and 7938 E), Sendhirakillai (SK- 1130 N and 7941 E),
Palvathunnan (PT-1132 N and 7941 E) and Kothattai (KT- 1130 N and 7942 E) are located 45 to 50 km
surrounding of Puducherry town. The forest area of each study site ranges from 1.2 to 10 ha. Fifty-year (1954 to
2014) climate data of the nine sites revealed a mean annual temperature of 28.3 C and the mean annual rainfall
of 1,171 mm (Parthasarathy 2015). The mean number of rainy days in the annual cycle is 55.5. The climate is
tropical dissymmetric type with the bulk of the rainfall received during the northeast monsoon (October
December). Soils are red ferralitic belonging to the Cuddalore sandstone formation of the Miocene period
(Meher-Homji 1974). The vegetation of this area is tropical dry evergreen forest type. These closed-canopy
forests are 23 layered, tree-dominated and liana-dense (Champion & Seth 1968, Parthasarathy et al. 2008). The
canopy is about 1012 m in height, dominated by large trees such as Pterospermum canescens and Lannea
coromandelica, while the sub-canopy is composed of smaller trees such as Memecylon umbellatum, Canthium
dicoccum and Garcinia spicata. Major lianas include Strychnos lenticellata Dennst., Combretum albidum G.
Don., Reissantia indica (Willd.) Halle, Pyrenacantha volubilis Wight and Capparis zeylanica L. Ecbolium
viride (Forsskal) Alston and Sansevieria roxburghiana Schultes & Schultes f. are the major native perennial
herbs present in this forest type (Parthasarathy et al. 2008).
Study design
Leaf traits and per cent leaf damage were studied from August 2012 to July 2014 by sampling a total of
13,200 (30 leaves 110 plant species 4 seasons) leaves from 606 individuals of 110 plant species (60 trees, 45
lianas and 5 herbs) of tropical dry evergreen forest along with their leaf-eaters (Appendix I). Over the study
period, per cent leaf damage was recorded four-times in a year. We classified each year into four seasons such
as monsoon (OctoberDecember), post-monsoon (JanuaryMarch), summer (AprilJune) and pre-monsoon
(JulySeptember) on the basis rainfall data (Selwyn & Parthasarathy 2006, Parthasarathy et al. 2015). A
minimum of 2 to 4 individuals for each species (for rare and sub-dominant species) to a maximum of 10
individuals (for common and dominant species) were sampled to evaluate leaf traits and foliar herbivores. Thirty
leaves were collected from each plant species and calculated the per cent of leaf damage by mapping on graph
sheet. Foliar herbivore damage was estimated as: folivory (%) = (total leaf area remnant leaf area / total leaf
53

.
area) 100. Leaf traits studied include leaf type, texture, surface, size and specific leaf area (SLA). Besides our
observations, relevant literature and regional flora books (Gamble & Fisher 19151935, Matthew 1991) were
referred to confirm the categorization of leaf traits (Appendix I).
Foliar herbivores of each species were observed for a minimum of one day (06.0018.00 hrs.) during the
study period with the aid of binocular and captured in digital camera. Leaf twig was carefully pushed inside a
polythene bag and then clipped with the insects feeding on them (except lepidopteran larvae). Insects were
reared on appropriate plant materials until their host status could be confirmed or until they reached the adult
stage. We taxonomically identified the collected foliar herbivores of each plant species and the collected
lepidopteran larvae were reared in the laboratory until the emergence of adult insects, which were later
identified and un-identified larvae classified as morpho-species. Insects that were reared to adult or that died in
the process of rearing and confirming their feeding habits were preserved by either pinning (for adult specimen)
or in alcohol (for larvae and beetles). One-way ANOVA was used to determine whether leaf damage and leaf
traits (type, texture, surface, leaf area) significantly differed among species or seasons sampled.
RESULTS
Foliar Herbivory Across Various Seasons
The mean leaf damage by foliar herbivores ranged from 1.8% to 21% (with an overall mean of 8.3%) for the
assessed 13,200 leaves from 606 individuals of the 110 plant species during two-year study period. In the four
seasons studied, the mean leaf damage was maximum (16.1910.44%) in monsoon followed by pre-monsoon
(9.666.66%), intermediate in post-monsoon (5.244.10%) and least (1.971.52 %) in summer (Table 1). Per
cent leaf damage differed significantly across the four seasons (F = 96.52, P < 0.05) in TDEF. Plant species
displayed differences in leaf damage: among tree species Memecylon umbellatum (with 21% of mean leaf
damage), Canthium dicoccum (19%) and Lepisanthes tetraphylla (18%) showed higher levels of leaf damage.
Among liana species, Combretum albidum (15%), Reissantia indica (14%), and Cissus vitiginea (13%) had
higher levels of folivory.
Table 1. Seasonal patterns of foliar herbivory in tropical dry evergreen forest on the Coromandel Coast of India.
Values are monthly average for the study period (August 2012-July 2014).
% leaf damage
2012-13
2013-14
Total (mean)
Monsoon
16.16 10.35
17.01 10.76
16.19 10.44
Post-monsoon
5.12 3.99
5.57 4.34
5.24 4.10
Summer
1.94 1.46
2.09 1.62
1.97 1.52
Pre-monsoon
9.51 6.59
10.20 7.04
9.66 6.66
Leaf traits and foliar herbivores
Figure 1. No. of plant species of different life-forms (tree, liana, herb) with respect to leaf traits of a total of 110 plant
species in tropical dry evergreen forest on the Coromandel Coast of India (for expansion of codes see Appendix I).
54

.
Figure 2. Proportion of plant species with respect to foliar herbivory among various leaf traits of tropical dry evergreen
forest (for expansion of codes sees Appendix I).
In tropical dry evergreen forest, eighty per cent of plant species have simple leaves and just 20% have
compound leaves (Fig. 1). The coriaceous leaf texture (45%) was common followed by sub-coriaceous (41%)
membranous (11%) and chartaceous (3%) category. The glabrous leaf surface was common (84%) and just 16%
of species have hairy leaves. In leaf area, microphyll dominated (50%) followed by notophyll (32%),
mesophyll (14%) and nanophyll (4%). Beetles and lepidopteran larvae were the dominant leaf-eaters in TDEF,
followed by grasshoppers (Fig. 2) and their details are dealt by groups: Beetles (Fig. 3B & J) - Eight species of
beetles consumed leaves of 63 plant species (57%) in TDEF. Among the total 110 plant species, fifty-eight
species are exclusively folivored by beetle species (Table 2). Beetles utilised leaves of both trees (52%) and
Table 3. List of foliar herbivores (common name, scientific name for identified species, and faunal code) that utilise leaves
of one or many of the 110 plant species in tropical dry evergreen forest on the Coromandel Coast of India.
Foliar herbivore
Lepidopteran larvae
Moth larva 1
Moth larva 2
Moth larva 3
Moth larva 4
Moth larva 5
Moth larva 6
Moth larva 7
Moth larva 8
Moth larva 9
Moth larva 10
Moth larva 11
Moth larva 12
Moth larva 13
Moth larva 14
Code
Foliar herbivore
(38 plant species were exclusively utilised)
L1
Moth larva 21
L2
Moth larva 22
L3
Moth larva 23
L4
Moth larva 24
L5
Moth larva 25
L6
Moth larva 26
L7
Moth larva 27
L8
Moth larva 28
L9
Angled castor (Ariadne ariadne)
L10
Yellow orange tip (Ixias pyrene)
L11
Common grass yellow (Eurema hecabe)
L12
Indian common Mormon (Papilio polytes)
L13
Lemon pansy (Junonia lemonias)
L14
Lemon emigrant (Catopsilia pomona)
Code
L21
L22
L23
L24
L25
L26
L27
L28
L29
L30
L31
L32
L33
L34
55

.
Mottled emigrant (Catopsilia pyranthe)
L35
Painted hand maiden moth (Euchromia polymena)
L36
(Papilio sp.)
L37
Plain tiger larva (Danus chrysippus)
L38
Tawny coster (Acraea violae)
L39
Moth larva 15
L15
Moth larva 16
L16
Moth larva 17
L17
Moth larva 18
L18
Moth larva 19
L19
Moth larva 20
L20
Beetles (58 plant species utilised exclusively)
Unknown
Bt1
Jewel beetle (Sternocera chrysis)
Leaf beetle
Bt2
June beetle (Phyllophaga sp.)
(Chrysochus
auratus)
Unknown
Bt3
Net winged beetle (Lycostomus praeustus)
Unknown
Bt4
Flower beetle (Clinteria coerulea)
Weevils
Straight-snouted
W1
Leaf rolling weevil (Apoderus scutellaris)
weevil
(Rhopalapion
longirostre)
Grasshoppers (4 plant species utilised exclusively)
Unknown
Gh1
Melanoplus sp.
Unknown
Gh2
Hooded grasshopper (Teratodes monticollis)
Unknown
Gh3
Bt5
Bt6
Bt7
Bt8
W2
Gh4
Gh5
lianas (43%) in closer proportion. They chose largely simple leaf type (84%) with glabrous surface (84%), subcoriaceous (46%) and coriaceous (41%) leaf texture followed by membranous (9%) and chartaceous (3%)
category. With regard to leaf size, microphyll (54%) leaves are largely utilised by beetles followed by notophyll
(27%), mesophyll (15%) and nanophyll (3%). Larvae (Fig. 3D, F, G, I, K & L) - In TDEF, 29 different morphospecies of lepidopteran larvae used leaves of 46 plant species (42%) as food source and largely preferred leaves
of lianas (61%) over tree species (37%). Leaves of thirty-eight plant species in TDEF are exclusively consumed
by lepidopteran larvae (Table 2). Like the other foliar herbivores (beetles, grasshoppers) lepidopteran larvae
mainly fed upon simple leaves (78%) with coriaceous (54%) and sub-coriaceous textures (35%). Grasshoppers
(Fig. 3C) - Five species of grasshoppers utilised leaves of 10 plant species (9%) of which four species are
exclusively fed upon by them (Table 2) in TDEF and majority of them chose leaves of lianas (60%).
Grasshoppers consumed leaves of sub-coriaceous (60%), coriaceous (20%) and membranous (20%) texture,
simple leaves (70%) with glabrous surface (80%) and microphyll (40%) leaf size.
Variation in leaf traits and per cent leaf damage
A significant relationship exists between per cent leaf damage and various leaf traits (Table 3). In tropical
dry evergreen forest, leaves of tree species are consumed more than those of liana species and the leaf damage
was significantly related during pre-monsoon (F = 10.06, P < 0.05) and post-monsoon season (F = 4.0, P <
0.05). Tree species exhibit high mean leaf damage in post-monsoon (6.414.86) and pre-monsoon (2.251.82)
seasons than those of liana species. Among tree species, Ficus hispida (19.3%) suffered maximum leaf damage
in post-monsoon and Memecylon umbellatum (39%) in pre-monsoon season. One-way ANOVA revealed that
there was no relationship between plant physiognomic type and per cent leaf damage (P > 0.05), so also
between leaf surface and per cent leaf damage. Leaf texture was significantly related to leaf damage in monsoon
season (F = 3.25, P < 0.05). Species with coriaceous leaf texture suffered high mean leaf damage (19.0410.96)
in monsoon season than those of sub-coriaceous (15.9510.22), membranous (8.925.71) and chartaceous
texture (189.26). Coriaceous leaves of Memecylon umbellatum recorded maximum leaf damage (43%) and
membranous leaves of Abrus precatorius had minimum leaf damage (2%) in monsoon season. Results of oneway ANOVA revealed a higher level of significance between leaf area and per cent leaf damage during postmonsoon (F = 17.36, P < 0.05) and summer (F = 12.06, P < 0.05). Mesophyll leaves were consumed largely in
post-monsoon (10.863.35) and summer (3.761.73) season. Mesophyll leaves of Ficus hispida (19.23)
recorded maximum leaf damage and nanophyll leaves of Abrus precatorius (0.23%) exhibit minimum leaf
56

.
damage in post-monsoon season also. In summer, notophyll leaves of Cassia fistula (7.32%) are largely
consumed by foliar herbivores (Appendix I).
Figure 3. Various foliar herbivores (beetle, larvae, and grasshoppers) utilized different plant life-forms in tropical dry
evergreen forest on the Coromandel Coast of peninsular India; A, Larva on coriaceous leaves of Atlantia monophylla; B,
Jewel beetle in membranous leaves of Acacia caesia; C, Grass hopper in sub-coriaceous leaves of Lantana camara; D,
Painted hand maiden moth larva in Argyreia cymosa; E, reared adult of Painted hand maiden moth larva; F, Moth larvae in
Memecylon umbellatum; G, Larvae of Tawny coster feeding leaves of Ecbolium viride; H, Reared adult of Tawny coster
larva; I, Larva in Diospyros ebenum; J, Beetle folivored notophyll leaves of Canthium dicoccum; K, Moth larva in
microphyll leaves of Cayratia pedeta and L, Moth larva in mesophyll leaves of Ficus hispida.
DISCUSSION
Foliar herbivory across various seasons
Our study documented the per cent of leaf damage caused by foliar herbivores in tropical dry evergreen
forest (TDEF) on the Coromandel Coast of peninsular India. In general, the mean annual per cent of leaf
57

.
damage in tropical dry evergreen forest is higher than temperate forest (7.1%) and lower than other tropical wet
evergreen forest (11.1%) as reported in various tropical forests by Coley & Aide (1991). Leaf damage averages
8.3% for 110 plant species of TDEF on the Coromandel Coast of India, and this is about one and a half fold
greater (14.9%) than that of 37 tree species of Chamela-Cuixmala biosphere reserve in Mexico (Cuevas-Reyes
et al. 2006) and almost double (16%) in Luquillo experimental forest on Puerto Rico for the 8 tree species
studied. In lowland tropical forest, mean per cent folivory ranged from 7 to 20.3 % (Coley & Aide 1991), but in
TDEF the variation was 1.8% to 21% for the studied 110 plant species. This may be because our study was
conducted on adult plants of different life-forms (60 trees, 45 lianas, 5 herbs), while Coleys study mainly
focussed on saplings of trees, where leaf damage is generally high (Coley & Barone 1996).
Table 3. Per cent leaf damage in four seasons with respect to leaf traits in tropical dry evergreen forest on the Coromandel Coast of
India. One- way ANOVA showed significant variation among various leaf traits (P < 0.05).
Monsoon
Post-monsoon
Summer
Pre- monsoon
Plant traits
MeanSd
F
P
MeanSd . F
P
MeanSd . F
P
MeanSd
F
P
Life-form T
18.2010.45 3.38 0.06
6.414.86 10.06 0.00
2.251.82 3.51 0.06 10.976.89 4.00 0.04
L
14.4310.27
3.922.28
1.690.95
8.366.24
Plant type E
15.6210.58 0.80 0.44
4.993.68 1.54 0.21
1.961.42 0.54 0.58
9.447.21 0.38 0.68
B
18.7710.31
5.114.61
1.891.66
10.845.59
D
17.0610.54
6.894.86
2.351.74
9.996.47
Leaf type C
15.8010.54 0.15 0.69
4.744.16 0.58 0.44
1.761.74 0.75 0.38 10.416.43 0.19 0.66
S
16.8010.54
5.504.14
2.081.47
9.716.82
Texture
M
8.925.71
3.25 0.02
4.674.08 0.98 0.40
1.631.54 0.37 0.771 6.224.12 2.02 0.11
Ch
189.26
8.213.25
2.331.15
12.075.71
Sc 15.9510.22
4.833.58
1.971.41
9.296.45
Co 19.0410.96
5.794.63
2.121.67
11.157.26
Surface
G
16.7510.52 0.09 0.75
4.993.57 2.97 0.08
2.031.52 0.05 0.81
9.926.95 0.04 0.83
H
15.9610.63
6.685.71
1.941.60
9.595.90
Leaf area Na
8.279.76
1.64 0.18
1.551.37 17.36 0.00
0.580.41 12.06 0.00
6.465.71 1.46 0.23
Mi 15.4810.66
4.544.17
1.631.31
8.947.35
No 18.9510.20
4.391.99
1.931.20
11.636.10
Me 17.1510.04
10.863.35
3.761.73
9.835.58
Note: For expansion of codes see Appendix 1.
Leaf traits and foliar herbivores
Leaves of tropical dry evergreen forest are fed upon by diverse foliar herbivore groups such as Coleoptera,
Lepidopteran larva and Orthoptera and these foliar herbivores are also the most important leaf eaters in various
other tropical forests (Angulo-Sandoval & Aide 2000, Arnold & Asquith 2002, Williams-Linera & Herrera
2003, Angulo-Sandoval et al. 2004, Mazia et al. 2012). Coleopterans are the major leaf-eaters (of 57% plant
species) in tropical dry evergreen forest as also reported in many other tropical forests (Murali & Sukumar 1993,
Varanda & Paise 2006). In our study, leaves of more than 50% of plant families are consumed by a single faunal
group and this pattern of leaf damage suggests that specialist foliar herbivores are more important than
generalists and it supports the assumption of Janzen-Connell model (Janzen 1970). The leaf damage caused by
twenty-nine species of Lepidopteran larvae exhibited a high degree of specialisation in TDEF and that is not the
case with beetles and grasshoppers. Arnold & Asquith (2002) also reported high diversity and density of
Lepidopteran larvae in the fragmented forest of Lago Gatun, Panama. The leaf damage of 38 plant species was
caused by 29 morpho-species of specialist larval herbivores in TDEF. This result suggests that specialist insect
herbivores are more important than generalist herbivores and this observation corroborates with the global
pattern of lepidopteran specialist feeders (Scriber 1973).
A noticeable trend in this study is the peak of insect abundance during the monsoon and pre-monsoon seasons
and this account for maximum leaf damage in these seasons. According to Coley & Barone (1996), foliar
herbivore populations are less during dry season, with a marked increase at the beginning of wet season and the
concordant rates of herbivory being lowest in the dry season and highest in rainy season. The higher leaf
damage in monsoon and pre-monsoon season in TDEF is probably due to the activity of Lepidoptera larvae,
which are more abundant in these seasons. Evidence from the literature (Raffa et al. 1992, Mopper & Simberloff
1995) provides a possible explanation that foliar herbivores respond to seasonal variation in food resources by
contemporizing their life-cycle with the phenology of their host plant species and in our observation most of the
58

.
plant species in TDEF show a peak in leaf-flushing during pre-monsoon and monsoon seasons. Lowman (1992)
also reported a higher level of leaf damage during the peaks of leaf production. Angulo-Sandoval & Aide (2000)
reported that when leaf production decreases an increase in predator density of foliar herbivores will reduce the
herbivore density and that could be the possible reason for less leaf damage in TDEF also during the postmonsoon and summer seasons.
Variation in leaf traits and per cent leaf damage
In the present study, tree species are highly folivored than liana species because liana leaves are more
exposed to sun and the level of herbivory in canopy usually less than the leaves at sub-canopy and understory
levels as also reported from the sub-tropical and temperate rain forest of New South Wales (Lowman 1985).
This study found that the extent of insect herbivory varied significantly with leaf traits and season (Table 1 & 3).
Overall, coriaceous and sub-coriaceous textures of leaves in tropical dry evergreen forest (TDEF) can be
considered as palatability enhancers to the foliar herbivores and this result also agrees with Lu & Lee (2010)
who found that leaf traits may play a key role in the host choice of foliar herbivores and the extent of leaf
damage. Our results also suggest that foliar herbivory rates across different seasons may not be strongly related
to single leaf trait. Evidently, dominant leaf traits and foliar herbivore interactions in tropical dry evergreen
forest are highly specific and also further influenced by season. In our results, leaf damage was high for those
plant species with high specific leaf area (SLA) and this agrees with the prediction of Poorter et al. (2004) that
trees with low SLA would host lesser number of herbivores. Thus, insect abundance and leaf damage are
dependent on the resource availability and quality such as palatability and total biomass content, to meet the
dietary need of foliar herbivores. The most dominant species of TDEF (Parthasarathy et al. 2008) exhibit
maximum foliar damage in wet season (pre-monsoon and monsoon) and this subscribes for the significant
relation obtained between foliar herbivory and plant density, but in dry season, the pattern differs. This result
also corroborates with the findings of other studies (Janzen 1970, Barone 1998, Angulo-Sandoval & Aide 2000)
explaining the higher rate of herbivory in plant species occurring in higher densities. In monsoon season the
numbers of foliar herbivores are high and they seem to exploit the most dominant favourable leaf traits that
prevail in TDEF species such as coriaceous and sub-coriaceous texture and glabrous leaves. In dry season (postmonsoon and summer), foliar herbivores of TDEF mainly depend on leaf biomass, because microphyll leaves
are dominant leaf size in our study, but mesophyll leaves exhibit maximum leaf damage in these seasons. The
positive relationship obtained between specific leaf area (SLA) and leaf damage also supports this view point.
Admittedly, foliar herbivory and leaf damage in TDEF are governed by various leaf traits, seasons and plant
density as well.
CONCLUSIONS
This study investigated the leaf traits and leaf resource use by faunal community in the under-studied
tropical dry evergreen forest (TDEF) of India and revealed the degree of specialisation between foliar herbivores
and individual leaf traits of this unique forest type, which are further influenced by seasonal variation, plant
density and herbivore abundance. The tropical dry evergreen forest supports a rich herbivore fauna of beetles,
lepidopteran larvae and grasshoppers and many of which help in ecosystem functioning of the TDEF. This study
underlines the importance of community-level approach in plant resource use by faunal communities for better
understanding of the forest biotic interactions useful for conservation.
ACKNOWLEDGEMENTS
K. Anil thanks Pondicherry University for financial support received through UGC-BSR fellowship.
REFERENCES
Adler LS, Wink M, Distl M & Lentz AJ (2006) Leaf herbivory and nutrients increase nectar alkaloids. Ecology
Letters 9: 960967.
Angulo-Sandoval P & Aide TM (2000) Leaf phenology and leaf damage of saplings in the Luquillo
Experimental Forest, Puerto Rico. Biotropica 32: 415422.
AnguloSandoval P, FernndezMarn H, Zimmerman JK & Alde TM (2004) Changes in patterns of understory
leaf phenology and herbivory following hurricane damage. Biotropica 36: 6067.
59

.
Arnold AE & Asquith NM (2002) Herbivory in a fragmented tropical forest: patterns from islands at Lago
Gatun, Panama. Biodiversity and Conservation 11: 16631680.
Barone JA (1998) Host-specificity of folivorous insects in a moist tropical forest. Journal of Animal Ecology 67:
400409.
Basset Y (1991) The spatial distribution of herbivory, mines and galls within an Australian rain forest
tree. Biotropica 23: 271281.
Basset Y (1992) Host specificity of arboreal and freeliving insect herbivores in rain forests. Biological Journal
of the Linnean Society 47: 115133.
Belsky AJ (1986) Does herbivory benefit plants? A review of the evidence. American Naturalist 127: 870892.
Carmona D, Lajeunesse MTJ & Johnson (2011) Plant traits that predict resistance to herbivores. Functional
Ecology 25: 358367.
Champion HG & Seth SK (1968) Revised Survey of the Forest Types of India. Manager of Publications, New
Delhi, pp. 404.
Coley PD & Aide TM (1991) Comparison of herbivory and plant defenses in temperate and tropical broadleaved forests. In: Price PW, Lewinsohn TM, Fernandes GW & Benson WW (eds) Plant-animal interactions
evolutionary ecology in tropical and temperate regions. John Wiley & Sons, Inc., New York, pp. 2549.
Coley PD & Barone JA (1996) Herbivory and plant defenses in tropical forests. Annual Review of Ecology and
Systematics 27: 305335.
Coley PD (1980) Effects of leaf age and plant life-history patterns on herbivory. Nature 284: 545546.
Cruz M & Dirzo R (1987) A survey of the standing levels of herbivory in seedlings from a Mexican rain forest.
Biotropica 19: 98106.
CuevasReyes P, Quesada M & Oyama K (2006) Abundance and Leaf Damage Caused by GallInducing
Insects in a Mexican Tropical Dry Forest. Biotropica 38: 107115.
Duffy JE (2002) Biodiversity and ecosystem function: the consumer connection. Oikos 99: 201219.
Edwards PB (1982) Do waxes on juvenile Eucalyptus leaves provide protection from grazing insects?
Australian Journal of Ecology 7: 347352.
Filip V, Dirzo R, Mass JM & Sarukhan J (1995) Within- and among year variation in the levels of herbivory on
the foliage of tress from a Mexican tropical deciduous forest. Biotropica 27: 7886.
Forkner RE, Marquis RJ & Lill JT (2004) Feeny revisited: condensed tannins as antiherbivore defenses in leafchewing herbivore communities of Quercus. Ecological Entomology 29: 17487.
Gamble JS & Fischer CEC (1915-1935) Flora of the Presidency of Madras. Vols. I-III, Adlard & Co., London.
Hoffman GD & Mcevoy PB (1985) The mechanism of trichome resistance in Anaphalis margaritacea to the
meadow spittlebug Philaenus spumarius. Entomologia Experimentalis et Applicata 39: 123129.
Janzen DH (1970) Herbivores and the number of tree species in tropical forests. American Naturalist 104: 501
528.
Janzen DH (1981) Patterns of herbivory in a tropical deciduous forest. Biotropica 15: 108111.
Janzen DH (1988) Ecological characterization of a Costa Rican dry forest caterpillar fauna. Biotropica 20: 120
135.
Karban R & Baldwin IT (1997) Induced Responses to Herbivory. The University of Chicago Press, Chicago, IL.
Loranger J, Meyer ST, Shipley B, Kattge J, Loranger H, Roscher C & Weisser WW (2012) Predicting
invertebrate herbivory from plant traits: evidence from 51 grassland species in experimental
monocultures. Ecology 93: 26742682.
Lowman MD (1985) Temporal and spatial variability in insect grazing of the canopies of five Australian
rainforest tree species. Australian Journal of Ecology 10: 724.
Lowman MD (1992) Herbivory in Australian rain forests, with particular reference to the canopies of Doryphora
sassafras (Monimiaceae). Biotropica 24: 263272.
Lu EY & Lee CY (2010) Insect Folivory and Leaf Traits of Seven Hardwood Species in the Subtropical
Rainforest of Fushan, Northeastern Taiwan. Taiwan Journal of Forest Science 25: 315326.
Marquis RJ (1991) Herbivore fauna of Piper (Piperaceae) in Costa Rican wet forest: Diversity, specificity, and
impact. In: Price PW, Lewinsohn TM, Fernandes GW & Benson WW (eds) Plant-animal interactions:
Evolutionary ecology in tropical and temperate regions. John Wiley and Sons Corporations, New York, pp.
179202.
60

.
Marquis RJ, Diniz IR & Morais HC (2001) Patterns and correlates of the interspecific variation in foliar insect
herbivory and pathogen attack in Brazilian Cerrado. Journal of Tropical Ecology 17: 12748.
Matthew KM (1991) An excursion flora of central Tamilnadu, India. CRC Press.
Mazia N, Chaneton EJ, Dellacanonica C, Dipaolo L & Kitzberger T (2012) Seasonal patterns of herbivory, leaf
traits and productivity consumption in dry and wet Patagonian forests. Ecological Entomology 37: 193203.
Meher-Homiji VM (1974) On the origin of the tropical dry evergreen forest in South India. Indian Journal of
Ecology and Environmental Sciences 44: 1939.
Mopper S & Simberloff D (1995) Differential herbivory in an oak population: The role of plant phenology and
insect performance. Ecology 76: 12331241.
Murali KS & Sukumar R (1993) Leaf flushing phenology and herbivory in a tropical dry deciduous forest,
southern India. Oecologia 94: 114119.
Novotny V, Basset Y, Miller, SE, Drozd P & Cizek L (2002) Host specialisation of leaf chewing in a New
Guinea rainforest. Journal of Animal Ecology 71: 400412.
Novotny V, Miller SE, Cizek L, Leps J, Janda M, Basset Y, Weiblen GD & Darrow K (2003) Colonizing aliens:
caterpillars (Lepidoptera) feeding on Piper aduncum and P. umbellatum in rainforests of Papua New Guinea.
Ecological Entomology 28: 704716.
Parthasarathy N (2015) Biodiversity of Lianas. Springer International Publishing, pp. 289.
Parthasarathy N, Vivek P & Anil K (2015) Biodiversity, Ecology and Conservation of Tropical dry Evergreen
forest. Lambert Academic Publishing, GmbH and Co. Germany.
Parthasarathy N, Selwyn MA & Udayakumar M (2008) Tropical dry evergreen forests of peninsular India:
ecology and conservation significance. Tropical Conservation Science 1: 89110.
Peeters PJ (2002) Correlations between leaf structural traits and the densities of herbivorous insect guilds.
Biological Journal of Linnean Society 77: 4365.
Poorter L, De Plassche MV, Willems S & Boot RGA (2004) Leaf traits and herbivory rates of tropical tree
species differing in successional status. Plant Biology 6: 746754.
Potter D & Kimmerer TW (1988) Do holly leaf spines really deter herbivory? Oecologia 75: 216221.
Raffa KF, Hall DJ, Kearby W & Katovich S (1992) Seasonal life history of introduced basswood thrips
(Thysanoptera: Thripidae) in Wisconsin, with observations on associated thrips species. Environmental
Entomology 21: 771779.
Schuldt A, Baruffol M, BoHnke M, et al. (2010) Tree diversity promotes insect herbivory in subtropical forests
of south-east China. Journal of Ecology 98: 917926.
Schuldt A, Bruelheide H, Durka, W, Eichenberg D, Fischer M, Krber W, Hrdtle W, Ma K, Michalski SG,
Palm WU & Schmid B (2012) Plant traits affecting herbivory on tree recruits in highly diverse subtropical
forests. Ecology Letters 15: 732739.
Scriber JM (1973) Latitudinal gradients in larval feeding specialization of the world Papilionidae (Lepidoptera).
Psyche 80: 35573.
Selwyn MA & Parthasarathy N (2006) Reproductive traits and phenology of plants in tropical dry evergreen
forest on the Coromandel coast of India. Biodiversity and Conservation 15: 32073234.
Unsicker SB & Mody K (2005) Influence of tree species and compass bearing on insect folivory of nine
common tree species in the West African savanna. Journal of Tropical Ecology 21: 22731.
Varanda EM & Paise MP (2006) Insect folivory in Didymopanax vinosum (Apiaceae) in a vegetation mosaic of
Brazilian Cerrado. Brazilian Journal of Biology 66: 671680.
Viola DV, Mordecai EA, Jaramillo AG, Sistla SA, Albertson LK, Gosnell JS, Cardinale BJ & Levine JM (2010)
Competition-defense tradeoffs and the maintenance of plant diversity. Proceedings of the National Academy
of Science USA 107: 1721717222.
WilliamsLinera G & Herrera F (2003) Folivory, herbivores, and environment in the understory of a tropical
montane cloud forest. Biotropica 35: 6773.
Wright SJ (2007) Plant diversity in tropical forests. In: Pugnaire FI & Valladares F (eds) Functional Plant
Ecology. CRC Press, Boca Raton, pp. 351367.
Xiang H & Chen J (2004) Interspecific variation of plant traits associated with resistance to herbivory among
four species of Ficus (Moraceae). Annals of Botany 94: 377384.
61

.
Appendix I. Leaf traits, foliar herbivores and per cent leaf damage of 110 plant species of tropical dry evergreen forest on the Coromandel Coast of India.
Per cent leaf damage (Mean Sd)
Plant species
Family
SS PT LT
LR LS LA SLA
FH
Monsoon
PostSummer
Premonsoon
monsoon
Trees
Aglaia elaeagnoidea (Juss.) Benth.
Meliaceae
4
E
C
Co G
Mi 105.31 L1
201.4
2.30.7 0.80.07 16.51.4
Albizia amara (Roxb.) Boivin
Mimosaceae
10 D
Cpi
M
G
Na 110
L9
22.80.4
1.40.2 0.60.07
141.4
Albizia lebbeck (L.) Benth.
Mimosaceae
4
D
Cpi
M
G
Mi 63.13
L29
17.52.1
0.40.1 0.80.07 12.50.7
Allophyllus serratus (Roxb.) Kurz
Sapindaceae
4
E
Ctri Sco G
Mi 137.36 L16
15.50.7
2.90.3 0.70.07 12.51.4
Atalantia monophylla (L.) Correa
Rutaceae
10 E
S
Co G
No 459.43 L32, L37
36.36.7
40.4
2.90.71
221.4
Azadirachta indica A. Juss.
Meliaceae
10 B
C
Sco G
Mi 234.43 L18
29.50.7
.20.1
0.80.14 16.50.7
Barringtonia acutangula (L.) Gaertner Barringtoniaceae 4
E
S
Sco G
Me 71.76
L20
50.7
13.51.5 3.60.71
3.60.1
Bauhinia racemosa Lam.
Caesalpiniaceae
4
D
S
Sco E
Mi 97.92
L21
4.52.1
18.31.4 4.50.35
2.70.2
Benkara malabarica (Lam.) Tirven.
Rubiaceae
4
E
S
Co E
Mi 42.47
L5
13.32.5 17.30.0 1.30.07
6.81.1
Breynia vitis-idaea (Burm. f.) Fischer Euphorbiaceae
4
E
S
M
G
Mi 32.61
Bt2
7.41.3
13.81.0 4.70.71
40.7
Butea monosperma (Lam.) Taubert
Papilionaceae
4
D
Ctri Co G
Me 106.26 L29
240.7
16.10.0 2.60.28
150.7
Calophyllum inophyllum L.
Clusiaceae
4
E
S
Co G
Me 115
Bt2
20.92.0 12.30.0 6.50.71
8.51.4
Canthium dicoccum (Gaertn.) Teijsm. Rubiaceae
10 E
S
Co G
No 141.94 L8, L34
40.21.7
8.21.3 3.90.71
232.1
& Binn.
Cassia fistula L.
Caesalpiniaceae
4
D
Cpi
Sco G
No 255.59 L3
23.31.1 12.80.5 7.30.71
170.7
Chionanthus zeylanica L.
Oleaceae
10 E
S
Co G
Mi 58.54
L2
14.50.7 13.60.6 5.60.42
90.7
Cordia obliqua Willd.
Cordiaceae
4
B
S
Co G
Mi 116.67 L21
30.31.2 11.41.3 3.60.21 15.70.4
Crateva magna (Lour.) DC.
Capparaceae
4
D
Ct
M
G
Me 219.35 Bt3
16.51.4
9.70.6 4.60.33 11.91.3
Diospyros ebenum Koen.
Ebenaceae
10 E
S
Co G
No 75.57
L19
21.90.6
40.3
1.60.71 12.71.0
Diospyros ferrea (Willd.) Bakh.
Ebenaceae
4
E
S
Co G
Mi 89.17
L19
110.7
2.80.8 0.30.14
6.70.5
Drypetes sepiaria (Wight & Arn.) Pax Euphorbiaceae
4
E
S
Co G
Mi 92.12
Bt2, L35
371.4
2.31.2 0.60.07 19.10.5
& Hoffm..
Ehretia pubescens Benth.
Boraginaceae
4
B
S
Sco Hp Mi 76
Bt1
17.81.9
2.80.7 0.30.07
130.7
Eugenia bracteata (Willd.) Roxb.
Myrtaceae
4
E
S
Co Hs Me 70.46
Bt4
41.4
11.21.4 0.50.07
2.60.1
Moraceae
4
B
S
Co Hs Me 75.48
Bt3
181.4
13.31.6 4.50.42
90.7
Ficus hispida L. f.
Moraceae
4
B
S
Co Hh Me 83.15
L26
14.00.8 19.22.7 6.70.07
8.51.4
Ficus religiosa L.
Moraceae
4
B
S
Co G
Me 105.22 Bt1
261.4
8.21.3 4.50.07 16.70.4
62
Flacourtia indica (Burm. f.) Merr.

Garcinia spicata (Wight & Arn.) J.D.
Hook.
Glycosmis mauritiana Yuich. Tanaka
Gmelina asiatica L.
Ixora pavetta T. Anderson
Lannea coromandelica (Houtt.) Merr.
Lepisanthes tetraphylla (Vahl.) Radlk.
Mallotus philippensis (Lam.) Muell.Arg.
Mallotus rhamnifolius Muell.-Arg.
Manilkara hexandra (Roxb.) Dubard
Maytenus emarginata (Wild.) Ding
Hou.
Memecylon umbellatum Burm. f.
Mimusops elengi L.
Morinda coreia Buch. -Ham.
Morinda pubescesns Sm.
Ochna obtusata DC.
Pamburus missionis (Wight) Swingle
Pleiospermium alatum (Wall. ex
Wight. & Arn.) Swingle
Pongamia pinnata (L.) Pierre
Premna latifolia Roxb.
Pterospermum canescens Roxb.
Pterospermum xylocarpum (Gaertn.)
Sant. & Wagh.
Salvadora persica L.
Sapindus emarginatus Vahl
Securenega leucopyrus (Willd.)
Muell.-Arg.
Semecarpus anacardium L. f.
Strychnos nux-vomica L.

.
252.8
10.72.1 3.50.42 13.60.6
171.4
11.21.3 4.80.71
8.51.4
Flacourtiaceae
Clusiaceae
4
4
B
E
S
S
Ch
Co
G
G
Mi
Me
280.5
79.89
Bt2
Bt1
Rutaceae
Verbenaceae
Rubiaceae
Anacardiaceae
Sapindaceae
Euphorbiaceae
4
4
4
4
10
4
E
E
E
D
E
E
S
S
S
Cpi
Cpi
S
Sco
M
Co
Sco
Sco
Sco
G
G
G
G
G
G
Mi
Mi
No
No
Me
Me
126.15
18.97
97.92
101.14
47.63
65
L32, L37, Bt8 25.35.9

Bt4
120.7
L15
162.8
Bt2
320.7
L1, L6, W2
362.1
Bt3
12.12.9
20.4
2.70.7
2.70.1
3.70.1
9.40.2
7.70.7
1.10.14
1.50.25
0.50.14
1.30.48
4.70.7
4.40.44
16.51.4
60.7
80.7
17
220.7
7.50.7
Euphorbiaceae
Sapotaceae
Celastraceae
4
4
4
E
B
E
S
S
S
Ch
Co
Co
G
G
G
Me
No
Mi
64.63
54.84
51.37
Bt2
Bt3
Bt4
7.51.4
13.52.8
141.4
9.40.1
4.10.7
1.40.2
1.20.07
1.60.14
0.30.07
5.80.4
6.51.4
7.650.2
Melastomataceae
Sapotaceae
Rubiaceae
Rubiaceae
Ochnaceae
Rutaceae
Rutaceae
10
4
10
4
4
4
4
E
E
E
B
D
E
E
S
S
S
S
S
S
S
Co
Co
Co
Co
Co
Co
Co
G
G
G
H
G
G
G
Mi
No
No
No
No
No
No
60.71
85.16
217.69
265.93
94.64
62.28
241.62
L23, L24, L33

L2
L8
Bt1
Bt4
L11
Bt2
430.7
12.51.4
31.62.3
31.81.1
21.31.8
70.7
18.60.5
1.40.1
3.60.1
4.60.3
50.2
3.60.2
6.40.1
3.40.3
0.40.03
1.60.21
2.20.07
1.30.14
1.30.07
3.30.71
1.40.07
397.1
70.7
15.80.4
15.52.1
130.7
5.54.2
12.51.4
Papilionaceae
Verbenaceae
Sterculiaceae
Sterculiaceae
4
4
10
4
B
E
B
B
Cpi
S
S
S
Sco
Sco
Co
Co
G
H
G
G
Mi
Mi
Mi
No
55.26
26.55
86.11
83.56
Bt3
Bt4
Bt5
Bt6
242.1
15.51.4
32.51.4
31.51.4
3.30.7
2.60.2
1.80.0
3.60.3
0.50.02
1.40.42
0.20.07
1.50.07
141.4
6.50.7
181.4
182.8
Salvadoraceae
Sapindaceae
Euphorbiaceae
4
4
4
B
B
E
S
Cpi
S
Co
Sco
M
G
G
G
Mi
Mi
Na
68.94
54.47
16.11
Bt2
Bt4
Bt7
6.81.1
120.7
5.51.4
1.90.1
2.70.1
1.10.5
1.40.07
1.50.04
0.30.02
20.7
6.91.2
3.51.4
Anacardiaceae
Moraceae
Loganiaceae
4
4
4
D
E
D
S
S
S
Co
Sco
Sco
G
Hs
G
No
Mi
No
6.46
96.58
78.33
Bt2
Bt3
Bt4
3.81.1
4.21.2
5.80.4
3.30.0
2.40.1
4.90.1
0.70.07
0.90.07
2.10.07
0.80.1
40.7
3.30.4
63
Syzygium cumini (L.) Skeels

Tarenna asiatica (L.) Kuntz ex
Schumann
Terminalia bellirica (Gaertn.) Roxb.
Tricalysia sphaerocarpa (Dalz.)
Gamble
Vitex altisima L. f.
Walsura trifolia (A.Juss.) Harms
Lianas
Abrus precatorius L.
Acacia caesia (L.) Willd.
Ampelocissus tomentosa (Heyne ex
Roth) Planch.
Argyreia cymosa (Roxb.) Sweet
Canavalia virosa (Roxb.) Wight &
Arn.
Cansjera rheedii Gmel.
Capparis brevispina DC.
Capparis rotundifolia Rottl.
Capparis sepiaria L.
Capparis zeylanica L.
Carissa spinarum L.
Cayratia pedata (Lam.) Juss. ex
Gagnep.
Cissampelos pareira L. var. hirsuta
(Buch.-Ham. ex DC.) Forman
Cissus quadrangularis L.
Cissus vitiginea L.
Coccinia grandis (L.) Voigt
Combretum albidum G.Don
Derris ovalifolia (Wight & Arn.)
Benth.
Dioscorea oppositifolia L.
Myrtaceae
Caesalpiniaceae
Rubiaceae
4
4
4
B
B
E
S
Cpi
S
Co
M
Co
G
G
G
Me
Na
No
145.68
62.5
53.67
Bt5
Bt6
L14

.
242.1
6.51.4 3.30.07 15.40.9
2.90.2
3.50.2 1.10.14
0.90.2
10.81.1
4.70.1 2.20.08
92.8
Combretaceae
Rubiaceae
4
10
D
E
S
S
Co
Sco
G
G
Me
No
69.55
33.11
GH1, L6
Bt7
2.81.1
220.7
8.70.1
3.80.0
3.80.07
1.70.14
1.70.3
14.54.2
Verbenaceae
Meliaceae
4
4
D
E
Ctri
Ctri
Sco
Co
G
G
Mi
No
19.58
88.64
GH3
L10, L30
50.7
170.7
2.40.0
3.80.0
1.20.07
1.40.02
20.7
11.60.6
Papilionaceae
Mimosaceae
Vitaceae
4
10
2
B
B
B
Cpi
Cpi
Cpal
M
M
Sco
Hp
G
H
Na
Mi
Me
23.43
18.87
134.3
GH2
Bt5
Bt2
20.7
2.70.5
12.80.4
0.20.0
1.20.0
8.50.0
0.20.02
0.30.07
2.80.21
7.58.5
3.41.3
6.80.3
Convolvulaceae
Papilionaceae
4
4
E
D
S
Ctri
Sco
Sco
G
G
No
Mi
46.13
28.67
Bt3
Bt4
2.70.4
5.80.4
3.50.2
2.20.1
1.30.03
1.30.09
1.80.4
2.60.6
Opiliaceae
Capparaceae
Capparaceae
Capparaceae
Capparaceae
Apocynaceae
Vitaceae
4
10
4
4
4
10
4
E
E
E
E
E
E
E
S
S
S
S
S
S
Cpal
Co
Sco
Sco
Sco
Sco
Co
Sco
G
G
G
G
G
G
H
No
No
Mi
Mi
No
Mi
Mi
76.08
90.63
79.29
43.13
171.37
30.47
106.22
Bt5
Bt6
Bt3
Bt1
Bt3
Bt6
Bt9
3.70.5
13.31.8
14.51.4
10.30.4
18.51.4
40.7
33.81.1
3.60.1
2.80.1
50.7
1.70.1
4.60.2
2.40.2
2.60.2
1.40.07
1.20.02
1.70.21
0.50.14
2.40.21
2.20.07
1.10.14
1.70.3
7.60.1
3.60.1
5.40.1
13.30.4
1.50.1
15.61.3
Menispermaceae
Sco
No
304.59
L3
21.21.7
2.50.0
1.30.07
12.10.6
Vitaceae
Vitaceae
Cucurbitaceae
Combretaceae
Papilionaceae
10
10
10
10
4
E
D
E
D
E
S
S
S
S
Cpi
Co
Co
Sco
Sco
Sco
G
H
G
G
G
Mi
No
Mi
Me
Mi
20.14
230.72
11.27
198
25
L2
L5
Gh3, L6
GH1, L7
Bt4, Gh2
2.10.9
272.1
2.70.2
34.53.5
2.50.7
3.50.3
6.50.4
2.80.7
8.80.1
3.21.4
1.50.07
2.50.07
0.60.07
1.60.14
1.80.07
0.80.0
17.82.5
1.50.0
13.92.3
1.70.3
Dioscoreaceae
No
262.85
Gh4
21.51.4
4.50.4
2.40.07
16.90.9
64
Grewia rhamnifolia Heyne ex Roth

Gymnema sylvestre (Retz.) R.Br. ex
Schultes
Hugonia mystax L.
Ichnocarpus frutescens (L.) R.Br.
Jasminum angustifolium (L.) Willd.
Lantana camara L.
Leptadenia reticulata (Retz.) Wight
& Arn.
Maerua oblongifolia (Forsk.) A.Rich.
Mukia maderaspatana (L.) M. Roem.
Olax scandens Roxb.
Pachygone ovata (Poir) Miers ex
Hook.
Plecospermum spinosum Trecul
Premna corymbosa (Burm.f.) Rottl.
& Willd.
Pyrenacantha volubilis Wight
Reissantia indica (Willd.) Halle
Rivea hypocrateriformis (Desr.)
Choisy
Salacia chinensis L.
Strychnos lenticellata Hill
Tiliacora acuminata (Lam.) Hook. f.
& Thoms.
Tinospora cordifolia (Willd.) Hook.
f. & Thoms.
Toddalia asiatica (L.) Lam.
Toxocarpus kleinii Wight & Arn.
Tylophora indica (Burm. f.) Merr.
Ventilago madraspatana Gaertn.
Wattakakka volubalis T. Cooke
Zizyphus oenoplia (L.) Mill.
Tiliaceae
Asclepiadaceae
10
10
B
E
S
S
Sco
Sco
H
G
No
Mi
182.42
26.79
L4
L31

.
32.52.8
3.90.1 1.30.14 15.40.1
8.51.4
1.60.5 0.50.07
7.10.9
Linaceae
Apocynaceae
Oleaceae
Verbenaceae
Asclepiadaceae
10
4
4
4
4
E
E
E
E
B
S
S
S
S
S
Ch
Co
Co
Sco
Co
G
G
G
H
G
Mi
Mi
Mi
No
Mi
9.15
5.08
121.88
57.43
11.94
L4
L12
L30
L21, Gh5
L38, Bt7
2.51.4
4.21.7
12.52.1
12.51.4
5.70.5
5.40.7
10.50.4
1.60.4
7.60.4
2.50.5
2.40.07
4.70.07
0.80.07
1.60.05
1.30.07
0.70.1
1.50.1
2.90.1
12.60.9
2.50.7
Capparaceae
Cucurbitaceae
Olacaceae
Menispermaceae
4
4
4
10
E
B
E
E
S
S
S
S
Sco
Sco
Co
Co
G
Hs
H
G
Mi
Mi
Mi
No
19.79
189
10.07
215.63
Bt1
Bt4
Bt3
Bt3
2.50.7
10.51.4
30.7
27.52.1
2.60.7
1.30.7
2.90.1
3.80.1
1.30.07
0.40.27
1.50.07
1.70.14
2.51.4
5.51.4
2.70.2
17.50.7
Moraceae
Verbenaceae
4
10
E
E
S
S
Co
Sco
G
G
No
No
165.85
423.2
Bt5
Bt6
190.7
110.7
30.7
2.40.1
1.20.05
1.50.14
142.8
4.31.1
Icacinaceae
Celastraceae
Convolvulaceae
10
10
4
E
E
E
S
S
S
Sco
Sco
Co
G
G
G
Mi
Mi
Mi
151.6
22.66
100
Bt7
Bt5
Bt6
23.51.4
26.51.4
31.50.7
8.70.7
5.50.5
3.60.2
4.40.49
2.60.28
1.40.14
101.4
20.51.4
152.8
Hippocrateaceae
Loganiaceae
Menispermaceae
4
10
4
E
E
E
S
S
S
Sco
Co
Sco
G
G
G
Mi
Mi
No
105
64.3
31.69
Bt1
Bt4
Bt2
22.50.7
232.1
10.71.0
2.50.5
2.40.1
2.80.1
1.50.14
1.40.07
1.30.07
14.50.7
16.51.4
71.4
Menispermaceae
Sco
Mi
27.05
Bt3
28.51.4
2.60.4
1.20.14
14.53.5
Rutaceae
Asclepiadaceae
Asclepiadaceae
Rhamnaceae
Asclepiadaceae
Rhamnaceae
4
4
4
4
4
10
E
E
D
E
E
B
Ctri
S
S
S
S
S
Sco
M
co
Sco
M
Sco
G
G
G
G
G
H
Mi
No
Mi
Mi
No
Mi
10.12
43.43
127
15.39
102.89
93.63
L12
L3
L36
Bt5
L3
L28, L31
211.4
3.50.7
111.4
131.4
23.51.4
24.50.7
3.80.1
3.40.1
6.50.1
7.50.0
4.60.4
3.50.0
1.80.12
1.50.52
2.50.31
3.50.64
3.40.29
1.50.07
8.51.4
20.7
3.60.6
40.7
192.8
172.1
65

.
Herbs
Dendrophthoe falcata (L. f.) Ettingsh Loranthaceae
4
E
S
Co G
Mi
Bt2
13.41.6
6.80.4 0.80.35 11.31.4
Ecbolium viride (Forssk.) Alston
Acanthaceae
4
E
S
Sco H
Mi
L39
3.90.7
1.90.3 1.40.21
1.80.4
Phoenix pusilla Gaertn.
Arecaceae
4
E
S
Co G
Mi
Bt1
72.1
2.90.9 0.80.35
5.51.1
Sanseveria roxburghii Schultes &
Agavaceae
10 E
S
Suc G
No
Gh2
70.7
1.20.1 0.90.49
4.80.6
Schultes
Theriophonum minutum (Willd.)
Araceae
4
E
S
Sco G
Mi
Gh1, Bt1
7.51.0
1.90.5 1.10.14
3.80.4
Baillon
Note: SS (Sample size): 30 leaf samples from each individuals of 10 plants for dominant species, 4 for sub-dominant and 2 for rare species. PT (Plant type): E =
Evergreen; B = Brevi-deciduous; D = Deciduous; LF (Life-form): T = Tree; L = Liana; H = Herb. LT (leaf type): S = simple; C = compound. LR (Leaf texture): M =
membranous; Ch = chartaceous; Co = coriaceous; Sco = sub coriaceous; LS (Leaf surface): G = Glabrous; H = Hairy (Hp = Pubescent; Hs = Scabrid, Hh = Hispid); LA
(Leaf area): Nanophyll (0.25-2.25 cm2); Microphyll (2.25-20.25 cm2); Notophyll (20.25-45 cm2); Mesophyll (45-182.25 cm2); SLA (Specific leaf area); FH (Foliar
herbivores): B = Beetle; W = Weevil; L = Larvae (Lepidoptera); Gh = Grasshopper; For expansion of codes see Supplementary table II); For per cent leaf damage data of
two years were averaged for each season.
66
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 6769, 2016
Research article
Three new combinations in Acmella (Asteraceae: Heliantheae)

Reshmi G. R.* and Rajalakshmi R.
Department of Botany, University of Kerala, Kariavattom, Thiruvananthapuram, Kerala, India
*Corresponding Author: grrechugr@gmail.com
Abstract: Three new combinations are proposed: Acmella vazhachalensis (Sheela) Reshmi &
Rajalakshmi, A. ghoshinis (Sheela) Reshmi & Rajalakshmi and A. tetralobata (Reshmi &
Rajalakshmi) Reshmi & Rajalakshmi, based on taxa originally described in Spilanthes.
Keywords: Acmella - Pappus - Chromosome number - Kerala - India.
[Cite as: Reshmi & Rajalakshmi (2016) Three new combinations in Acmella (Asteraceae: Heliantheae). Tropical
Plant Research 3(1): 6769]
INTRODUCTION
Spilanthes was first described by Jacquin (1760) with two species, Spilanthes incipida and S. urens. Richard
(1807) described Acmella as a genus of five species that differ from species of Spilanthes in having ray florets
and lack of pappus. Cassini (1822) suggested that Acmella might be treated better as a section within Spilanthes.
De Candolle (1836) followed Cassini's suggestion and recognized two sections, namely sect. Salivaria DC. and
sect. Acmella (Rich.) DC. Moore (1907) in his revision of the genus Spilanthes described section Salivaria with
13 species and section Acmella, with 26 species. Jansen (1981) provided convincing evidences for the
recognition of Acmella and Spilanthes as distinct genera based on morphological, chromosomal and molecular
evidences. He characterized Spilanthes with stiff awned pappus, monomorphic achenes, sessile leaves, discoid
heads and white to purplish-white corolla. On the other hands, Acmella consists of soft pappus bristles,
dimorphic achenes, petiolate leaves, radiate and discoid heads and usually orange-yellow to yellow or
occasionally white corolla. He (Jansen 1985) transferred some of the taxa from Spilanthes to Acmella and finally
recognized 30 species and 9 infraspecific taxa in Acmella.
While revising the genus Spilanthes in India, Sivarajan & Remesan (1987) overlooked the detailed
morphological and chromosomal evidences provided by Jansen (1981) and merely followed Moore (1907).
After the revision of Sivarajan and Remesan, detailed taxonomical revision was not reported for Spilanthes. Still
some of the Indian treatises (Ramsewak et al. 1999, Saraf & Dixit 2002, Thomas 2011, Shefali Arora et al.
2011, Kishan et al. 2011, Veda et al. 2012, Anuradha Sharma et al. 2012) have followed the broader concept of
the genus Spilanthes.
Three taxa endemic to Kerala, India were recently reported; include Spilanthes vazhachalensis Sheela, S.
ghoshinis Sheela and S. tetralobata Reshmi & Rajalakshmi (2007, 2010, 2014) following Moore (1907).
However detailed taxonomic study revealed that these three species are shared the characters of Acmella rather
than Spilanthes. Jansen described a base number of 12 or 13 for Acmella, and 16 for Spilanthes (Jansen &
Stuessy 1980, Jansen 1981). Cytological study of these three taxa, conducted by Reshmi & Rajalakshmi (2015)
also proved the basic chromosome number, x = 13. Therefore, in the light of available chromosome data
(Reshmi & Rajalakshmi 2015) and evaluation of morphological characters these taxa should be included in
Acmella, requiring the following new combinations,
1. Acmella ghoshinis (Sheela) Reshmi & Rajalakshmi comb. nov.
(Fig.1 A& B)
Spilanthes ghoshinis, Sheela, JETB 34(4): 798800; 2010. Type: India, Kerala, Ernakulam, Desom, Aluva,
25.02.2009, Sheela 00615 (Holo: KFRI!).
67
Reshmi & Rajalakshmi (2016) 3(1): 6769

.
2. Acmella tetralobata (Reshmi & Rajalakshmi) Reshmi & Rajalakshmi, comb. nov.
(Fig.1 C& D)
Spilanthes tetralobata Reshmi & Rajalakshmi, IJAR 2(11): 10921097; 2014. Type: India, Kerala,
Ernakulam district, Koothattukulam, 28 m, 11.09.2012, G. R. Reshmi 7106 (Holo TBGT! Iso KUBH!).
3. Acmella vazhachalensis (Sheela) Reshmi & Rajalakshmi comb. nov.
(Fig. 1 E &F)
Spilanthes vazhachalensis, Sheela, JETB 31(2): 474477; 2007. Type: India; Kerala, Trichur District,
Vazhachal, 25.04.1991, Sheela 00400 (Holo: KFRI!).
Figure1. A, Habit of Acmella ghoshinis; B, Twig of A. ghoshinis; C, Habit of A. tetralobata; D, Twig

of A. tetralobata; E, Habit of A. vazhachalensis; F, Twig of A. vazhachalensis.
68
Reshmi & Rajalakshmi (2016) 3(1): 6769

.
ACKNOWLEDGEMENTS
The first author thanks Kerala State Council for Science, Technology and Environment (KSCSTE),
Government of Kerala, Thiruvananthapuram, India, for its financial support to carry out Research programme.
We thank the curators of KUBH, KFRI, and TBGT for making specimens available for examination.
REFERENCES
DeCandolle AP (1836) Spilanthes. In: DeCandolle AP (ed) Prodromus systematis naturalis regni vegetables.
Treuttel and Wurtz, Paris, pp. 620626.
Cassini H (1822) Spilanthes. In: Cassini H (ed) Dictionorie des sciences naturelles. Le Normant, Paris. pp. 328
331.
Jacquin NJ (1760) Enumeratio Systematica Plantarum quas in insulis Caribaeis. Inter Documentation Company
AG, Leiden, pp. 28
Jansen RK & Stuessy TF (1980) Chromosome counts of Compositae from Latin America. American Journal of
Botany 67: 585594.
Jansen RK (1981) Systematics of Spilanthes (Compositae: Heliantheae). Systematic Botany 6: 231257.
Jansen RK (1985) The systematics of Acmella (Asteraceae- Heliantheae). Systematic Botany Monographs 8: 1
115.
Kishan LT, Shailesh KJ & Veenu J (2011) An updated review on medicinal herb genus Spilanthes. Journal of
Chinese Integrated Medicine 9 (11): 11701178.
Moore AH (1907) Revision of genus Spilanthes. Proceedings of the American Academy of Arts and Sciences 42:
521569.
Ramsewak RS, Erickson AJ & Nair MG (1999) Bioactive N-isobutylamides from the flower buds of Spilanthes
acmella. Phytochemistry 51(6): 729732.
Reshmi GR & Rajalakshmi R (2014) Spilanthes tetralobata sp. nov. (Asteraceae): a new species from Kerala,
India. International Journal of Advanced Research 2: 10921097.
Reshmi GR & Rajalakshmi R (2015) Chromosome number and polyploidy in Spilanthes Jacq. (Asteraceae:
Heliantheae). Nucleus (Springer) 58(2): 107109.
Richard LC (1807) Acmella. In: Persoon C (ed) Synopsis Plantarum, Paris, 42: pp. 472473.
Saraf DK & Dixit VK (2002) Spilanthes acmella Murr.: study on its extract spilanthol as larvicidal
compound. Asian Journal Experimental Science 16: 919.
Sheela D (2007) Spilanthes vazhachalensis: a new species from Kerala, India. Journal of Economic and
Taxonomic Botany 31 (2): 474477.
Sheela D (2010) Spilanthes ghoshinis: a new species from Kerala, India. Journal of Economic and Taxonomic
Botany 34 (4): 798800.
Sivarajan VV & Remesan C (1987) The genus Spilanthes Jacq. (Compositae- Heliantheae) in India. Journal of
Economic and Taxonomic Botany 10(1): 141148.
Thomas T (2011) Antibacterial action of gradient extracts of flower heads of Spilanthes paniculata Wall. ex.
DC. Plant Sciences Feed 1 (11): 186189.
Veda P, Supaluk P, Somsak R & Virapong P (2013) High therapeutic potential of Spilanthes acmella: a review.
Experimental and Clinical Sciences International online journal for advances in science 12: 291312.
69
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 7077, 2016
Research article
Prospects of organic farming in hill farms of Nepal

Sabita Aryal Khanna
Kathmandu University, School of Science, Department of Environmental Science and Engineering, Nepal
*Corresponding Author: sabita@ku.edu.np
Abstract: Agriculture is the most important industry contributing about 37.0% to national GDP
and provides livelihood for over 65.7% of the population of Nepal. Chemical based agriculture has
destroyed the ecosystem, declined natural resource base, provided diminishing returns, reduced
food self-sufficiency, forced for migration, disrupted food security and export opportunities. The
landscape with diversified flora, fauna, ecosystem, and manpower availability gives opportunity to
organize movement of organic agriculture thus some products of Nepal like tea and coffee have
already been certified as organic products and have good export returns. The practice of producing
organic vegetables, fruit, milk, meat, fishes and their product are also increasing. Thulodurlung
VDC of Lalitpur district is the one which is becoming popular for producing organic coffee. Hence
the study is conducted at this site and also at nearby village Chamranbesi VDC of Kavre district.
The study was aimed to examine the existing status of farming as well as prospects of other
organic crops including vegetables, cereals and dairy products. Different approaches such as PRA,
FGD, Household survey and Key informants interviews were used to collect the information.
Data collected were analyzed quantitatively and qualitatively. The overall findings showed that
organic farming can be successfully done in both sites. However, it is easier to achieve the result in
Thulodurlung as compared to Chamranbesi. There is a growing attitude of sound biological and
livestock based farming. Lack of functional road, unavailability of irrigation water, disease and
pest infestation on crops are major challenges. On a commercial scale, apart from organic coffee,
other farm and organic dairy products such as ghee, cheese, butter need to be marketed. There is
an instant need of attention on sound marketing strategies, technology of processing the harvest
and overall improvement in the capacity of farmer to produce organic crops.
Keywords: Agriculture - Livelihood - Natural resources - Commercial scale production - Nepal.
[Cite as: Khanna SA (2016) Prospects of organic farming in hill farms of Nepal. Tropical Plant Research 3(1):
7077]
INTRODUCTION
Agriculture is the primary source of nutrition for the global population. Almost three quarters of the
population in developing countries subsists on agriculture. It is one of the major aspects of human development.
Agriculture is most important for Nepal as it provides livelihood resources for over 65.7% of the population and
contributes about 37.0% to the national GDP (Factfish 2015). In the past decades, green revolution has brought
some significant changes in the worlds food production systems such as increased food production and
productivity, income from agriculture has risen and employment opportunities have been diversified in both
developed and developing countries (Joshi et al. 2007). At the same time the green revolution has also brought
several agro ecological consequences. (Tilman et al. 2002, Pretty et al. 2011) It has had less impact on
resources poor farmers (Rosset et al. 2000); it has contributed in natural resource declination (Espinel 2015)
thus having created several environmental problems (Singh 2000, Tilman et al. 2001). It has shown diminishing
returns in intensive agricultural areas (Naylor 1996). This has helped to widen the gap between rich and poor. It
has greatly contributed in reducing food self-sufficiency for most of the poor people and resource less countries.
With this realization, the organic movement was started and widened in 1972 in Europe and USA in the
global context (Raynolds 2000). In Nepal, the organized movement of Organic agriculture was started in 1986
(Paudyal 2015). In the present context, this concept is gradually increasing and some products of Nepal like tea
70
Khanna (2016) 3(1): 7077

.
and coffee have been already certified as organic products. But those crops which have daily consumption like
vegetables, cereals are yet to be declared as organic food. Through the practice of producing organic vegetables,
fruit, milk, meat, fishes and their product have existed for more than twenty years in some places of Nepal like
Gamcha of Bhaktapur and Fulbari of Chitwan. There are also some other emerging places where farmers have
started doing organic farming (Tripathi & Khanna 2010). Unless the valued crop production with the approach
of organic farming is promoted in these areas the land will remain unattended. Nepal being a mountainous
country, having an area of 1,47,181 km2 out of which only 42,590 km2 is used for agricultural practice.
Moreover the fortune of integrated small scale subsistence farming using only traditional agriculture practices
and its virginity towards chemicals add more scope of valued production.
Thulodurlung VDC is the one which is becoming popular for producing organic coffee. Hence the study is
conducted at this site to examine the prospects of other organic crops including vegetables, cereals and dairy
products. In addition, the nearby village of Chamranbesi is also selected for the same purpose.
Objectives:
To know the current status of farm activities.
To examine the possibility of prospective crops and products.
To investigate the prospects of organic farming.
Different approaches such as participatory rural appraisal (PRA), focus group discussion, household survey
and key informants interviews were used to collect the required information.
PRA tools such as resource/social mapping, ethno history, pair matrix and seasonal calendar used to collect
qualitative information to fulfil the research objectives. PRA tools were exercised among groups of both men
and women (at least 25) in the selected sites.
Key informants were voluntarily chosen, to facilitate participatory exercise, particularly to undertake written
participation. Only that information came through group consensus was considered as findings. For this
exercise, researcher helped participants as a facilitator. In focus group discussions, all the participants were
allowed free discussion and valuable information was picked up and recorded.
A household survey was also conducted to collect quantitative information on demographic characteristics
and their farming activities. 30 samples were taken in each site through random sampling. Questionnaires were
prepared and the data were taken through direct interviews.
Data analysis and interpretation: Data collected from household survey were analysed through X-cel and
interpreted in average. All the information collected from PRA, household survey and focus group discussion
were put together and qualitatively analysed to produce concrete results.
Demographic characteristics
Thulodurlung VDC covers 300 households and Chamranbesi covers 312 households. Detail of the household
ethnicity wise is given in table 1. In both sites agriculture is the main source of livelihood. Apart from this
occupation few people are involved in teaching services, business, overseas employment and other services as
secondary source of income.
Table 1. Education status of Thulodurlung and Chamranbesi.
Literacy (%)
Number of schools
Illt.
Lit.
Sec. Hig.
Prim. Low. Sec.
Sec.
Thulodurlung
Male
33.3 100
25
33.3
2
1
1
Female
66.7 0
75
66.6
Total
29.6 29.6 33.3 7.4
Chamranbesi
Male
25
87.5 44.4 50
3
1
1
Female
75
12.5 55.5 50
Total
55.6 18.5 14.8 11.1
Note: Illt. = Illiterate, Lit. = Literate, Prim. = Primary (5th), Low. Sec. = Lower secondary (6th
Sec. = Secondary (9th10th Class), Hig. = High school (10th).
Study Sites
Gender
Hig.
-
8th),
71
Khanna (2016) 3(1): 7077

.
The education status of these VDCs is improving. Education of male is better than female with higher
literacy % as compared to female. Most of the female groups above 25 were illiterate. In present context,
children go to school in their own village but due to lack of higher schools, many of them stopped their further
education which is especially prevalent among female groups. Some of them continue their studies either nearby
schools in Banepa, Panauti or in Kathmandu.
Land under cultivation
Durlung and Chamranbesi VDCs comprise both upland and lowland type of farming area. Generally farmers
have their own land for cultivation (Table 2). However, in Durlung, about 7.7% and 15.38 % are cultivating on
leased lands of upland and lowland respectively. The average upland area under cultivation is comparatively
more than area of lowland in both VDCs. Between two VDCs, upland area owned by the households is more in
households of Chamranbesi (96.3 %) and lowland is more in Thulodurlung (61.5%).
Table 1. Area and type of land under cultivation in Thulodurlung and Chamranbesi.
Study Sites
Thulodurlung
Chamranbesi
Attributes
Own (% households)
Lease(% households)
Average land area / household (ropani)
% households
Own (% households)
Lease(% households)
Average land area / household (ropani)
% households
Upland
84.61
15.38
16.12 (range: 4-28)
92.3
100
0
7.55 (range:2-27)
96.3
Lowland
92.3
7.7
4.4 (range:2-12)
61.5
100
0
3.46 (range:1-9)
48
Livestock
In both places, livestock normally included buffalo, cow and goat along with their young ones (Table 3).
Likewise, they have also bulls for ploughing purpose which is more in Thulodurlung. Normally people like to
keep more buffaloes than cows as the buffalo milk contains more fat than cow milk. The average number of
buffalo in sampled households is 3.36 in Thulodurlung and 3.24 in Chamranbesi. In Thulodurlung, almost all
have kept goat that are sold for meat purposes. About 30.76 % households keep hens in Thulodurlung; which is
comparatively a big size than in Chamranbesi (7.4).
Table 3. Livestock in Thulodurlung and Chamranbesi VDCs
Site
Thulodurlung
Chamranbesi
Attributes
Avg no. / household
% of owned household
Avg no./ household
% of owned household
Cow/Bull
2
54
1.3
37
Buffalo
3.36
96
3.24
98.15
Goat
9.8
100
4
88.9
Hen
4.7
30.76
5.5
7.4
Crops, crop productivity and cropping pattern

Maize and Rice are the main crops of people in both VDCs and these crops are mainly grown for living but
most of the households find it insufficient for year round. Apart from these they also grow finger millet, wheat
and different seasonal vegetables. But wheat is not found in Thulodurlung. Generally vegetables are grown in
small area only for home consumption in most of the households as they have no transportation facilities for
marketing. Among different vegetables, cauliflower, onion, garlic, gourds are the major ones. Besides these,
some portion of the people in both VDCs generate additional income from broom grass.
The average rice productivity of Thulodurlung and Chamranbesi is 2.27 (range: 16) t/haand 3.57 (range:
1.97) t/ha respectively. As compared to the national average (2.7t/ha), the productivity of rice is more in
Chamranbesi whereas it is lower in Thulodurlung. From this survey it was also noted that the productivity of
maize (1.02t/ha) of Chamranbesi is higher than in Durlung (0.65t/ha) but this productivity is lesser than the
national average i.e. 2.03 t/ha. This might be due to the disease like blight which has appeared in maize since
three years in almost all the area of both VDCs.
The main cropping pattern in Durlung involved two crops in a year in case of lowland whereas in
Chamranbesi it included three crops in a year.
Thulodurlung:Lowland: Maize + Rice
Upland: Maize/Finger millet + Coffee or Maize + Finger millet
72
Khanna (2016) 3(1): 7077

.
Chamaranbesi:Lowland: Maize-Rice-Wheat
Upland: Maize + Finger millet
Prospective farming
Coffee and silk worm farming are newly introduced farming which are found to have good source of income
generation. Coffee production is more popular in Durlung and silk worm in Chamranbesi. There are also several
other farming that can raise the livelihood of the people. It includes bee rearing, fish farming, asparagus
cultivation, and poultry. The status of these prospective farming is presented briefly in the table 4.
Table 4. Prospects of new farming with their status in Thulodurlung and Chamranbesi VDCs
Bee farming
Thulodurlung
Chamranbesi
Silkworm
farming
Thulodurlung
Chamranbesi
Coffee
farming
Study
Sites
Thulodurlung
Farming
Status
Supportive Reasons
First introduced by CDHP in 1985

Again introduced 5 years back (2003)
with an aim of organic production by
Cope program of Helvitas, a nongovernmental organization
Except ward no. 1, all the area of Durlung
comprised coffee production along with
other crops
93% sampled households are involved in
this farming
people earning 10,000 to 60,000 rupees
annually or even higher
There is a processing unit established
People are totally inclined with organic
coffee farming
Four coffee co-operatives existed in ward
no. 7, Gumrang
Coffee farming at initial stage; only 12
households of the whole VDCs involved
Suitable climate for coffee

cultivation
Less use of chemicals from
very beginning; easier to
move
towards
organic
production
Good income generating source
High level of farmers interest
Know about silkworm farming but not yet

involved
Suitable climate for mulberry

cultivation
The village is very near to
Khopasi, the highest cocoon
producing area of Nepal
Suitable climate for mulberry
cultivation
Started mulberry cultivation from four

years ago (2004)
Nawa Jagriti Resam farmers group
established in 2004 with 25 members
Only 1-2 bee hives in the village; only for
home consumption
Promoting people to keep hives in coffee
farm by Cope
Suitable climate
May have good contribution
from Durlung (where coffee is
already established) in terms
of sharing knowledge and
source
Farming about to turn into

totally organic
Adequate
rearing
place
including coffee farm
73
Chamranbesi
Thulodurlung
Chamranbesi
Poultry
Thulodurlung
Asparagus
Thulodurlung
Chamranbesi
Chamranbesi
Fish
farming
20 households have kept bee hive only for

home consumption
Not started in commercial scale
Khanna (2016) 3(1): 7077

.
Adequate rearing
place as
mustard farms and nearby
forests
People are avoiding chemical
fertilizers and about half of
the households have stopped
using chemicals
-
Not yet started for selling

Good prospects in 9, 2 and 3 ward no.
Farmers involved in planning and
discussion for fish farming
Water
resources
available
especially
fresh
water
streams, ponds and small
springs
High market demand in nearby
by town
and cities viz;
Panauti, Banepa, Kathmandu
and Lalitpur
Lots of wild asparagus available in nearby

forest
Root part, only used for medicinal value
in local level
Not yet cultivated
Has initiated searching asparagus seeds
for cultivation
Favourable climatic condition

Distance between the village to
nearby city market is less;
very high price and market
demand
Farmers interest
About 30.7% sampled households involved

in keeping poultry
High demand
About 7.4% of sampled households

keeping poultry with them
Social belief; not to keep poultry farming
in Brahmins house still existed
Very high demand of egg and

meat; Demand even not
fulfilled in the village
Others
Other prospects included strawberry farming and mushroom cultivation. There are many wild strawberry
plants growing in most of the areas in Chamranbesi. Likewise, some respondents have thought about mushroom
cultivation because of its growing demand in the market in recent times.
Manuring and plant protection
The scenario of manuring practice is different between the two study sites. Almost all used only farm yard
manure as a source of fertilizer in both VDCs but in Chamranbesi significant number of households (48.15%)
also use chemical fertilizers like urea, DAP and MoP along with farm yard manure according to their own
knowledge. However, the trend of using chemical fertilizer is decreasing mainly due to expensive prices,
decreasing productivity and higher disease attack. Their growing attitude of keeping more livestock could be
better to have sufficient manure rather than buying expensive fertilizers has also made them avoid chemicals.
Likewise, some of them are also aware about the harmful effect of chemicals and significance of organic
fertilizers.
74
Khanna (2016) 3(1): 7077

.
For plant protection about 40.74 % households in Chamranbesi buy chemical fungicides and pesticides
mainly for potato, vegetables, and rice. They use these chemicals either asking with the seller or according to
their own knowledge.
Concept of organic agriculture
People of Durlung became familiar with the concept of organic agriculture after the establishment of Cope
program under Helvitas. This program was established with an aim of producing organic coffee and people were
trained for different practices of organic farming. Since 5 years they have stopped using chemicals in their farm.
They have also learnt about making fertilizers and pesticides from the available biological resources. Almost all
people are doing organic farming and they are very much interested in this type of farming. They are hoping for
certification of their coffee as an organic coffee very soon and also planning to declare their area as an organic
village.
In Chamranbesi, about half of the respondents (48.15%) said that they know about organic farming through
friends, neighbour and market but they lack doing so due to inadequate knowledge. However, about half of
these respondents want to do in future. For this, they need to have special training and some sort of support.
Marketing
People mainly sell some vegetables, milk, khuwa, goat either in their own village or nearby towns like
Panauti and Banepa. Milk product for selling includes only khuwa, because farmers are likely to get more profit
from this as compared to ghee and curd. In Durlung, normally group of farmers collect their milk in one place
where milk is processed into khuwa; whereas in Chamranbesi, they prepare khuwa themselves in their home.
Though agriculture is the main source of livelihood, people are very much disadvantaged with marketing
process of their products due to lack of road facilities. Transportation of the products from the production sites
to market involves long route of walk which ultimately restricts the marketing of fresh vegetables and fruits.
Though they take some amount to the market, they have to bear a huge loss from postharvest spoilage and low
return.
Problems related with agriculture
From farming to marketing, people face several problems like irrigation, seed, transportation/marketing,
disease, pest, technical ignorance and animals. In Durlung, the greatest problem was irrigation problem followed
by marketing and technical ignorance. Due to irrigation problem they couldnt grow vegetables on a commercial
scale. Though transportation is also a serious problem, people are hopeful for the road construction which is
about to touch their village.
People of Chamranbesi are facing great difficulties in transporting their products to the market. It requires
about 4 hours of walk to reach the road. Hence they have to pay much for the delivery of the products. Another
major problem faced by the farmers of both sites is lack of technical assistance. They think if they could get
technical guidance from technicians, they could improve their production and also get rid of loss caused by
various disease and pest attacks.
Available natural resources
Natural resources in two villages comprise land, biodiversity and water resources. These resources are
detailed in the socio-resource map. The land under cultivation is already been described.
Biodiversity
In Durlung, there are two community forests, one natural forest and one pine forest. In Chamranbesi, forest
has surrounded the Eastern and northern boundary of the village. These forests are rich in medicinal plants,
herbs and different kind of trees. Likewise there are several other valuable plants available in the village. Except
for medicinal purpose, none of the plants are beeing used for household and other income generating purposes.
Some of the plants available in these areas are:
Siltimur, pipala, khirro, utis, pine, rhododendron, Chiraito, amala, kafal, Harro, Barro, asparagus, lemon
grass, bakaino, nigalo, lapsi etc.
Water resources
Both villages are naturally facilitated with water resources. Big and small streams and rivers are flowing
inside and outside the villages. These resources are only utilized for drinking purpose in Durlung and they are
not benefited for irrigation and other purposes. Irrigation facility has not reached in all areas of Chamranbesi
75
Khanna (2016) 3(1): 7077

.
expect a few. Some years back, people in Chamranbesi had their own electricity supply generated from
Sukumkhola hydropower that supplied electricity in 40 households of 9 wards and also shared electricity in 12
households of 4 wards in Durlung. Now the electricity is supplied from Kathmandu; however they still use their
own at the time of load shedding. There is also another hydropower of sukumkhola and both of these
hydropowers are of 3 KW. Total 85 households are benefited from sukumkhola hydropower.
Supporting organizations
VDC office is situated only in Chamranbesi but in Durlung, the office was destroyed by Maoist during
conflict period. People want to have VDC office very much active and should support them both in agriculture
and non-agriculture sector.
The current supporting non-governmental organization in Durlung is Helvitas, Nepal, which has changed the
livelihood of the people to some extent by promoting organic coffee production and marketing. In 1985, another
organization CDHP came here for five years with an aim of improving health status, promoting goat and cattle
keeping, and several other developmental activities. Jaldhara, a project that was started in 1995 worked in
Durlung for 7 years. This program mainly aimed for the construction of infrastructures like bridge, road, toilet
but also brought improved terracing. Likewise, this project was also active in Chamranbesi with similar aims
and activities. At present, there are no other organizations that are supporting them in promoting their livelihood
except UNICEF.
With regard to agricultural activities, people can raise their livelihood if they can upgrade this occupation
from subsistence level to income generating level. Among different perspective of agricultural activities, organic
farming can be established in both VDCs in long run. About 88.9 % of the respondents in Chamranbesi and
almost all in Durlung were interested in doing organic farming but they need support in terms of skill
development and marketing.
Community, Co-operatives and groups
The concept of community formation was first introduced in Durlung by CDHP during 19851990. Since
then, there are several communities and groups and about 50% households of the total are involved in different
groups in Durlung. Basically, Groups and Co-operatives were formed based on similar farming activities like
coffee producing group, silk farming group and veterinary group. Their names are presented in Annex 4.
Likewise women/mothers group was also formed with an aim of empowering female group in the village in
each VDC.
Members of a group are linked with each other through regular meetings, informal discussions and
gatherings. Generally, a regular formal meeting is held on a particular date each month. For other meetings,
members are informed through direct contact and generally these meetings fail to inform the members living in
distant places. During meetings; discussion regarding problems, current situation, and future planning for
improvement take place. Members of co-operatives utilize money in farm investment, health treatment and also
some kind of household activities.
These groups and co-operatives have strong positive impact upon livelihood of the people that is why almost
all love doing farming and other activities in a group.
CONCLUSION AND RECOMMENDATIONS
The study was conducted in two particular VDCs with an aim of finding the prospect of organic farming and
marketing of organic products. The overall findings showed that organic farming can be successfully done in
both sites. However, it is easier to achieve the result in Thulodurlung as compared to Chamranbesi since this
VDC is about to turn into a completely organic village. In spite of chemicals in use, the growing attitude of
avoiding chemicals and keeping livestock has added the prospect of organic farming in Chamranbesi. In
addition, the available biological resources and farmers interest together contribute the possibility of organic
farming in future. Since both of the sites lack road facilities, production of vegetables, cereals and fruits are
limited to only home consumption. Problems associated with irrigation and disease have further restricted the
farming on a commercial scale. However, there are several aspects that can be considered to improve the
farming status of these places. Following recommendations are made to achieve results. On the basis of this
study, following points have been recommended,
76
Khanna (2016) 3(1): 7077

.
According to marketing perspective, apart from organic coffee, other farm products need to be explored in
Durlung village.
Before proceeding for organic farming, people especially in Chamranbesi need to be trained regarding the
concept and skill development. Likewise, they need to be aware about the harmful effects of chemical
fertilizers and pesticides.
Special marketing strategies need to be formed or developed to sustain organic farming in the long run.
There are several challenges and problems which need to be identified and assessed so as to develop
suitable strategies.
Dairy products like ghee, cheese, butter could be a good source of income generation if the village is
facilitated with processing unit.
The prospective farming described above need to be initiated or expanded for which farmers should be
provided with special support in terms of skill development and marketing.
Modification of high volume fresh vegetables and fruits into small volume processed form can be an
opportunity to minimize the problem of road access. For this several processing units need to be
established within the village.
For sustainable management, people should be facilitated with technical guidance at every step of farm
activities.
REFERENCES
Espinel RL (2015) Multifunctionality in Peasant Agriculture: a means of Insertion into Globalization. Available
from: http://www.agter.asso.fr/IMG/pdf/Espinel_2008_Multifunctionality_in_Peasant_Agriculture.pdf (accessed:
05 Nov. 2015).
Factfish (2015) Nepal Statistics. Available from: http://www.factfish.com/imprint. (accessed: 10 Nov. 2015).
Joshi PK, Gulati A & Birthal PS (2007) Agricultural Diversification in India. In: Agricultural diversification
and smallholders in South Asia. International Food Policy Research Institute, Washington, DC, USA, pp.
219242.
Naylor RL (1996) Energy and resource constraints on intensive agricultural production. Annual
Review of Energy and the Environment 21: 99123.
Paudyal KP (2015) Organic Agriculture in Nepal. Available from: http://www.afaci.org/file/anboard2/Nepal
(word).pdf (accessed: 10Oct. 2015).
Pretty J, Toulmin C & Williams S (2011) Sustainable intensification in African agriculture. International
Journal of Agricultural Sustainability 9(1): 524.
Raynolds LT (2000) Re-embedding global agriculture: The international organic and fair trade movements.
Agriculture and Human Values 17: 297309.
Rosset P, Collins J & Lappe FM (2000) Lessons from the Green Revolution. Tikkun 15 (2): 5256.
Singh RB (2000) Environmental consequences of agricultural development: a case study from the Green
Revolution state of Haryana, India. Agriculture, Ecosystems & Environment 82(13): 97103.
Tilman D, Cassman KG, Matson PA, Naylor R & Polasky S (2002) Agricultural sustainability and intensive
production practices. Nature 418 (6898): 671677.
Tilman D, Fargione J, Wolff B, D'Antonio C, Dobson A, Howarth R, Schindler D, Schlesinger
WH, Simberloff D & Swackhamer D (2001) Forecasting agriculturally driven global environmental change.
Science 292(5515): 281284.
Tripathi L & Khanna SA (2010) Areport on Study the Development of Organic Farming and Certification in
Nepal, A Case Study of Chamrangbesi Valley. Kathmandu University Library.
77
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 7886, 2016
Research article
Low Na/K ratio in the leaves of mangroves mitigates salinity

stress in estuarine ecosystem
Anjum Farooqui1*, Ranjana1 and Yogesh Joshi2
1
Birbal Sahni Institute of Palaeobotany, 53, University Road, Lucknow, India

2
Department of Botany, Kumaun University, S.S.J. Campus, Almora, India
*Corresponding Author: ranjanamsc9@gmail.com
Abstract: Plants have their own genotype which allows them to absorb, accumulate or exclude
nutrients particularly sodium (Na) and potassium (K) through their root system that are essential
for physiological activities and growth in coastal wetlands. Dominant mangrove species such as
Avicennia alba, A. officinalis, A.maritima, Suaeda monoica, S. nudiflora, S. maritima and
Acanthus ilicifolius were selected from Andhra Pradesh coast to observe the sodium (Na) and
potassium (K) accumulation in the leaves and the salinity in the aqueous soil solution in their
respective rhizosphere. Results reveal that only Acanthus ilicifolius grew in low soil salinity of
~0.5 to 1.2 ppt. and are moderate accumulators of Na and K in the leaves. Avicennia alba is highly
sensitive to salinity due to its high Na/K ratio in leaves. A. officinalis and A. marina, S. nudiflora
and S. maritima show low Na/K ratio and are tolerant to salinity stress. Acanthus ilicifolius and
Suaeda monoica show moderate ratio of Na/K in their leaves. The leaf epidermal modifications
through salt glands (Avicennia and Acanthus spp.) and prevention of water loss by non-gladular
trichomes as in case of Avicennia species helps in mitigation of salinity stress but their affinity
towards K uptake and translocation to leaves primarily regulates the physiology and growth of
plants to tolerate or succumb to salinity stresses. Except for Suaeda monoica both S. nudiflora and
S. maritima are non-accumulators of Na and K showing their low ratio in leaves. K is an essential
component in the leaves to mitigate increased salinity in the ecosystem despite the leaf epidermal
morphological adaptations to exude excess salt. Plants which are efficient in K absorption are
more tolerant to salinity stress while others are comparatively sensitive.
Keywords: Mangroves - Sodium - Potassium - Leaf epidermis - Salinity.
[Cite as: Farooqui A, Ranjana & Joshi Y (2016) Low Na/K ratio in the leaves of mangroves mitigates salinity
stress in estuarine ecosystem. Tropical Plant Research 3(1): 7886]
INTRODUCTION
Coastal wetlands are vulnerable to direct impacts of changing climate that induces sea level fluctuations and
salinity related ecological disturbances. Worldwide coastal biodiversity is sensitive to these dynamic frequent
changes and several species are at the verge of extinction. Anthropogenic activities enhance the deterioration of
coastal ecology not conducive for different mangrove species. Mangroves and associates confined to intertidal
areas are subjected to high salinity in two ways. Firstly, through sea water inundation (Morrow & Nickerson
1973, Tomlinson 1986) and secondly, at the leaf level through the deposition of airborne salt spray (Boyce
1954). Adaptability, tolerance and susceptibility are the three major physiological processes (Farooqui et al.
1995) that the biotic forms have in their genes to abate stress conditions. The total concentration of salts
translocated through root system needs to be regulated within the growing plant tissues (Paliyavuth et al. 2004).
The roots of Avicennia species have the ability to filter over 90% of salt from the water they uptake (Scholander
et al. 1962, Drennan & Pammenter 1982). In addition to this, the ions are stored in the leaf hypodermis,
followed by active excretion through glands on the leaf surface (Waisel et al. 1986, Smith et al. 1989, Dschida
et al. 1992, Balsamo & Thomson 1995). Salt glands are the special adaptive structures (multicellular trichomes)
which are predominantly found on the leaves and stems of halophytic species in particular. These are efficient
desalination devices capable of salt excretion from plant tissues (Fahn 2000). The function of stomatal
78
Farooqui et al. (2016) 3(1): 7886

.
conductance in any plant species is regulated through the osmotic processes during which potassium plays a
major role in maintaining the turgor pressure of guard cells (Dietrich et al. 2001).
Plants exhibit various adaptive strategies in response to different abiotic stresses such as salinity, which
limits the plant growth and productivity (Munns 2002). Avicennia grows in the slightly elevated and mostly in
adverse and frequently changing part of the intertidal habitat. The surface could be often dry for extended
periods and salts increase in the substratum due to its upward movement in the soil column through capillary
action. Three species of Avicennia such as A. alba, A. marina and A. officinalis are in general encountered along
the Indian coastal zone along with Suaeda species (S. maritima, S. monoica and S. nudiflora) and Acanthus
ilicifolius. However, A. marina, A. oficinalis, S. nudiflora and Acanthus ilicifolius are common in the coastal
belt of India and form pure stands. The east coast of India is characterized by shallow coastal wetlands which
are more affected by even the slightest change in climate induced sea level changes enhanced by anthropogenic
activities. It estimated that about 1.76 lakhs hectares of land in Andhra Pradesh are affected by salinity, in the
districts of Prakasam, Guntur, Krishna and East Godavari. The soil along the coast (about 1015 km from the
sea) is mostly sandy in nature and of marine origin. The sub-soil water is also found to be generally rich in salt
content in certain areas. The salt tolerance potential varies from genotype to genotype and species to species
within the plant kingdom (Moisender et al. 2002). The present study highlights the leaf epidermal morphology
to mitigate salinity stress and Na/K ratio in the leaves responsible for species zonation of three dominant plant
genera such as Avicennia, Acanthus and Suaeda in coastal wetlands.
Sediment soil of 50 cm depth from ground surface was collected from around the rhizosphere of Avicennia
alba, A. officinalis, A. marina, Suaeda monoica, S. nudiflora, S. maritima and Acanthus ilicifolius. These were
sub-sampled at an interval of 10 cm to measure the total dissolved salts (salinity). The salinity was measured in
the aqueous soil solution in order to know the concentration of salts available for the absorption by plants. For
this ten sites were selected to collect the soil from rhizosphere of the respective plant species. Since the salinity
variation was not much in a 50 cm sediment profile, the salinity given in table 1 is the average of 5 sub-samples
in 50 cm deep sediment. The sample numbers 110 (Table 1) are 10 different soil cores. The salinity was
measured in aqueous soil solution. For this 10g air dried soil sample was dissolved in 100ml of deionized water
and kept overnight after rigorous shaking for 1hr. Samples were homogenized for 30 minutes prior to salinity
measurement in ppt (parts per thousand) by using Orion-5 Star (Thermo-Orion, Scientific Equipment, USA) at
standardized 20C temp.
Table 1. Average salinity (5 sub-samples of 50 cm core) in the soil rhizosphere of studied plants (*Salinity in ppt- parts
per thousand).
Core
Numbers
1
2
3
4
5
6
7
8
9
10
Average (*ppt)
Avicennia
alba
2.2
2.8
1.2
1.5
1.9
3.0
2.3
2.1
1.2
1.6
1.98
Avicennia
officinalis
1.5
1.8
1.0
0.9
1.2
1.3
1.6
1.9
2.0
1.3
1.45
Avicennia
marina
3.0
3.2
3.5
2.9
2.5
1.9
3.8
2.9
1.9
2.0
2.76
Suaeda
monoica
2.1
1.2
1.5
1.2
1.4
1.5
1.0
2.0
2.8
1.7
1.64
Suaeda
nudiflora
3.0
3.2
4.0
4.1
3.2
3.8
4.5
4.3
4.2
3.1
3.74
Suaeda
maritima
2.4
2.6
2.0
2.8
3.7
2.8
3.6
4.0
3.1
2.5
2.85
Acanthus
ilicifolius
1.0
0.7
0.8
0.9
1.1
1.0
0.9
1.2
0.5
0.7
0.88
Ten mature leaf samples each of Avicennia alba, A. officinalis, A. marina, Suaeda monoica, S. nudiflora and
S. maritima along with Acanthus ilicifolius were collected randomly from the coastal areas of Krishna- Godavari
delta (Fig. 1 & 2). The Sodium (Na) and Potassium (K) was measured by Flame photometer (ELICO-CL-360)
in 10 g air dried leaf samples which was acid digested (nitric acid and perchloric acid). The residue was made up
to 100 ml using deionized water before the analysis. For morphological study the leaves were boiled in water
and glycerine (60:40) with few drops of conc. nitric acid until the peel appeared separated. The peel was
brushed off in clear water to remove the tissues and later mounted on slides in glycerinated medium. The upper
79
Farooqui et al. (2016) 3(1): 7886

.
and lower epidermal surface was observed under Olympus BX-51 and the photographs taken with DP-26
camera. The contributions of AF include microscopic observations, chemical analysis of leaves/soil and
compilation of the manuscript. Ranjana contributed by processing of the samples and microscopic observations.
YJ contributed in giving suggestions and providing partial input in the analysis of results and discussion.
Figure 1. Map showing south-east coast and the percentage of salt tolerant mangroves.
Figure 2. A, Avicennia marina and A. officinalis along with Suaeda nudiflora and S. maritima on highlands that are often
inundated by tides but not regularly. Inset, showing interspersed salt accumulation on the surface; B, Avicennia officinalis
along the back water channel towards land and Acanthus ilicifolius on highlands; C, Avicennia alba, regularly inundated by
high tides twice a day (near shore).
80
Farooqui et al. (2016) 3(1): 7886

.
RESULTS
Salinity in aqueous soil solution
The soil samples in the vicinity of roots of Avicennia alba shows minimum 1.2 and maximum 3.0 ppt (Table
1). The average soil salinity in 10 sites where A. alba was growing shows 1.98 ppt in the aqueous soil solution.
The average soil salinity was 1.45 ppt in the region where A. officinalis was growing. The minimum salinity was
1 and the maximum was 2.0. Out of all the Avicennia species the highest soil salinity was in the soil where A.
marina was growing. The average salinity in soil was 2.76. The minimum was 1.9 and the maximum was 3.8.
Among the three species of Suaeda the highest average salinity (3.74) in aqueous soil solution was recorded
where S. nudiflora was growing followed by S. maritima (2.85) and S. monoica (1.64). The minimum salinity
was recorded in the rhizosphere of S. monoica (1 ppt) followed by S. maritima (2) and S. nudiflora (3). The
maximum salinity was recorded in S. nudiflora (4.5 ppt) followed by S. maritima (3.7) and S. monoica (2.8).
The lowest average salinity was recorded in the vicinity of Acanthus ilicifolius (0.88 ppt). The minimum
concentration in the soil was 0.7 ppt and the maximum salinity recorded was 1.2 ppt.
Table 2. Sodium, Potassium in the leaves (10 samples) of dominant plant taxa recorded along the Krishna Godavari
delta, east coast of India.
Na/K
Potassium (K)
Sodium (Na)
1
2
3
4
5
6
7
8
9
10
Average
1
2
3
4
5
6
7
8
9
10
Average
1
2
3
4
5
6
7
8
9
10
Average
Avicennia
alba
98.5
112.8
120.8
99.0
100.5
125
123
97.0
89.0
90.8
105.64
10.1
11.9
12.0
9.5
9.0
12.0
12.0
9.0
9.0
10.0
10.45
9.8
9.5
10.1
10.4
11.2
10.4
10.3
10.8
9.9
9.1
10.15
Avicennia
officinalis
160.0
165.8
170.7
165.0
175.0
165.0
168.0
155.0
162.0
155.0
164.15
32.0
35.8
37.2
32.0
38.0
35.0
35.5
30.0
32.0
32.0
33.95
5.0
4.6
4.6
5.2
4.6
4.7
4.7
5.2
5.1
4.8
4.85
Avicennia
marina
48.8
51.8
54.8
55.0
37.0
49.0
52.0
55.0
38.0
56.0
49.74
21.2
25.2
30.6
30.0
22.0
23.0
26.0
26.0
23.0
26.0
25.30
2.3
2.1
1.8
1.8
1.7
2.1
2.0
2.1
1.7
2.2
1.98
Suaeda
monoica
83.0
92.0
74.0
83.0
89.0
96.0
92.0
72.0
79.0
89.0
84.9
11.0
20.0
11.0
14.0
12.0
14.0
12.0
11.0
13.0
15.0
13.30
7.5
4.6
6.7
5.9
7.4
6.9
7.7
6.5
6.1
5.9
6.52
Suaeda
nudiflora
7.9
12.0
20.0
19.0
21.0
8.0
7.9
9.0
10.0
14.0
12.88
1.9
3.0
6.0
5.0
8.0
2.0
2.0
4.0
7.0
7.0
4.59
4.0
4.0
3.3
3.8
2.6
4.0
4.0
2.3
1.4
2.0
3.14
Suaeda
maritima
12.0
8.0
7.0
13.0
8.0
9.0
10.0
12.0
9.0
13.0
10.10
5.0
2.0
2.0
5.0
5.0
3.0
5.0
4.0
4.0
6.0
4.10
2.4
4.0
3.5
2.6
1.6
3.0
2.0
3.0
2.3
2.2
2.66
Acanthus
ilicifolius
82.3
98.0
76.0
78.0
89.0
100.0
95.0
99.0
81.0
72.0
87.03
17.8
28.0
15.0
15.0
18.0
19.0
24.0
23.0
19.0
15.0
19.38
4.6
3.5
5.1
5.2
4.9
5.3
4.0
4.3
4.3
4.8
4.60
Sodium and potassium in leaves

The leaf samples of A. alba show the minimum 99 mg g-1 and maximum 125 mg g-1of sodium concentration.
The minimum concentration in A. officinalis was 155 mg g-1 and maximum 175 mg g-1. The high sodium
concentration in the leaves of Avicennia officinalis and A. alba was an average of 164 and 105 mg g-1,
respectively followed by A. marina which shows three times lower concentration (average 50 mg g-1). The
minimum concentration of sodium in A. marina was 38 mg g-1 and maximum 56. The potassium concentration
81
Farooqui et al. (2016) 3(1): 7886

.
was however, highest in A. officinalis followed by A. marina and A. alba (Table 2). The Na/K ratio varied
between these three species. While it was highest in A. alba (~10.15), it was low in A. officinalis (~4.85)
followed by A. marina (~1.98). The Highest Na accumulation was in Suaeda monoica followed by several
times low Na in S. nudiflora and S. maritima. Similar trend was recorded in K concentration in the leaves of
these three species. Acanthus ilicifolius has the affinity to absorb and accumulate Na and K (87 and 19 mg g -1,
respectively) in the leaves but the high concentration of K (low Na/K ratio) in the leaves along with the
adaptability feature such as salt glands to exude excess salt enables it to tolerate salinity stresses.
Leaf epidermal morphological adaptability
The salt glands observed in Avicennia species and Acanthus ilicifolius are a distinctive
multicellular trichome. In both the genera, glandular hairs are found on the upper leaf surface and much more
densely in the abaxial indumentums (Metcalfe & Chalk 1957). On the upper leaf surface they are sunken in
shallow pits (Fig. 3A, B). In Avicennia species the non-glandular trichomes are many in number so much so that
the stomata become obscure and are hardly visible (Fig. 3C). In the lower surface they occur scattered among
long non-glandular hairs composed of three or four cells (Fig. 3D). The non-glandular hairs /trichomes are not
present in Acanthus ilicifolius. It is only the glandular trichomes that are present on the upper and lower leaf
surface (Fig. 3EH). The salt gland has two to four vacuolated cells at the level of the epidermis, the stalk cell
with an almost completely cutinized wall, and four to eight terminal cells. The terminal cells have a thin,
perforated cuticle which separates from the cell walls apically, leaving an enclosed cavity between them. The
secreted salt evaporates or is washed during rainy season. The Suaeda species do not show any glandular or
non-glandular trichomes (Farooqui et al. 2009) either to excrete salt or to resist excess transpiration through the
non-glandular trichomes. However, the leaves of Suaeda are needle like and succulent in nature, particularly S.
nudiflora. The succulent leaves and low salt accumulation in Suaeda species helps them to adapt in salt stressed
conditions when the physiologically active water becomes low in the pore water of the substrate.
DISCUSSION
All along the south- east coast of India, one can encounter pure forms of Avicennia officinalis, A. marina,
Suaeda maritima and S. Nudiflora in most of the intertidal zones (Orissa, Tamil Nadu, Andhra Pradesh). These
are in general scrubby bushes that are spread in large areas (Fig. 3) interspersed between salty substrates devoid
of any vegetation. Avicennia officinalis and A. alba record high Na in their leaves that shows its affinity for Na
absorption and translocation in the shoot system. Out of these Avicennia marina, is the lowest Na accumulator.
However, the accumulation of K was least in A. alba and it was three times higher in A. officinalis followed by
slight decrease in the values of K in A. marina. Thus, results show that both A. officinalis and A. marina have
the affinity for potassium uptake and its translocation in the shoot system and in the leaves. The Na/K ratio was
highest in A. alba and comparatively it is 52 % low in A. officinalis and 80 % low in A. marina. These two
species with low Na/K ratio are recorded in abundance in the coastal wetlands of India, thereby, indicating that
K is an essential component in the shoot system to mitigate increased salinity in the ecosystem despite the
similar leaf epidermal morphological features adapted to exude excess salt accumulation in all the three species.
The tolerance of all halophytes to salinity depends on controlled uptake and compartmentalization of Na and K
and the synthesis of compatible solutes, even where salt glands are functional to exude excess salts (Flowers &
Colmer 2008). Potassium plays an essential role in many physiological processes and the main function to
regulate the turgor in stomatal cells helps mitigate the water stresses caused by high salinity when the
physiologically active water becomes low for plants in wetlands.
Therefore, in India, Avicennia alba occurs only in mangrove forests from Sunderbans of West Bengal,
Orissa, Andhra Pradesh (Coringa) but are either stray or absent from other mangrove areas in the south-east
coast with increased salinity (Farooqui 2010, Srivastava et al. 2012). It is however not recorded from Saurashtra
and Kutch areas where the high salinity exists in the coastal land due to dry and arid climatic conditions.
Avicennia marina and A. Officinalis are distributed throughout in the mangrove areas due to its wide salinity
tolerance perhaps due to its affinity for K absorption.
The association of Acanthus ilicifolius along with Avicennia officinalis and A. marina is restricted to
landward zones. In general, Acanthus tends to grow in the areas of good fresh water input in the coastal
wetlands with low salinity. Results show that its Na/K ratio is similar to A. officinalis. Its affinity for potassium
uptake is similar to A. marina. Thus, low Na/K ratio coupled with salt glands for salt exudation enables the
82
Farooqui et al. (2016) 3(1): 7886

.
Figure 3. A, The Upper leaf epidermal illustration of salt glands (G) in Avicennia alba; B, The upper leaf epidermis
of A. officinalis showing salt glands (G); C, Lower leaf epidermis showing non-glandular trichomes (tr) and
obscured stomata (st); D, Enlarged view of Non-glandular trichomes in A. officinalis (stained with Safranin); E,
Upper leaf epidermis of Acanthus ilicifolius showing multicellular peltate trichome known to secrete salts (salt
glands).
83
Farooqui et al. (2016) 3(1): 7886

.
species to survive the low to moderate salinity stress. But unlike Avicennia, lack of non-glandular trichomes in
Acanthus does not help this species to restrict excess water loss during transpiration under salinity stress
conditions. Therefore, it prefers to grow in low salinity zones and not in high salinity areas despite having salt
glands for excess salt exudation and low Na/K ratio.
The affinity for Na and K uptake was highest in S. monoica with high Na/K ratio. Thus, this plant species is
therefore, not found in abundance in the study area perhaps due its high salt accumulation without any
functional adaptability features in the leaves to exude excess salt. The only adaptation is that it tends to increase
the volume of the needle like succulent leaf. The experimental analysis showed that increased salinity in the
substrate tends to increase the thickness of the cuticle in Suaeda which helps in the restriction of water
evaporated during the transpiration (Hajibaghera et al. 1983) Uphof (1941) noted that the epidermis of xerosucculents and coastal halophytes is characterized by a thick cuticle and a cover of waxy layers. Thus cuticular
resistance plays a major role in decreasing transpiration rates, a phenomenon frequently observed under saline
conditions (Waisel 1972, Yeo 1981) and a decreased flux of water through the root system would mitigate
against excessive ion transport to the shoot.
The study reveals that all the three genera (Avicennia, Acanthus and Suaeda) adapted to saline coastal areas
behave differently with respect to Na and K affinity and accumulation in their leaves. While Avicennia has two
features (glandular trichomes and non-gladular trichomes), the Acanthus has only glandular trichomes for salt
exudation and no other adaptability to mitigate excess water evaporation through stomata. Suaeda species show
increased leaf volume with the increasing salinity in the substrate. Among all the physiological processes in
plants, the salinity shows an inhibiting influence on the photosynthesis. This is not only attributed to stomatal
closure leading to a reduction of intercellular CO2 concentration, but also to non-stomatal factors like reduction
in green pigments and leaf area. It is also depicted from literature that salt affects photosynthetic enzymes,
chlorophylls and ionic contents (Misra et al. 1997). Salinity limits the growth and production by affecting ion
balance, water status, mineral nutrition, etc. (Munns 1993). Salt glands are specialized adaptive structures found
predominantly on the leaves and stems of halophytic species. They are considered to be efficient desalination
devices capable of removing salts from the plant tissues via an energy-dependent secretion process. High
concentrations of salts in the root zone decrease soil water potential and the availability of water (Lloyd et al.
1989). This deficiency in available water under saline conditions causes dehydration at cellular level and
ultimately osmotic stress occurs. The excessive amounts of toxic ions like Na + and Cl- create an ionic imbalance
by reducing the uptake of beneficial ions such as K+, Ca2+, and Mn2+ (Hasegawa et al. 2000). Therefore, the
present study reveals that plants which are efficient in K absorbtion are more tolerant to salinity stress while
others are sensitive despite the leaf epidermal modifications to mitigate such stresses.
Figure 4. Relative affinities of dominant mangrove species to Na and K accumulation in the leaves.
Figure 4 shows that Avicennia alba, S. monoica and Acanthus ilicifolius are salt-sensitive
mangroves/associates as the Na/K ratio is quite high but A. officinalis, A. marina, S. nudiflora and S. maritima
are salt tolerant mangroves having affinity for K uptake as their Na/K ratio in the leaves is low. The K
concentrations in the physiological processes of these species perhaps help in the mitigation of the salt stress in
84
Farooqui et al. (2016) 3(1): 7886

.
saline substrate and therefore, have become more abundant along the south-east coast competing with other
species leading to local extinction of several other mangroves.
CONCLUSION
Salt stress has a significant effect on growth and distribution/zonation of mangroves relative to salinity in the
substrate. Out of all the studied species, Avicennia officinalis, A. marina, S. nudiflora and S. maritime
maintained the beneficial K ions in their leaf tissues, therefore, exhibit its dominance in saline substrate. A. alba
and S. monoica are sensitive to salinity stresses as these have low affinity for K absorption. Acanthus ilicifolius
too moderately sustains the salinity stress as the Na/K ratio is low but lacks leaf epidermal modifications to
prevent water loss during transpiration except the glandular trichomes for salt excretion. Thus, potassium and
sodium ions have a strong correlation with the salt tolerance potential in different plant species of studied
mangroves. The current study also proves that Na/K ratios are the useful screening tools for salt tolerance at
specific level in mangroves. The study can be used during the coastal zone management and strategies adopted
for reforestation of mangroves in varied coastal ecosystems world over.
ACKNOWLEDGEMENT
The authors are thankful to Director BSIP, Lucknow India for providing necessary facilities to accomplish
this work and to Dr. B. Sekar for helping in the chemical analysis of leaves. One of the authors Ranjana is
thankful for financial help as BSRS.
REFERENCES
Balsamo RA & Thomson WW (1995) Salt effects on membranes of the hypodermis and mesophyll cells of
Avicennia germinans (Avicenniaceae) a freeze-fracture study. American Journal of Botany 82: 435440.
Boyce SG (1954) The salt spray community. Ecological Monograph 24: 2967.
Dietrich P, Sanders D & Rainer Hedrich (2001) The role of ion channels in light-dependent stomatal opening.
Journal of Experimental Botany 52 (363): 19591967.
Drennan P & Pammenter NW (1982) Physiology of salt excretion in the mangrove Avicennia marina (Forsk.)
Vierh. New Phytology 91: 597606.
Dschida WJ, Platt-Aoloia KA & Thomson WW (1992) Epidermal peels of Avicennia germinans (L.) Stearn: A
useful system to study the function of salt glands. Annals of Botany 70: 501509.
Fahn A (2000) Structure and function of Secretory cells. Advances in Botanical Research 31: 3775.
Farooqui A, Srivastava J & Hussain SM (2009) Comparative Leaf epidermal morphology and Foliar Na: K
accumulation in Suaeda species: A case study from coastal ecosystem, India. Phytomorphology 59 (3&4):
102111.
Farooqui A (2010) Salt water intrusion, metal accumulation and mangroves along the pednapatnam
Machlipatnam coastline, Andhra Pradesh. Indian Journal of Applied Geochemistry 12 (1): 126138.
Farooqui A, Kulshreshtha K, Singh SN, Pandey V & Ahmad KJ (1995) Photosynthesis, stomatal response and
metal accumulation in Cineraria maritima and Centuaria moschata grown in metal rich soil. Science of
Total Environment 164: 203207.
Flowers TJ & Colmer TD (2008) Salinity tolerance in halophytes. New Phytology 79(4): 94563.
Hajibagheri MA, Hall JL & Flowers TJ 1983 The structure of the cuticular transpiration in leaves of the
halophyte Suaeda maritima (L.) DUM. New Phytology 94: 125131.
Hasegawa PM, Bressan RA, Zhu JK & Bohnert HJ (2000) Plant cellular and molecular responses to high
salinity. Annual Review of Plant Physiology and Plant Molecular Biology 51: 463499.
Lloyd J, Kriedemann PE & Aspinall D (1989) Comparative sensitivity of Prior Lisbon lemon and Valencia
orange trees to foliar sodium and chloride concentrations Plant Cell Environment 12: 529540.
Metcalfe CR & Chalk L (1957) Anatomy of the Dicotyledons, Systematic Anatomy of the leaf and stem.Vol. I.
2nd Ed. Clarendon Press, Oxford.
Misra A, Sahu N, Misra M, Singh P, Meera I, Das N, Kar M & Sahu P (1997) Sodium chloride induced changes
in leaf growth, and pigment and protein contents in two rice cultivars. Biologia Plantarum 39 (2): 257262.
Moisender PH, Mcclinton E & Paerl HW (2002) Salinity effects on growth, photosynthetic parameters and
nitrogenase activity in estuarine planktonic cyanobacteria. Microbiologia Ecology 43: 432442.
85
Farooqui et al. (2016) 3(1): 7886

.
Morrow L & Nickerson NH (1973) Salt concentrations in ground waters beneath Rhizophora mangle and
Avicennia germinans. Rhodora 75: 102106.
Munns R (1993) Physiological processes limiting plant growth in saline soils: some dogmas and hypotheses.
Plant Cell and Environment 16: 1524.
Munns R (2002) Comparative physiology of salt and water stress. Plant Cell and Environment 25: 239250.
Murphy L, Kinsey S & Durako M (2003) Physiological effects of short-term salinity changes on Ruppia
maritima. Aquatic Botanica 75: 293309.
Paliyavuth C, Clough B & Patanaponpaibon P (2004) Salt uptake and shoot water relations in mangroves.
Aquatic Botanica 78: 349360.
Scholander PF, Hammel HT, Hemmingsen E & Garey W (1962) Salt balance in mangroves. Plant Physiology
37: 722729.
Smith JAC, Popp M, Luttge UWJ, Cram M, Diaz H, Griffiths HS, Lee J, Medina E, Schafer C, Stimmel KH &
Thonke B (1989) Ecophysiology of xerophytic and halophytic vegetation of a coastal alluvial plain in
northern Venezuela. New Phytologist 111: 293307.
Srivastava J, Farooqui A & Hussain SM (2012) Sedimentology and salinity status in Pichavaram mangrove
wetland, south-east coast of India. International Journal of Geology Earth and Environmental Science 2(1):
715.
Tomlinson PB (1986) The Botany of Mangroves Published by the Press Syndicate of the University of
Cambridge, First edition.
Uphof JCT (1941) Halophytes. Botanical Review 7: 157.
Waisel Y, Ethel A & Sagami M (1986) Salt tolerance of leaves of mangrove Avicennia marina Physiologia
Plantarum 67: 6772.
Yeo AR (1981) Salt Tolerance in the Halophyte Suaeda maritima L. Dum.: Intracellular Compartmentation of
Ions. Journal of Experimental Botany 32(3): 487497.
86
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 87101, 2016
Review article
Allelopathic invasion of alien plant species in India and their

management strategies: A review
Vijaya Yadav, N. B. Singh*, Himani Singh, Ajey Singh and Imtiyaz Hussain
Plant Physiology Laboratory, Department of Botany, University of Allahabad, Allahabad, Uttar Pradesh, India
*Corresponding Author: singhnb166@gmail.com
Abstract: Invasion of alien plant species is a persuasive threat to the native plant diversity and
caused habitat loss around the world. Invasions not only harm the native flora but have adverse
impact on economic status as well as health of the country. A detailed study on allelopathy,
allelochemicals and allelopathic mechanism of exotic invasive plants are reviewed. Main emphasis
is given on the modes of invasion, reproductive characters, transmission modes, establishment,
adaptability of weeds in the environment and the major exotic plants in India and their allelopathic
effects on native vegetation. The allelopathy of most common exotic invasive plants in India such
as Ageratum conyzoides, Eupatorium adenophorum, Parthenium hysterophorus, Lantana camara,
Mikania micarantha, Argemone mexicana and Eichhornia crassipes has been described.
Mechanical, chemical, biological and cultural control methods have been less effective
individually, so integrated management with the participation of native people and proper land
management have been proved beneficial. The various measures and management strategies to
overcome and control the invasion of weeds have been discussed.
Keywords: Invasion - Exotic - Weeds - Allelochemicals - Management.
[Cite as: Yadav V, Singh NB, Singh H, Singh A & Hussain I (2016) Allelopathic invasion of alien plant species
in India and their management strategies: A review. Tropical Plant Research 3(1): 87101]
INTRODUCTION
An invasive species are introduced as an alien, exotic, and non-indigenous species non-native to that location
but very aggressive leading to damage to the other plant species, human health and economic structure, or the
organisms from their native place immigrating to a new locality are referred as exotic species (Mack et al.
2000). Invasion of exotic plant species has emerged as a global problem causing adverse impact on the
ecosystems, economy, and human health. Invasion is raised as one of the major causes of biodiversity loss
(Inderjit et al. 2008, Rastogi et al. 2015). The invasive alien species is considered as one of the reasons of
habitat destruction. In past, many losses to biodiversity have not been analysed but now a days they become
necessary to record the biological invasion for conservation of biodiversity and to take effective measures for
their control. More than 40% of the species are in the threatened and endangered lists due to invasive species
(Wilcove et al. 1998). It is also estimated that about 20% of plant species are non-indigenous in many continents
and more than 50% on several islands (Rejmanek et al. 1994). Thousands of alien plant species are known to
invade globally and several exotic species are still unrecognised (Ruiz et al. 2000).
In India, among all alien flora 10% are Asian, 20% Asian and Malaysian, 15% of Europe and Central Asia
and 55% of America (Nayar 1977). Due to enormous growth of invasive exotic species, India is facing
significant environmental as well as economic problems. In India about 42% of the weeds in crop fields are
aliens (Khuspe et al. 1982, Nandpuri et al. 1986). It is estimated that alien weeds have caused 30% loss in crop
production (Singh 1996). In a study of the invasive alien flora in Uttar Pradesh, India comprises of 44 families
including 109 genera and 152 species (Singh et al. 2010). The most common exotic species of India which has
been discussed include Ageratum conyzoides, Eupatorium adenophorum, Parthenium hysterophorus, Lantana
camara, Mikania micarantha, Argemone mexicana and Eichhornia crassipes (aquatic plant). Among which
Lantana camara, Parthenium hysterophorus, Ageratum conyzoides are the worst, highly invasive and
challenging (Kohli et al. 2004).
87
Yadav et al. (2016) 3(1): 87101

.
For a successful invasion in the new habitat a plant species must defeat adversities and interact with native
plants as competitors and have wide adaptability and physiological plasticity (Levine et al. 2004, Richardson et
al. 2000). According to enemy release hypothesis (Colautti et al. 2004) the invasive plants are less prone to
herbivores and pathogens hence attain large scale distribution and abundance. The mutualist facilitation
hypothesis (Richardson et al. 2000) shows the replacement of the native mutualist species with several new
mutual species in their range help invaders in proper establishment and invasion. The pollinators, seed
dispersing organisms and mycorrhizal symbiosis sometimes help invasion of plants. Eltonian empty niche
hypothesis, states that sometimes invaders make use of unused resources and occupy an empty niche when they
are introduced into a new community (Elton 1958). The novel weapon hypothesis (Callaway et al. 2004) states
that several invasive species have specific biochemical compounds which have allelopathic effects and act on
plant and soil microbial interactions. The exotic plants get advantage in invasion due to the release of
phytochemicals to which the native plants and soil microbes are not adapted.
This paper is based on the research done on the exotic plants worldwide. The major objective of this article
is to explore invasive alien plants, reason of their fast invasion, establishment and effective growth harming
native biodiversity and economy of the country. The measures of control of invasive flora including physical,
biological and chemical modes have been also reviewed.
Allelopathy
The term allelopathy was first given by Molisch (1937), consist of two Greek words, allelon meaning
mutual and pathos meaning to suffer, harmful effects on each other . Allelopathy is a natural phenomenon in
which different plant species affect the physiology of other plants existing in their vicinity, either negatively or
positively (Rice 1984).
Allelopathy is defined as the adverse effect of a plant on another plant through the release of several
secondary metabolites by plant parts into the soil (Inderjit & Callaway 2003). Allelopathy is a useful mechanism
for alien plant invasion. Chemical exudates released from roots and other plant parts play crucial role which
arbitrate mutualistic, competitive and pathogenic effects on native flora (Inderjit et al. 2005, Mitchell et al.
2006).
Causes of rapid dispersal of invasive plants
The small sized seeds weighing less than 50 mg, less than 10 years of juvenile life and short interval
between crops are one of the reasons of their propagation and invasiveness (Sharma et al. 2005). Parthenium
hysterophorus produces large number of small seeds which can travel long distance and cause fast invasion
(Rejmanek & Richardson 1996). Most of alien invasive species have small seeds with wings or pappus, which
facilitate them to cover a long distance through anemochory (Wan & Wang 1990). Sometimes C3 plants adapted
to C4 mechanism and CAM (facultative CAM plants) if they invade arid areas with high temperature thus helps
in successful invasion (Sage 2004). Dispersal through animals is also a major cause for the rapid invasion in
disturbed and undisturbed habitats (Rejmanek & Richardson 1996, Binggelli 1996).
Geographical range of flora is also responsible for the invasion of the plant. The seeds and propagules of a
species with widespread distribution have more chance of being transported and established in other countries
and continents. (Forcella et al. 1984, Rejmanek 1995, Goodin et al. 1998).
The exotic flora with different modes of vegetative reproduction, along with sexual reproduction helps in
fast invasion in which allelopathy plays a key role (Fig. 1).Vegetative propagation increases compatibility of
exotic plants in the environmentand invasion in both terrestrial as well as in aquatic habitat (Pieterse & Murphy
1990). The invasiveness of Eichhornia crassipes mainly occurs by means of stolon (Barrett 1989). All exotic
genera have a strong coping capacity against abiotic and biotic barriers in the invaded region which may be
through various associations like root symbionts and non-specific mutualism (Richardson et al. 2000).
Furthermore they have large tolerance in lower resources and thus make them more competitive and invasive
(Noble et al. 1980).
Allelopathy is considered as the major cause of invasion of exotic plants. There is wide range of
allelochemicals which are known to have both positive and negative effects on neighbouring plants. Secondary
metabolites are classified as organic acids, alcohols, aliphatic compounds, cinnamic acid and its derivatives,
terpenoids, aldehydes, ketones, lactones, fatty acids, alkynes, quinone compound, simple phenols, benzoic acid,
88
Yadav et al. (2016) 3(1): 87101

.
steroids, amino acids, peptides, alkaloids, sulfide, glucosinolates, nucleotides. Among these phenolic acids and
terpenoid compounds are the most common forms of allelochemicals (Song 1990, Sun & Yu 1992).
Allelochemicals released from invasive species harm native species through inhibition of nutrients uptake,
disturbing cell division and root and shoot elongation (Cruz et al. 1998, Cruz et al. 2007), alteration in
membrane permeability (Li et al. 2010), inhibition of chlorophyll formation and protein synthesis (Chen et al.
2002, Li et al. 2010, Wein et al. 2004) and inactivation of some hormones and enzymes (Li et al. 2010,
Muzaffar et al. 2012). Allelochemicals also affect process of photosynthesis by interrupting photosystem II
(Yang et al. 2002). Allelopathy has also been acknowledged as a mechanism which promotes plant to dominate
and establish ecologically (Narwal et al. 2005).
Parthenium hysterophorus checks the germination and growth of other plant species by releasing certain
allelochemicals or allelopathic interaction (Adkins & Sowerby 1996). Lantana camara is proved as a noxious
exotic species which is capable to disturb the regeneration process of the other plant species by affecting their
germination and survival through the allelopathy (Gentle & Duggin 1997). Eupatorium adenophorum is known
to inhibit germination of seeds of other species like rye grass, maize and clover (Zang et al. 1993). Mikania
micarantha has allelopathic effect on several plant species (Zang et al. 2002).
Small
seeds
Modes of
dispersal of
seeds
Phenotypic
plasticity
Introduction of alien
species
Germination
and
establishment
Growth and
reproduction
Sexual
reproduction
Habitat
prone to
invasion
Widespread
distribution
Allelopathy
Asexual
reproduction
Production of
propagules
Production of
seeds
Population
blast
Biodiversity and
economic loss
Figure 1. A Schematic representation showing the process of exotic plant invasion, establishment and reproduction.
89
Yadav et al. (2016) 3(1): 87101

.
Table 1. List of some major invasive flora and their impact.
S.No. Species and Common

name
1.
Ageratum conyzoides
(Goat weed)
2.
Eupatorium adenophorum
(Crofton weed)
3.
Eichhornia crassipes
(Water hyacinth)
4.
Lantana camara
(Lantana)
5.
Mikania micrantha
(Mile-a-minute)
6.
Parthenium hysterophorus
(Congress Grass or Carrot
Grass)
7.
Argemone Mexicana
(Mexican Prickly Poppy )
Family and
Habit
Asteraceae
Herb
Native
Threats
Tropical America Allelopathic, highly

invasive, threat to
croplands of Himalayan
region.
Asteraceae
Mexico
Allelopatic effect causes
Shrub
serious threats to native
flora.
Pontederiaceae Tropical South
Serious aquatic weed,
Aquatic herb
America
allelopathic in nature,
causes hindrance in
navigation, reduces water
quality and algal growth.
Verbeneaceae Tropical America Strongly allelopathic,
Shrub
serious threat to medicinal
plants, responsible for
forest fire.
Asteraceae
Sub-tropical zone Known for its allelopathic
Herb
of America
potential, highly invaded
forest areas.
Asteraceae
Tropical America Aggressive colonizer,
Herb
highly allelopathic,
allergic to animals and
human being, threat cause
to crops and other native
flora.
Papaveraceae Tropical Central Harm native flora through
Herb
and
allelopathy.
South America
References
Dogra et al. 2009,
Roder et al. 1998
He & Liu 1990
Raghubanshi et al. 2005,

Sun et al. 1988
Sharma et al. 2005,

Raghubanshi et al. 2005
Raghubanshi et al. 2005,

Zhang et al. 2002
Kohli & Rani 1994,
Kanchan & Chandra 1980,
Kohli & Batish 1994.
Reddy 2008
Certain plants have ability to amend their growth and development in association with changes in the
environment (Dorken & Barrett 1981). This phenomenon is known as phenotypic plasticity. Parthenium
hysterophorus has ability to grow in any soil because it shows wide phenotypic plasticity to any type of soil
which leads to its establishment as a successful invader. Generally there are two types of invaders, one is tall,
fast growing with small seeds suitable for rapid dispersal and the second is short heighted plants with high seed
mass showing slow invasion but with high seed spread (Annapurna & Singh 2003). Habitat also plays a
significant role in conferring invasiveness to exotic plants, however, it does not imply that any invasive species
reaching that habitat will establish successfully. Habitat which is susceptible to invasion may have poor species
diversity, poorly adapted native species, absence of predators and empty niches (Mantri et al. 2002). Some
allelopathic invasive plants which are covering most parts of India have been discussed and listed in table 1.
Common invasive exotic species, their allelochemicals and allelopathic effects
Eupatorium adenophorum
Eupatorium is commonly called as crofton weed which is native of Central America (Song et al. 2000). It is
a noxious invasive species with profusely branched stem. Crofton weed depends on few factors i.e. humidity,
light, ecology and biodiversity of a particular area to invade (Meng et al. 2003). Allelopathy plays a vital role in
its invasion. There are many allelochemicals identified and isolated from Eupatorim adenophorum. The
essential oil of crofton weed contains 1-methyl-2(1-methyl) benzene, hedycaryol, cetral, bornyl acetate, pcymene, lonipinene, copaene (Wu & Yang 1994). Ding et al. (1999) reported sesquiterpene lactoneeuprtoranolide from the flowers of E. adenophorum.
It has been observed that it has severe allelopathic effect on the other neighbouring plants which is the key
factor of its easy invasion. It inhibits the germination of seeds of several crop plants like clover, rye grass, and
maize which are commercially very important crops (He & Liu 1990, Zhang et al. 1993). E. adenophorum
leachate affects not only physiological characters but it also creates anatomical abnormalities in the plant by
90
Yadav et al. (2016) 3(1): 87101

.
destroying root cells of corn (Zhang et al. 1993). The allelopathic effects by using chloroform extracts of aerial
parts of Eupatorium. Cadinenes and b-sitosterol were isolated and their effects were seen on the Allium cepa,
Raphanus sativus and Cucumis sativus seeds (Baruah et al. 1994).
Mikania micrantha
Mikania micrantha is a perennial plant with small compact florets, invasive weed commonly known as plant
killer and mile-a-minute which is a native of neotropical region (Xie et al. 2010, Tripathi et al. 2012) and now a
days became in majority in natural habitats including forests, agricultural systems in north east India. It entered
India at the time of World War II to mask airfields and for covering tea plantations (Tripathi et al. 2012).
Mikania is known to grow in forests, on bank of rivers and streams, in disturbed areas (Kong et al. 2000,
Zan et al. 2000, Feng et al. 2002). It has covered most parts of Asia including India, Sri Lanka, Mauritius,
Bangladesh, etc. (Deng et al. 2004). Raghubanshi et al. (2005) reported about 61% invasion in forests, teak
plantations and disturbed forests. In north-east India, it has been observed that it climbs both small and tall trees,
covering their canopy. It is the worlds worst weed and is known for its allelopathic potentials. Mikania have a
good reproductive capacity; can grow through its plant debris and underground rhizomes vegetatively
(Rejmanek & Richardson 1996). Several allelochemicals like mikanin, eupafolin, luteolin, eupalitin. The
essential oil of M. micrantha contains terpenolene, limonene, ocimene, caryophylene, etc. (But et al. 2009, Feng
et al. 2004). The volatile oils of Mikania micrantha have allelopathic and inhibitory effects on several plants
(Zhang et al. 2002). The water leachates of Mikania have allelopathic potential on various economically
important crop plants like Raphanus sativus, Lolium multiflorum and Trifolium repens (Shao et al. 2003). Weng
(1964) found that the biomass and nitrogen content of tomato seedlings and some legume crops are also
inhibited by Mikania micrantha.
Lantana camara
Lantana camara is the one of the ten worst weeds of the world. It is an evergreen aromatic shrub which is
native of Central and South America (Raghubanshi et al. 2005). Lantana has been introduced in many countries
as ornamental plant and now a day has become serious problem for the native plants (Bever 1982). In India it
was introduced during 18091810 as an ornamental plant in Calcuttas gardens (Kohli et al. 2006) and is now
found all over India. This plant spreads fast due to human interference and disturbances (Sharma et al. 2005). It
has invaded more than 13.2 million ha pasture land and other areas in India (Singh et al. 1996). Lantana is
harmful to herbivores and the cost of its management is US$ 70 per hectare (Singh et al. 1996). Lantana is
highly invasive due to fitness homeostasis, phenotypic plasticity, widespread geographical range, modes of
reproduction and ultimately none and the most potent, the phenomenon of allelopathy (Sharma et al. 2005).
Various allelopathic compounds like salicylic acid, gentisic acid, coumarin, ferulic acid, p-hydroxybenzoic
acid and 6-methyl coumarin were analysed in Lantana camara (Yi et al. 2006). Some other allelochemicals
identified in Lantana camaraare cytotoxic in nature found in the leaves are lantadene A and lantadene B (Ma et
al. 2004). Pan et al. (1993) have reported presence of lantadene A, B, oleanolic acid, lantalonic acid, icterogenin
from the leaves and lantolonic, ursolic acid and oleonolic acid from the roots of Lantana camara. Lantana
density in forest, increases due to allelopathy which results in decline of species richness (Day et al. 2003).
Allelochemicals present in Lantana decrease the vigour of native plants of region and results ultimately poor
productivity (Sharma et al. 1988, Sharma & Sharma 1989). They are also responsible for wild fire in many
forest rich parts of India (Raghubanshi et al. 2005). The leaf extract of Lantana camara inhibits the emergence
and growth of leaf bud of water hyacinth and increases the superoxide dismutase activity, H 2O2 accumulation
and increase in membrane peroxidation (Zheng et al. 2006). Growth of commercially important vegetable crops
like tomato, radish and cucumber has been inhibited by leachates of Lantana camara. (Liu & Jia 2002). The
stem, leaf and fruit leachates of Lantana inhibit seed germination and seedling growth of some terrestrial plants
(Quan et al. 2009.)
Eichhornia crassipes
Eichhornia is an aquatic floating plant which is popularly known as water hyacinth. It is the native of
tropical region of South America. It was introduced in many countries as animal fodder and planted for
purification of water bodies (Raghubanshi et al. 2005). Water hyacinth mainly invades polluted and nutrient rich
water bodies particularly rich in nitrogen, phosphorus and potassium. This plant has become serious weed in
most of tropical, warm and fresh water habitats. In India it was introduced during 19141916 AD, from Brazil.
91
Yadav et al. (2016) 3(1): 87101

.
It is floating hydrophytes which causes hindrance in navigation, block rivers, lakes and irrigation system and
reduces the quality of water bodies (Raghubanshi et al. 2005).
The major allelochemicals found in water hyacinth are linoleic acid, glycerol -1, 9-12 (ZZ)-octa decadienoic
acid and N-phenyl-2-napthylamine (Yang et al. 1992). N-phenyl-2-napthylamine is known to inhibit the growth
of several aquatic floras. It increases the protein content and decreases superoxide dismutase activity in some
aquatic flora. Hydroponic water of E. crassipes reduces the growth of alga (Sun et al. 1988). It is known for its
anti-algal activity even more than that of popular algaecide copper sulphate (Sun et al. 1993).
Parthenium hysterophorus
Parthenium hysterophorus is an exotic species native of Tropical America and has invaded most of parts of
India now a days. It bears a strong invasive potential and known for its strong allelopathic effects. Parthenium
hysterophorus L. (commonly known as Congress grass or Carrot weed) and belongs to family Asteraceae.
Parthenium is known to enter India in 19501960 A.D. along with common staple grains imported from United
States of America. It was first seen in 1955 in Maharashtra. It is said that it has entered India accidently in 1810
and lived hidden till 1956 until Rao reported it in Pune, Maharashtra for the first time (Roxburgh 1984, Bennet
et al. 1976).
It is a wasteland weed and aggressive colonizer with a higher productivity, plasticity, and high fecundity i.e.
a single plant can produce thousands of viable seeds and successful invagination of these seeds found along
wastelands, overgrazed areas, road sides, agricultural areas, railway tracks. Parthenium has first started invading
upon extreme hilly regions of India and encroached in lower and middle Himalayan region and now engulfed
almost whole of India (Dogra et al. 2009). ). In last two decades Parthenium become major and common weed
which has been covered urban as well as natural habitats and replaced native plant species (Dogra et al. 2009).
The major phenolic compounds found in P. Hysterophorus are, o-coumaric, gentisic, ferulic, p-coumaric,
caffeic, vallinic, salicylic acid, trans-cinammic and p-hydroxybenzoic acids and sesquiterpene lactone etc.
(Kohli & Rani 1994). Allelopathy has played an important role in successful and massive invasion of the weed
and helps this weed to colonize the native area successfully (Bais et al. 2003, Heirro & Callaway 2003). Native
plant species are not habitual to the chemical released by these new plants hence they fail to survive, establish
resulting in low density, stunted growth, and decrease in population. The microbes present in soil also are
unable to detoxify these allelochemicals (Callaway & Aschehoug 2000). Parthenium affects the crops and other
native flora by releasing phenolics and sesquiterpenes resulting into inhibition of their growth and development
(Kanchan & Jayachandra 1980, Kohli & Batish 1994).
The pollen and dust material of Parthenium cause allergic dermatitis in human being (Gunaseelan 1987,
Morin et al. 2009). It has been related to cytotoxicity of the allelochemical, sesquiterpene lactone found in
Parthenium (Narasimban et al. 1984). It is also known to cause diarrhoea, breathlessness and choking (Maishi
et al. 1998). Excess and close exposure to P. Hysterophorus pollen grains leads to allergic bronchitis (Towers
& SubbaRao 1992).
Ageratum conyzoides
Ageratum conyzoides is popularly known as billy goat weed or goat weed or tropical white weed, belongs to
family Asteraceae and is widely distributed on road sides, fields, cultivated areas, tropical and subtropical areas
of the world, pastures interfering with the native vegetation, including crops, grasses and forage crops (Marks &
Nwachuku 1986). Ageratum is highly adaptable and produces a huge number of seeds (800010000/plant). Its
seeds are achenes which are dispersed easily and acquiring favourable conditions flourish freely (Marks &
Nwachuku 1986). The seeds are photoblastic and remain viable for a year. It also spreads vegetatively (stolons).
It has been found before 1882 in India which is evident from The British Flora of India (Hooker 1882). It has
been found that it affects the native plant and crops by releasing several phenolic compounds (Kong et al. 2003).
The allelochemicals reported in Ageratum conyzoides are ageratochromene (Wei et al. 2004), precocene I
and precocene II have strong insecticidal effects (Lu 1982). Along with it, endo-borneol, farnesol,
hexadecanoid-acid, linoleic acid, nerolidol and quercetin, kaempferol, and its glucosides are present (Sharma &
Sharma 1995, Kong et al. 2002, Okande 2002). Precocene I and II are also known for insecticidal as well as
anti-juvenile hormone activity (Okande 2002). In cultivated lands, Ageratum has reduced the yield of staple
crops like wheat, corn, rice etc. Rice yield inversely proportional to the density of Ageratum (Roder et al. 1998).
The farmers of Himalayan region (Shivaliks) have left their fields due to loss of croplands (Kohli et al. 2006).
92
Yadav et al. (2016) 3(1): 87101

.
Argemone mexicana
Argemone mexicana L. is native to Central America (Mexico) commonly known as Mexican prickly poppy,
is one of invasive alien plant species invaded most parts of India (Reddy 2008). A. mexicana is widely
distributed herbaceous plant mainly found by road sides, fields and croplands. A. mexicana is a member of
family Papaveraceae with enormous seed production around 60 to 90 capsules per plant and each capsule
carrying 300 to 400 seeds, can remain dormant for several weeks or months (Karlsson et al. 2003). Most seeds
germinate to form seedlings, some seeds not even germinate the year after shedding and get accumulated to
form seed bank (Karlsson et al. 2003, Sanaa 2012).
Study revealed the presence of several allelochemicals like salicylic acid, p-hydroxybenzoicacid, vanillic
acid, cinnamic acid (Burhan & Shaukat 1999).The aqueous extract of A. mexicana checks the germination of
Lens culinaris (Paul & Begum 2010). Chandra et al. (2007) found that germination and seedling growth of other
plants were negatively affected by salicylic acid. It has been also reported that treatment with A. mexicana leaf
aqueous extracts causes significant decrease in fresh and dry weight of sorghum (Alagesaboopathi 2013). Thus,
the studies revealed that allelochemicals found in A. mexicana inhibit the growth of other plants and ultimately
cause threat to native flora and biodiversity.
Impact of invasive plant species on environment and economy
The invasive plants have a perilous effect on biodiversity and ecosystem. They are responsible to destroy
native biodiversity by decreasing the density and frequency of the native flora. The species above discussed
have their allelopathic effect and are the worst weeds in India. They produce several allelochemicals which are
not only eradicating native flora but also cause serious health hazards in livestock and humans. They are
poisonous to cattles and small children if taken accidently. Lantana affects sandal wood forest and supports
spreading of spike disease in sandal (Holm et al. 1997).
Other invasive plant like Parthenium is very injurious to plant species, livestock and human health. It is
known to cause dermatitis, skin irritation, nausea, and several respiratory problems. It is also responsible for
harsh taste of milk of cattles if feeded, due to presence of hepatotoxic compound parthenin (Kohli & Rani 1994)
and if this milk is consumed by human beings it may cause deleterious problems.
There are two types of invasive species on economy; direct as well as indirect (Bigsby & Whyte 2001). The
effect called direct when effect caused by the invader and the indirect effects produced by the presence of
invader and its impact on human health. It has been identified that there are several major impacts of invasion,
i.e. production, price and market effects, trade, food security and nutrition, human health and environment,
financial cost. The cost estimated for the invasive species annually ranges from million to billion dollars
(Pimentel et al. 2000).
Management strategies
The biggest challenge now a day is how to manage noxious weeds which are highly invasive and
allelopathic enough to harm the native flora, vegetation, cropland, animals and ultimately human beings too.
There are number of control measures known through which the invasive species can be managed up to some
extent if applied properly. They may be mechanical, cultural, biological and chemical methods. Mechanical
control includes hand picking, hoeing, mowing, tilling, etc. which are effective when soil is moist, and the roots
of the weed are not very deep (Sheley et al. 1999a). Mowing can control harsh weeds effectively by reducing
production of seeds, by preventing reserve food storage. Mowing has been done during flowering.
Other mechanical methods like chopping, cutting, ploughing can be used for shrubs or trees (Cross &
Wiedmann 1985, Mchenry & Murphy 1985, Rasmussen 1991). Tillaging also controls growth of annual species
(Young et al. 1998).
Biological control
Biological control is mainly used to reduce number and dominancy of a particular plant (Wilson &
McCaffrey 1999). Insects, nematodes, pathogens can play important role in controlling noxious weeds.
According to Blossey, biological control is cost effective and self-sustained option for controlling weeds
(Blossey et al. 1994). These biological agents reduce the seed production hence control weeds to spread on large
scale (Balciunas & Villegas 1999). In India, Lantana is known to be controlled by Teleonemia scurpulosa
commonly popular as Lantana lace bug (Sharma 1988). There are various biological methods to be used to
93
Yadav et al. (2016) 3(1): 87101

.
eradicate these weeds, like Mexican beetle Zygogramma bicolorata is used to control Parthenium (Kohli et al.
2006). A weevil Eochetina spp. can be used for the removal of water hyacinth with a positive approach (Mandal
2011). M. micrantha can be inhibited by Helopeltis theivora the tea mosquito bug (Abraham et al. 2002).
Cultural control
There are several other practises are being used since many decades to control the invasion in which grazing,
fire, planting of competitive plant species are the major cultural practises. Grazing is useful only upto some
extent, depending on the nature of the invasive plant. If the species is palatable for cattle like Cyperus, Cynodon
etc. can be consumed as forage then grazing works but in case of toxic weeds like Parthenium, Lantana, etc.
which have adverse and harmful effects on livestock if consumed. Plant species producing allelochemicals are
highly toxic to livestock (Kingsbury 1964).
Grazing is also affected by behaviour of cattle i.e. the pattern, habit and period of grazing. Cattle mostly
feed on soft and non-flowering plants only while few of the cattle like goat also use to feed on plants with spines
as well as flowers also (Thomsen et al. 1993). Grazing can be effective when done just before flowering period
and defoliation (Kennet et al. 1992). Rotational grazing is practised to manage invasive species (DiTamaso
2000a). In this intensive grazing for few days is conducted.
Fire is another way to maintain and check invasion ultimately helps in maintaining the ecosystem (Hatch et
al. 1991). Burning is proved to be successful in controlling the non- woody invasive species (Ueckert et al.
1988). The burning may be time dependent, which will help to check the dispersal of seeds and to destroy the
viable seeds (DiTomaso et al. 1999a, Sheley et al. 1999a).
Another cultural practise which is employed to control the noxious weeds is re-vegetation which means
planting a species which establishes strong competition between that species and the invasive flora. It has been
proved a good long term scheme to curb the invasion of foreign flora and their dominancy (Borman et al. 1990).
The limitation of the scheme is the selectivity of the plants. They must be more aggressive to the plant to be
wiped out. Broad leaf plants can suppress the smaller weeds easily (Lee 1986). Various eco-friendly methods
have also been used like antagonistic plants such as Cassia sericea (Joshi et al. 1991). Essential oil of
Eucalyptus has potential to control Parthenium hysterophorus (Kohli et al. 1998, Singh et al. 2005).
In agriculture the inhibitory effects of allelochemicals itself are utilized for weed control (Kohli et al. 1998).
The allelochemicals have both stimulatory and inhibitory effects that had on different crops which are
concentration dependent. At low concentration, they have positive effect and they proved beneficial for the
plants, but in higher concentration they have toxic effect which may be used to control foreign plants (Narwal
1994).
So allelopathy has broad prospect in increasing crop plant protection, biological control, etc. The research
and application of allelopathy itself have the great significance on the prevention of exotic invasive noxious
weeds.
Chemical control
The herbicides and weedicides are the most commonly used since decades to stifle the invasive species. The
uses of chemicals are quite expensive but have rapid and satisfactory effect on unwanted plants. They can be
applied on the vegetation by aircrafts, helicopters, sprayers and herbicide applicators. A number of synthetic
chemicals like herbicides and weedicides are being used like paraquat, glyphosate, simazine, 2,4-D and 2,4,5T,dicamba, triclopr, etc. which are usually growth regulators but when used in high dosages they kill the plants.
Other herbicides which act on amino acid synthesis of plants are glyphosate, imazapyr, metsulfuron, etc. are
also effective to control weeds by checking photosynthetic process of the plant (Bussan & Dyer 1999).
Prevention of invasion of alien species can be done with a proper management approach. For the purpose, to
control and manage the invasive flora some continuous and effective steps should be taken for example, by
preventing the introduction of seeds and propagules of invasive species, spreading awareness regarding invasion
and its detrimental effects, early detection and their management through a proper and effective management
strategy (DiTomaso 2000b). Invasion may occur through seed dispersal by water, wind, animals, insects and
human activities. Alien seeds and propagules can also be imported through the seed purchase and a new species
can be imported for ornamental purpose and for the programs to get rid of pollutants and pollution. The purity of
the seeds and propagules should be checked and assured. The invasion also takes place through the soil. The soil
94
Yadav et al. (2016) 3(1): 87101

.
contains several seeds and propagules which invaded through roadsides and construction work (DiTomaso
2000b). The animals which feed on them and the seeds dispersed through their excretory wastes and produces
viable seeds. Invasion also occurs through vehicles andtransportation through water, roadways and railways, etc.
The disturbed and barren areas are more prone to invasion (Forcella & Harvey 1983, Tyser & Key 1988). The
invasive species are more virulent for the disturbed areas to form dominant vegetation (Sheley et al. 1998). So it
can be managed by re-vegetation of competitive species to control over invasive plants through allelopathy.
General awareness is another way to make people educate about the invasion, invasive species and their
harmful effects. People should be informed through web, posters, papers and articles (DiTomaso 2000b). The
farmers of the country should be aware to invasive flora and it is necessary to make them realise about the loss
of vegetation and crop field through the effect of alien flora so that they can protect their fields and crops on
their own level. Several education events should be organised to make aware about the economic and
biodiversity loss. The most valuable and best method to control the invasive species is early monitoring. The
weed should be detected as soon as possible and should be removed before it proliferate, reproduce and spread
to become invasive and unmanageable. Ecologically exotic plants can be managed by developing a resistant
plant community of various species which can cover several niches (Jacobs et al. 1999, Sheley et al. 1998).
Proper detection and monitoring can be done by professionals, management experts in a systematic way to
control over invasion (Zamora & Thrill 1999). Regular visits, field surveying, photography and removal of
particular weed from that particular area before establishment are convenient ways to control exotic species
(Sheley et al. 1999b).
The biggest challenge now a day is to manage noxious weeds which are highly invasive and allelopathic
enough to harm the native flora, vegetation, cropland, animals and ultimately human beings too. Once the plant
species establishes itself in a particular habitat, it is hard to manage and remove permanently, then periodic
strategy should be needed to manage them.
Earlier various screening systems have been developed independently all over the world. A screening system
was developed for woody invasive plants (Reichard & Hamilton 1997). Developing screening system in an
applied initiative to different regions may produce effective result if used properly (Curtis et al. 1999). A proper
monitoring of invasion can be done through well- timed and quantitative approach using mapping methods (like
map overlays or GPS). Along with it images can be taken through remote sensing satellites to estimate the level
of invasion (Reddy 2008). Studies shows that first juvenile plants should be cleared then high density large
plants should be eradicated (Higgins et al. 2000).
CONCLUSION
The exotic plants invasion is the major problem these days across the world and has adverse effect on
vegetation and agricultural system. There are several factors which influence the invasion process of alien flora.
Various hypotheses have been proposed through which invasion of exotic plants takes place. Studies around the
world highlighted allelopathy which has major role in establishment and rapid invasion of alien plants. Studies
also reveal that the allelochemicals found in these exotic plants are the major strength to compete them with any
type of habitat and environment with successful invasion. Various strategies have been also discussed regarding
control of the invasive plant species including biological, cultural and chemical practises. Sometimes allelopathy
itself can be proved as a control measure against invasion. It is a long term process to completely eradicate and
control over invasive alien flora but with proper management and time to time screening can help to overcome
this problem in India as well as on global scale. The consequences of invasion are, however, still miserable and
there is an immediate need of studies on biological invasions on large scale in India.
Public awareness is necessary regarding environmental change and biodiversity loss. People should make
aware of sustainable use of land and the effects of invasive species on the native flora and biodiversity.
According to Robertson et al. (2003) scientists should mark alien species according to the level of threat and
rate of spread in each of the climatic zones. There is also a need to provide adequate sources and strategies
through which proper management can be done to control the invasion process in future.
REFERENCES
Abraham M, Abraham CT & Joy PJ (2002) Natural enemies on Mikania micrantha H.B.K. in Kerala. Journal of
Tropical Agriculture 40: 3941.
95
Yadav et al. (2016) 3(1): 87101

.
Adkins SW & Sowerby MS (1996) Allelopathic potential of the weed Parthenium hysterophorus L. in
Australia. Plant Protection Quarterly 1: 2023.
Alagesaboopathi C (2013) Allelopathic Effect of Different Concentration of Water Extract of Argemone
mexicana L. on Seed Germination and Seedling Growth of Sorghum bicolor (L.) Moench. Journal of
Pharmacy and Biological Sciences 5: 5255.
Annapurna C& Singh JS (2003) Phenotypic plasticity and plant invasiveness: Case study of congress grass.
Current Science 85: 197201.
Bais HP, Vepachedu R, Gilroy S, Callaway RM & Vivanco JM (2003) Allelopathy and exotic plant invasion:
from molecules and genes to species interactions. Science 301: 13771380.
Balciunas J & Villegas B (1999) Two new seed head flies attack yellow starthistle. California Agriculture 53(2):
811.
Barrett SCH (1989) Waterweed invasions. Scientific American 260: 9097.
Baruah NC, Sarma JC, Sarma S & Sharma RP (1994) Seed germination and growth inhibitory cadinenes from
Eupatorium adenophorum Spreng. Journal of Chemical Ecology 20: 18851892.
Bennet SSR, Naithani HP & Raizada MB (1978) Parthenium L. in India -A review and history. Indian Journal
of Forestry 1(2): 128131.
Bever BO (1982) Medicinal plants in West Africa. Cambridge University Press, London, pp. 118.
Bigsby H & Whyte C (2001) Quantifying phytosanitary barriers to trade. In: Hooker N & Murano E (eds)
Interdisciplinary Food Safety Research. CRC Press, New York.
Bingelli P (1996) A Taxonomic, biogeographical and ecological overview of invasive woody plants. Journal of
Vegetation Science 7: 121124.
Blossey B, Schroeder D, Hight SD & Malecki RA (1994) Host specificity and environmental affect of the
weevil Hylobius transversovittatus, a biological control agent of purple loosestrife (Lythrum salicaria).
Weed Science 42: 128133.
Borman MM, Krueger WC & Johnson DE (1991) Effects of established perennial grasses on yields of
associated annual weeds. Journal of Range Management 4: 318326.
Burhan N & Shaukat SS (1999) Allelopathic Potential of Argemone mexicana L.A Tropical Weed. Pakistan
Journal of Biological Sciences 2: 12681273.
Bussan AJ & Dyer WE (1999) Herbicides and rangeland. In: Sheley RL & Petroff JK (eds) Biology and
Management of Noxious Rangeland Weeds. Corvallis, OR: Oregon State University Press, pp. 116132
But PPH, He ZD, Ma SC, Chan YM, Shaw PC, Ye WC & Jiang RW (2009) Antiviral constituents against
respiratory viruses from Mikania micrantha. Journal of Natural Products 72: 925928.
Reddy CS (2008) Catalogue of invasive alien flora of India. Life Science Journal 5(2): 8489.
Callaway RM & Aschehoug ET (2000) Invasive plants versus their new and old neighbours: a mechanism for
exotic invasion. Science 290: 521523.
Chandra A, Anand A & Dubey A (2007) Effect of Salicylic Acid on Morphological and Biochemical Attributes
in Cowpea. Journal of Environmental Biology 28: 193196.
Chen L, Liao L, Wang S, Huang Z & Xiao F (2002) Effect of Vanillin and P-hydroxybenzoic acid on
Physiological Characteristics of Chinese Fir Seedlings. The journal of Applied Ecology 13: 1291.
Colautti RI, Ricciardi A, Grigorovich IA & MacIssac HJ (2004) Is invasion success explained by the enemy
release hypothesis? Ecology letters 7: 721733.
Cross BT & Wiedemann HT (1985) Grubbing for control of blackbrush acacia (Acacia rigidula) invading root
plowed rangeland. Weed Science 33: 263266.
Cruz-Ortega R, Anaya AL, Hernndez-Bautista BE & Laguna-Hernndez G (1998) Effects of Allelochemical
Stress Produced by Sicyosdeppeion Seedling Root Ultrastructure of Phaseolus vulgaris and Cucurbita
ficifolia. Journal of Chemical Ecology 24: 20392057.
Cruz-Ortega R, Lara-Nez A & Anaya AL (2007) Allelochemical Stress Can Trigger Oxidative Damage in
Receptor Plants: Mode of Action of Phytotoxicity. Plant Signaling & Behaviour 2: 269270.
Curtis CD & Carino DA (1999) Threats of invasive plants to the conservation of biodiversity. In: Chou CH,
Waller GR & Reinhardt C (eds) Biodiversity and Allelopathy: From Organism to Ecosystem in the Pacific.
Academia Sinica, Taipei, pp. 2127.
96
Yadav et al. (2016) 3(1): 87101

.
Day MD, Wiley CJ, Playford J & Zalucki MP (2003) Lantana: Current Management, Status and Future
Prospects. Australian Centre for International Agricultural Research 5: 120.
Deng X, Ye WH, Feng HL, Yang QH, Cao HL, Xu KY & Zhang Y (2004) Gas exchange characteristics of the
invasive species Mikania micrantha and its indigenous congener M. cordata (Asteraceae) in South China.
Botanical Bulletin of Academia Sinica 45: 213220.
Ding ZH, Guo YB & Ding JK (1999) Chemical constituents from the flower of Eupatorium adenophorum. Acta
Botanica Yunnanica 21: 505511.
DiTomaso JM (2000a) Irrigated pastures. In: Colbert F (ed) Principles of Weed Control in California. 3rd ed.
Fresno, CA: Thomson Publishing.
DiTomaso JM (2000b) Invasive weeds in rangelands: Species, impacts, and management. Weed Science 48(2):
255265.
DiTomaso JM, Kyser GB & Hastings MS (1999a) Prescribed burning for control of yellow starthistle
(Centaurea solstitialis) and enhanced native plant diversity. Weed Science 47: 233242.
Dogra KS, Kohli RK & Sood SK (2009) An assessment and impact of three invasive species in the Shivalik
hills of Himachal Pradesh, India. International Journal of Biodiversity and Conservation 1(1): 410.
Dorken ME& Barrett SCH (2004) Phenotypic plasticity of vegetative and reproductive traits in monoecious and
dioecious populations of Sagittaria latifolia (Alismataceae): A clonal aquatic plant. Journal of Ecology 2:
3244.
Einhellig FA, Rasmussen JA, Hejl AM & Souza IF (1993) Effects of Root Exudate Sorgoleone on
Photosynthesis. Journal of Chemical Ecology 19: 369375.
Elton CS (1958) The ecology of Invasions by animals and plants. Methuen, London.
Feng HL, Cao HL, Liang XD, Zhou XX & Ye WH (2002) The distribution and harmful effects of Mikania
micrantha in Guangdong. Journal of Tropical and Suubtropical Botany 10: 263270.
Feng HL, Li Y, Ye WH, Yu Z & Huang ZR (2004a) The chemical component of the stem and leaves of Mikania
micrantha. Chinese Traditional and Herbal Drugs 35: 1718.
Feng HL, Yang CJ, Zhang X & Ye WH (2004b) Preliminary studies on the bioactivity of crude extract from
Mikania micrantha on insect and plant, pathogen. Acta Scientiarum Naturalium Universitatis Sunyatseni 43:
8285.
Forcella E & Wood JT (1984) Colonization potentials of alienweeds are related to their native distributions:
Implications for plant quarantine. Journal of the Australian Institute of Agricultural Science 50(1): 3640.
Forcella F & Harvey SJ (1983) Eurasian weed infestation in western Montana in relation to vegetation and
disturbance. Madrono 30: 102109.
Gentle CB & Duggin JA (1997) Allelopathy as a competitive strategy in persistent thickets of Lantana camara
L. in three Australian forest communities. Plant Ecology 132: 8595.
Goodin BJ, McAllister AJ & Fahrig L (1998) Predicting invasiveness of plant species based on biological
information. Conservation Biology 13: 422426.
Gunaseelan VN (1987) Parthenium as an additive with cattle manure in biogas production. Biological Wastes 1:
195202.
Hatch DA, Bartolome JW & Hillyard DS (1991) Testing a management strategy for restoration of Californias
native grasslands. In: Proceedings of the Symposium on Natural Areas and Yosemite: Prospects for the
Future. Denver, CO: U.S. National Park Service, pp. 343349.
He AJ & Liu LH (1990) Effect of water extract of Eupatorium adenophorum on the germination of several
plants. Chinese journal of Weed Science 4: 3538.
Heirro JL & Callaway RM (2003) Allelopathy and exotic plant invasion. Plant and Soil 256: 2939.
Higgins SI, Richardson DM & Cowling RM (2000) Using a dynamic landscape model for planning the
management of alien plant invasions. Ecological applications 10: 18331848.
Holm LG, Plucknett DL, Pancho JV & Herberger JP (1977) The Worlds Worst Weeds: Distribution and
Biology. The University Press of Hawaii, Honolulu USA, pp. 609.
Inderjit (2005) Soil microorganisms: an important determinant of allelopathic activity. Plant and Soil 274: 227
236.
Inderjit & Callaway RM (2003) Experimental designs for the study of allelopathy. Plant and Soil 256: 111.
97
Yadav et al. (2016) 3(1): 87101

.
Inderjit & Seastedt TR, Callaway RM & Kaur J (2008) Allelopathy and plant invasions: traditional, congeneric,
and biogeographical approaches. Biological Invasion 10: 875890.
Jacobs JS, Carpinelli MF & Sheley RL (1999) Revegetating noxious weed-infested rangeland. In: Sheley RL &
Perroff JK (eds) Biology and Management of Noxious Rangeland Weeds. Oregon State University Press,
Corvallis, pp. 133141.
Joshi S (1991) Biological control of Parthenium hysterophorus L. (Asteraceae) by Cassia uniflora Mill.
(Leguminosae) in Bangalore, India. Tropical Pest Management 37: 182184.
Kanchan SD & Jayachandra (1980) Allelopathic effects of Parthenium hysterophorus L. IV.Identification of
inhibitors. Plant and Soil 55: 6775
Karlsson LM, Tamado T& Milberg P (2003) Seed Dormancy Pattern of the Annuals Argemone ochroleuca and
A. mexicana (Papaveraceae). Flora-Morphology, Distribution, Functional Ecology of Plants 198: 329339.
Kennett GA, Lacey JR, Butt CA, Olson-Rutz KM & Haferkamp MR (1992) Effects of defoliation, shading and
competition on spotted knapweed and bluebunch wheatgrass. Journal of Range Management 5: 363369.
Kingsbury JM (1964) Poisonous Plants of the United States and Canada.Prentice-Hall, Englewood Cliffs, pp.
296397.
Kohli R, Batish KD & Singh HP (1998) Allelopathy and its implications in agro ecosystems. Journal of Crop
Production 1(1): 169202.
Kohli RK, Batish DR, Singh HP & Dogra KS (2006) Status, invasiveness and environmental threats of three
tropical American invasive weeds (Parthenium hysterophorus L., Ageratum conyzoides L., Lantana camara
L.) in India. Biological Invasions 8: 15011510.
Kohli RK & Batish DR (1994) Exhibition of allelopathy by Parthenium hysterophorus L. in agroecosystems.
Tropical Ecology 35: 295307.
Kohli RK & Rani D (1994) Parthenium hysterophorus a review. Research Bulletin (Science) of Panjab
University 44: 105149.
Kohli RK, Batish D & Singh HP (1998) Eucalypt oils for the control of Parthenium (Parthenium hysterophorus
L.). Crop Protection 17: 119122.
Kohli RK, Dogra KS, Batish DR & Singh HP (2004) Impact of invasive plants on the structure and composition
of natural vegetation of northwestern Indian Himalayas. Weed Technology 18: 12961300.
Kong C, Hu F & Xu X (2002) Allelopathic potential and chemical constituents of volatile oil from Ageratum
conyzoides under stress. Journal of Chemical Ecology 28: 11731182.
Kong CH, Xu T & Hu F (1998) Allelopathy of Ageratum conyzoides (II): Releasing mode and activity of main
allelochemicals. Chinese Journal of Applied Ecology 9: 257260.
Kong GH, Wu QG & Hu QM (2000) Appearing of exotic weed Mikania micrantha H.B.K. in China. Journal of
Tropical and Subtropical Botany 8: 27.
Lee GA (1986) Integrated control of rush skeleton weed (Chondrilla juncea) in the western U.S. Weed Science
34: 26.
Levine JM, Alder PB & Yelenik SG (2004) A meta-analysis of biotic resistance to exotic plant invasions.
Ecology Letters 7: 975989.
Li Z-H, Wang Q, Ruan X, Pan C-D & Jiang D-A (2010) Phenolics and Plant Allelopathy. Molecules 15: 8933
8952.
Lu RD (1982) Studies on anti-insect pheromone: chemical components of Ageratum conyzoides and their effects
on insects. Entomological Knowledge 19: 2225.
Ma WJ, Xiao DJ & Deng SZ (2004) Terpenoids constituents of the leaves of Lantana camara. Guangzhou
Chemistry 29: 1419.
Mack RN, Simberloff D, Lonsdale WM, Evans H, Clout M & Bazzaz FA (2000) Biotic invasion: Causes,
epidemiology, global consequences and control. Ecological Applications 10: 689710.
Maishi AI, Ali PKS, Chaghtai SA & Khan G (1998) A proving of Parthenium hysterophorus L. British
Homoeopathic journal 87: 1721.
Mandal FB (2011) The management of alien species in India. International Journal of Biodiversity and
Conservation 3(9): 467473.
Mantri A, Annapurna C & Singh JS (2002) Terrestrial plant invasion and global change. In: Tripathi G &
Tripathi YC (eds) Bioresource and Environment.Campus Book International, New Delhi, pp. 2544.
98
Yadav et al. (2016) 3(1): 87101

.
Marks MK & Nwachuku AC (1986) Seed bank Characterstics in a group of Tropical weeds. Weed Research 26:
151157.
McHenry WB & Murphy AH (1985) Weed management of California rangeland. In: Kurtz EA & Colbert FO
(eds) Principles of Weed Control in California. Thomson Publishing, Fresno, pp. 413423.
Meng XX, Feng JC, Zhou YJ, Wang FM, Qu M, Lin G, Zhao CJ, Zhu YJ, Ying J & Yang H (2003) Ecological
factor analysing of the invasion of crofton weed in southwestern Shichuan province. Journal of central
university for nationalities (Natural Sciences Edition) 12: 293300.
Mitchell CE, Agrawal AA, Bever JD, Gilbert GS, Hufbauer RA, Klironomos JN, Maron JL, Morris WF, Parker
IM, Power AG, Seabloom EW, Torchin ME & Vazquez DP (2006) Biotic interactions and plant invasions
.Ecology Letters 9: 726740.
Morin L, Reid AM, Sims-Chilton NM, Buckley YM, Dhileepan K, Hastwell GT, Nordblom TL & Raghu S
(2009) Review of approaches to evaluate the effectiveness of weed biological control agents. Biological
control 5: 115.
Muzaffar S, Ali B & Wani NA (2012) Effect of Catechol, Gallic Acid and Pyrogallic Acid and the Germination,
Seedling Growth and the Level of Endogenous Phenolics in Cucumber (Cucumis sativus L.). International
Journal of Life Science Biotechnology and Pharma Research 1: 5159.
Narasimban TR, Murthy BSK, Harindramath N & Rao PVS (1984) Characterization of a toxin from Parthenium
hysterophorus and its mode of excretion in animals. Journal of Biosciences 6: 729738.
Narwal SS (1994) Allelopathy in crop production. Scientific Publishers, Jodhpur, India, 288 p.
Narwal S, Palaniraj R & Sati S (2005) Role of Allelopathy in Crop Production. Herbologia 6: 327332.
Nayar MP (1977) Changing pattern of the Indian flora. The Bulletin of the Botanical Survey of India 19: 145
154.
Noble IR & Slatyer RO (1980) The use of vital attributes to predict successional changes in plant communities
subject to recurrent disturbances. Vegetation 43: 521.
Okunade AL (2002) Ageratum conyzoides L. (Asteraceae). Fitoterapia 73: 116.
Pan WD, Mai LT, Li YJ, Xu XL & Yu DQ (1993) Studies on chemical constituents of the leaves of Lantana
camara. Acta Pharmaceutica Sinica 28: 3539.
Paul N & Begum N (2010) Allelopathic Effect of Argemone Mexicana L. on Germination and Seedling Growth
Characteristics of Lentil (Lens culinaris). Journal of Bio-Science 18: 146147.
Pieterse AH & Murphy KJ (eds) (1990) Aquatic Weeds, Oxford University Press, Oxford.
Pimentel D, Lach L, Zuniga R & Morrison D (2000) Environmental and economic costs of non-indigenous
species in the United States. Bioscience 50: 5365.
Quan G, Zhang J, Xu H, Mao D & Qin Z (2009) Allelopathic effects of different parts of invasive plant Lantana
camara. Chinese agricultural Science Bulletin 25(12): 102106.
Raghubanshi AS, Rai LC, Gaur JP & Singh JS (2005) Invasive alien species and biodiversity in India. Current
Science 88(4): 539540.
Rasmussen GA (1991) Oakbrush: classification, ecology, and management. In: James JF, Evans JO, Ralphs MH
& Child RD (eds) Noxious Range Weeds. Westview Press, San Francisco, pp. 352363.
Rastogi J, Rawat DS & Chandra S (2015) Diversity of invasive alien species in Pantnagar flora. Tropical Plant
Research 2(3): 282287.
Reichard SH & Hamilton CW (1997) Predicting invasions of woody plant introductions into North America.
Conservation Biology 11: 193203.
Rejmanek M & Richardson DM (1996) What attributes make some plant species more invasive? Ecology 77(6):
16551661.
Rejmanek M & Randall JM (1994) Invasive alien plants in California: 1993 Summary and comparison with
other areas in North America. Madrona 41: 161177.
Rice EL (1984) Allelopathy, 2nd edition. Academic Press, New York, USA.
Richardson DM, Allsopp N, DAntonio CM, Milton SJ & Rejmanek M (2000) Plant invasions - the role of
mutulasims. Biological Reviews of the Cambridge Philosophical Society 75: 6593.
Rimando AM, Dayan FE, Czarnota MA, Weston LA & Duke SO (1998) A New Photosystem II Electron
Transfer Inhibitor from Sorghum bicolor. Journal of Natural Products 61: 927930.
99
Yadav et al. (2016) 3(1): 87101

.
Robertson MP, Villet MH, Fairbanks DHK, Henderson L, Higgins SI, Hoffmann JH, Le Maitre DC, Palmer
AR, Riggs I, Shackleton CM & Zimmermann HG (2003) A proposed prioritisation system for the
management of weeds in South Africa. South African Journal of Science 99: 3743.
Roder W, Keobulapha B, Phengchanh S, Prot JC & Matias D (1998) Effect of residue management and fallow
length on weeds and rice yield. Weed Research 38: 167174.
Roxburgh W (1884) Hortus bengalensis. Mission Press, Calcutta.
Ruiz GM, Fofonoff PW, Carlton JT, Wonham MJ & Hines AH (2000) Invasion of coastal marine communities
in North America: Apparent patterns, processes, and biases. South African Journal of Science 31: 481531.
Sage RF (2004) The evolution of C4 photosynthesis. New Phytologist 161: 341370.
Sanaa A (2012) Perspectives on the Relationship between Invisibility, Richness, Plant Size, Seed Production,
Seed Bank and Community Productivity of Invasive Argemone ochroleuca Sweet in Taif, Saudi Arabia. Life
Science Journal 9: 953958.
Shao H, Peng SL, Zhang C, Xiang YC & Na P (2003) Allelopathic potential of Mikania micrantha. Chinese
Journal of Ecology 22: 6265.
Sharma GP, Raghubanshi AS & Singh JS (2005) Lantana invasion: An overview. Weed Biology Management 5:
157167.
Sharma OP, Makkar HPS & Dawra RK (1988) A review of the noxious plant Lantana camara. Toxicon 26:
975987.
Sharma OP & Sharma PD (1989) Natural products of the Lantana plant -the present and prospects. Journal of
Scientific & Industrial Research 48: 471478.
Sharma PD & Sharma OP (1995) Natural products chemistry and biological properties of Ageratum plant.
Toxicological and Environmental Chemistry 50: 213232.
Sharma OP (1988) How to combat Lantana (Lantana camara L.) menace? A current perspective. Journal of
scientific and Industrial research 47: 611616.
Sheley RL, Jacobs JS & Carpinelli ME (1998) Distribution, biology, and management of diffuse knapweed
(Centaurea diffusa) and spotted knapweed (Centaurea maculosa). Weed Technology 12: 353362.
Sheley RL, Manoukian M & Marks G (1999b) Preventing noxious weed invasion. In: Sheley RL & Petroff JK
(eds) Biology and Management of Noxious Rangeland Weeds. Oregon State University Press, Corvallis, pp.
6972.
Sheley RL, Kedzie-Webb S & Maxwell BD (1999a) Integrated weed management on rangeland. In: Sheley RL
& Petroff JK (eds) Biology and Management of Noxious Rangeland Weeds. Oregon State University Press,
Singh CM, Angras NM & Kumar S (1996) Weed management. MD publications, New Delhi.
Singh HP, Batish DR, Setia N & Kohli RK (2005) Herbicidal activity of volatile oils from Eucalyptus citriodora
against Parthenium hysterophorus. Annals of Applied Biology 146: 8994.
Singh KP, Shukla AN & Singh JS (2010) State-level inventory of invasive alien plants, their source regions and
use potential. Current Science 90(1): 107114.
Song J (1990) Allelopathy Among Plants. Chinese Journal of Ecology 9(6):4347.
Song QS, Fu JJ, Tang JW, Feng LZ & Yang CR (2000) Allelopathic potential of Eupatorium adenophorum.
Acta Phytoecologica Sinica 24: 362365.
Sun WH & Yu SW (1992) Allelopathy and Its Potential Application. Plant Physiology Communications 28(2):
8187.
Sun WH, Yu SW, Yang SY, Zhao BW, Yu ZW, Wu HM, Huang SY & Tang CS (1993) Allelochemicals from
root exudates of water hyacinth (Eichhornia crassipes). Acta Phytophysiologica Sinica 19: 9296.
Thomsen CD, Williams WA, Vayssieres M, Bell FL & George MR (1993) Controlled grazing on annual
grassland decreases yellow starthistle. California Agriculture 7(6): 3640.
Towers GHN & SubbhaRao PV (1992) Impact of the pan-Tropical weed, P. hysteroporus L. on human affairs.
In: Richardson RG (ed) Proceedings of the first international weed control congress, Melbourne, Australia.
Weed science society of Victoria, pp. 134138.
Tripathi RS, Khan Ml & Yadav AS (2012) Biology of Mikania micrantha H.B.K.: a Review In: Bhatt JR, Singh
JS, Singh SP, Tripathi RS & Kohli RK (eds) Invasive Alien Plants: An Ecological Appraisal for the Indian
Subcontinent. CAB International, MoEF, New Delhi, India, pp. 99107.
100
Yadav et al. (2016) 3(1): 87101

.
Tyser RW & Key CH (1988) Spotted knapweed in natural area fescue grassland: an ecological assessment.
Northwest Science 2: 151160.
Ueckert DN, Petersen JL, Potter RL, Whipple JD & Wagner MW (1988) Managing prickly pear with
herbicides and fire. In: Texas Agricultural Experiment Station Report No. 4570, pp. 1015.
Wan F H & Wang R (1990) The biology and ecology characteristic of the malignancy harm grass Ambrosia
artemisiifolia. Journal of Weed Science 4(1): 4548.
Wei XY, Haung HJ, Wu P, Cao HL & Ye WH (2004) Phenolic constituents from Mikania micrantha.
Biochemical Systematics and Ecology 32: 10911096.
Weir TL, Park S-W & Vivanco JM (2004) Biochemical and Physiological Mechanisms Mediated by
Allelochemicals. Current Opinion in Plant Biology 7: 472479.
Weng WP (1964) Evidence for the presence of growth inhibitory substances in Mikania cordata (Brum. f.) B.L.
Robinson. Journal of Rubber Research Institute Malaya 18: 231242.
Wilcove DS, Rothstein D, Dubow J, Phillips A & Losos E (1998) Quantifying threats to imperilled species in
the United States. Bioscience 48: 607615.
Wilson LM & McCaffrey JP (1999) Biological control of noxious rangeland weeds. In: Sheley RL & Petroff JK
(eds) Biology and Management of Noxious Rangeland Weeds. Oregon State University Press, Corvallis, pp.
97116.
Wu TJ & Yang GZ (1994) Chemical constituents of the essential oil of Eupatorium adenophorum Spreng.
Jounral of Central China Norman University (Natural Sciences Edition) 28: 8797.
Xie LJ, Zeng RS, Bi HH, Song YY, Wang RL, Su YJ, Chen M, Chen S & Liu YH (2010) Allelochemical
mediated invasion of exotic plants in China. Allelopathy Journal 25(1): 3150.
Yan F, Yan Z-M & Han L-M (2000) Review on research methods for allelopathy and allelochemicals in plants.
Acta Ecologica Sinica 20(4): 692696.
Yang SY, Yu ZW, Sun WH, Zhao BW, Yu SW, Wu HM, Huang SY, Zhou HQ, Ma K & Lao XF (1992)
Isolation and identification of antialgal compounds from root system of water hyacinth. Acta
Phytophysiologica Sinica 18: 399402.
Yang C-M, Lee C-N& Chou C-H (2002) Effects of Three Allelopathic Phenolics on Chlorophyll Accumulation
of Rice (Oryza sativa) Seedlings: I. Inhibition of Supply-Orientation. Botanical Bulletin of Academia Sinica
43: 299304.
Yi Z, Zhang MX, Ling B, Xu D & Ye JZ (2006) Inhibitory effects of Lantana camara and its contained
phenolic compounds on Eichhornia crassipes growth. Chinese Journal of Applied Ecology 17: 16371640.
Young JA, Palmquist DE & Blank RR (1998) The ecology and control of perennial pepperweed (Lepidium
latifolium L.). Weed Technology 12: 402405.
Zamora DL & Thill DC (1999) Early detection and eradication of new weed infestations. In: Sheley RL &
Petroff JK (eds) Biology and Management of Noxious Rangeland Weeds. Oregon State University Press,
Zan QJ, Wang YJ, Wang BS, Liao WB & Li MG (2000) The distribution and harm of the exotic weed Mikania
micrantha. Chinese Journal of Ecology 19: 5861.
Zhang XL, Wang JW & Zheng LP (1993) The allelopathic effects of maceration exctrat of Eupatorium
adenophorum on roots of Zea Mays- Preliminary studies with electron microscopy and electron probe X-ray
microanalyser. Jounral of Yunnan University 15(Suppl.2): 112117.
Zhang MX, Ling B, Kong CH & Pang XF (2003) Chemical components of volatile oil from Mikania micrantha.
Chinese Journal of Applied Ecology 13: 13001302.
Zheng HQ, Wei N, Wang LF & He P (2006) Effects of Lantana camara leaf extract on the activity of
superoxide dismutase and accumulation of H2O2 in Water hyacinth leaf. Journal of Plant Physiology and
Molecular Biology 32: 189194.
101
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 102104, 2016
Short communication
Ipomoea triloba L. (Convolvulaceae): A New Angiosperm Record

for the Flora of Bangladesh
Md Sharif Hossain Sourav
Department of Environmental Science and Management, North South University, Plot-15, Block-B,
Bashundhara, Dhaka 1229, Bangladesh
*Corresponding Author: nature.sourav@gmail.com
[Cite as: Sourav MSH (2016) Ipomoea triloba L. (Convolvulaceae): A New Angiosperm Record for the Flora
of Bangladesh. Tropical Plant Research 3(1): 102104]
Family Convulvulaceae consist of about 50 genera having 1500 species, mainly distributed in tropics and
also in subtropical regions of the world. In Bangladesh, this family is represented by 15 genera and 55 species.
The genus Ipomoea is represented by 24 species in Bangladesh (Ahmed et al. 2008). A twining climber with
pinkish flowers was collected from Dhaka district of Bangladesh in the month of November 2015 which has
been identified as Ipomoea triloba is described and illustrated for the first time from Bangladesh. In November
2015, this species was found in Dhaka which associated with Merremia hederacea (Burm.f.) Hallier f., Coccinia
grandis (L.) Voigt. & Tinospora cordifolia (Willd.) Hook. f. & Thom. Specimens of this plant were collected
from this undisturbed natural habitat (latitude 2346'18.75; longitude 9024'45.27) for further study. After
details examination of the specimens, carefully consulting relevant literature (Ordetx 1949, Holm et al. 1979,
Wagner et al. 1999, Ahmed et al. 2008, Perera & Nilanthi 2015) and seeking expert opinion, the collected
material was identified as Ipomoea triloba L., which forms a new record for angiospermic flora of Bangladesh.
Hence, the species is hereby presented as a new angiosperm record for Bangladesh.
During an early morning short visit to the fallow fields of Dhaka, Bangladesh in November 2015 conducted
to observe flowering of morning glories and other members of the family, this taxon was observed. This species
was found to have very small and pink coloured flowers as well as fruits, apparently not matching with any
species known till date in Bangladesh. The specimens of the same are deposited at Bangladesh National
Herbarium (DACB). The detailed description and illustration of the species based on herbarium material are
given below.
Species description
Ipomoea triloba L. Sp. Pl. 1: 161. 1753.
(Figs. 12)
An annual climber with 13 meters long, somewhat angled stems, about 1.53.0 mm wide, milky. Leaves
are cordate, acuminate, mostly 510 cm long (can reach up to 12 cm), longer than wide, not always three-lobed
as the specific epithet suggests. Petiole slender, 4.6 to 10 cm, glabrous or sometimes minutely tuberculate,
glabrous or pubescent. Inflorescence axillary, peduncle shorter to longer than the petiole, angular toward the
apex, one-flowered or cymosely few to several-flowered, branches of the cyme very short. Flowers aggregate,
pedicels glabrous, 2.5 to 8 mm, closing before noon, sepals slightly unequal, 8 to 10 mm long, the outer ones
little shorter, oblong to narrowly elliptic-oblong, glabrous or sparsely hairy on the back, corolla 5lobed, funnelshaped, 2022mm long, glabrous, pinkish, with or without white markings, bell-shaped, outer wide 9mm, inner
wide 8mm when opened. Stamens 5, 8 mm long, stigma 14 mm long, ovary 2 to 4-celled, conical, densely
pubescent. Fruit a capsule, about 610 mm wide, depressed globose with sharp point, bristly hairy. Seeds
glabrous or with a few minute hairs, 4-seeded, 3 mm long, 2.5mm wide, hard, shiny, chocolate brown.
Flowering & Fruiting: Flowering observed early November, but starts from September to continue till
December; fruiting occurs from October to December and during the filed study, both immature and mature
fruits were found.
Received: 02 January 2016
102
Sourav (2016) 3(1): 102104

.
Figure 1. A, Ipomoea triloba, from its current habitat; B, Twining stems with leaves and unopened flowers; C,
Bloomed flower; D, Fruit bearing twigs.
Figure 2. A, Stem with Leaf and Inflorescence; B, Flower showing pedicel and sepals; C, Leaf and axillary
inflorescence; D, A flower; E, Corolla dissected; F, Sepals; G, Stigma; H, Fully opened flower; I, Stamens; J,
Seeds; K, Fruit. (Illustrations by: MSH Sourav & Mashuda Pervin)
103
Sourav (2016) 3(1): 102104

.
Specimen Examined: BANGLADESH, Dhaka, Near Bannai Lake, adjacent to Hatirjheel, 11.11.2015, M.S.H.
Sourav 25 (DACB).
Habitat: The species was recorded from a wasteland (Fig. 1) adjoining the road. The stem scrambles over the
ground and twines into other plants for support. As this grows in close association with other similar-leaved
vines, it is very difficult to distinguish in vegetative phase. It can be told apart only after flowers appear. It
grows profusely in shade and wet conditions.
Distribution: This species is a native climber of Tropical America (Wagner et al. 1999), now naturalized
throughout the tropics (Stone 1970). It was introduced into East and South-East United States. Elsewhere
Bangladesh, the nearest occurrence records are found from India, China, Nepal, Myanmar, Pakistan and
Srilanka (Perera & Nilanthi 2015). In India, it was recorded from Gujarat, Kerala, Karnataka, Maharashtra,
Rajasthan, Uttar Pradesh and West Bengal (Magesh et al. 2012).
Uses: The leaves are cooked and eaten as a vegetable and decoction of the leaves is used against stomach ache
in Benin, West Africa, where they are also said to be sold sometimes in local markets (Achigan-Dako et al.
2010). Because of the attractive flower and habit, this species can be used as wild ornamental species (Divya &
Thomas 2015). It is considered to be an important plant in honey production in Cuba and other Central
American countries (Ordetx 1949).
ACKNOWLEDGEMENTS
The author expressed his deep gratitude to Nidhan Singh for revising the papers and Surajit Koley for e-flora
of India especially. The author also wants to show his gratitude to J. M Garg for identification and Md. Amanat
Ullah for other technical help.
REFERENCES
AchiganDako EG, Pasquini MW, AssogbaKomlan F, Ndanikou S, Ydomonhan H, Dansi A&AmbroseOji B
(2010) Traditional vegetables in Benin. Institut National des RecherchesAgricoles du Bnin. Imprimeries du
CENAP, Cotonou.
Ahmed ZU, Hassan MA, Begum ZNT, Khondker M, Kabir SMH, Ahmad MATA, Rahman AKA & Haque EU
(eds) (2008) Encyclopedia of Flora and Fauna of Bangladesh. Vol.7. Angiosperms: dicotyledons
(Balsminaceae-Euphorbiaceae). Asiatic Society of Bangladesh, Dhaka, pp. 247273.
Magesh CR, Lakshminarasimhan P & Ven P (2012) New plant records for Jharkhand. ZOOs PRINT 27(5): 25.
Divya & Thomas B (2015) Potential ornamental Convolvulacean members of Kottayam district, Kerala, India.
European Journal of Environmental Ecology 2(3): 156161.
Holm LG, Pancho JV, Herberger JP & Plucknett DL (1979) A Geographical Atlas of World Weeds. New York,
USA: John Wiley and Sons.
Ordetx G (1949) The Aguinaldos, major bee plants of Cuba. American Bee Journal 89: 7273.
Perera PCD & Nilanthi D (2015) Review of major abundant weeds of cultivation in Sri Lanka. International
Journal of Scientific and Research Publications 5(5): 22503153.
Stone BC (1970) The flora of Guam. Micronesica 6: 1659.
Wagner WL, Herbst DR & Sohmer SH (1999) Manual of the flowering plants of Hawaii. Revised edition.
Bernice P. Bishop Museum special publication. University of Hawaii Press/Bishop Museum Press,
Honolulu, pp. 1919.
104
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 105111, 2016
Review article
Endogenous glycine betaine accumulation mediates abiotic stress

tolerance in plants
Aryadeep Roychoudhury* and Aditya Banerjee
Post Graduate Department of Biotechnology, St. Xaviers College (Autonomous),
30, Mother Teresa Sarani, Kolkata-700016, West Bengal, India
*Corresponding Author: aryadeep.rc@gmail.com
Abstract: Abiotic stresses like salinity, drought, cold, high temperature, etc. are largely
responsible for a considerable degree of annual crop losses worldwide. Till date, several effectors
have been reported which confer stress tolerance to the plants. Glycine betaine (GB) is one such
important regulator which accumulates in the cell as a crucial osmolyte and alleviates the cell from
damages due to salinity, drought, temperature and oxidative stresses. This has been verified
through several investigations at the physiological, biochemical and molecular levels. Apart from
acting as an important compatible solute, GB has a prominent role in maintaining cellular
homeostasis and mediating chaperone activity to prohibit undesirable protein folding under stress.
The protective role of exogenous applications of GB in crops is also a well-studied fact. In
addition, several detailed literatures are available on the transgenic technology and on-field tests
which have depicted the increasing stress tolerance developed in plants accumulating higher levels
of endogenous GB. All these issues have been reviewed and documented in the present
communication.
Keywords: Abiotic stresses - Stress tolerance - Glycine betaine - Transgenic technology.
[Cite as: Roychoudhury A & Banerjee A (2016) Endogenous glycine betaine accumulation mediates abiotic
stress tolerance in plants. Tropical Plant Research 3(1): 105111]
INTRODUCTION
Environmental stresses, particularly salinity and drought, are the major constraints which limit the global
distribution and production of crop plants. In response to such abiotic stresses, plants have evolutionarily
developed a plethora of stress responsive cascades which aid them in developing stress tolerance. The
accumulation of certain organic metabolites of low molecular weight, collectively called the compatible solutes,
is one such ubiquitous mechanism in plants (Bohnert et al. 1995). These compatible solutes act as crucial
osmoprotectants and help the plant system to survive severe osmotic stress. Accumulation of compatible
osmolytes aid plant cells to regain turgor and resume growth during ionic stress. The osmotic potential of the
cytosol is reduced in order to facilitate the uptake as well as the retention of water molecules. Being small
organic molecules of low molecular weight, these compatible solutes have the tendency to accumulate to high
levels without interfering with normal intra and intercellular homeostasis (Roychoudhury et al. 2008).
Compatible solutes have been depicted to prevent ion entry into the sensitive plant parts and also to increase the
ion exclusion from them. These metabolites are polar, highly soluble and typically hydrophilic and hence exhibit
their protective functions by maintaining the hydration sphere of proteins under desiccating conditions.
Compatible solutes at high concentrations prohibit misfolding of proteins, thus acting as low molecular weight
chaperones, stabilize macromolecules or molecular assemblies, increase the thermal stability of enzymes and
prevent dissociation of enzymatic complexes (Roychoudhury et al. 2013).
Compatible solutes often act as the scavengers of reactive oxygen species (ROS; extremely toxic, short lived
active oxygen species) and thus preserve membrane and cellular integrity. Some common compatible solutes
accumulated under desiccation stress are reducing sugar and major carbohydrates like sucrose, fructose, glucose;
sugar alcohols (pinitol, ononitol, cyclitol); polyols (either straight chain compounds like adonitol, sorbitol and
mannitol or cyclic polyols as myo-inositol); complex sugars (trehalose, raffinose and fructans); total free amino
acids especially proline (Pro) and glycine betaine (GB); organic acids like lactate, malate, citrate, succinate,
105
Roychoudhury & Banerjee (2016) 3(1): 105111

.
fumarate, benzoate, salicylate, malonate and -amino butyric acid (GABA); free ammonia and quaternary
ammonium compounds (-alanine-betaine, proline-betaine and hydroxyprolinebetaine); tertiary sulfonium salts
(dimethylsulfoniopropionate, choline o-sulfate) etc. (Roychoudhury & Chakraborty 2013, Roychoudhury et al.
2015).
GB (N, N, N-trimethylglycine) is an important member among the compatible solutes with widely ranging
protective functions under salinity, drought and extreme temperature stresses (Chen & Murata 2008). GB is
dipolar at physiological pH, but electrically neutral in nature. The essential role carried out by GB in plants
exposed to salinity stress is the protection of plant tissues via osmotic adjustment (OA), stabilization of proteins
like RuBisCO, photosynthetic apparatus protection and scavenging of ROS (Wani et al. 2013). In many
halophytic plants and cyanobacteria, GB accumulates to an osmotically significant level (Rhodes & Hanson
1993). Natural accumulators of GB have shown the accumulation of betaine in response to salt, drought and
cold. Water stress, induced by polyethylene glycol showed enhanced GB accumulation in rice; however, the
level was much higher in the tolerant variety Pokkali, as compared to the sensitive varieties IR-29 and Pusa
Basmati (Basu et al. 2010), showing that GB could act as a better osmoprotectant against dehydration stress in
the tolerant variety. The taxonomically distant plant species referred to as the natural accumulators of GB,
normally contain low levels of GB under control conditions. However, they accumulate considerable levels of
GB on exposure to stress (Giri 2011).
It has been suggested that compatible solutes like GB at low levels protect macromolecules like nucleic
acids, proteins and lipids and also act as reservoirs of carbon and nitrogen sources (Umezawa et al. 2006). The
major crops like potato and tomato are unable to accumulate GB. Thus, these species have been regarded as the
targets for engineering betaine biosynthesis (Wani et al. 2013). In these review, we present a picture of the
thorough development of abiotic stress tolerance in plants mediated by GB accumulation.
BIOSYNTHESIS OF GLYCINE BETAINE (GB)
A brief discussion of the biosynthetic pathway of GB is essential, since these genes and hence enzymes can
be cloned and expressed in crop plants which do not accumulate GB. GB can be synthesized via two pathways.
The starting metabolites in these pathways are different. One is initiated by choline, while the other with
glycine. The common pathway in higher plants starts with choline, which is catalyzed by choline
monooxygenase (CMO) into the hydrated form of betaine aldehyde. Betaine aldehyde is acted upon by NAD +
dependent betaine aldehyde dehydrogenase (BADH) to ultimately form GB. Both CMO and BADH are
localized within the stroma of chloroplasts (Sakamoto & Murata 2002). Choline dehydrogenase (CDH) and
BADH regulates the formation of GB in Escherichia coli, while choline oxidase A (codA) in the soil bacterium,
Arthrobacter globiformis catalyses a single step conversion of choline to GB and hydrogen peroxide (Giri
2011).
The alternate pathway for GB biosynthesis has been reported in only two extreme halophilic
microorganisms, Ectothiorhodospira halochloris and Actinopolyspora halophilia. In this recently reported
pathway, glycine undergoes three successive N-methylations catalysed by glycine sarcosine methyltransferase
(GSMT) and sarcosine dimethylglycine methyltransferase (SDMT). Both GSMT and SDMT are actually two Sadenosylmethionine-dependent methyl transferases (Takabe et al. 2006).
DEVELOPING STRESS-TOLERANT PLANTS BY EXOGENOUS APPLICATION OF GB:
AGRONOMICAL ASPECTS
Exogenous treatment of GB has been reported as a potential and traditional approach for crop tolerance
against multiple abiotic stresses. Thus, this strategy can be used as an easy method for generating abiotic stresstolerant crops directly in the field by foliar spray of solutions containing GB at the optimum concentrations. The
correlation between GB accumulation in plants and their economical productivity under stress has been
indicated (Smirnoff & Stewart 1985). The natural accumulation of GB is widespread across plant families like
Asteraceae, Chenopodiaceae, Poaceae and Solanaceae in response to stress (Jones & Storey 1981). Exogenous
foliar treatment of soybean plants (low accumulator of GB with average accumulation of around 5 mol g-1 dry
weight) enhanced the GB accumulation by 12 folds to 60 mol g-1 dry weight (DW). This led to an overall
increased photosynthetic yield, nitrogen fixation, leaf area expansion and seed production (Makela et al. 1996).
The effectiveness of exogenous foliar application of GB depends on plant species, developmental stage at the
106

.
time of application, concentration of GB used and the number of applications (Ashraf & Harris 2004). It is
quintessential to determine an optimum GB concentration depending on the type of crop species to achieve the
best possible stress tolerance. For instance, at higher concentrations, GB is more sensitive for the broadleaved
species like bean, tomato and grape than for the cereals. Hence, the proper concentration of GB should be used
with caution, so as to derive the maximum benefit of OA in different plant species (Muhammad et al. 2006,
Wani et al. 2013).
Another recent instance of GB acting as a scavenger of ROS has been reported (Sui et al. 2012). Production
of ROS as a result of wounding is enhanced when yeast antagonists are used as biocontrol agents in the wounds.
It was found that pre-exposure of some yeast strains with GB ameliorated the ROS-mediated oxidative stress in
the antagonistic yeast. The exogenous GB application to the antagonistic yeast strain Candida oleophila
improved their adaptation to apple fruit wounds. The GB-treated yeasts also showed up regulation of major
antioxidant genes like peroxisomal catalase, peroxiredoxin TSA1 and glutathione peroxidase. Thus, such
improved biocontrol efficacy can be achieved via the activation of antioxidant responses in the biocontrol
yeasts.
GB as an important osmoprotectant has been reported in the perennial grasses Holcus lanatus and
Alopecurus pratensis (Gargallo-Garriga et al. 2015). The role of GB in decreasing the level of malondialdehyde
(MDA) to protect the membrane systems has been recently explained in the leaves of Aegiceras corniculatum
and Kandelia obovata exposed to drought stress (Guan et al. 2015). Other positive effects of GB in developing
drought tolerance have also been depicted in tobacco, wheat, barley, sorghum and soybean (Ashraf & Harris
2004). The foliar application of GB to the tomato plants under field condition at a dose of 3.36 Kg ha -1 during
mid-flowering period increased fruit yield to 36% and 39% during salt and heat stress respectively, as compared
to control (Makela et al. 1998). Generally, under normal field conditions, rice has been reported as a GB nonaccumulator. However, some basal levels of GB have been found in rice cultivars like KDML105, Annapurna
and Dongjin, exposed to salinity stress (Wani et al. 2013). A change in the transcript levels of WCOR410 and
catalase in wheat and tomato plants was observed after the exogenous application of GB. WCOR410 is an
acidic dehydrin which improved the freezing tolerance of the tomato plants during cold acclimation. Catalase,
on the other hand, exhibited imparted antioxidant potential by scavenging hydrogen peroxide, a member of the
reactive oxygen species (Allard et al. 1998).
MAJOR TRANSGENIC APPROACHES IN DEVELOPING ABIOTIC STRESS TOLERANCE VIA
INCREASED ACCUMULATION OF GB
Several transgenic approaches have been undertaken to emphasize the alleviation of salinity and drought
stress through accumulation of GB. The most investigated among these transgenics in terms of the
morphological and physiological aspects are those overexpressing the codA gene, which we have mentioned
under the biosynthesis section. These transgenics exhibited stress tolerance at almost all stages of development
along with improved photosynthetic activity and greater yield of fruits and seeds. The transgenic rice plants
showed accumulation of 5.3 mmol g-1 fresh weight (FW), whereas the wild type plants were found to be nonaccumulators of GB (Sakamoto et al. 1998). Maize, being a natural accumulator of GB, exhibited highly
increased accumulation of 5.7 mmol g-1 FW of GB in the transgenic lines (Quan et al. 2004). The codA gene
from Arthrobacter spp. was also overexpressed in the model plant Arabidopsis thaliana, Eucalyptus globulus,
Japanese persimmon (Diospyros kaki), Brassica campestris L. spp. chinensis, Solanum tuberosum and
Lycopersicon esculentum to generate tolerance against salinity, drought, chilling and low relative humidity (Giri
2011). A G-protein named RabAc4 involved in membrane trafficking has been reported for mediating GBmediated chilling tolerance in plants (Wani et al. 2013).
Both GB and Pro have the capability to destabilize the DNA double helix by lowering the helix melting
point in vivo. As a result, under stress conditions, GB acts as an effective activator of replication and
transcription by promoting the melting of DNA helices and thus the stress-responsive genes are easily accessed
by the transcription machinery (Rajendrakumar et al. 1997). Transformation of Arabidopsis by GSMT and
SDMT from Aphanothece halophytica resulted in higher GB accumulation than in the transgenic lines
overexpressing the choline oxidizing enzymes. This obviously indicates that under stress conditions, choline
oxidizing enzymes have limited applications in genetic engineering program, whereas transgenic approaches
involving GSMT and SDMT in combination led to better and higher endogenous GB content. However, such
107

.
high endogenous GB accumulation was not recorded when the plants were exposed to 0.1 M NaCl stress,
supplemented with 5 mM glycine (Waditee et al. 2005). The choline oxidase (COX) gene from Arthrobacter
pascens fused to a chloroplast targeting sequence was expressed in rice under the control of a salt-inducible
promoter and a constitutive ubiquitin promoter. Under salinity stress, the total GB accumulation was obviously
higher in the lines expressing the transgene via the constitutive promoter. However, the inducible lines showed a
swapping 89% increase in the GB content under salt stress, whereas the increase in the constitutively expressed
lines was 44% (Su et al. 2006). Until recent times, it was unknown whether the ortholog of CMO was functional
or a pseudogene in rice was functionally orthologous to BADH. When the CMO from sugarbeet was
overexpressed in the chloroplast of tobacco, the transplastomic lines showed foliar accumulation of 0.2 to 0.5
mmol g-1 FW and the plants were tolerant to oxidative stress (Zhang et al. 2008). The OsCMO gene was isolated
from Oryza sativa L. spp. japonica cv. Nipponbare using RT-PCR. The up regulation of OsCMO by salt stress
was also corroborated by Northern Blot analyses. Transgenic tobacco plants overexpressing OsCMO exhibited
increased GB content with enhanced salt tolerance. Immunoblotting analysis demonstrated that a functional
OsCMO protein with correct size was present in transgenic tobacco, but rarely accumulated in wild-type
rice plants. Surprisingly, a large amount of truncated proteins derived from OsCMO was induced in the rice
seedlings in response to salt stresses. This suggested that it is presumably the non-functionality at the protein
level of OsCMO which hinders the synthesis of GB in rice plants exposed to stress (Luo et al. 2014). On the
other hand, the importance of BADH1 in the accumulation of GB in rice has also been shown (Tang et al. 2014).
The down regulation of OsBADH1 by RNA interference (RNAi) exhibited much lower salinity, drought and
cold tolerance in transgenic rice. The decrease of stress tolerance occurring in the OsBADH1-RNAi repression
lines was associated with an elevated level of malondialdehyde content and hydrogen peroxide. The transgenepositive and transgene-negative lines derived from heterozygous transgenic T 0 plants did not show any
accumulation of GB. Moreover, transgenic OsBADH1-RNAi repression lines showed significantly reduced seed
set and yield. Hence, the down regulation of OsBADH1 resulted in the reduction of ability to dehydrogenate the
accumulating metabolism-derived aldehydes and subsequently gave rise to decreased stress tolerance and crop
productivity, without changing the GB content. The authors suggested that OsBADH1 possessed an enzymatic
activity to catalyze other aldehydes in addition to betaine aldehyde (the precursor of GB) and thus alleviated
their toxic effects under abiotic stresses (Tang et al. 2014).
The heterologous expression of BADH gene from the xerophytic leguminous plant Ammopiptanthus nanus in
E. coli conferred salt and heat tolerance under the stress conditions of 700 mM NaCl and 55 oC temperature (Yu
et al. 2014). This report actually emphasizes the fact that AnBADH is a crucial mediator of abiotic stress
tolerance in A. nanus. Hence, this gene can be engineered in heterologous stress-sensitive cultivars to increase
their stress tolerance. The plant tolerance to various abiotic stresses is improved without any strong phenotypic
changes (Fan et al. 2012). Thus, targeting the GB-synthesizing genes give a feasible correlation of the
transgenic load and the crop yield. This view was justified in transgenic sweet potatoes (Ipomoea batatas cv.
Sushu-2) overexpressing the chloroplastic BADH gene from Spinacia oleracea (SoBADH). The genetically
engineered cultivars exhibited higher accumulation of GB which conferred tolerance against salinity, low
temperature and oxidative stresses. Such tolerance was achieved via increased protection against cell damage by
a steady maintenance of cell membrane integrity, higher photosynthetic activity and elevated rates of ROS
scavenging. The transgenics also showed increased accumulation of Pro with a synchronized up regulation of
ROS-scavenging genes, possibly through some unknown interconnecting cross-talk pathway. This experiment
has also led to the development of a novel germplasm for sweet potato production on marginal lands under
suboptimal conditions (Fan et al. 2012). When the SoBADH was overexpressed in the chloroplastic genome of
carrot, the genetically modified carrots accumulated high levels of GB up to 100 mmol g -1 DW, which conferred
high salinity tolerance to the transgenic lines (Kumar et al. 2004).
The scarcity of endogenous choline and the transport of choline across the chloroplast envelope are the two
most significant hurdles for enhancing GB accumulation in the transgenic plants overexpressing various
enzymes of the GB biosynthetic pathway. The enzymes oxidising choline into GB was targeted in Arabidopsis,
Brassica napus and tobacco (Huang et al. 2000). Exogenous supply of choline in these plants led to a significant
increase in GB content. A model was designed for the labelling kinetics of choline metabolites, which proved
the importance of choline import into chloroplasts in the biosynthesis of GB (McNeil et al. 2000). The activity
of phosphoethanolamine N-methyltransferase (PEAMT) was found to be 30 to 100 times higher in spinach than
108

.
in tobacco (Nuccio et al. 2000). This can also be a reason for the limitation in the endogenous choline supply of
GB non-accumulators, like tobacco. So, it is likely that the targeting of PEAMT can increase the choline supply
and hence promote GB accumulation in the GB non-accumulators.
CONCLUSION AND FUTURE PERSPECTIVES
The present review highlights quite thoroughly the roles and instances of GB, acting as an osmolyte in plants
exposed to multiple abiotic stresses. The greatest advantage in targeting GB-associated genes is that the
phenotypic characters, especially the crop productivity, is least affected. Thus, this approach can be extremely
useful for generating crops under suboptimal climates showing the same production capacity. Recently, the
accumulation of GB was reported in the plant growth-promoting bacteria, isolated from the rhizosphere of the
plantation crops like coconut, cocoa and arecanut exposed to stress (Gupta et al. 2014). It was shown that the
presence of GB, along with other essential anti-stress factors, ultimately enabled the bacteria to survive the
abiotic stress. These obviously indicate that along with its capacity as a potent osmolyte, GB might also be
involved in directly regulating transcriptomic changes during stress and also adjusting the cellular metabolic
homeostasis. Further researches in these directions are required to establish the overall roles of GB in altering
the chromatin architecture for initializing the predicted transcriptomic adjustments under stress (Gupta et al.
2014).
ACKNOWLEDGEMENTS
Financial support from Science and Engineering Research Board (SERB), Government of India through the
research grant (SR/FT/LS-65/2010) and from Council of Scientific and Industrial Research (CSIR), Government
of India through the major grant [38(1387)/14/EMR-II] to Dr. Aryadeep Roychoudhury is gratefully
acknowledged.
REFERENCES
Allard F, Houde FM, Krol M, Ivanov A, Huner NPA & Sarhan F (1998) Betaine improves freezing tolerance in
wheat. Plant and Cell Physiology 39: 11941202.
Ashraf M & Harris PJC (2004) Potential biochemical indicators of salinity tolerance in plants. Plant Science
166: 316.
Basu S, Roychoudhury A, Saha PP & Sengupta DN (2010) Comparative analysis of some biochemical
responses of three indica rice varieties during polyethylene glycol-mediated water stress exhibits distinct
varietal differences. Acta Physiologiae Plantarum 32: 551563.
Bohnert HJ, Nelson DE & Jensen RG (1995) Adaptations to environmental stresses. Plant Cell 7: 10991111.
Chen THH & Murata N (2008) Glycinebetaine: an effective protectant against abiotic stress in plants. Trends in
Plant Science 13: 499505.
Fan W, Zhang M, Zhang H & Zhang P (2012) Improved tolerance to various abiotic stresses in transgenic sweet
potato (Ipomoea batatas) expressing spinach betaine aldehyde dehydrogenase. PLoS ONE 7: e37344.
Gargallo-Garriga A, Sardans J, Prez-Trujillo M, Oravec M, Urban O, Jentsch A, Kreyling J, Beierkuhnlein
C, Parella T & Peuelas J (2015) Warming differentially influences the effects of drought on stoichiometry
and metabolomics in shoots and roots. New Phytologist 207: 591603.
Giri J (2011) Glycinebetaine and abiotic stress tolerance in plants. Plant Signaling & Behavior 6: 17461751.
Guan GF, Wang YS, Cheng H, Jiang ZY & Fei J (2015) Physiological and biochemical response to
drought stress in the leaves of Aegiceras corniculatum and Kandelia obovata. Ecotoxicology 24: 16681676.
Gupta A, Gopal M, Thomas GV, Manikandan V, Gajewski J, Thomas G, Seshagiri S, Schuster SC, Rajesh P &
Gupta R (2014) Whole genome sequencing and analysis of plant growth promoting bacteria isolated from
the rhizosphere of plantation crops coconut, cocoa and arecanut. PLoS One 9: e104259.
Huang J, Hirji R, Adam L, Rozwadowski KL, Hammerlindl JK, Keller WA & Selvaraj G (2000) Genetic
engineering of glycinebetaine production toward enhancing stress tolerance in plants: metabolic limitations.
Plant Physiology 122: 747756.
Jones RGW & Storey R (1981) Betaines. In: Paleg LG & Aspinal D (eds) The physiology and biochemistry of
drought resistance in plants. Academic Press, New York, pp. 171204.
Kumar S, Dhingra A & Daniell H (2004) Plastid-expressed betaine aldehyde dehydrogenase gene in carrot
cultured cells, roots, and leaves confer enhanced salt tolerance. Plant Physiology 136: 28432854.
109

.
Luo D, Niu X, Yu J, Yan J, Gou X, Lu BR & Liu Y (2014) Rice choline monooxygenase (OsCMO) protein
functions in enhancing glycine betaine biosynthesis in transgenic tobacco but does not accumulate in rice
(Oryza sativa L. ssp. japonica). Gene 549: 7784.
Makela P, Jokinen K, Kontturi M, Peltonen-Sainio P, Pehu E & Somersalo S (1998) Foliar application of
glycine betaine - a novel product from sugar beet as an approach to increase tomato yield. Industrial Crops
and Products 7: 139148.
Makela P, Peltonen-Sainio P, Jokinen K, Pehu E, Setala H, Hinkkanen R & Somersalo S (1996) Uptake and
translocation of foliar applied glycine betaine in crop plants. Plant Science 121: 221230.
McNeil SD, Rhodes D, Russell BL, Nuccio ML, Shachar-Hill Y & Hanson AD (2000) Metabolic modeling
identifies key constraints on an engineered glycine betaine synthesis pathway in tobacco. Plant Physiology
124: 153162.
Muhammad I, Ambreen A & Nabeela K (2006) Four foliar application of glycine betaine did not alleviate
advers effects of salt stress on growth of sunflower. Pakistan Journal of Botany 38: 15611570.
Nuccio ML, Ziemak MJ, Henry SA, Weretilnyk EA & Hanson AD (2000) cDNA cloning of
phosphoethanolamine N-Methyltransferase from spinach by complementation in Schizosaccharomyces
pombe and characterization of the recombinant enzyme. The Journal of Biological Chemistry 275: 14095
14101.
Quan R, Shang M, Zhang H, Zhao Y & Zhang J (2004) Improved chilling tolerance by transformation with betA
gene for the enhancement of glycinebetaine synthesis in maize. Plant Science 166: 141149.
Rajendrakumar CS, Suryanarayana T & Reddy AR (1997) DNA helix destabilization by proline and betaine:
possible role in the salinity tolerance process. FEBS Letters 410: 201205.
Rhodes D & Hanson AD (1993) Quaternary ammonium and tertiary sulfonium compounds in higher plants.
Annual Review of Plant Physiology 44: 357384.
Roychoudhury A & Chakraborty M (2013) Biochemical and molecular basis of varietal difference in plant salt
tolerance. Annual Review & Research in Biology 3: 422454.
Roychoudhury A, Banerjee A & Lahiri V (2015) Metabolic and molecular-genetic regulation of proline
signalling and its cross-talk with major effectors mediates abiotic stress tolerance in plants. Turkish Journal
of Botany 39: 887910.
Roychoudhury A, Basu S, Sarkar SN & Sengupta DN (2008) Comparative physiological and molecular
responses of a common aromatic indica rice cultivar to high salinity with non-aromatic indica rice cultivars.
Plant Cell Reports 27: 13951410.
Roychoudhury A, Paul S & Basu S (2013) Cross-talk between abscisic acid-dependent and abscisic acidindependent pathways during abiotic stress. Plant Cell Reports 32: 9851006.
Sakamoto A & Murata N (2002) The role of glycine betaine in the protection of plants from stress: clues from
transgenic plants. Plant Cell & Environment 25: 163171.
Sakamoto A, Alia & Murata N (1998) Metabolic engineering of rice leading to biosynthesis of glycinebetaine
and tolerance to salt and cold. Plant Molecular Biology 38: 10111019.
Smirnoff N & Stewart GR (1985) Stress metabolites and their role in coastal plains. In: Beeftink WG, Rozema J
& Huiskes AHL (eds) Ecology of coastal vegetation. Vegetatio 61&62: 273278.
Su J, Hirji R, Zhang L, He C, Selvaraj G & Wu R (2006) Evaluation of the stress-inducible production of
choline oxidase in transgenic rice as a strategy for producing the stress-protectant glycine betaine. Journal of
Experimental Botany 57: 11291135.
Sui Y, Liu J, Wisniewski M, Droby S, Norelli J & Hershkovitz V (2012) Pretreatment of the yeast antagonist,
Candida oleophila, with glycine betaine increases oxidative stress tolerance in the microenvironment of
apple wounds. International Journal of Food Microbiology 157: 4551.
Takabe T, Rai V & Hibino T (2006) Metabolic engineering of glycinebetaine. In: Rai A & Takabe T (eds)
Abiotic stress tolerance in plants: toward the improvement of global environment and food. Springer,
Dordrecht, Netherlands, pp. 137151.
Tang W, Sun J, Liu J, Liu F, Yan J, Gou X, Lu BR & Liu Y (2014) RNAi-directed down regulation of betaine
aldehyde dehydrogenase 1 (OsBADH1) results in decreased stress tolerance and increased oxidative markers
without affecting glycine betaine biosynthesis in rice (Oryza sativa). Plant Molecular Biology 86: 443454.
110

.
Umezawa T, Fujita M, Yamaguchi-Shinozaki K & Shinozaki K (2006) Engineering drought tolerance in plants:
discovering and tailoring genes to unlock the future. Current Opinion in Biotechnology 17: 113122.
Waditee R, Bhuiyan MNH, Rai V, Aoki K, Tanaka Y, Hibino T, Suzukim S, Takanom J, Jagendorf AT, Takabe
T & Takabe T (2005) Genes for direct methylation of glycine provide highlevels of glycinebetaine and
abiotic-stress tolerance in Synechococcus and Arabidopsis. Proceedings of the National Academy of Sciences
USA 102: 13181323.
Wani SH, Singh NB, Haribhushan A & Mir JI (2013) Compatible solute engineering in plants for abiotic stress
tolerance - role of glycine betaine. Current Genomics 14: 157165.
Yu HQ, Wang YG, Yong TM, She YH, Fu FL & Li WC (2014) Heterologous expression of betaine aldehyde
dehydrogenase gene from Ammopiptanthus nanus confers high salt and heat tolerance to Escherichia coli.
Plant Molecular Biology 86: 443454.
Zhang J, Tan W, Yang XH & Zhang HX (2008) Plastid expressed choline mono-oxygenase gene improved salt
and drought tolerance through accumulation of glycine betaine in tobacco. Plant Cell Reports 27: 1113
1124.
111
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 112119, 2016
Research article
Antibiotic sensitivity of bacterial and fungal isolates from tomato

(Solanum lycopersicum L.) fruit
O. B. Bello1*, I. S. Bello2, D. Aminu3, O. J. Olawuyi4, N. B. Afolabi-Balogun5, A. O.
Lawal1, A. H. Azeez6 and U. Habib7
1
Department of Biological Sciences, Fountain University, Osogbo, Nigeria

Department of Clinical Pharmacy and Pharmacy Practice, University of Ilorin, Nigeria
3
Department of Crop Production, University of Maiduguri, Nigeria
4
Department of Botany, University of Ibadan, Ibadan, Nigeria
5
Department of Chemical Sciences, Fountain University, Osogbo, Nigeria
6
Department of Crop, Soil and Pest Management, Federal University of Technology, Akure, Ondo State, Nigeria
7
Department of Plant Breeding and Genetics, University of Agriculture, Peshawar, Pakistan
*Corresponding Author: obbello2002@yahoo.com
2
Abstract: Decayed ripened tomato fruit contaminated with spores and toxins with relatively heat
resistant could poised food poisoning in humans and animals. This research investigated the effect
of antibiotic sensitivity of fungi and bacteria isolated from tomato (Solanum lycopersicum) fruit in
Osogbo markets, Nigeria. One hundred decayed fruit of tomato were procured from three main
markets (Igbonna, Oja Oba and Sabo) within the metropolis. Fungi and bacteria were cultured on
Sabourand dextrose, MacConkey and Tomato juice agar media. Eight species of bacteria
(Pseudomonas aeruginosa, Bacillus cereus, Bacillus subtilis, Proteus mirabilis, Salmonella typhi,
Escherichia coli, Klebsiella aerogenes and Staphylococcus aureus) and six fungi (Rhizopus
stolonifer, Fusarium spp., Mucor spp., Aspergillus niger, Saccharomyces cerevisiae and
Penicillium spp.) were isolated and characterized. Fungal isolates were highly virulent compared
with bacteria in the decayed tomato fruit. Sabo market had the most prevalence fungi and bacteria
isolates, while Igbonna and the OjaOba markets followed in that trend. Mucor spp. and Bacillus
subtilis exhibited the highest fungal and bacterial counts of 4210 4 cfu g-1 each in the Sabo market.
Chloramphenicol was the most suitable antibiotic for controlling both micro flora. Except B.
subtilis, varied degrees of antibiotic sensitivities and resistances were observed on all the bacteria.
Technological improvement of harvesting, packaging, handling, storage and preservation could
reduce tomato fruit losses and invariably enhance shelf life and quality.
Keywords: Decayed tomato - Bacteria - Fungal isolates - Antibiotics.
[Cite as: Bello OB, Bello IS, Aminu D, Olawuyi OJ, AfolabiBalogun NB, Lawal OA, Azeez AH & Habib U
(2016) Antibiotic sensitivity of bacterial and fungal isolates from tomato (Solanum lycopersicum L.) fruit.
INTRODUCTION
Tomato (Solanum lycopersicum L.) is a berry, annual, shortlived herbaceous plant of the Solanaceae
family. It is usually sprawls on the ground, and could reach about 15 m height (Wogu & Ofuase 2014). It has a
weak woody stem covered with glistering yellow to reddish glandular hairs, rarely vine over other plants. The
leaves are between 10 and 25 cm long with 59 leaflets on the petioles, which are odd and pinnate. Each leaflet
is about 8cm long with serrated margins. Flowers are yellow from 12 cm with fine and pointed lobes on its
corolla (Ijato et al. 2011). The fruit is edible with a smooth epicarp, and varies in shape and size. Immature fruit
is green and becomes yellow or bright red as it ripens (Chinedu & Enya 2014). Tomato plant is cultivated in the
savannah agro-ecological zone of Nigeria during cropping season and dry season under furrow irrigation. The
plant usually produces higher yield and better fruit qualities with minimal foliar diseases under irrigation
compared to those cultivated during the cropping season.
112
Bello et al. (2016) 3(1): 112119

.
Tomato fruit ranks 7th as the most important staple crop worldwide, after wheat, maize, rice, soybeans,
cassava and potatoes, with production estimate of approximately 160 million tonnes, cultivated on 4.8 million
hectares in the year 2011 (FAOSTAT 2011, Ogunbanwo et al. 2014). The fruit is consumed as vegetable,
dietary supplement, eaten raw as salad and for garnishing assorted cooked food or condiment, contributing to a
healthy wellbalanced diet. It is also valuable in the food industries (Fatima et al. 2015, Bello et al. 2016a).
Ripened raw tomato fruit of 100 g constituents are carbohydrate 4 g, energy 75 kg (18 k), dietary fiber 1 g,
sugar 2.6 g, fat 0.2 g, vitamin C (22%, 13 mg), protein 1g, and water 95 g (Ijato et al. 2011). Nutritionally, the
fruit contains calcium, niacin, flavonoids, lycopene, beta-carotene, derivatives of hydroxycinnamic acid, high
amount of water and vitamins, specifically A, C, and E which are very vital in metabolic activities of humans
(Gerszberg et al. 2015). The deep red coloration of the fruit has been attributed to lycopene, a form of
carotenoid pigment with a powerful antioxidant that protects humans against diabetes, cardiovascular diseases
and anti-cancer as well as preventing blood clotting (Wu et al. 2011, Murray 2012, Raiola et al. 2014, Abdul
Hammed et al. 2015).
High pH (4.96.5), water and nutrient contents enhance microbial growth such as bacteria and fungi, which
degrade the nutrients through enzymes production (Trias et al. 2008, Matthew 2011, Ogunbanwo et al. 2014),
and heighten spoilage susceptibility, thereby reducing the nutritional and market values. Contamination of
tomato fruit by microbes is due to poor handling during the production chain, transportation, distribution,
marketing and storage (Akinyele & Akinkunmi 2012). Environmental factors such as temperature, frost and
rainfall constituted adverse effects on quality of the fruit and their storage shelf life (Akinyele & Akinkunmi
2012). Besides the damage to the fruit, microbial infections poise potential health hazards to animals and
humans, as some of the organisms are pathogenic, producing toxins capable of causing diseases such as
diarrhoea, gastroenteritis, respiratory infections and meningitis, if ingested (Barth et al. 2009).
The Centre for Disease Control and Prevention estimated 76 million cases of food borne diseases yearly, and
the etiology is predominantly of microbial origin (Wokoma 2008). Ghosh (2009) reported that fungi were more
virulent than bacteria in tomato fruit spoilage. The main tomato diseases in the forest and savanna ecologies of
Nigeria are Aspergillus niger, Pseudomonas solanacearum, Sclerotium rolfsii and Fusarium oxysporum
(Ogunbanwo et al. 2014). The need for an elaborate study of contaminating pathogens of the tomato fruit
becomes essential, considering tomato fruit as a readytoeat food with minimal processing or eaten raw and
can possess serious threats to food safety (Ofor et al. 2009). Similarly, the high price of fresh ripened tomato
fruit sold in the Nigerian markets is a major concern. A relatively cheaper spoiled fruit consumed by the poor
contain microbial infectious diseases. Based on these, a study was conducted to characterize and determine
antibiotic sensitivity of fungi and bacteria isolated from tomato (Solanum lycopersicum L.) fruit in Osogbo
markets, South Western Nigeria.
MATERIALS AND METHODS
Sample collection
One hundred decayed fruit of tomato were procured from the three main markets of Igbonna, Oja Oba and
Sabo markets in Osogbo, Nigeria. The samples were collected with hand gloves into sterile polythene bags
aseptically and immediately conveyed for analysis in the laboratory.
Isolation, cultural and morphological identification of fungi
A scalpel sterile blade was employed to cut off a segment of the tomato fruit samples of about 35 cm. The
samples were stored in a clean chamber at room temperature for nine days. In the Petri dishes, the emerged
fungi isolated aseptically were placed on the sterilized Potato Dextrose Agar and then incubated at 25 oC for 6
days, as described by Amusa et al. (2002). The related pathogens were isolated using morphological and cultural
characteristics by comparing with the culture that was collected from the seed health pathology laboratory,
International Institute of Tropical Agriculture (IITA), Ibadan, Nigeria, as identified by the International
Mycological Institute, CABI Bioscience, Egham, UK (Amusa et al. 2002).
Isolation and morphological identification of bacteria
The fruit samples were cultured by swabbing the cut interior aseptically and they were plated on 2.7 mL
Tomato juice and 4.5 mL MacConkey broth. The microbial growth detected as turbidity in the broth was
thereafter sub-cultured on Cysteine Lactose Electrolyte Deficient agar (CLED) and incubated at 37 oC for 24
hours (Amusa et al. 2002). Tentative classifications of isolates were conducted by gram staining, Oxidase and
113
Bello et al. (2016) 3(1): 112119

.
motility tests as well as sub-cultures were carried out on CLED. These include deep yellow and opaque colonies
of S. aureus, mucoid yellow to whitish blue colonies of Klebsiella spp., greenish blue or blue colonies of P.
aeruginosa, greenish colour colonies of Proteus spp. and yellow coloured colonies of lactose fermenting E. coli.
(HiMedia manual 2003). Substantiation of bacterial pathogens diversity were carried out by sub-culturing on
Xylose Lysine Deoxycholate agar (XLD agar; M1108, Himedia, Cetrimide Agar for Pseudomonas spp.,
Mannitol salt agar for Staphylococcus aureus and SalmonellaShigellaagar (SS agar M108, Himedia,
Mumbai) for Salmonella, Mumbai). MacConkey agar was also sub-cultured for additional enteric pathogens and
different biochemical tests. Furthermore, the swab samples were emulsified in normal saline (0.85% sodium
chloride), covered and assessed for protozoa.
Biochemical Tests
Biochemical tests such as catalase, sugar, indole and coagulase tests were carried out as described by Holt et
al. (1994). Enzymatic assays for catalase and coagulase as well as tests for constituent sugar and indole were
also carried out, as described by Holt et al. (1994).
Assessment of colony form units (CFU ml-1)
Bacterial strains were earlier preserved on nutrient agar at 4C (Noor et al. 2013). After two hours of growth
in 5 ml of the nutrient broth as preculture, the OD of the culture broth was measured at 600 nm (OD 600) and
thereafter adjusted to 0.1. Thirty (30) L was introduced into three different sets of 30 ml of the nutrient broth
and incubated at 37C. These were then shaken at 0, 100 and 200 rotations per minute (rpm) (Noor et al., 2013).
At 12hour interval, the growths were monitored by measuring OD 600. The colony form units (CFU)/ml were
estimated by counting colonies on nutrient agar at 24 hours (Noor et al. 2013).
Antibiotic sensitivity test
The standardized disc diffusion method was adopted by applying the zone size interpretation chart to
evaluate bacteria sensitivity on the selected antibiotics. The sensitivity tests were carried out following M2A6
disc diffusion method using nutrient agar, as suggested by the National Committee for Clinical Laboratory
Standards (NCCLS 1997). Identified bacteria were cultured overnight with nutrient agar, then suspended in a
sterile physiological saline (0.9%w/v NaCl) to achieve an equivalent of 0.5 McFarland turbidity standards. A
sterile, nontoxic cotton swab was plunged into the standardized innocula and used to spread the entire surface
of Mueller Hinton agar plates (NCCLS 2002). Antibiotic discs (PS003GVE, polytes Laboratory, Enugu,
Nigeria) were position aseptically on the surface of the agar plates using sterilized forceps, and thereafter
incubated at 37oC for 24 hours, while zones of inhibition were measured and classified as either sensitive or
resistant, as described by CLSI (2012). The antibiotics screened were Ampicilin (SamAce Ltd., AkodaEde,
Nigeria, 30g mL-1), Ampiclox (Mecure Industries Ltd., Lagos, Nigeria, 30g mL-1), Ciprofloxacin (Tuyil
Pharmaceutical Industries Ltd., Ilorin, Nigeria, 10g mL-1), Amoxicillin (Michelle Laboratories Ltd., Enugu,
Nigeria, 30g mL-1), Streptomycin (North China Pharmaceutical Co., Ltd., Shijiangzhuang, China, 10g mL-1),
Septrin (Tuyil Pharmaceutical Industries Ltd., Nigeria, 30g mL-1), Augumentin (Tuyil Pharmaceutical
Industries Ltd., Ilorin, Nigeria, 10g mL-1), Chloramphenicol (Maxheal Pharmaceuticals, India, 30g mL-1), and
Gentamycin (Furen Pharmaceutical Group Co., Ltd., Henan, China, 10g mL-1).
Fungal culture
Morphological and cultural features of fungal isolates revealed Fusarium spp., Rhizopus stolonifer,
Aspergillus niger, Saccharomyces cerevisiae, Mucor spp., and Penicillium spp. in the decayed tomato fruit
samples (Table 1). This corroborates with the findings of several researchers (Ghosh 2009) who reported the
presence of Fusarium spp., Penicillium spp. and Aspergillus niger as the prominent isolates in tomato fruit
spoilage. These results also agree with the reports of an independent researcher, Akinmusire (2011) who
reported that Rhizopus stolonifer, Mucor spp. and Fusarium oxysporumas as major spoilage organisms of ripe
tomato fruit from selected markets in Maiduguri, North Eastern Nigeria. On the other hand, Oyemaechi et al.
(2014) isolated Aspergillus phoenicis in Onitsha, Nigeria.
In the decayed tomato samples assessed, fungi isolates were more prevalent than bacteria. However, there
were wide variations in microbial population, with Mucor spp. being predominant (28 isolates), followed by A.
niger (11 isolates) and Penicillium spp. (8 isolates) (Fig. 1). These results corroborate previous studies of
114
Bello et al. (2016) 3(1): 112119

.
Ogunbanwo et al. (2014) and Fatima et al. (2015) who reported that fungi were more prevalent compared to
bacteria in Nigeria. Amongst the three major markets investigated however, Sabo market had the highest fungi
and bacteria isolation rates, while Igbonna and the OjaOba markets followed in that order (Table 2). Mucor
spp. had the highest average fungal value of 42104 in Sabo market compared to other two markets. Wogu &
Ofuase (2014) and Oyemaechi et al. (2014) opined that the storage conditions, poor handling, transportation,
distribution and marketing practices could increase the level of microbial contamination of tomato fruit. These
products are very rich in carbohydrates content and poor in the level of proteins with a pH value ranging from
slightly acidic to neutral, which enhance enabling niche to several pathogenic bacteria and fungi (Trias et al.
2008, Ogunbanwo et al. 2014). The microbial spoilage of tomato fruit has also been obsereved to contain
Aflatoxin which possibly being a source of potential health hazards to humans, as it produces mycotoxins which
induce mycotoxicosis disease (Oyemaechi et al. 2014).
Table 1. Cultural and morphological characters of fungal isolates in tomato fruit samples from three major markets in
Osogbo, Nigeria.
Fungal isolates
Rhizopus stolonifer
Fusarium spp.
Saccharomyces cerevisiae
Penicillium spp.
Mucor spp.
Aspergillus niger
Morphological characteristics
Hyphae were nonseptate and there were
enlarged condiophores at the tip, producing
round vesiclelike chains.
Condiophores produced conidia in clusters
or single forms. Septate hyphae with
canoeshaped macroconidia.
Hyphae were absent, but Saccharomyces
spp. produced ascospores, particularly in
the V8 medium of acetate ascospore agar.
Multilateral
budding
was
usually
rudimentary Pseudohyphae.
From a specialized conidiogeny, chains of
singlecelled conidia (Ameroconidia) were
produced in basipetal succession called a
phialide.
Hyphae were nonseptate and branched.
Long sporangiophores with nonseptate
terminal spore sporangia.
Branched and septatehyphae condiophore

with secondary branches. Enlarged
condiophore at the tip, producing round
vesiclelike chains.
Cultural characteristics
Profuse proliferation of filamentous
condiophores.
At first, the conidia were cottony
and white, and thereafter changed to
pink.
Very fast growing colonies with
moist, flat, smooth, dull, tannish
cream or glistening in color.
Prolific production of colonies with

filamentous, cottony, velvety, flat or
woolly in texture.
Profuse
proliferation
of
sporangiophores that covered agar
surface with white fluff that shortly
turned grey. The reverse side was
white.
Greenish, filamentous condiophore,
with rapid growing of black velvety
spores.
Figure 1. Distribution of fungal isolates in tomato fruit samples from three major markets in Osogbo, Nigeria.
115
Bello et al. (2016) 3(1): 112119

.
Table 2. Mean fungal count of tomato fruits from three major markets in Osogbo, Nigeria.
Isolated fungi
Sabo
Igbonna
Oja Oba
Rhizopus stolonifer
16 104
12 104
11 104
4
4
Penicillium spp.
41 10
31 10
30 104
4
4
Saccharomyces cerevisiae
15 10
10 10
9 104
4
4
Mucor spp.
42 10
40 10
38 104
4
4
Aspergillus niger
35 10
32 10
30 104
4
4
Fusarium spp.
38 10
30 10
23 104
Based on the incidence and characterization of bacterial isolates in tomato fruit samples (Table 3), eight
probable bacteria identified were S. aureus, S. typhi, P. mirabilis, P. aeruginosa, K. aerogenes, E. coli, B.
subtilis and B. cereus. The occurrence and distribution of the bacterial isolates from decayed tomato fruit
samples in the three major markets of Osogbo metropolis (Fig. 2) revealed that out of 61 isolates observed, B.
subtilis was prominent with the highest value of 30 isolates, followed by P. aeruginosa with 8 isolates, while P.
mirabilis and K. aerogenes had one isolate each.
Table 3. Characterization of bacterial isolates in tomato fruit samples from three major markets in Osogbo, Nigeria.
Features
Isolate descriptions
Cultural
Cocci
Rod
Rod
Rod
Rod
Rod
Shape
Smooth
Smooth
Entire
Entire
Entire
Entire
Margin
Yellow
Creamy
White
White
White
Pink
Color
Rod
Smooth
White
Rod
Smooth
White
Morphological
Motility test
Gram reaction
Shape
Cell arrangement
Rod
Single
Rod
Single
Rod
Single
Rod
Single
Rod
Single
+
+
Rod
Single
+
+
Rod
Single
Sugar fermentation test

Lactose
A
Glucose
+
AG
+
AG
Biochemical test
Indole
Oxidase
Catalase
Coagulase
+
+
+
+
Feasible bacteria
+
Cocci
Cluster
Staphylococcus Salmonella Proteus Pseudomonas Klebsiella Escherichia Bacillus Bacillus

aureus
typhi
mirabilis aeruginosa
aerogenes coli
cereus
subtilis
Note: = Negative, + = Positive, AG = Acid and gas production, A = Acid production only.
Figure 2. Distribution of bacterial isolates in tomato fruit samples from three major markets in Osogbo, Nigeria.
116
Bello et al. (2016) 3(1): 112119

.
Regarding the three markets, Sabo market exhibited the highest bacterial and fungal counts, while by
Igbonna and Oja Oba followed in that order (Table 4). B. subtilis was prominent with the value of 42104 cfu g-1
in Sabo market, while the lowest count of 1104 cfu g-1 was obtained for B. cereus. Observations made under
this context signified that high rate of B. subtilis probably due to opportunistic contaminations through poor
handling processes of tomato fruit. The occurrence of S. aureus that is usually associated with faecal matter also
affirmed poor hygiene in the markets. This is consistent with the studies of Oyemaechi et al. (2014), who
suggested that the presence of S. aureus in the decayed tomato fruit possibly due to contamination with organic
manure and/ or faecal matter. It is noteworthy that the observations in this study negate some other researchers
reports outside Nigeria. For instance, Garg et al. (2013) isolated lactic acid bacteria, Vibrio furnissii, Serratia
marcescens and Aeromonas hydrophila in India. The reasons adduced to these are that varied geographical and
seasonal weather as well as inconsistencies in the agronomic practices (cultivation, harvesting, handling and
packaging) involved in tomato production could result to these differences in the microbial isolates.
Table 4. Mean bacterial count of tomato fruit from three major markets in Osogbo, Nigeria.
Isolated bacteria
Pseudomonas aeruginosa
Salmonella typhi
Proteus mirabilis
Klebsiella aerogenes
Bacillus cereus
Bacillus subtilis
Escherichia coli
Staphylococcus aureus
Sabo
21 104
10 104
3 104
25 104
1 104
42 104
13 104
19 104
Markets/ CFU/g
Igbonna
11 104
8 104
Nil
3 104
Nil
34 104
7 104
11 104
Oja Oba
5 104
2 104
Nil
Nil
Nil
15 104
3 104
9 104
With the exception of B. subtilis isolates, the other seven bacteria had heterogeneous degrees of sensitivity
and resistance to antibiotics, similar to the observation of Ghosh (2009) in Nigeria (Table 5). Wogu & Ofuase
(2014) reported that existence of bacteria with multiple antibiotic sensitivities and resistances in the tomato
spoilage indicated high risks and potential hazards on consumption. Bruises and damages inflicted on fruit
Table 5. Patterns of antibiotic sensitivity of bacterial isolates in tomato fruit samples from three major markets in Osogbo,
Nigeria.
Isolated
bacteria
CH
Antibiotics
Total
GE
ST
SE
AU
AX
AN
Markets
A B C A B C A B C A B C A B C A B C A B C A B C S No (%) R No (%)
R R R S S S S R R S S R S R R S S S S S S S S S 17(47)
19 (53)
CP
AM
ABC
Pseudomonas
SRR
aeruginosa
Salmonella
S R R R R R S S S R R R S R R S R R S S S S S S S S S 15 (42) 21 (58)
typhi
Proteus
S S R S S R S S S S S R S S S S S S S S S S S S S S S 24 (67) 12 (33)
mirabilis
Klebsiella
S R R S S S S S S S R R S R R S R R S S S S S S S S S 19 (53) 17(47)
aerogenes
Bacillus
S S R S S R S S S R R S S S R S S R S S S S S S S S S 21 (56) 15 (56)
cereus
Bacillus
S S S S S S S S S S S S S S S S S S S S S S S S S S S 36 (100) 0 (0)
subtilis
Escherichia
S R R S S S S S S S S S S S R S S S S S S S S S S S S 33 (92) 3 (8)
coli
Staphylococcus
R R R S S S S S S S S S S S R R R R S S S S S S S S S 29 (81) 7 (9)
aureus
Note: Antibiotics: CH= Chloramphenicol, CP= Ciprofloxacin, AM= Amoxicillin, GE= Gentamycin, ST=
Streptomycin, SE= Septrin, AU= Augumentin, AX= Ampiclox, AN= Ampicilin; Test results: R= Resistant,
S= Sensitive; Markets: A= Sabo, B= Oja Oba, C= Igbonna.
during harvest and handlings could enhance proliferation of microbes as a vehicle of infections of such damaged
tissue, thereby causing fruit decay. This confirmed the assertion of Matthew (2011) that spoilage microbes often
gain entry into the fruit through wounds. Ghosh (2009) also suggested that the prevalence of microbial
contamination could be aggravated by poor sanitation including crosscontamination with other products in
117
Bello et al. (2016) 3(1): 112119

.
transit. Tomato traders always display ripened tomato fruit in the open, and the heat of sun rays increase the rate
of rotting. Previous researchers suggested that high temperatures may encourage deterioration of tomato fruit
(Matthew 2011), and speed up the physiologic processes, leading to accumulation and suboxidation of
metabolic byproducts (Fatima et al. 2015). A range of temperatures between 7.2oC and 10oC were
recommended for the storage of ripened tomato fruit, while a range of 12.821.1 oC is appropriate for matured
green fruit. Matthew (2011) also observed that supply of tomatoes all year round may not be attainable through
production alone, but an integrated approach of preservation and storage of excess at harvest could improve the
shelf life. The quality of vegetables and fruit can only be maintained after harvest; thus it is absolutely
imperative to harvest promptly especially at the peak quality period (Bello et al. 2016b). This is because
overripe or immature fruit may have short shelf life in storage compared with those picked at appropriate
maturity levels (Eni et al. 2010).
CONCLUSION
Fungi isolates were more prevalent than bacteria in the decayed tomato samples. Sabo market had the most
prevalence of both fungi and bacteria isolates, while Igbonna and the OjaOba markets followed in that order.
Mucor spp. exhibited the highest average fungal value in the Sabo market. Chloramphenicol was the most
suitable antibiotic for controlling both microflora. Except B. subtilis, varied degrees of antibiotic sensitivities
and resistances were observed on all the bacteria. Technological improvement of harvesting, packaging,
handling, storage and preservation could reduce tomato fruit losses and invariably enhance shelf life and quality.
REFERENCES
AbdulHammed M, Bello OS, Azeez MA & Adedeji NO (2015) Bioformation of carotenoids in tomatoes
(Solanum lycopersicum) under two ripening conditions: A Kinetic study. International Journal of Science
and Engineering Resources 6(8): 19.
Akinmusire OO (2011) Fungal species associated with the spoilage of some edible fruits in Maiduguri, Northern
Eastern Nigeria. Advance Environmental Biology 5(1): 157161.
Akinyele BJ & Akinkunmi CO (2012) Fungi associated with the spoilage of berry and their reaction to magnetic
field. Journal of Yeast and Fungal Resources 3(4): 4957.
Amusa NA, Kehinde IA & Ashaye OA (2002) Biodeterioration of bread fruit in storage and its effects on the
nutrient composition. African Journal of Biotechnology 1(2): 5760.
Barth M, Hankinson TR, Zhuang H & Breidt F (2009) Microbiological spoilage of fruits and vegetables. In:
W.H. Sperber, M.P. Doyle (eds.), Compendium of the Microbiological Spoilage of Foods and Beverages,
Food Microbiology and Food Safety. C Springer Science Business Media, LLC, pp. 135183.
Bello OB, Habib U, Olawuyi OJ, Opeyemi AS, Alafe AH & Owoade TA (2016a) Microorganisms causing postharvest tomato (Solanum lycopersicum L.) fruit decay in Nigeria. Journal of Entomology and Zoology
Studies 4(1): 374377.
Bello OB, Olawuyi OJ, Opeyemi AS, Alafe AH & Oladimeji DT (2016b) Genotypic pathogenic microbes
associated with deterioration of sweet orange (Citrus sinensis L) fruit in Nigeria. Scientia Agriculturae
13(1): 1922.
Chinedu SM & Enya E (2014) Isolation of Microorganisms associated with deterioration of tomato (Solanum
lycopersicum) and pawpaw (Carica papaya) fruits. International Journal Curriculum Microbiolology and
Applied Sciences 3(5): 501512.
CLSI (2012) Performance standards for antimicrobial susceptibility testing. Clinical and Laboratory Standards
Institute 4(1): 4555, Wayne, PA. USA.
Eni AO, Ibokunoluwa O & Oranusi U (2010) Microbial quality of fruits and vegetables. African Journal of
Food Sciences 4(5): 291296.
Fatima D, Mebrouk K & Miloud H (2015) Biopreservation of tomato paste and sauce with Leuconostoc spp.
metabolites. African Journal of Food Sciences 9(6): 359366.
FAOSTAT (2011) http://faostat3.fao.org/faostat-gateway/go/to/down load/Q/QC/E
Garg RK, Batav N, Silawat N & Singh RK (2013) Isolation and identification of pathogenic microbes from
tomato puree and their delineation of distinctness by molecular techniques. Journal of Applied Biology and
Biotechnology 1(4): 2431.
Gerszberg A, Hnatuszko-Konka K, Kowalczyk T & Kononowicz AK (2015) Tomato (Solanum lycopersicum
L.) in the service of biotechnology. Plant Cell Tissue and Organ Culture 120: 881902.
Ghosh A (2009) Identification of microorganisms responsible for spoilage of tomato (Lycopersicum esculentum)
fruit. Journal of Phytology 1(6): 414416.
118
Bello et al. (2016) 3(1): 112119

.
HiMedia manual (2003) HiMedia manual for Microbiology and Cell Culture Laboratory Practices. Hi
media Laboratories, Pvt. Ltd, Mumbai, pp. 69.
Holt JG, Krieg NR, Sneath PHA, Stanley JT & Williams ST (1994) Bergeys Manual of Determinative
Bacteriology, 9th Edn., Williams and Wilkins, Baltimore, pp. 4446.
Ijato JY, Adebiyi AO & Ijadunola JA (2011) Antifungal effects of four tropical plant aqueous and ethanol
extracts on post-harvest rot of tomato (Lycopersicum esculentum) in AdoEkiti, Nigeria. New York Science
Journal, 4(1): 6468.
Matthew T (2011) Post harvest microbial deterioration of tomato (Lycopersicon esculentum) fruits. Report and
Opinion 3(4): 5257.
NCCLS (1997) Performance standards for Antimicrobial susceptibility Tests. 6th edn. Document M2A6.
National Committee for Clinical Laboratory Standards, Wayne, Pa.
NCCLS (2002) Performance Standards for antimicrobial susceptibility testing. 8th Informational Supplement.
M100 S12. National Committee for Clinical Laboratory Standards, Villanova.
Noor R, Islam Z, Munshi SH & Rahman F (2013) Influence of temperature on Escherichia coli growth in
different culture media. Journal of Pure and Applied Microbiology 7(2): 899904.
Ofor MO, Okorie VC, Ibeawuchi II, Ihejirika GO, Obilo OP & Dialoke SA (2009) Microbial contaminants in
fresh tomato wash water and food safety considerations in SouthEastern Nigeria. Life Science Journal 6(3):
8082.
Ogunbanwo ST, Fadahunsi IF & Molokwu AJ (2014) Thermal stability of lactic acid bacteria metabolites and
its application in preservation of tomato pastes. Malaysian Journal of Microbiology 10(1): 1523.
Oyemaechi, CU, Chukwuezi, FO & Ozougwu VEO (2014) Microbial agents of tomato spoilage in Onitsha
metropolis. Advance Biological Research 8(2): 8793.
Raiola A, Rigano MM, Calafiore R, Frusciante L & Barone A (2014) Enhancing the human-promoting effects
of tomato fruit for biofortified food. Mediators of Inflammation 2014: Article ID 139873.
Trias R, Baeras L, Montesinos E & Badosa E (2008) Lactic acid bacteria from fresh fruit and vegetables as
biocontrol agents of phytopathogenic bacteria and fungi. International Journal of Microbiology 11(4): 231
236.
Wogu M & Ofuase O (2014) Microorganisms responsible for the spoilage of tomato fruits, Lycopersicum
esculentum, sold in markets in Benin City, Southern Nigeria. Scholars Academic Journal of Biosciences
2(7): 459466.
Wokoma ECW (2008) Preliminary report on disease of tomatoes in Choba, Rivers State. Journal of Applied
Sciences and Environment 12(3): 178121.
Wu Z, Sun S, Wang F & Guo D (2011) Establishment of regeneration and transformation system of
Lycopersicum esculentum Microtom. British Biotechnology Journal 3: 5360. www.sciencedomain.org/
download.php ?f=13 11487212-Guo.pdf.
119
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 120130, 2016
Research article
Inventory of invasive alien plants in Bethuadahari wildlife

sanctuary in Nadia district, West Bengal, India
Dibyendu Talukdar1* and Tulika Talukdar2
1
Department of Botany, R.P.M. College, University of Calcutta, Uttarpara 712258, WB, India
Department of Botany, APC ROY Govt. College, University of North Bengal, Siliguri, Darjeeling, WB, India
*Corresponding Author: dibyendutalukdar9@gmail.com
2
Abstract: A survey was performed in Bethadahari wildlife sanctuary, Nadia district, West
Bengal during the span of 2010-2015 to invent the invasive alien plant species in 121 hectare
forest area and adjoining five villages. Study revealed occurrence of 103 alien angiosperm plant
species under 32 families, with four monocot families (Araceae, Poaceae, Cyperaceae,
Pontederiaceae). Fabaceae leads with 20 taxa and followed by Asteraceae with 17 plant species.
Amaranthaceae, Solanaceae, and Euphorbiaceae were some other important families which
possessed eight, seven, and five taxa, respectively. Year-wise quadrat studies revealed increasing
number of alien species in the study area. Parthenium hysterophorus, Ageratum conyzoides, A.
haustonianum, Eupatorium odoratum, Chromolaena odorata of Asteraceae; Cassia sophera and
Leucaena leucocephala of Leguminosae; Amaranthus spinosus and Alternanthera sessilis of
Amaranthaceae; Lantana camara of Verbenaceae and Trema orientralis of family Urticaceae were
the major invasive species. Remarkably, several alien species have been used in diverse economic
purposes by villagers, showing use of nearly 49% plants in local health-care systems.
Keywords: Invasive alien plants - Biodiversity - Bethuadahari wildlife sanctuary - Folk use.
[Cite as: Talukdar D & Talukdar T (2016) Inventory of invasive alien plants in Bethuadahari wildlife sanctuary
in Nadia district, West Bengal, India. Tropical Plant Research 3(1): 120130]
INTRODUCTION
Invasive alien species have potential to spread and established themself outside their native ranges which
affect the native natural ecosystems or local human-mediated systems (Mooney & Hobbs 2000, Lockwood et al.
2007). Biological invasions by alien taxa are the second worst threat to native ecosystem and become a recurrent
cost for agriculture and forestry. Due to rapid increase and extent of invasive species, diversity of worlds flora
and fauna is becoming homogenized (Lockwood et al. 2007) and is recognized as a primary concern for loss of
biodiversity (Sax et al. 2002, Davis 2003). Yet, a large number of invasive and alien plant species are regularly
used for wood fuel, sheltering, fishing, medicinal, and other purposes (Singh et al. 2010, Talukdar & Talukdar
2012a, b, 2013).
At least 10% of the worlds vascular plants (~3,00,000) can invade native ecosystems and its flora and fauna
in direct and indirect ways (Raghubanshi et al. 2005). About 40% of the Indian flora is alien and 25% of which
are invasive alien species predominantly of neotropic origin (Raghubanshi et al. 2005, Reddy 2008, Mandal
2011). As India possesses rich biodiversity, reports on invasive and alien taxa may pave the way for
comprehensive regional and national data base for effective management and utilization of exotic floras
(Srivastava & Singh 2009, Rastogi et al. 2015).
The state of West Bengal is located between 8550 and 8950 E and 2138 and 2710 N, and one of
populous as well as biodiversity rich states of India. The lower Indo-Gangetic basin constitutes fertile land for
diverse types of flora and fauna, introduced by anthropogenic activities since time immemorial. The Nadia
district (situated between 225230 and 400540 N latitude and 880810 and 884815E longitude) is an
important part of this basin, possessing forested areas, wetlands, and agricultural lands. Bhagirathi is the major
river and with Jalangi, Churni, Ichhamati and some small rivers constitute the riverine and floodplain (Baor)
systems in this district. Among the 14 wildlife sanctuary in West Bengal, Bethuadahari wildlife sanctuary is
120
Talukdar & Talukdar (2016) 3(1): 120130

.
situated in this district. It is 122 Km from state capital Kolkata and represents middle part of lower Gangetic
basin. The tropic of cancer passes through this district. The climate of the district is tropical monsoon with three
distinct seasons, summer (Marchearly June), rainy (JuneSeptember) and winter (OctoberFebruary), and
mean annual rainfall ca. 1800 mm. While maximum summer temperature may soar to 43C, winter is extremely
chilled with temperature may plummet to 23 C. The sanctuary is very rich in biodiversity and famous for
spotted deer. As biological invasions are frequently influenced by ecosystem functioning (Sausa et al. 2011),
climate change (Thuiller et al. 2007, Bradley et al. 2009, Biswas et al. 2014), environmental pollution (Crooks
et al. 2011), and other physico-chemical mechanisms, a proper first hand inventory in protected areas is
absolutely essential to measure threats imposed by alien species on indigenous resources.
Although one report is available regarding the floral diversity of Bethuadahari reserve forest (Das & Lahiri
1990), no investigation was carried out to document the invasive alien plants in this forest area. This sanctuary
is 121 hectare man-made deer sanctuary, situated close to National Highway (NH) 34, linking state capital
Kolkata with North Bengal and Bhutan. In recent years, widening of NH 34 accompanied by vehicle and rail
transport and constructions of inhabitants in and around the forest has greatly impacted the forest ecosystem.
The sanctuary has a deer park, a wetland inside and is an attractive destination of migratory birds during winter.
As invasive species has huge ecological impacts and preference over native species in forest ecology,
documentation of alien plants in this important sanctuary is necessary. The objectives of the present study are,
thus, to make an inventory of the alien flora, their classification and use by local people in and around the
sanctuary.
Study site
The present investigation was carried out by extensive field survey during the last six years (20102015) in
different intervals (MarchJune, SeptemberJanuary) in sanctuary area of 121 hectares and 5 villages adjacent it
(Fig. 1).
Figure 1. A map of study areas (red dots) in the position of Nadia district, West Bengal, India.
Collection of data and methods of inventory

Plant samples were collected either in flowering or in fruiting stage, and voucher specimens were deposited
in departmental herbaria, R.P.M. College, Uttarpara, Hooghly. Invasiveness of the alien species, enlisted by
previous works (Lowe et al. 2000, Huang et al. 2009), was studied using techniques of Baider & Florens (2011).
121

.
Accordingly, a combination of random walks through the area along with a more quantitative sampling of the
seedlings and larger woody plants (flowering or fruiting stage) in a series of square quadrats (1 1 m for
seedlings and 10 10 m for tree) was followed. Frequency (F) of particular plant species was calculated by
dividing the number of quadrats in which a particular species occur with total number of quadrats laid down.
The identity of specimens was verified with existing literatures, monographs, and was also confirmed by IPNI
(International Plant Names Index) data base (www.IPNI.org). Economic use of different flora was investigated
through interviews of knowledgeable people like village elders, medicinemen, farmers, teachers, etc. Collected
information was critically cross-checked by structured questionnaires, and documented thereafter. Nativity of
the species was documented from the existing literatures (Rao & Murugan 2006) and the works done in the
region (Das & Lahiri 1990).
RESULTS
Documentation and classification of alien taxa
Present inventorization of the alien invasive flora in the Bethuadohori wildlife sanctuary pointed out
presence of 103 species which is belonging to 83 genera under 32 families (Table 1). Regarding plant growth
type, 85% of total documented plant species were herbs, and it was followed by shrub (9%), tree (4%) and
climbers (2%). Several genera were found to possess three or more species (Table 1). Major proportion (94%) of
alien flora is dicotyledonous which is grouped under 96 species and 28 families (Table 1). It was followed by
Monocotyledons (6%) distributed in seven genera of four families. Among the total 32 families, Fabaceae
dominated with 20 species, followed by Asteraceae (17 species), Amaranthaceae (8), Solanaceae (7) and then
others (Table 1; Fig. 2).
Table 1. Invasive alien plant species in Bethuadohori Wildlife Sanctuary at Nadia district of West Bengal, India
S.No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
Species
Aerva javanica (Burm. f.) Juss.
ex Schult
Achyranthes aspera L.
Aeschynomene americana L.
Ageratum conyzoides L.
Ageratum houstonianum Mill.
Alternanthera philoxeroides
(Mart.) Griseb.
Alternanthera pungens Kunth
Alternanthera sessilis (L.) R.Br.
ex DC
Amaranthus spinosus L.
Argemone mexicana L.
Bidens pilosa L.
Blumea lacera (Burm. f.) DC.
Boerhaavia erecta L.
Calotropis gigantea (L.) R.Br.
Calotropis procera (L.) R.Br.
Cassia alata L.
Cassia javanica L.
Cassia occidentalis L.
Cassia sophera L.
Catharanthus pusillus
(Murray)Don
*Chromolaena odorata (L.) King
& Robinson
Chrozophora rottleri (Geis.)
Spreng.
Chenopodium album L.
Cleome gynandra L.
Cleome monophylla L.
Cleome rutidosperma DC.
Coix lacryma-jobi L.
Crotalaria pallida Dryand
Family
Amaranthaceae
Life form
Herb
Nativity
Trop. America
Use
M
Amaranthaceae
Fabaceae
Asteraceae
Asteraceae
Amaranthaceae
Herb
Herb
Herb
Herb
Herb
Trop. America
Trop. America
Trop. America
Trop. America
Trop. America
M
Co, shola
NU
NU
Veg
Amaranthaceae
Amaranthaceae
Herb
Herb
Trop. America
Trop. America
M
M, Veg
Amaranthaceae
Papaveraceae
Asteraceae
Asteraceae
Nyctaginaceae
Asclepiadaceae
Asclepiadaceae
Fabaceae
Fabaceae
Fabaceae
Fabaceae
Apocynaceae
Herb
Herb
Herb
Herb
Herb
Shrub
Shrub
Shrub
Tree
Herb
Herb
Herb
Trop. America
Trop. America
Trop. America
Trop. America
Trop. America
Trop. Africa
Trop. Africa
West Indies
S.E. Asia
Trop. S. America
Trop. S. America
Trop. America
M
NU
M
M, veg
M, veg, Cf
M
M
M, Thatching
Or, M
M, Bf
M, Bf
M
Asteraceae
Herb
Trop. America
NU
Euphorbiaceae
Herb
Trop. Africa
Chenopodiaceae
Cleomaceae
Cleomaceae
Cleomaceae
Poaceae
Fabaceae
Herb
Herb
Herb
Herb
Herb
Herb
Europe
Trop. America
Trop. America
Trop. America
S. E. Asia
Trop. America
Veg, Cf
M
M
M
Pearl, fishing
Bf
122
29
30
31
Crotalaria retusa L.
Croton bonplandianum Boil.
Cryptostegia grandiflora R.Br.
Fabaceae
Euphorbiaceae
Asclepiadaceae
32
33
34
35
36
37
38
39
40
41
Cuscutaceae
Cuscutaceae
Cyperaceae
Fabaceae
Solanaceae
Solanaceae
Rubiaceae
Amaranthaceae
Verbenaceae
Poaceae
Asteraceae
Asteraceae
Pontederiaceae
Asteraceae
Euphorbiaceae
Euphorbiaceae
Convolvulaceae
Asteraceae
Asteraceae
Herb
Herb
Aq. Herb
Herb
Herb
Herb
Herb
Herb
Herb
Europe
Trop. America
Trop. America
Europe
Trop. America
Trop. America
Trop. America
Trop. America
Trop. America
NU
M
NU
NU
Cf
Cf
Cf
NU
NU
51
52
53
54
55
56
57
58
59
Cuscuta chinensis Lam.

Cuscuta reflexa Roxb
Cyperus rotundus L.
Cytisus scoparius (L.) Link
Datura innoxia Mill.
Datura metel L.
Dentella repens (L.) Forst
Digera muricata (L.) Mart.
Duranta repens L.
Echinochloa crusgalli (L.)
P.Beauv.
Echinacea paradoxa Britton
Eclipta prostrata (L.) Mant.
*Eichhornia crassipes Kunth
*Eupatorium odoratum L.
Euphorbia hirta L.
Euphorbia heterophylla L.
Evolvulus nummularius (L.) L.
Gnaphalium coarctatum Willd.
Gnaphalium pensylvanicum
Willd.
Gomphrena serrata L.
Hyptis suaveolens (L.) Poit.
Impatiens balsamina L.
Indigofera astragalina DC.
Indigofera linifolia (L. f.) Retz.
Ipomoea quamoclit L.
Ipomoea aquatica Forsk
*Lantana camara L.
Lathyrus aphaca L.

.
Herb
Trop. America
Bf
Herb
Temp.S. America
M
Woody
Trop. Africa
M
Climber
(Madagascar)
Herb
Mediterranean
NU
Herb
Mediterranean
NU
Herb
Africa, S. Europe
M
Herb
Europe
M
Shrub
Trop. America
M
Shrub
Trop. America
M
Herb
E. Asia, Australia Veg
Herb
S. W. Asia
Veg
Shrub
Trop. America
Or
Herb
Trop. S. America
M
Amaranthaceae
Lamiaceae
Balsaminaceae
Fabaceae
Fabaceae
Convolvulaceae
Convolvulaceae
Verbenaceae
Fabaceae
Herb
Herb
Herb
Herb
Herb
Herb
Aquatic
Herb
Herb
Trop. America
Trop. America
Trop. America
Trop. America
Trop. America
Trop. America
Trop. America
Trop. America
Mediterranean
Or
Aromatic
Or
Cloth washing
NU
Bf
M, veg
Bf
M, Cf,
mulching
60
Lathyrus sativus L.
Fabaceae
Herb
Mediterranean
61
62
Leonotis nepetiifolia (L.) R.Br.

*Leucaena leucocephala (Lam.)
de Wit
Ludwigia perennis L.
Malachra capitata (L.) L.
Mecardonia procumbens (Mill.)
Small
Melilotus alba Desv.
*Mikania micrantha Kunth
Mimosa pudica L.
Monochoria vaginalis (Burm.f.)
C. Presl.
Nicotiana plumbaginifolia Viv.
Ocimum basilicum L
*Opuntia stricta (Haw.) Haw.
Oxalis corniculata (DC.)
Raeusch.
Parthenium hysterophorus L.
Pennisetum purpureum Schum.
Peperomia pellucida (L.) Kunth
Peristrophe paniculata (Forssk.)
Brummitt
Lamiaceae
Fabaceae
Herb
Tree
Trop. Africa
Trop. America
Onagraceae
Malvaceae
Scrophulariaceae
Herb
Herb
Herb
Trop. America
Trop. America
Trop. N. America
Pulse, Fd,
besan, Cf, veg
M
Bf, basket
making,
M
M
NU
Fabaceae
Asteraceae
Fabaceae
Pontederiaceae
Europe
Trop. America
Trop. S. America
Trop. America
Insecticide
NU
M
M
Solanaceae
Lamiaceae
Cactaceae
Oxalidaceae
Herb
Climber
Herb
Aquatic
herb
Herb
Herb
Herb
Herb
Trop. America
Trop. America
Trop. America
Europe
NU
M
NU
M
Asteraceae
Poaceae
Piperaceae
Acanthaceae
Herb
Herb
Herb
Herb
Trop. N. America
Trop. N. America
Trop. America
Trop. America
NU
Cf
Folk play
NU
42
43
44
45
46
47
48
49
50
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
123
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
Phaseolus aureus L.
Phyllanthus fraternus Webster
Physalis angulata L.
Pilea microphylla (L.) Liebm.
Pistia stratiotes L.
Polygonum barbatum L.
Polygonum hydropiper L.
Portulaca oleracea L.
Prosopis juliflora (Sw.) DC.
Ruellia tuberosa L.
Scoparia dulcis L.
Sesbania grandiflora (L.) Pers.
Sida acuta Burm.f.
Solanum torvum Sw.
Solanum xanthocarpum Schrad.
& H. Wendl.
Solanum nigrum L.
Sonchus oleraceus L.
Spilanthes radicans Jacq.
Tephrosia purpurea (L.) Pers.
Torenia fournieri Linden ex E.
Fournier
Trema orientralis (L.) Blume
Fabaceae
Euphorbiaceae
Solanaceae
Urticaceae
Araceae
Polygonaceae
Polygonaceae
Portulacaceae
Fabaceae
Acanthaceae
Scrophulariaceae
Fabaceae
Malvaceae
Solanaceae
Solanaceae

.
Herb
Trop. America
Fd, Cf, veg
Herb
Trop. America
M
Herb
Trop. America
Folk play
Herb
Trop. America
NU
Herb
Trop. America
M
Herb
S. E. Asia
M
Herb
S. E. Asia
M
Herb
Trop. S. America
Or
Tree
Trop.S. America
Wood works
Herb
Trop. America
Or
Herb
Trop. America
M
Shrub
Trop. America
Bf, veg, M, Or
Herb
Trop. America
M
Shrub
Trop. America
M
Shrub
Trop. America
NU
Solanaceae
Asteraceae
Asteraceae
Fabaceae
Scrophulariaceae
Herb
Herb
Herb
Herb
Herb
Trop. America
Mediterranean
Trop. America
Trop. America
Australia
M
NU
M
M
NU
Ulmaceae
Tree
S. E. Asia
Tridax procumbens L.
Urena lobata L.
Wedelia chinensis (Osbeck)
Merr.
Vernonia cinera L.
Vigna sublobata (L.) Wilczek
Asteraceae
Malvaceae
Asteraceae
Herb
Herb
Herb
Trop. America
Trop. America
S. E. Asia
M, Bf, fishing,
thatching
M
M
M
Asteraceae
Fabaceae
Herb
Herb
Temperate America NU
S. E. Asia
Pulse, bori,
besan, Cf
Note: *-enlisted in database of worlds 100 worst invasive; M-medicinal, Co-compost, Or -ornamental, Bf-biomass
fuel, Cf-cattle feed, Fd-Food, Veg-Vegetables, NU-not in use.
Figure 2. Distribution of alien plant species in different angiopsperm families; A-Family Fabaceae, B-Asteraceae, CAmaranthaceae, D-Solanaceae, E-Euphorbiaceae, F-Malvaceae / Scrophulariaceae / Lamiaceae / Convolvulaceae / Asclrpia-daceae / Poaceae (three species each), G-Cuscutaceae / Verbenaceae / Pontederiaceae / Acanthaceae / Polygonaceae (two
species each), H-rest of the 16 families with one species each, as mentioned in table 1.
Habitat distribution
About 38% of invasive species identified in the present study were most abundant in roadside (NH 34 and
rural roads) close to forest. On the other hand 32% taxa were reported from forest area and 20% and 10% plant
species preferred to grow in cultivated fields and banks of water bodies respectively. Ecological studies of last
six years exposed the high frequency of Parthenium hysterophorus, Eupatorium odoratum, Ageratum
conyzoides, A. houstonianum, Chromolaena odorata along the roadside than the interior of the forest (Fig. 3).
124

.
The ratio of number of plants (cumulative of 400 quadrats/year in six years) between forest area and roadside
varied between 0.530.88 for these five aster members, while it was near to 1.0 for Cassia sophera (0.98), >1.0
for Leucaena leucocephala, Trema orientalis, Alternanthera sessilis, and Amaranthus spinosus, and was 2.15
for Lantana camara (Fig. 3). While, Tridax procumbens, Eclipta prostrata and Wedelia chinensis exhibited
higher frequency in interior of the forest (F=8086%) than the roadside (F=6572%). Within the forest,
members of Amranthaceae (Achyranthes aspera, Alternanthera philoxeroides, Alternanthera sessilis and
Amaranthus spinosus) dominated intermingling with numbers of legumes (Aeschynomene americana, Leucaena
leucocephala and Mimosa pudica) different species of Crotalaria, Cassia, and species of other families in
different magnitudes. The frequency of Parthenium has been found to be reducing interestingly, in plots where
species of family Amaranthaceae dominated. The tree Trema orientralis has been found flourishing in the forest
area better than the roadside (Fig. 3). Members of family Polygonaceae, Araceae, Cyperaceae, Pontederiaceae
preferred wetland areas, while Solanaceae, Euphorbiaceae, Malvaceae, Cactaceae, Convolvulaceae and
Asclepiadaceae were more frequent in dry land.
Figure 3. Forest/roadside ratio of number of plants for eleven alien taxa.
Documentation of spreading of alien flora over the last six years (20102015) pointed out a sharp increase of
Ageratum, Chromolaena, Parthenium, Eupatorium, Leucaena, Cassia, Amaranthus, Alternanthera, Lantana,
and Trema species (Fig. 4).
Figure 4. Frequency % of 11 invasive plants as recorded from six consecutive years (2010-2015) with 400 square quadrats
laid down/year, A- Parthenium hysterophorus L., B- Eupatorium odoratum L., C- Ageratum conyzoides L., D- Ageratum
houstonianum Mill., E- Chromolaena odorata (L.) King & Robinson, F- Leucaena leucocephala (Lam.) de Wit, G-Cassia
sophera L., H- Alternanthera sessilis (L.) R.Br. ex DC, I- Lantana camara L., J- Trema orientralis (Linn.) Blume, and KAmaranthus spinosus L.
125

.
Resource utilization of alien plants by local people
Local population in the present study area used the documented 103 plant species as food, fodder,
ornamental, medicinal, religious, and other commercial purposes (Fig. 5). Fabaceae dominated as the most
valuable source of food, fodder, manure, fuel, folk play and different other objectives (Table 1). According to
local people, beans (Phaseolus spp), mung (Vigna spp), khesari (Lathyrus sativus L.) and jangli matar (Lathyrus
aphaca L.) have been used considerably in their daily life. Seed flour has been used as food supplement,
adulterant, the whole plant as fodder, soil fertilizer (mulching), and tender pods as vegetables and cattle feeds.
About 49% of total alien plants were used as medicinal purposes, while 11.6% plants were utilized as food and
9.7% used as cattle feed. Among the small-scale cottage industries, preparation of beads on string using seeds of
Coix (Poaceae) and commercial shola using Aeschynomene americana (Fabaceae) were regular jobs for local
people. Another financially viable activity within the sanctuary area was different types of wood works, for
which Prosopis julifera (Fabaceae) tree was mainly used. Along with these, different plant parts have been used
as insecticide and aromatic purposes (Table 1; Fig. 5).
Figure 5. Utilization of resources of alien plants by local people; others include folk play, aromatic, insecticide.
Nativity of documented alien flora

Tropical America contributed nearly 60% plants, followed by share of South-East Asia, Europe and tropical
South America, tropical Africa, the Mediterranean and other regions (Fig. 6).
Figure 6. Nativity of 103 alien taxa in 12 world geographic regions with lions share is from tropical (Trop) America, SSouth, E-east, N-North, W-West.
126

.
DISCUSSION
The present investigation reported that the floral diversity of Bethuadahari Wildlife Sanctuary consist
57.11% invasive species. However, rest of the flora was made up of native species. The results indicated
dominance of alien flora in the forest areas. Herbs constituted major portions (85%) of this alien species and
except only seven species, all others belong to 28 different dicot families. The result exhibited dramatic increase
in number of herbaceous flora from the documentation of about 49.2% herbs in this forest area during 1990 (Das
& Lahiri 1990), indicating introduction of more alien species in the forest areas in recent times. The presence of
higher number of leguminous plants is also supported by earlier reports from Sub-Himalayan North Bengal
(Talukdar 2013e, Talukdar & Talukdar 2012a, b), and other areas of Himalaya (Joshi 2002, Bajpai et al. 2015).
Although Fabaceae contained largest number of alien species in the present study, the dominance of invasive
alien species of Asteraceae was also found in some other regions of India (Rao & Murugan 2006) and other
countries (Heywood 1989, Huang et al. 2009, Feng & Zhu 2010). The higher frequency of some asters along the
roadside than the interior of the forest indicated uneven distribution of Asteraceae weeds in the present study
area, resulting in forest/roadside ratio in number of plants below 1.0 for the family. Ecological study revealed
that in contrast to asters, species of Amaranthaceae and Fabaceae were more evenly distributed. Larger number
of Trema orientralis and Lantana camara in forest wasteland indicated aggressiveness and invasion potential of
these taxa in nutrient-deficient areas in expense of growth of native taxa, leading to forest/roadside ratio for
these two species over 1.0. The result is in partial agreement with earlier record, showing occurrence of Lantana
camara, but not Trema orientalis in this reserve forest area during 1990 (Das & Lahiri 1990). This suggested
introduction of Trema after 1990 in the study area, probably to meet the growing demand of fuel plants by local
people. Besides abiotic and biotic constrains, relative degree of disturbances in habitats may profoundly affect
physical environment, which can create permissive conditions for introduction and gradual establishment of
alien species to invade native systems in this forest area (Huang et al. 2009, Singh et al. 2010). Habitat
disturbances in the present study areas may be one of the prime reasons for rapid establishments of some of the
worst invasive species in the present study area.
Reduction in frequency of one of the worlds worst noxious weeds, Parthenium hysterophorus in plots
where members of family Amaranthaceae dominated suggested antagonistic/allelopathic effect of species
belonging to the family Amaranthaceae on spreading of Parthenium. Inhibitory or allelopathic effect has been
found in many plant species interactions including effect of aster, Blumea lacera L. on cereals and common
kharif weeds (Oudhia et al. 1989, Biswas et al. 2014, Sarkar et al. 2015). The inhibitory effects of one of the
worlds worst invasive plants Lantana camara has been reported regarding chromotoxicity and severe oxidative
imbalance on target crop legumes in present (Talukdar 2013a). However, role of chromosomal rearrangements,
ploidy level variations and other intrinsic biochemical mechanisms have been suspected behind aggressiveness
of alien invasions, as polyploid species and favorable chromosomal rearrangements, reported in legumes like
Lathyrus (Talukdar 2010a, b, 2012a, b) may have better fitness than common native plants (Pandit et al. 2011).
Furthermore, altered morphological, biochemical and molecular make-up in aneuploid genomes and diploid
mutated genotypes may confer new strategy for adaptations in stressful environments (Talukdar 2009a, b, 2011,
2012b, Talukdar & Biswas 2007), and thus, origin of new invasive flora cannot be ruled out.
Ecological study pointed out steep rise in population of 11 plants species (Parthenium hysterophorous,
Ageratum conyzoides, Eupatorium odoratum, Chromolaena odorata, Cassia sophera, Leucaena leucocephala,
Lantana camara, Trema orientalis, Amaranthus spinosus, Alternanthera sessilis and Eichhornia crassipes) in
the present forest area during the last six years. Thus, these 11 taxa, were selected as indicator of invasion by
non-indigenous species in the present forest area, and their distribution data manifested as ratio of forest and
roadside was presented. The four taxa of asters (Parthenium, Ageratum, Eupatorium, Chromolaena) with 5
species, ascertained by this criteria, have no economic utilization by locals. However, both Cassia and
Leucaena were extensively utilized as anti-diabetic medicine and fuel, respectively. The small tree, Trema
orientalis was mainly used as cheap fuel. Three taxa exhibiting high aggressiveness in degraded forest areas
were Lantana camara, Amaranths spinosus and Alternanthera sessilis which have been partially used by local
people. It was found that majority (~49%) of the invasive plant species were used by village medicine men for
primary health care systems, followed by food and feed. Many leguminous plants were used as both food and
forage. This has enormous importance as legumes are cheap and easily available source of plant-origin protein
with many essential amino acids, antioxidant flavonoids and minerals (Dixon & Sumner 2003, Talukdar 2013b).
127

.
Some recent surveys in Sikkim Himalayas (India) revealed extraordinary potential of legumes in formulation of
diverse types of ethnic food and medicines (Talukdar 2013d, Talukdar & Talukdar 2012a, b). Conservation of
legume germplasm is absolutely essential to prevent their dwindling genetic diversity throughout the world
including India. Due to its remarkable hardiness against abiotic (salinity, heavy metals etc.) and biotic stress,
Lathyrus spp. which have potential to grow well in low input and marginal farming condition and can sustain
soil nutrition in degraded areas has been recognized in the present study areas. This assumes importance as
potential of legume-based farming has been studied in recent decade and genetic improvement programs have
been undertaken (Talukdar 2008, 2009a, b, 2010a, b,c, 2011, 2012a, b). A recent study revealed arsenic and
other heavy metal bioaccumulation in photosynthetic and edible parts of crop legumes like Phaseolus vulgaris,
Lens culinaris, Cicer arietinum, and Lathyrus sativus in the lower Gangetic basin led severe agronomic loss due
to alteration in antioxidant defense mechanisms and severe impairment in plant growth (Talukdar 2012c, 2013b,
c).
Among the 103 taxa documented in the present study, seven species (Eupatorium odoratum, Chromolaena
odorata, Eichhornia crassipes, Lantana camara, Leucaena leucocephala, Mikania micrantha, Opuntia stricta)
have been conscripted as worlds 100 worst invasive species (Lowe et al. 2000). Tropical American flora was
found dominated as alien and the strong allelopathic effects on native species may be one of the reasons of their
dominance over indigenous flora (Huang et al. 2009, Talukdar 2013a). However, it is noteworthy that after
introduction, tough competition exists among alien flora in the invaded areas which can be exploited for their
better utilization, management and prevention of extinction of indigenous flora. Also, loss of diversity of native
flora due to invasive species cannot be straightforward in a dynamic and functional ecosystem as increasing
biotic and abiotic stress factors may impede growth and reproduction of native species with simultaneous
introduction, establishment and successful invasion of more hardy alien species, better utilizing the rapidly
depleting soil fertility, habitat fragmentation and other adverse conditions to colonize (Thuiller et al. 2007,
Talukdar 2013e). Digitization of regional native flora in global scientific data bases may have significant impact
in this regard (Talukdar 2015).
CONCLUSION
The result presented here is the cumulative of studies of six consecutive years. It is alarming to note that
number of alien species in this protected wildlife sanctuary constitutes a major share of forest biodiversity, of
which 11 species are enlisted as most aggressive invasive plants within the study area. It is also important to
note that the alien plants have been used by local people for medicinal and other economic purposes, although
they can impose considerable threat to the growth and reproduction of native flora in the forest area. Thus, the
present study can be used as reference work for threat assessment, management and utilization of alien and alien
invasive flora in this ecologically sensitive protected area.
ACKNOWLEDGEMENT
The authors are grateful to the local villagers and forest authorities for giving necessary supports during the
entire course of the study.
REFERENCES
Baider C & Vincent Florens FB (2011) Control of invasive alien weeds averts imminent plant extinction.
Biological Invasions 13: 26412646.
Bajpai O, Kumar A, Srivastava AK, Kushwaha AK, Pandey J & Chaudhary LB (2015) Tree species of the
Himalayan Terai region of Uttar Pradesh, India: a checklist. Check List 11(4): 1718.
Biswas S, Maity M, Srimany S, Chatterjee S, Karmakar T, Datta R, Patra J, Koley M & Talukdar D (2014)
Compositions, distributions and status of economic plants among invasive floras of Uttarpara, West Bengal,
India. International Journal of Pharmacognosy 1(12): 800809.
Biswas S, Maity M, Bhandari G, Batabyal R, Patra J, Bhuiya A, Ojha B, Halder N & Talukdar D (2014) Floral
diversity and ecology in Kalyani area of Nadia district, West Bengal, India. Plant Science Today 2(1): 38
42.
Bradley BA, Oppenheimer M & Wilcove DS (2009) Climate change and plant invasions: restoration
opportunities ahead? Global Change Biology 15: 15111521.
128

.
Crooks JA, Chang AL & Ruiz GM (2011) Aquatic pollution increases the relative success of invasive species.
Das AP & Lahiri AK (1990) Angiospermic flora of Bethuadahari Reserve Forest, Nadia (India). Indian Forester
116: 871882.
Davis MA (2003) Biotic globalization: does competition from introduced species threaten biodiversity?
Bioscience 53: 481489.
Dixon RA & Sumner LW (2003) Legume natural products: understanding and manipulating complex pathways
for human and animal health. Plant Physiology 131: 878885.
Feng J & Zhu Y (2010) Alien invasive plants in China: risk assessment and spatial patterns. Biodiversity and
Conservation 19: 34893497.
Heywood VH (1989) Patterns, extents and modes of invasions. In: Drake JA, Mooney HA, diCastri F, Groves
RH, Kruger FJ, Rejmnek M, & Williamson (eds) Biological Invasion: A Global Perspective. John Wiley,
pp. 3160.
Huang QQ, Wu JM, Bai YY, Zhou I & Wang GX (2009) Identifying the most noxious invasive plants in China:
role of geographical origin, life form and means of introduction. Biodiversity and Conservation 18: 305316.
Joshi BD (2002) A brief review on the flora of medicinal importance and prospects of developing a sustainable
network of small scale pharmaceutical industries in Uttaranchal. Himalayan Journal of Environment and
Zoology 16(2): 233.
Lockwood JL, Hoopes MF & Marchetti MP (2007) Invasion ecology. Blackwell, Oxford.
Lowe S, Browne S, Boudjela SM & De poorter SM (2000) 100 of the worlds worst invasive alien species. A
selection from the Global Invasive Species Database published by The Invasive Species Specialist Group
(ISSG) a specialist group of the Species Survival Commission (SSC) of the World Conservation Union
IUCN, pp. 12.
Mandal FB (2011) The management of alien species in India. International Journal of Biodiversity and
Mooney HA & Hobbs RJ (2000) Invasive species in a changing world. Island Press, Washington, DC.
Oudhia P, Kolhe SS & Tripathi RS (1998) Allelopathic effect of Blumea lacera L. on rice and common Kharif
weeds. Oryza 35: 175177.
Pandit MK, Michael, Pocock JO & Kunin WE (2011) Ploidy influences rarity and invasiveness in plants.
Journal of Ecology 99:11081115.
Raghubanshi AS, Rai LC, Gaur JP & Singh JS (2005) Invasive alien species and biodiversity in India. Current
Science 88: 539540.
Rastogi J, Rawat DS & Chandra S (2015) Diversity of invasive alien species in Pantnagar flora. Tropical Plant
Research 2(3): 282287.
Rao RR & Murugan R (2006) Impact of exotic adventives weeds on native biodiversity in India: implications
for conservation. In: Rai LC & Gaur JP (eds) Invasive Alien Species and Biodiversity in India. Banaras
Hindu University, Banaras, pp. 93109.
Reddy CS (2008) Catalogue of invasive alien flora of India. Life Science Journal 5(2): 84-89.
Sarkar A, Chakraborty P, Chakrabarti S, Sengupta A, Das P, Roy B, Majumder A, Kundu D, Halder D &
Talukdar D (2015) Inventorying floral diversity, compositions, and resource utilization capacity by local
people in Khan Garden of Hooghly district, West Bengal, India. Journal of Biology and Nature 3: 6779.
Sausa R, Morais P, Dias E & Antunes C (2011) Biological invasions and ecosystem functioning: time to merge.
Sax DF, Gaines SD & Brown JH (2002) Species invasions exceed extinctions on islands worldwide: a
comparative study of plants and birds. American Naturalist 160: 766783.
Singh KP, Shukla AN, & Singh JS (2010) State-level inventory of invasive alien plants, their source regions and
use potential. Current Science 90: 107114.
Srivastava A & Singh R (2009) Key management issues of Forest-invasive species in India. Indian Journal of
Environmental Education 9: 1624.
Talukdar D (2008) Cytogenetic characterization of seven different primary tetrasomics in grass pea (Lathyrus
sativus L.). Caryologia 61: 402410.
129

.
Talukdar D (2009a) Dwarf mutations in grass pea (Lathyrus sativus L.): Origin, morphology, inheritance and
linkage studies. Journal of Genetics 88: 165175.
Talukdar D (2009b) Recent progress on genetic analysis of novel mutants and aneuploid research in grass pea
(Lathyrus sativus L.). African Journal of Agricultural Research 4: 15491559.
Talukdar D (2010a) Reciprocal translocations in grass pea (Lathyrus sativus L.). Pattern of transmission,
detection of multiple interchanges and their independence. Journal of Heredity 101: 169176.
Talukdar D (2010b) Cytogenetic characterization of induced autotetraploids in grass pea (Lathyrus sativus L.).
Caryologia 63: 6272.
Talukdar D (2010c) Allozyme variations in leaf esterase and root peroxidase isozymes and linkage with
dwarfing genes in induced dwarf mutants of grass pea (Lathyrus sativus L.). International Journal of
Genetics and Molecular Biology 2: 112120.
Talukdar D (2011) Isolation and characterization of NaCl-tolerant mutations in two important legumes, Clitoria
ternatea L. and Lathyrus sativus L.: Induced mutagenesis and selection by salt stress. Journal of Medicinal
Plants Research 5: 36193628.
Talukdar D (2012a) Ascorbate deficient semi-dwarf asfL1 mutant of Lathyrus sativus exhibits alterations in
antioxidant defense. Biologia Plantarum 56: 675682.
Talukdar D (2012b) Meiotic consequences of selfing in grass pea (Lathyrus sativus L.) autotetraploids in the
advanced generations: Cytogenetics of chromosomal rearrangement and detection of aneuploids. Nucleus 55:
7382.
Talukdar D (2012c) Changes in neurotoxin, -N-OXALYL- L , -diaminopropionic acid (-ODAP), level in
grass pea (Lathyrus sativus L.) genotypes under arsenic treatments. Journal of Applied Biosciences 38:180
185.
Talukdar D (2013a) Allelopathic effects of Lantana camara L. on Lathyrus sativus L.: Oxidative imbalance and
cytogenetic consequences. Allelopathy Journal 31:7190.
Talukdar D (2013b) Bioaccumulation and transport of arsenic in different genotypes of lentil (Lens culinaris
Medik.). International Journal of Pharma and Bio Science 4(B): 694701.
Talukdar D (2013c) Arsenic-induced oxidative stress in the common bean legume, Phaseolus vulgaris L.
seedlings and its amelioration by exogenous nitric oxide. Physiology and Molecular Biology of Plants 19:
6979.
Talukdar D (2013d) In Vitro antioxidant potential and type II diabetes related enzyme inhibition properties of
traditionally processed legume-based food and medicinal recipes in Indian Himalayas. Journal of Applied
Pharmaceutical Science 3: 026032.
Talukdar D (2013e) Species richness and floral diversity around Teesta Barrage Project in Jalpaiguri district of
West Bengal, India with emphasis on invasive plants and indigenous uses. Biology and Medicine 5: 0114.
Talukdar D (2015) Digitization of regional plant flora: Step towards global biodiversity information service.
Journal of Biotechnology, Bioinformatics and Bioengineering 2: 712.
Talukdar D & Biswas AK (2007) Seven different primary trisomics in grass pea (Lathyrus sativus L.). I
Cytogenetic characterization. Cytologia 72: 385396.
Talukdar D & Talukdar T (2012a) Floral diversity and its indigenous use in old basin (Khari) of river Atreyee at
Balurghat block of Dakshin Dinajpur district, West Bengal. NeBIO 3: 2632.
Talukdar D & Talukdar T (2012b) Traditional food legumes in Sikkim Himalayas: Preparation of foods, uses
and ethnomedicinal perspectives. International Journal of Current Research 4: 6473.
Talukdar T & Talukdar D (2013) Ethno-medicinal uses of plants by tribal communities in Hili block of Dakshin
Dinajpur district, West Bengal. Indian Journal of Natural Products and Resources 4: 110118.
Thuiller W, Richardson DM & Midgley GF (2007) Will climate change promote alien plant invasions? In:
Nentwig W (eds) Ecological Studies: Biological Invasions. Springer-Verlag, Berlin, Heidelberg, pp. 197
211.
130
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1):131135, 2016
Research article
Impact of seasonal changes on air layering and rooting hormone

in Spondias pinnata (J. Koenig ex L. f.) Kurz.
Anita Tomar
Centre for Social Forestry and Eco-rehabilitation, Allahabad, U.P, India
*Corresponding Author: anitatomar@icfre.org
Abstract: Air layering trials were conducted in Spondias pinnata during four different seasons,
winter (January), rainy (July), spring (March) and autumn (October). Juvenile branches with 1.00
to 2.00 cm diameter were girdled using Indole Acetic Acid (IAA) and Indole Butyric Acid (IBA)
with the rooting hormones (100, 300 and 500 ppm) along with control. The impact of seasons and
rooting hormone were investigated. Callus was formed at the girdled portions of all the air layers
with or without hormones. Result revealed that July (Rainy season) proved to be better season for
making air layers than other months viz. October (autumn), January (winter), and March (spring)
in Spondias pinnata and IBA 500 ppm was more effective in promoting root formation.
Keywords: Rooting hormones - Air layers - Treatments - Season - Growth.
[Cite as: Tomar A (2016) Impact of seasonal changes on air layering and rooting hormone in Spondias pinnata
(J. Koenig ex L. f.) Kurz. Tropical Plant Research 3(1): 131135]
INTRODUCTION
Air-layering is a method of reproducing plants by inducing roots to form a plant stem without cutting off the
stem from the parent plant. It is an excellent way to replicate existing plant without disturbing the parent plant
bearing fruit or flowering. Air-layering can produce larger plants which are readily mature, much faster than
growing them from seed or cuttings. Air layering is used to reproduce a number of tropical fruit trees and
shrubs. Airlayering is a method of producing a new plant that is identical to the parent plant in all respects, like
fruit taste, colour and size. The new plant is formed while still attached to the parent plant upon which it
depends for water and nutrients until roots develop. In this type of propagation a large plant can be developed in
a relatively short period of time. Airlayering outdoor is performed best during spring and summer, although, it
can be done during any season of the year. Spring and summer layers are usually rooted and ready for
transplanting in winter (Dewayne & Thomas 2010). Air layering was first discovered by Chinese about 20
centuries ago and long known to horticulturists as a method of reproducing ornamental and cultivated plants
(Mergen 1953). It is oldest techniques used by the gardeners to propagate many woody plants (Hossain 2007).
Vegetative propagation through air layering has an advantage over other methods, since reserve food of the
parent branch induces the formation of well-developed root system. Also, the air layered branches in general,
have balanced root system than cuttings, and develop rapidly on planting out. Season is the important factor for
successful layering in woody plant because of rooting on layers enhanced by light and presence of sufficient
moisture and optimum temperature (Bose et al. 1986). In India, this method of vegetative propagation has not
been tried on forest trees on an appreciable scale. There are several vegetative methods for multiplication of the
quality stock in forest tree species but air layering is often used as a method of propagation where the formation
of roots from cuttings is slow (Hartmann & Kester 1975).
Hog-plum {Spondias pinnata (J. Koenig ex L. f.) Kurz.} is a deciduous, glabrous tree with edible fruit,
growing up to 25 m in height. The tree is found wild or cultivated throughout the tropical Indian subcontinent.
Despite a valuable and threatened plant, S. pinnata is not cultivated on a large scale in its native habitat. Due to
the limited distribution of S. pinnata commonly known as Amra of family Anacardiaceae and their inadequate
regeneration in nature the present study was conducted to standardize air layering, one of the vegetative methods
of propagation to facilitate ex-situ conservation of this species.
131
Tomar (2016) 3(1): 131135

.
Spondias pinnata (J. Koenig ex L. f.) Kurz. trees for air layering were selected from sites situated between
latitude 2507' to 2510' N and longitude 8154' to 8158' E and at 98 m elevation during the year 2012. Air
layering was conducted in randomized block design with three replications each. Each experiment consisted of
seven treatments i.e. Control (lanolin paste only), IAA and IBA with 100, 300 and 500 ppm each prepared in
lanolin paste. For each species 105 air layers (15 for every treatment) were made in four different seasons i.e.
(i) Winter (January) (ii) Rainy (July) (iii) Spring (March) (iv) Autumn (October).
Juvenile branches with 1.00 to 2.00 cm diameter were selected for tying the air layers. The bark of the twig
(approximately 1 inch wide ring) was removed with the help of knife and 100, 300 and 500 ppm IBA and IAA
in the form of powder was applied to the wounded surface. Untreated layers served as control. Sphagnum moss
about two handfuls moistened with water and thereafter squeezed to remove excess water was placed around the
treated area and wrapped with a polyethylene sheet and finally tied at both ends with plastic thread to avoid the
escape of moisture. In this experiment, the first observation on air layered branches to confirm root initiation
was recorded after 40 days of setting the experiment and subsequently other observations were made after every
tenth day for a period of two months to monitor the development of roots. When roots were visible through the
transparent polythene sheet the air layers were detached from mother plant just below the girdle. After air-layers
were cut down, plastic film and moss were removed and the roots were counted. A black polythene cover is said
to be better than white one, though the latter is better to observe the progress of rooting from time to time. Some
roots were broken when the tightly packed moss was removed and many root systems dried during the time,
they were exposed for root count (Nautiyal 2002). Roots upto 1 mm and more in length were counted because
some shorter protuberances have the appearance of emerging roots but consist of parenchymatous tissue
(Mergen 1955). These rooted air layers were transplanted in polythene bags filled up with growing media
containing sand, soil and FYM (1:1:2 ratio). These polybags were kept in shade for about one weeks and
watered regularly until the root system was well established in the soil.
RESULTS
Observations recorded for the airlayers on root growth characters after been presented in Appendix I. Air
layers have initiated callus development followed by root formation at the base of incision. However, the callus
and root initiation varied considerably for different seasons.
Figure 1. Air layers of Spondias pinnata.
The air layer tests in Spondias pinnata have revealed that IBA significantly affected the rooting, number of
primary and secondary roots per layer and mean length of the roots in air layers, whereas, other treatments did
not affect the callus significantly (Fig. 1). IBA 500 ppm stimulated maximum rooting in air layers in all the four
seasons of the year. The rainy seasons exhibited maximum rooting (57.7 %) in the air layers, followed by
autumn (45.3%) winter (42.6) and spring (36.9 %) in IBA 500 ppm. The next maximum rooting in air layers
after IBA 500 ppm was observed in IAA 500 ppm treatments. The number of primary and secondary roots per
layer was also influenced by various concentrations of IBA and IAA treatments. The minimum rooting in air
layers was recorded with IAA and IBA in control in all the seasons studied. The number of primary and
secondary roots produced by the air layers was recorded as the maximum again with IBA 500 ppm. The rainy
season produced the maximum number (18.0 2.80 and 5.20 1.23 ) of primary and secondary roots in IBA
500 ppm followed by autumn (12.12 2.29 and 3.76 0.31) in IAA 500 ppm, winter (12.011.51 and 4.23
0.81) in IBA 500 ppm and spring (9.19 1.34) in IAA 500 ppm and 3.12 0.32 in IBA 500 ppm respectively).
The mean length of roots was found maximum with IBA 500 ppm ,which ranged from 5.40 0.91 cm in rainy
season in IBA 500 ppm followed by 4.100.45cm, 4.95 0.31cm , 3.72 0.21 (in IAA 500 ppm in Autumn,
132
Tomar (2016) 3(1): 131135

.
Winter and Spring respectively). It was also observed that shade is essential for the success of air layers, as good
results were observed in layers, which were under shade of the crown.
DISCUSSION
The rainy (July) season was more favourable for rooting of air layers in Spondias pinnata due to the fact that
constant moisture is one of the essential conditions for successful air layering (Nautiyal 2002, Kadami & Dabral
1954). Callus formation was observed in January, March, July and October in all the treatments. However, the
root initiation was more in July, compared to other months, due to higher temperature coupled with higher
humidity (Nautiyal 2002). These results are in conformity with the findings of Chandra (1967) for Magnolia
grandiflora; Shrivastava et al. (1994) for Albizia lucida and Nautiyal (2002) for Ficus spp. Rainy season with
rooting hormone provides a favourable environment for inducing roots because of high humidity and rains. The
rooting percentage varies in response to season, the high humidity (RH >7595 %) and optimum temperature
(>2532 C) in rainy season favor maximum growth as sphagnum absorbs humidity while temperature helps in
root initiation (Kumar et al. 2013). Kanwar & Kahlon (1986) reported that layering in Guava was successful
when carried out between mid-July and early-October in India. The best rooting and survival in Guava were
obtained in July and August in Bangladesh (Akhter 2002). Dhillon & Mahajan (2000) reported that August was
the best time for air layering in litchi in respect of rooting success and survivability.
In the winter season the falling temperature might have an adverse effect on root formation in air layers.
Better rooting response of air layers was observed with IBA 500 ppm during the month of July. These results
have verified the findings of Chauhan & Dua (1982) and Puri & Nagpal (1988) in Morus alba and Dalbergia
sissoo, respectively. Nagpal & Singh (1986) reported that IAA and IBA treatments enhanced the rooting
capability in Bauhinia variegata, during pre-monsoon and monsoon period. The seasonal changes in the rooting
response appears to be regulated by a balance of internal translocation of substances including carbohydrates,
nitrogenous substance, hormonal growth regulators and co-factors acting synergistically with auxins (Khosla et
al. 1982).
Scientists of the various parts of the globe engaged in tree improvement programmes, achieved poor to good
success of air layering in different tree species. Good success on rooting and root characteristics of air layering
in Chebulic mycobalum has been recorded by Misra & Jaiswal (1994) after treatments with Indole Butyric Acid
(IBA). IBA has been found to stimulate root initiation in air layers of many plant species like Carissa carandas
and Dalbergia sissoo (Puri & Nagpal 1988). Air layering has been reported in many other forest tree species viz.
Ficus krishnae & Ficus auriculata (Tomar & Singh 2011), Bombax ceiba (Venkatesh et al. 1978), Gmelina
arborea (Arya & Haque 1982), Prosopis cineraria (Solanki et al. 1984). Acacia nilotica (Sharma et al. 2004),
Eucalyptus microtheca (Hussain & Ponnuswami 1964), Guadua angustifolia (Verma et al. 2013) etc.
CONCLUSION
The results of the study show that air layering method is a potential, viable and economical method of
vegetative propagation for Spondias pinnata. Air layering is relatively simple and very easily adopted by
farmers due to high success rate and low mortality. From the results obtained by air layering , it can be
concluded that July (Rainy season) proved to be better season for making air layers than other months viz.
October (autumn), January (winter), and March (Spring) in S. pinnata. The hormone treatments proved
successful in inducing the root initiation in comparison to control. IBA 500 ppm was more effective in
promoting root formation and growth while minimum rooting was recorded with IAA and IBA (control). The
number of primary and secondary roots produced was recorded as maximum with IBA 500 ppm.
ACKNOWLEDGEMENT
The author gratefully acknowledged the financial assistance received from Indian Council of Forestry
Research and Education (ICFRE), Dehradun under project FRI-581/CSFER-18.
REFERENCES
Akhter AA (2002) Effect of time and methods on the success of layering in Guava, M.S. Thesis. Dept. Hort.,
Bangladesh Agricultural University, Mymensingh.
Arya RS & Haque M S (1982) Grafting and budding in Gmelina arborea. Indian Forester 108 (7): 497500.
133
Tomar (2016) 3(1): 131135

.
Bose TK, Mitra SK & Sadhu MK (1986) Guava In: Propagation of tropical and sub-tropical horticultural
crops. Naya Prokash, Calcutta, pp. 291301.
Chandra J P (1967) Preliminary observations on air layering of Magnolia grandiflora L. Indian Forester 93:
120122.
Chauhan PS & Dua IS (1982) Improvement of Forest Biomass. Pragati Press Delhi, pp. 187191.
Dhillon BS & Mahajan BVC (2000) Effect of different dates on the air layering performance of litchi cultivars
in Punjab. Agricultural Science Digest 20(3): 207208.
Hartmann HT & Kester DE (1975) Plant Propagation Principles and Practices. 3rd Edition. Printice Hall, New
Jersey.
Hossain AMD (2007) Effect of time of operation & variety on the rooting success and survivability of air layer
in Guava, M.S. Thesis. Agricultural University, Mymensingh, Bangladesh.
Hussain A M & Ponnuswami PK (1964) Preliminary observations on air layering in Eucalyptus microtheca
F.v.M. Indian Forester 90(7): 486487.
Kadambi K & Dabral SN (1954) Air layering in Forestry practice. Indian Forester 81: 721724.
Kanwar JS & Kahlon GS (1986) Propagation studies in litchi. Journal of Research Punjab Agricultural
University 23(1): 3339.
Kedarnath S & Dhaundiyal RP (1963) Preliminary observations on air layering in Eucalyptus microtheca.
F.v.M. Indian Forester 90(7): 486487.
Khosla PK, Nagpal R & Puri S (1982) Propagation of some agroforestry species by air layering. Indian Journal
of Forestry 5: 171174.
Mergen F (1953) Air layering as a possible method to reproduce selected Slash Pine. Naval Stores Review
63(21): 1920.
Mergen F (1955) Air layering of slash pines. Journal of Forestry 53: 265270
Misra KK & Jaiswal H R (1994) Effects of growth regulators on rooting and survival of air layers of Karaunda
(Carissa carandas L.) Annals of Agricultural Research 11(2): 208210.
Nagpal R & Singh K (1986) Propagation of some social forestry species by air layering. In: Khosla PK & Kohli
RK (eds) Social Forestry for Rural Development. pp. 222227.
Nautiyal DP (2002) Vegetative Propogation in three important fodder tree species of family Moraceae from
Garhwal Himalaya, Ph. D. Thesis. H.N.B. Garhwal University, Srinagar, Uttarakhand, India.
Puri S & Nagpal R (1988) Effects of auxins on air layers of some agroforestry species. Indian Journal of
Forestry 11(1): 2832.
Sharma SK, Verma SK & Bhojvad PP (2004) Cloning of Acacia nilotica for multiplication and establishment of
clonal seed orchard. In: Multipurpose trees in the Tropics: Management and Improvement strategies. AFRI,
Jodhpur, pp. 691700.
Solanki KR, Kackar NL & Jindal SK (1984) Propagation in Prosopis cineraria (L) Mac bride by air-layering.
Current Science 53(21): 11661167.
Srivastava SC, Kumar P & Misra YD (1994) Air layering in galwang (Albizia lucida Benth.): A Lac host plant.
Indian Forester 120: 524528.
Tomar A & Singh VRR (2011) Effect of Air Layering time (Season) with the Aid of Indole Butyric acid in
Ficus krishnae and Ficus auriculata. Indian Forester 137(12): 13631365.
Venkatesh CS, Arya RS & Emmanuel CJSK (1978) A note on air - layering and budding in Semul. Indian
Forester 104(2): 142144.
Verma PK, Das N, Kaushik PK, Kumar V & Yadav A (2013) Vegetative Propagation through air layering of
Guadua angustifolia Kunth - A commercially important bamboo. Indian Forester 139(12): 10881091.
134
Tomar (2016) 3(1): 131135

.
Appendix I. Effect of air layering time (season) and rooting hormone on root initiation in Spondias pinnata.
RAINY
A
AUTUMN
D
CONTROL 60.2 5.34 1.20 0.03 0.88 0.15 0.91 0.30 60.0 2.78 0.97 0.09
D
0.00
WINTER
E
SPRING
D
1.01 0.06 50.8 1.66 2.01 0.11 0.32 0.04 1.41 0.21 50 0.83 1.59 0.30
0.00
1.01 0.08
IBA 100
82.1 28 6.20 0.70 1.53 0.24 3.91 0.69 79.2 22.6 3.91 0.12 0.96 0.01 2.17 0.22 80.3 23.8 2.13 0.34 0.25 0.07 1.80 0.21 83.4 20.6 2.13 0.20 1.02 0.01 1.31 0.21
IBA 300
92.6 40.2 12.0 1.73 3.73 0.30 4.09 0.56 90.2 31.6 6.24 0.32 1.29 0.16 3.26 0.25 81.2 41.3 6.91 0.82 1.93 0.21 3.31 0.21 91.6 27.7 5.20 0.41 2.00 0.21 2.14 0.39
IBA 500
97.1 57.7 18.0 2.80 5.20 1.23 5.40 0.91 91.6 45.3 10.01 1.24 2.29 0.19 3.92 0.31 88.7 42.6 12.01 1.51 4.23 0.81 4.31 0.40 92.8 36.9 7.79 0.80 3.12 0.32 2.97 0.21
IAA 100
83.0 22.0 3.40 0.43 0.61 0.12 3.40 0.61 81.7 21.6 3.46 0.56 0.92 0.06 3.01 0.40 79.6 17.1 4.21 0.21 1.65 0.11 2.21 0.07 90.9 14.9 4.21 0.30 0.98 0.02 3.13 0.21
IAA 300
91.3 38.4 10.0 1.08 2.02 0.23 4.24 0.62 94.6 27.9 5.13 0.21 1.56 0.04 1.98 0.11 88.1 17.9 5.91 0.34 2.13 0.14 3.11 0.21 90.3 20.1 3.19 0.52 0.86 0.10 1.75 0.12
IAA 500
95.9 47.2 14.2 3.42 3.97 0.81 4.70 0.86 95.9 39.2 12.12 2.29 3.76 0.31 4.10 0.45 96.2 40.3 10.41 3.22 3.41 0.11 4.95 0.31 96.2 30.6 9.19 1.34 2.23 0.22 3.72 0.21
Note: A = Percent callused layer, B = Rooting percent, C= No. of Primary roots, D = No.of Secondary roots, E = Root Length.
135
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1):136141, 2016
Research article
Sacred plants from ancient to modern era: Traditional

worshipping towards plants conservation
Deepti Pandey1* and Vimal Chandra Pandey2
1
Department of Ancient Indian History & Archaeology, University of Lucknow, Lucknow, India
Plant Ecology and Environmental Science Div., CSIR-National Botanical Research Institute, Lucknow, India
*Corresponding Author: deeptipandey@hotmail.com
2
Abstract: The present research paper describes the sacred plants of the Indo-Gangetic plain and
their associated deity and festivals. Several intensive surveys were carried out to find the definite
role and importance of nine sacred plant species in the Indo-Gangetic plain, India in the life style,
religious activities and health care. These sacred plants are used in variety of ceremonies in
various ways throughout the year by the people of study area. Furthermore, these plants are
considered as sacred due to their medicinal, aesthetic and natural qualities. Thus, our ancestors
linked various God and Goddess with several plants for their conservation and named as sacred
plants. These ancient beliefs show the human relation with plants which are also helpful in the
conservation of plant species for their valuable qualities.
Keywords: Ancient beliefs - Plants conservation - Sacred plants - Traditional worshiping.
[Cite as: Pandey D & Pandey VC (2016) Sacred plants from ancient to modern era: Traditional worshipping
towards plants conservation. Tropical Plant Research 3(1): 136141]
INTRODUCTION
The ancient beliefs show the relation of human beings and plants. The plant worshipping was quite common
in a highly evolved Harappan culture, dating the third or fourth millennium B.C. It was also present among the
seals of Mohenjo-daro, one seal depicted a stylised Peepal (Ficus religiosa L.) tree with two heads of unicorns
emerging from its stem. Tree worshipping was also present during the Vedic period (Bhatla et al. 1984). In
India, many religious festivals are celebrated by the people from Kashmir to Kanyakumari as India is known for
its diversity like religion, customs, myths, languages, culture etc. Furthermore, all people celebrate religious
festivals with scientific background and use one or several plants or plant parts in their ceremonies. The various
parts of plants have been used as a source of medicine by man from ancient to modern era (Bisht & Badoni
2009, Mehra et al. 2014, Kumaran & Citarasu 2015, Truyen et al. 2015, Bajpai et al. 2016). Our ancestors lived
and spent their life in nature. They had a very strong belief on the basis of their knowledge about the valuable
qualities of plants (Bajpai et al. 2016). Man secured his life from diseases by using various parts of medicinal
plants. So, probably this became the basis of conserving plants and might have started worshipping plants
(Sharma & Joshi 2010, Mehra et al. 2014). Folklore, culture, food and medicinal practices are deeply linked and
influenced by plants (Badoni & Badoni 2001). On the basis of ancient scriptures, a wide variety of plants like
Ficus religiosa L., Azadirachta indica A. Juss., Ocimum tenuiflorum L. etc. has divine qualities, therefore used
in number of religious activities, marriages and other ceremonies (Robinson & Cush 1997). India has deeprooted traditional worshiping of plants, which provide base for the grass root conservation practices (Gadgil
1987, 2000, Gadgil & Rao 1998). In this paper some of the plants species which have divine qualities for human
health and medicinal practices but held sacred in the Indo-Gangetic plain of India are discussed.
Study site description
The Indo-Gangetic plain was selected as the present study site (Fig. 1). This region is known for the Indus
Valley Civilization, which was liable for the birth of ancient culture. According to ancient Indian history, this
region was also referred to as Aryavarta (Land of Aryans) during Vedic and Epic eras. The Indo-Gangetic plain
is also known as Indus-Ganga and the North Indian River Plain (Taneja et al. 2014). The Ganga is the leading
136
Pandey & Pandey (2016) 3(1): 136141

.
river of this region after whose name this plain is named. The Ganga and its tributaries have brought large
quantities of alluvium soil from the mountains and deposited it here to build this plain.
Figure 1. Location map of study site: Indo-Gangetic plain.
Methodology
The present study is based on intensive field surveys during 20102011. Identification of the collected
sacred plant specimens were done at Herbarium of National Botanical Research Institute, Lucknow. The present
information regarding sacred plant was collected through consulting the local people, villagers, traditional
medicine practitioners and priests of the temples to know the local name, sacred value and medicinal importance
of mentioned plants.
The present study reveals nine sacred plants species are of great medicinal value, associated God and
Goddess and have religious significance (Table 1). It also provides the information on sacred plants based
festivals, their month, deity and sacred beliefs (Table 2). These sacred plants are worshiped by the local people
of the Indo-Gangetic plain of India in various religious activities, marriages and traditional medicine practices.
The details of sacred plants and their associated God and Goddess as well as medicinal importance are described
in table 1. The medicinal importance of the plants is mentioned by several researchers in their studies (Kumar et
al. 2012, 2013). From ancient time to present day, the people depend upon plants for food, fodder, fibre, fuel,
shelter, shade and medicine. The Vedas has described the close relationship between man, nature and religion.
But the religious and festival aspects of sacred plants of the Indo-Gangetic plain are given a few attentions but
not much explored. These nine sacred plants symbolize a specific God or Goddess because of their medicinal,
aesthetic and natural qualities.
Table 1. List of sacred plants and associated God and Goddess who residing in these plants.
S.No. Local Name Scientific Name
Family
Associated God & Goddess
Medicinal Value
1
Kela
Musa balbisiana Colla
Musaceae
Lord Brihaspati, Vishnu
Fruits are taken with milk
as remedy for body
weakness.
2
Bel
Aegle marmelos (L.) Correa
Rutaceae
Lord Shiva
Digestive disorders have
been cured by ripe fruits
3
Neem
Meliaceae
Goddess Sheetala Mata
Bark extracts, leaves and
seeds shows antifungal and
antibacterial properties and
used in pimples,
rheumatism, ringworm,
wounds and cuts.
137
Pandey & Pandey (2016) 3(1): 136141

.
Tulsi
Ocimum tenuiflorum L.
Lamiaceae
Goddess Lakshmi
Leave are used in toothache
with common salt and also
used in fever and cold.
Anvala
Phyllanthaceae Lord Vishnu and Shiva
Fresh fruit juice has been
taken to improve eye sight
and to cure anaemia.
Kamal
Nelumbo nucifera Gaertn.
Nelumbonaceae Goddess Lakshmi
Plant is used in the
treatment of diarrhoea,
sunstroke, blood vomiting,
uterine disorders, burning
sensation, cough and
epistaxis.
Sweta Madar Calotropis gigantea (L.) Dryand. Apocynaceae
Lord Shiva
Leaves juice is used in skin
affections; roots are used in
old ulcers.
Shami
Prosopis cineraria (L.) Druce Leguminosae
God Sani
Bark is used in leprosy,
dysentery, bronchitis,
asthma, leukoderma,
hemorrhoids and also in
muscle tremors.
Peepal
Ficus religiosa L.
Moraceae
Triad Brahma, Vishnu &
Moderately warm fresh leaf
Mahesh (Shiva)
juice is used as the ear drop;
gum diseases are prevented
by root chewing.
Table 2. Sacred plants associated Religious festivals.
S.No.
1
2
3
4
Festivals
Sheetala Ashtami
Nimb Saptami
Vat Savitri
Somvari Amavasya
Amla Navmi
(Akshay Navmi)
Tulsi Vivah
Month
March
April
MayJun.
In all Amavasya
(New Moon)
Oct.Nov.
Plant species
Ficus religiosa L.
Local Name
Neem
Neem
Bargad
Peepal
Family
Meliaceae
Meliaceae
Moraceae
Moraceae
Anvala
Phyllanthaceae
Oct.Nov.
Ocimum tenuiflorum L.
Tulsi
Lamiaceae
The Bel tree {Aegle marmelos (L.) Correa} is believed to be associated with Lord Shiva (Fig. 2A). The Bel
tree is generally planted near to temples and garden. Its leaves and fruits are used in the worship of Lord Shiva.
The traditional devotees write the name of Rama on its leaves by sandal paste and worship the Lord with them.
It gives endless virtue on the devoted person. The women of the Indo-Gangetic plain worship this tree in order
to get their desires fulfilled.
Many ancient beliefs centre on the Neem tree (Azadirachta indica A. Juss.). It is associated with Sheetala
Mata (Cool one) - the goddess of smallpox. It is believed that the Sheetla Mata live in this tree. The leaves of
this tree are used in the treatment of person who suffers from smallpox. He is fanned by the leafy twigs of this
tree. Furthermore, the leaves are used in several methods to lessen and relieve this disease. There are many
folksongs, folktales and folk proverb in which an inspiring appeal is made to the Sheetla Mata to free the patient
from the smallpox.
Tulsi (Ocimum tenuiflorum L.) is the most holy plants growing in front of almost all Indian houses as an
auspicious point of view or a symbol of peace and worshipped by women (Fig. 2B). It is worshipped as Goddess
(wife of Lord Vishnu) and also known as Vishnupriya (the beloved of Vishnu). It is also considered to be an
incarnation of Goddess Lakshmi. Its associated religious festival is Tulsi Vivah which is the ceremonial
marriage of the Tulsi with Lord Vishnu. This festival is helpful in removing obstacle if delay in marriage. It is
used in most of the religious ceremonies. It has great medicinal value to mankind. Its leaves give relief in stress
and cold. It enhances the concentration power of the person and also sharpens the memory. Besides, its leaves
are often kept in water for purification. Tulsi plant enriches atmosphere through its divine fragrance and purifies
air (Kumari & Charantimat 2011). Hence, it is known as Miracle or Queen of Herbs.
138
Pandey & Pandey (2016) 3(1): 136141

.
Figure 2. Some sacred plants: A, Aegle marmelos (L.) Correa; B, Ocimum tenuiflorum L.; C, Ficus religiosa L.; D, Musa
balbisiana Colla; E, Phyllanthus emblica L.; F, Prosopis cineraria (L.) Druc; G, Nelumbo nucifera Gaertn.; H, Calotropis
gigantea (L.) Dryand.; I, Ficus benghalensis L.
Peepal (Ficus religiosa L.) is the most sacred tree in India (Fig. 2C). It is believed as the residence place of
the triad ~Brahma, Vishnu and Mahesh (Shiva). Its roots, trunk and leaves represent Lord Brahma, Vishnu and
Mahesh (Shiva), respectively. Worshipping the Peepal tree helps in controlling the thoughts, removes obstacles
in marriage and financial growth and brings multiple source of income to the believer as well as good for
children and fertility. According to astrological point of view, it is believed that if a person has manglik dosh,
marrying a Peepal tree, removes the dosh and a person can marry a non-manglik person. The women worship
this tree on the 15th of all months which falls on Monday, i.e. Somvari Amavasya. They pour water and milk on
it roots. The sandal paste, vermillion, akshat (wet rice) and flowers are also offered to Peepal tree. They tie
thread round the trunk of Peepal tree 108 times. It is ancient belief that these threads bother the tree spirit, which
consequently grants the boon to worshiper.
Banana tree (Musa balbisiana Colla) is a very pious tree and represents Lord Brihaspati and Vishnu (Fig.
2D). It is worshiped on Thursdays to get the benefits of Jupiter. Its fruit is also offered to Lord Vishnu and
Laksmi for happy married life and financial condition. Its leaves are also used in many religious ceremonies and
festivals. Its leaves are used as plates for food due to being long and broad.
Anvala (Phyllanthus emblica L.) is named amalak in Sanskrit (Fig. 2E). It is worshipped by women
especially in the month of Kartik (OctoberNovember) with a view to be favoured with male progeny. On the
ninth day of the bright half of the month of Kartik~which is known as Akshaya Navami (the immortal ninth)-a
special offering is made to this tree. On this day Brahmanas are fed while sitting under the shadow of this tree.
This brings unlimited punya (fortune) to the host. In absence of big trees, saplings are used for this purpose. It is
also believed that eating food under the anvala tree in the month of Kartik absolves one from the Anna doshas
for a year.
139
Pandey & Pandey (2016) 3(1): 136141

.
Shami tree {Prosopis cineraria (L.) Druce} is a sacred tree and deciduous in nature (Fig. 2F). It is
mythological important in local communities. Shami tree represents God Sani. It is sacred to Indian culture
especially by Hindus who worship it before going on a main journey and on the occasion of Dushehra festival. It
is believed that Shami tree worshiping is helpful to check bad impacts of Sani. Religious Hindu women worship
the tree regularly. In addition, it is mentioned as representative of all five F viz. Forest, Fiber, Fuel, Fodder and
Food in ancient literature. The ancient literature also describes the importance of the medicinal value of this tree.
Its bark has folkloric repute to possess anti-inflammatory, antirheumatic, tonic, and vermifuge properties. It is
also used in the treatment of anxiety, asthma, bronchitis, dyspepsia, fever, dysentery, leprosy, piles, wandering
of the mind, and tremors (Rani et at. 2013). It is well regarded by environmentalists, practitioners, farmers and
villagers for inclusion in afforestation projects in extremely dry areas.
Kamal (Nelumbo nucifera Gaertn.) is known as Indian lotus or sacred lotus (Fig. 2G). It is associated to
Goddess Lakshmi. She is the Hindu Goddess of wealth, fortune and prosperity (both spiritual and material). It is
used in Diwali festival and other religious occasion and has cultural significance in Hindus. All parts of the
Kamal plants are edible. Its rhizomes are widely used for vegetable as well as other purposes like chips and
known as Kamal Kakdi.
Sweta Madar {Calotropis gigantea (L.) Dryand.} is a sacred plant and associated to Lord Shiva (Fig. 2H). It
can be found in most of the Hindu houses. The leaves of Sweta Madar warmed in oil applied in inflammatory
part of the body. It is also used by the people who are suffering from rheumatism.
The Bargad tree (Ficus benghalensis L.) symbolizes Lord Shiva (Fig. 2I). It also depicts the Trimurti
Brahma (roots), Vishnu (bark), and Shiva (leaves). Additionally, it represents life and fertility in Indian cultures.
According to Hindu calendar, the Amavashya of the Hindi month Jyeshtha (May-June) is called as Vat Savitri
Amavashya or Bargadahi Amavashya. Married women offer their worship to this tree and tying raw cotton
thread around the tree on Amavashya of Hindi month Jyeshtha to celebrate Vat Savitri. This brings unlimited
and continued good fortunes on this day to the worshipper. This tree is reported extensively in ancient Indian
medicine systems for various diseases. Several parts of this tree were used to cure many deadly diseases such as
diarrhoea, dysentery, diabetes, menorrhagia, leucorrhoea, nervous disorders, tonic and astringent (Gopukumar &
Praseetha 2015). It is also considered as Indias National Tree and denotes spiritual knowledge. The Bargad tree
is often being planted around temples and a place of religious importance.
Thus, the above results and discussion proved the relation of human and the nature towards plants
conservation. The traditional worshipping has protected many plants which have tremendous medicinal value
and made them as sacred, so that with the fear of deity nobody eradicates it. So we have to protect these sacred
plants for us and our next generation for better survival. On the basis of this study, we have to follow our
ancestors belief for human and nature sustainability.
ACKNOWLEDGEMENT
The authors are thankful to the local people, villagers, traditional medicine practitioners and priests of the
temples of the Indo-Gangetic plain of India for their help to find out the local name, sacred value and medicinal
importance of plants.
REFERENCES
Badoni A & Badoni K (2001) Ethnobotanical heritage. In: Kandari OP & Gusain (eds) Garhwal Himalaya:
Nature, Culture and Society. Trans Media Srinagar (Garhwal). pp. 125147.
Bajpai O, Pandey J & Chaudhary LB (2016) Ethnomedicinal Uses of Tree Species by Tharu Tribes in the
Himalayan Terai Region of India. Research Journal of Medicinal Plant 10(1): 1941.
Bhatla N, Mukherjee T & Singh G (1984) Plants: Traditional worshipping. Indian Journal of History of Science
19(1): 3742.
Bisht C & Badoni A (2009) Distribution and indigenous uses of some medicinal plants in district Uttarkashi,
Uttarkhand, India. Researcher 1: 160.
Gadgil M & Rao S (1998) Nurturing Biodiversy an Indian Agenda. Center for Environemnt Education,
Ahmadabad, 157 p.
Gadgil M (1987) Diversity: cultural and ecological. Trends in Ecology & Evolution 2: 369373.
140
Pandey & Pandey (2016) 3(1): 136141

.
Gadgil M (2000) Grass roots conservation practices: Revitalizing the traditions. In: Kothari A et al. (eds)
Communities and conservation Natural Resources Management in South and Central Asia. Sage
Publication, New Delhi, pp. 220237.
Gopukumar ST & Praseetha PK (2015) Ficus benghalensis Linn. - The Sacred Indian Medicinal Tree with
Potent Pharmacological Remedies. International Journal of Pharmaceutical Sciences Review and Research
32(1): 223227.
Kumar A, Pandey VC & Tewari DD (2012) Documentation and determination of consensus about
phytotherapeutic veterinary practices among the Tharu tribal community of Uttar Pradesh, India. Tropical
Animal Health and Production 44: 863872.
Kumar A, Pandey VC, Singh AG & Tewari DD (2013) Traditional uses of medicinal plants for dermatological
healthcare management practices by the Tharu tribal community of Uttar Pradesh, India. Genetic Resources
and Crop Evolution 60: 203224.
Kumaran T & Citarasu T (2015) Ethnopharmacological investigation and antibacterial evaluation of the
methanolic extract of Asparagus racemosus (Shatavari). Tropical Plant Research 2(3): 175179.
Kumari DBS & Charantimath A (2011) Sacred plants- their role in religion and uses in health care system of
Savangere district. The Socioscan 3(1&2): 14.
Mehra A, Bajpai O & Joshi H (2014) Diversity, utilization and sacred values of Ethno-medicinal plants of
Kumaun Himalaya. Tropical Plant Research 1(3): 8086.
Rani B, Singh U, Sharma R, Gupta A, Dhawan NG, Sharma AK, Sharma S & Maheshwari RK (2013) Prosopis
cineraria (L) Druce: a desert tree to brace livelihood in Rajasthan. Asian Journal of Pharmaceutical
Research and Health Care 5: 5864
Robinson C & Cush D (1997) The Sacred Cow: Hinduism and ecology. Journal of Beliefs & Values: Studies in
Religion & Education 18(1): 2537.
Sharma V & Joshi BD (2010) Role of Sacred plants in religion and health care system of local people of Almora
district of Uttarkhand state (India). Academic Arena 2(6): 57 60.
Taneja G, Pal BD, Joshi PK, Agarwal PK & Tyagi NK (2014) Annual Report. International Food Policy
Research Institute. 44 p.
Truyen DM, Mansor M & Ruddin AS (2015) A note on Aroids Ethnobotany in Hau River, Vietnam. Tropical
Plant Research 2(1): 5863.
141
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 142152, 2016
Research article
Biomass extraction impact on vegetation community structure in

Kaimur wildlife sanctuary, Uttar Pradesh, India
Azram Tahoor1, Azra Musavi2 and Jamal A. Khan1*
1
Department of Wildlife Sciences, Aligarh Muslim University, Aligarh, Uttar Pradesh, India
2
Department of Economics, Aligarh Muslim University, Aligarh, Uttar Pradesh, India
1
Department of Wildlife Sciences, Aligarh Muslim University, Aligarh, Uttar Pradesh, India
*Corresponding Author: secretarywsi@gmail.com
[Accepted: 11 March 2016]
Abstract: The objective of this study was to assess the impacts of biomass extraction on vegetation community
structure. The area selected for this study was Kaimur wildlife sanctuary situated in Mirzapur and Sonbhadra
district of Uttar Pradesh, India. Area was stratified into high, medium and low disturbed area on the basis of
presence of human induced disturbance indicators. Within each stratified area, 10 m radius circular plots were
laid to record the vegetation, habitat and disturbance variables. Results showed that tree density and diversity
indices were recorded highest in least disturbed area. The overall highest Importance Value Index is of Sal
(116), recorded from medium disturbed area. Human trail and grazing cover was negatively correlated with
density and diversity indices of tree and sapling. Canopy cover was positively correlated to herb diversity
indices. Tree, shrub and herb diversity indices are positively correlated to distance from human habitation.
Present study concludes that grazing and lopping are the prime disturbance factor for changes in vegetation
community structure. The density of some of the sampled plant species is very low which in the coming future
will face local extinction. For future and long term aspects, urgent initiatives are required to conserve and
protect vegetative species.
Keywords: Vegetation composition - IVI - Canopy cover - Human induced disturbance - Grazing.
[Cite as: Tahoor A, Musavi A & Khan JA (2016) Biomass extraction impact on vegetation community structure
in Kaimur wildlife sanctuary, Uttar Pradesh, India. Tropical Plant Research 3(1): 142152]
INTRODUCTION
Protected areas (PAs) are home to variety and variability of living organisms. There are around millions of
people living around these PAs and are dependent on the forest resources for their basic needs (Kothari et al.
1995). These nearby residing villagers extract biomass in the form of livestock grazing, lopping, fuelwood
collection, extraction of NTFPs (Non Timber Forest Products). At a large scale, the extraction of forest resource
exerts impact on vegetation composition (Silori & Mishra 2001, Seng et al. 2004, Shahabuddin & Prasad 2004,
Rabha 2014). It is because of unsustainable extraction of forest and increase in human population day by day.
Harvesting or extraction of forest resource from the PAs is illegal but still it is going on. Earlier studies in India
and abroad found that disturbance due to biomass extraction has negative impact on vegetation (Bhuyan et al.
2003, Sagar et al. 2003, Arjunan et al. 2005, Raghubanshi & Tripathi 2009, Biswas & Mallik 2010, Hoang et al.
2011, Sutomo et al. 2015), diversity (Singh et al. 2003, Karkee 2004, Mishra et al. 2004, Kumar & Shahabuddin
2005, Mehta et al. 2008, Singh et al. 2008, Nagendra 2010, Sarkar & Devi 2014), richness (Mishra et al. 2004,
Kumar & Shahabuddin 2005, Tousignant et al. 2010, Tripathi et al. 2010, Sherma et al. 2014) and density
(Silori & Mishra 2001, Schwartz & Caro 2003, Mishra et al. 2004, Lalfakawma 2009, Sundarapandian &
Subbiah 2015). Because on individual basis, floristic composition is considered as one of the major
distinguishing feature of a community (Dansereau 1960) therefore any kind of depletion in biodiversity is bound
to change community structure (Mishra et al. 2004).
Kaimur wildlife sanctuary (KWS) is surrounded by more than 125 villages. Out of these, 20 villages are
present inside the sanctuary having human and cattle population 12,327 and 10265 respectively (Chandra 2010).
Because of biomass extraction, these types of forest are being converted to dry deciduous scrub, dry savannah
and dry grasslands (Champions & Seth classification 1968). This calls for an urgent need to research the
background factor responsible for species depletion and change in vegetation community structure. This
Sanctuary is home to a variety of wild species which are accorded different levels of protection according to the
142
Published online: 30 April 2016
Tahoor et al. (2016) 3(1): 142152

.
Indian Wildlife Protection Act, 1972. Changes in vegetation composition due to biomass extraction will
adversely affect the wildlife also. So the main objective of this study was to assess the factors responsible for
changes in vegetation community and the impacts thereof.
Study area
Kaimur wildlife sanctuary (KWS) is situated in Kaimur hills of Mirzapur and Sonbhadra district of Uttar
Pradesh (Fig. 1). It covers an area of 500 km2 in the semi-arid zone of northern India (Rodgers et al. 2000) with
geographical extent of 8220'15" E to 2452'00" N and 8308'23" E to 2427'51" N. For administrative
purpose, sanctuary is divided into four ranges namely Halia, Ghorawal, Robertsganj and Gurma. Geologically,
mostly part of the sanctuary is hilly and undulating terrains. Soil found in this area is red clay which is stiff and
ferruginous in nature. This Sanctuary faces three seasons namely summer (MarchJune), Monsoon (June
September) and winter (NovemberFebruary). The maximum/minimum temperature is 46.8 and 4C
respectively. According to Champions & Seth (1968) sanctuary comes under dry deciduous type forest. There
are four major types of forest forms in this sanctuary namely Sal forest, Bamboo, Scrub and Deciduous forest.
Sal forest comes under dry peninsular and enriched by Shorea robusta. The Bamboo forest consists of dry
Bamboo brades. The Scrub forest is accompanied by open dry scrubby vegetation. While, deciduous forest
consists of dry deciduous mixed woody species.
Figure 1. Map of India showing location of Kaimur wildlife sanctuary in Mirzapur and Sonbhadra districts of Uttar Pradesh.
Sampling sites
Through a reconnaissance survey, the area was stratified into high, medium and low disturbed categories on
the basis of indicators reflecting biomass extraction. Highly disturbed term was used for that area which faced
high biomass extraction on the basis of presence of human induced disturbance indicators namely cattle dung,
lopping, fuel wood collection and extraction of NTFPs. Low disturbed area termed on the basis of fewer signs of
biomass extraction. Medium disturbed term was used for that area faced neither high nor low biomass
extraction. These terms (high, medium and low disturbed area) which was used in this study was only for
analytical and research purpose.
Data collection
Within each stratified area (high, medium and low disturbed) line transects of 2 km was laid. On each
transect a circular plot of 10 meter radius at every 200 m (statistically independent samples) was placed on the
transect line. Tree layer was sampled within the 10 m radius of plot. Individuals of tree species with>30cm girth
at breast height and >3m height was considered as trees (Mueller-Dombois & Ellenbergh 1974). In each plot
tree species, tree individuals, girth at breast height of each tree was recorded. Canopy cover and tree height was
measured by ocular estimation. A nested 5 m circular plot was used to assess shrub layer and regeneration
pattern. Woody species with GBH <30cm and height <3m were considered as shrub (Mueller-Dombois &
Ellenbergh 1974). Species and number of individual were recorded. Regeneration pattern was assessed by
143
Tahoor et al. (2016) 3(1): 142152

.
recording individuals and species of seedling and sampling. The quantification of species and individuals of
herb and grass was measured by randomly laying 1m square quadrate at four locations within the larger one.
Point-Intercept method was used to assess ground cover (Mueller-Dombois & Ellenbergh 1974). For that, a rod
of 1m length having 20 marked points was laid randomly on the ground for five times by covering a total of 100
standard points, to record herb, grass, bare ground, litter and stone.
Data analysis
The characterization and dominance of vegetation is revealed by Importance Value Index (IVI) (Keel et al.
1993). The IVI was computed for each of the tree species by adding the relative values of frequency, density and
dominance (basal area) or dominance (Curtis & McIntosh 1950, Krebs 1989). Sorensons similarity index
(community coefficient) for similarity of plant species in the three categories of disturbed area was calculated
with the help of following formula (Jaccard 1912):
Cj = j(a+b-j)
where, Cj= Jaccard similarity coefficient, a=number of species in A area, b=number of species in
B area and j=number of species common to both the area.
Index of Dissimilarity = 1-Index of Similarity
Diversity indices of vegetation like species diversity, richness and evenness was analysed using
Paleontological Statistics Software Package for Education and Data Analysis (Hammer et al 2001, Hall 2005,
Bajpai et al. 2012). One way Analysis of Variance (ANOVA) was applied on different categories of disturbed
area (high, medium and low) to test the significant differences in community attributes of vegetation. Pearsons
Product Moment Correlation was used on habitat and disturbance variables with vegetation. Principal
Component Analysis (PCA) was used to reduce the dimensionality of vegetation variables by developing 2
uncorrelated Principal components. All the statistical analysis was performed on SPSS ver17.0 (Statistical
Package for Social Sciences).
RESULTS
A total of 57 plant species were found during vegetation sampling. Out of 57, there are only 30 tree species
recorded during the research period in the study area. In high, medium and low disturbed area, 28, 26 and 31
plant species were recorded including tree, shrub, herb and grass respectively. For woody species, IVI was
calculated. This index gives a clear idea about the importance of tree species in any area or habitat with the help
of three parameters namely dominance, frequency and density. In high disturbed area only 12 tree species were
recorded with highest and lowest IVI of Butea monosperma and Nyctanthes arbortristis (74.39 and 6.125
respectively). In the same area, highest density is of Holoptelea integrifolia whereas Bambusa arundinacea was
recorded highest for dominance. In medium disturbed area, the highest IVI along with dominance, frequency
and density was recorded for Shorea robusta. While in the same area, lowest IVI was recorded for Rewa. In low
disturbed area, the highest and lowest IVI were calculated for Cassia fistula and Ficus religiosa (84.19 and 5.74
respectively). On individually mentioning, Cassia fistula was the most dominant tree species in low disturbed
area (78.48) whereas Tectona grandis was recorded highest on population basis. Among the tree species
common to the three disturbed areas, the highest IVI is of Shorea robusta (116.46) from area facing medium
biomass extraction. The IVI of all the woody species in all the disturbed area is given in table 1.
Table 1. Importance Value Index (IVI) of tree species in high, medium and low disturbed area of Kaimur wildlife sanctuary.
Tree species
Cassia fistula
Terminalia arjuna
Terminalia elliptica
Acacia nilotica
Terminalia bellirica
Bambusa arundinacea
Bamboo sp.
Ficus benghalensis
High disturbed area

Dom
Fre
Den
IVI
0.26 12.723 12.723 25.712
0.549 9.087 9.0878 18.724
16.656 11.744 11.744 40.145
8.532 2.936 2.936 14.404
Medium disturbed area

Dom
Fre
Den
IVI
0.245 17.867 17.867 35.980
2.526 4.466 4.466 11.460
5.883 5.955 5.955 17.794
2.526 4.466 4.466 11.460
Low disturbed area

Dom
Fre
Den
IVI
78.486 2.855 2.856 84.198
0.062 2.855 2.856 5.774
1.1198 5.711 5.713 12.544
0.156 3.426 3.427 7.011
8.668 5.425 5.427 19.521
0.209 2.855 2.856 5.922
144
Holoptelea integrifolia 0.319 14.925 14.925

Buchanania lanzan
Delonix regia
Nyctanthes arbortristis 0.252 2.936 2.936
Abutilon indicum
#
Kaima
Bauhinia purpurea
Acacia catechu
Kurayya#
Madhuca indica
3.263 4.404 4.404
Makoicha#
Azadirachta indica
Butea monosperma
53.032 10.681 10.681
Ficus religiosa
11.922 2.936 2.936
Rewa#
#
Rimjim
Tectona grandis
Shorea robusta
0.448 6.850 6.850
Bombax ceiba
Cassia siamea
0.483 7.340 7.340
Albizia lebbeck
Diospyros melanoxylon 4.273 13.434 13.434
Note: Dom= Dominance, Fre= Frequency, Den=
Density are in percentage; # common name.
30.169
6.125
12.071
74.394
17.794
14.149
15.163
31.143
Density,
Tahoor et al. (2016) 3(1): 142152

.
1.188 4.2262 4.227 9.642
0.556 2.977 2.977 6.512
0.101 3.807 3.770 7.679
0.204 2.855 2.856 5.916
0.071 2.855 2.856 5.783
0.257 8.965 8.968 18.190
0.217 4.283 4.284 8.785
4.006 2.977 2.977 9.962
0.088 2.855 2.856 5.800
0.052 2.855 2.856 5.764
0.197 2.855 2.856 5.909
0.426 10.493 10.493 21.414 0.147 5.491 5.493 11.132
4.145 2.977 2.977 10.100 0.030 2.855 2.856 5.742
0.469 2.977 2.977 6.425
0.210 2.855 2.856 5.922
0.187 15.229 15.235 30.651
75.517 20.473 20.473 116.463 0.124 2.855 2.856 5.836
3.649 7.444 7.444 18.538 0.159 2.855 2.856 5.872
0.065 2.855 2.856 5.777
0.092 2.855 2.856 5.804
7.900 3.456 3.458 14.815
IVI= Importance value index; *Values of Dominance, Frequency and
The density, diversity, richness and evenness of tree species was found to be significantly different in high,
medium and low disturbed area (F2,198=31.770,p<0.01; F2,198=390.51,p<0.01 F2,198=450.56,p<0.01and
F2,198=332.34,p<0.01) respectively. The density, diversity, richness and evenness of tree species were found to
be highest in low disturbed area (131.487.777, 0.190.024, 0.180.024 and 0.120.015 respectively). The
shrub density was calculated lowest in the low disturbed area. The density of shrub was found to be significantly
in high, medium and low disturbed area (F2,198=33.334,p<0.01). The diversity and its attribute for herb and grass
layer were calculated highest in low disturbed area. The population of regeneration composition (sapling and
seedling) was also calculated highest in high disturbed area. The detail of density and diversity indices of plants
is provided in table 2.
Table 2. Density, diversity, richness and evenness of plants (trees, shrub, herb, grass, sapling and seedling) in high, medium and
low disturbed area of Kaimur wildlife sanctuary. (Values are in MeanStandard Error)
Area
Vegetation Variable
Tree density
Tree diversity
Tree richness
Tree evenness
Shrub density
Shrub diversity
Shrub richness
Shrub evenness
Herb diversity
Herb richness
Herb evenness
Grass diversity
Grass richness
High disturbed
area
131.487.777
0.190.024
0.180.024
0.120.015
281.8140.494
00
00
00
0.050.035
0.050.035
0.050.034
0.220.021
0.100.011
Medium disturbed
area
143.7812.316
0.230.029
0.220.032
0.150.017
1.901.90
0.010.004
0.010.006
00.003
0.010.005
0.010.002
00.002
0.100.018
0.050.009
Low or minimal
disturbed area
230.417.198
1.270.039
1.750.062
0.640.016
86.4611.956
0.040.012
0.050.0017
0.020.008
0.310.025
0.170.012
0.200.016
0.030.007
0.020.005
Overall
166.895.278
0.540.027
0.690.038
0.290.013
125.5995.49
0.010.004
0.020.006
0.010.003
0.120.015
0.0070.013
0.080.013
0.120.010
0.060.005
145
Grass evenness
Sapling density
Sapling diversity
Sapling richness
Sapling evenness
Seedling density
Seedling diversity
Seedling richness
Seedling evenness
0.150.0015
1299.03122.156
0.200.023
0.150.019
0.130.015
610.5864.943
0.140.020
0.160.025
0.090.014
0.060.012
350.2525
0.030.01
0.030.0010
0.020.007
579.5347.844
0.130.018
0.130.019
0.080.012
Tahoor et al. (2016) 3(1): 142152

.
0.020.004
0.080.013
222.1125.384
638.2549.039
0.230.032
0.150.014
0.250.034
0.140.014
0.160.026
0.100.010
134.0813.296
449.5329.318
0.160.026
0.140.012
0.230.033
0.170.015
0.120.016
0.100.008
The Sorensons Index of dissimilarity between high-medium, medium-low and high-low disturbed areas
were found to be 0.7, 0.75 and 0.8125 respectively (Table 3). A total of 9 habitat variables were selected to
observe association with density and diversity attributes of vegetation layers. The density and diversity aspects
of tree species showed significant negative correlation with elevation (r=-0.299, p=0.01; r=-0.724.p=0.01; r=0.718, p=0.01 and r=-0.718, p=0.01 respectively). With increase in elevation; the density, diversity, richness,
evenness and mean tree height reduced. On the other hand, density of shrub and regenerating composition
(sapling and seedling) had significant positive correlation with elevation (r=0.182, p=0.01; r=0.299, p=0.01 and
r=0.237, p=0.01 correspondingly). But the diversity, richness and evenness of regenerating constituents
decreased as the elevation level rises. The diversity indices of ground canopy i.e. grass and herb showed positive
and as well as negative association with the elevation respectively. The vegetation layer showed positive
association with the distance from human settlements. As the distance from villages increased the population of
tree increased. The diversity components of tree, shrub, herb and sapling showed significant positive association
with the village distance. As the distance from water-body increased, population of tree decreased along with
diversity aspects. The population of shrub was negatively associated with the canopy cover (r=-0.180, p=0.01).
The grass cover showed positive association with top canopy and ground canopy (herb diversity and richness).
But for regeneration component, grass showed negative association especially with population of sapling (r=0.108, p=0.01).
Table 3. Sorensons Index of similarity and dissimilarity of plant species among high, medium and low disturbed areas in
Kaimur wildlife sanctuary.
Areas
Index of Dissimilarity
High and Medium disturbed area 0.7(70)
Medium and Low disturbed area
0.75(75)
High and Low disturbed area
0.8125(81.25)
Note: Values in parenthesis are in percentage.
Index of Similarity
0.3(30)
0.25(25)
0.1875(18.75)
The correlation between vegetation and habitat variables is given in table 4. The disturbance variables
selected in the present study are human trail, cattle dung density, grazing cover, weed density, weed cover,
lopping density, mean lop score and fire. Human trail showed negative association with the diversity, richness
and evenness of tree species (r=-0.204, p=0.01; r=-0.208, p=0.01 and r=-0.192, p=0.01 respectively). Similarly,
the density of sapling and seedling was positively associated with human trail (r=0.281, p=0.01 and r=0.210,
p=0.01 respectively). The density of sapling and seedling was positively associated with human trail (r=0.281,
p=0.01 and r=0.210, p=0.01 respectively). Whereas; diversity, richness and evenness of sapling species showed
significant negative correlation with human trail (r=-0.096, p=0.05; r=-0.096, p=0.05 and r=-0.095, p=0.05)
respectively. The grazing cover was significantly negatively associated with density, diversity, richness and
evenness of tree species (r=-0.111, p=0.01; r=-0.147, p=0.001; r=-0.155, p=0.01 and r=-0.136, p=0.01
respectively). But for ground layer, grazing cover was significantly positively associated especially with the
diversity, richness and evenness of grass (r=0.211, p=0.01; r=0.233, p=0.01 and r=0.200, p=0.01 respectively).
A significant negative correlation was calculated between diversity, richness and evenness of sapling and
livestock grazing (r=-0.85, p=0.05; r=-0.89, p=0.05 and r=-0.083, p=0.05) correspondingly. Weed cover showed
a negative association with the diversity, richness and evenness grass (r=-0.011, p=0.01; r=-0.095, p=0.05 and
r=-0.119, p=0.05 respectively). The lopping density showed a positive association with the population of sapling
(r=-0.09, p=0.05). Fire is also another threat to fragile plants especially seedlings, grass and h erb. But in the
146
Tahoor et al. (2016) 3(1): 142152

.
Table 4. Correlation of density, diversity, richness and evenness of plants (trees, shrub, herb, grass, sapling and seedling) with habitat
variables in Kaimur wildlife sanctuary.
Habitat
variable
Vegetation variable
Elevation
(m)
DFHH
(km)
DFWB
(km)
Canopy
cover
(%)
Shrub
cover
(%)
Herb
cover
(%)
Grass Mean tree Mean tree

cover
height
GBH
(%)
(m)
(m)
Tree density
-0.299**
0.078
-0.277** 0.548**
0.007
0.214** 0.267**
Tree diversity
-0.724**
0.583** -0.314** 0.637** -0.091* 0.500** 0.450**
Tree richness
-0.718**
0.646** -0.315** 0.583** -0.08** 0.524** 0.460**
Tree evenness
-0.718**
0.530** -0.312** 0.667**
-0.069
0.478** 0.452**
Shrub density
0.182**
-0.065
0.288** -0.180** -0.319**
-0.042
-0.044
Shrub diversity
-0.145**
0.126**
-0.037
0.102*
-0.037
0.023
-0.097*
Shrub richness
-0.142**
0.126**
-0.037
0.102*
-0.036
0.023
-0.096*
Shrub evenness
-0.150**
0.126**
-0.036
0.101
-0.038
0.028
-0.1*
Herb diversity
-0.278**
0.124** -0.0131** 0.185**
-0.048
0.230** 0.228**
Herb richness
-0.147**
0.323** -0.083** 0.108**
-0.041
0.136** 0.121**
Herb evenness
-0.051
0.192**
-0.026
0.026
-0.011
0.014
0.023
Grass diversity
0.199**
-0.210** 0.261**
0.032
-0.010
-0.136**
-0.011
Grass richness
0.170**
-0.160** 0.204**
0.031
-0.017
-0.109**
0.001
Grass evenness
0.214**
-0.216** 0.283**
0.030
-0.006
-0.142**
-0.015
Sapling density
0.299**
-0.284** 0.188**
-0.057
0.043
-0.162** -0.108**
Sapling diversity
-0.104*
0.104*
0.124**
0.010
0.179**
0.023
0.043
Sapling richness
-0.182**
0.158**
0.076
0.041
0.162**
0.067
0.081*
Sapling evenness
-0.101*
0.098*
0.108**
0.011
0.166**
0.029
0.039
Seedling density
0.237**
-0.367** 0.103*
-0.037
-0.018
-0.204** -0.092*
Seedling diversity
-0.085*
-0.020
-0.083*
0.063
0.030
-0.023
0.026
Seedling richness
-0.115**
0.029
-0.059
0.64
0.026
0.004
0.069
Seedling evenness
-0.103*
-0.001
-0.081
0.67
-0.032
-0.012
0.040
Note: DFHH= Distance from human habitation, DFWB= Distance from water body, GBH= Girth at
kilometres and m= metres; ** Correlation significant at 0.001 level, *Correlation significant at 0.05 level.
0.465** 0.129**
0.485** 0.072
0.408** 0.068
0.544** 0.072
-0.131** 0.106**
0.116** 0.026
0.115** 0.029
0.127** 0.031
0.041
0.037
0.017
0.011
0.017
0.013
0.108** -0.065
0.078
-0.077
0.119** -0.061
-0.111** 0.048
0.031
0.050
0.050
0.049
0.025
0.048
0.087* -0.007
0.153** 0.039
0.143** 0.036
0.151** 0.034
Breast height, km=
Figure 2. Presentation of habitat variables by first two components in PCA in Kaimur wildlife sanctuary. (Component Plot
in Rotated Space)
147
Tahoor et al. (2016) 3(1): 142152

.
present findings, fire was positively correlated with diversity indices of tree, sapling and seedling layer. The
correlation between vegetation and disturbance variables is given in table 5. The outcome of PCA shows that
Principal Component 1 and 2 explained 17.40% and 11.39% of variance (Fig. 2). Thus cumulatively total
variance explained by first two components is 28.81%. The first component showed heavy loadings for density,
diversity, richness and evenness of tree, tree height, herb density, herb diversity, herb cover, grass cover, canopy
cover and distance from human habitation coupled with low lopping density and cow dung density. The
component 2 showed heavy loadings for sapling diversity, sapling richness, sapling evenness, seedling diversity,
seedling richness, seedling evenness together with low weed density and herb diversity (Table 6).
Table 5. Correlation of density, diversity, richness and evenness of plants (trees, shrub, herb, grass, sapling and
seedling) with disturbance gradients in Kaimur wildlife sanctuary.
Disturbance
Human Cattle
gradients
Trail
dung
density
Vegetation
(ha)
variables
Tree density
-0.035
-0.069
Tree diversity
-0.204** -0.075
Tree richness
-0.208** -0.070
Tree evenness
-0.192** -0.078
Shrub density
-0.124** -0.018
Shrub diversity
-0.078
-0.016
Shrub richness
-0.078
-0.016
Shrub evenness
-0.081* -0.016
Herb diversity
-0.079
-0.067
Herb richness
-0.011
0.032
Herb evenness
-0.023
-0.005
Grass diversity
0.284** 0.040
Grass richness
0.262** 0.035
Grass evenness
0.279** 0.036
Sapling density
0.281** -0.035
Sapling diversity
-0.096* -0.032
Sapling richness
-0.096* -0.031
Sapling evenness
0.095*
-0.031
Seedling density
0.210** -0.011
Seedling diversity
0.020
0.004
Seedling richness
0.01
0.005
Seedling evenness 0.014
0.005
Note: %= Percentage, ha= Hectare; **
0.05 level.
Grazing
cover
(%)
Weed
density
(ha)
Weed
cover
(%)
Lopping
Mean
density Lopping
(ha)
score
Fire
-0.111**
0.066
0.041
0.140** 0.135**
0.007
-0.147** 0.166**
0.085*
-0.102* -0.115** 0.151**
-0.155** 0.205** 0.116** -0.117** -0.130** 0.179**
-0136** 0.149**
0.072
-0.093* -0.104* 0.155**
-0.089* -0.101* -0.081*
0.0
0.005
-0.039
-0.052
-0.060
-0.048
-0.058
-0.058
-0.016
-0.051
-0.060
-0.048
-0.058
-0.057
-0.016
-0.053
-0.062
-0.050
-0.060
-0.060
-0.017
-0.058
0.134**
0.086*
-0.001
-0.069
-0.037
-0.018
0.078
0.047
0.047
-0.039
-0.026
-0.015
-0.018
-0.014
-0.017
-0.017
-0.005
0.211** -0.0135** -0.0115** 0.029
0.033
-0.055
0.233** -0.107** -0.095*
0.039
0.043
-0.054
0.200** -0.142** -0.119**
0.029
0.033
-0.054
0.028
-0.172** -0.135* 0.124** 0.117**
0.011
-0.085* -0.090*
-0.079
0.006
-0.011
0.239**
-0.089*
-0.077
-0.069
0.025
0.006
0.278**
-0.083*
-0.58
-0.055
-0.008
-0.021
0.193**
0.092*
-0.053
-0.046
0.127** 0.125**
0.006
0.017
-0.081*
-0.066
-0.048
-0.047
0.097*
0.008
-0.096*
-0.078
-0.011
-0.010
0.100*
0.014
-0.078
-0.064
-0.042
-0.041
0099*
Correlation significant at 0.001 level, *Correlation significant at

In the present study the density and diversity of woody species in highly disturbed area were recorded
lowest. The present findings are similar to the studies done previously (Bhuyan et al. 2003, Sagar et al. 2003,
Arjunan et al. 2005, Mishra et al. 2008, Tripathi et al. 2008 Sahoo & Davidar 2013, Bajpai et al. 2015). Though
the above authors had done their studies in different types of forests stands but the findings are similar to the
present one. Like Kaimur, other studies in different PAs of same habitat had similar findings (Mueller-Dombois
& Ellenbergh 1974, Pandey & Shukla 2001, Kumar & Shahabuddin 2005, Shahabuddin & Kumar 2006, Sharma
& Raghubanshi 2006, Mueller-Dombois & Ellenbergh 1974, Tripathi et al. 2010). However some authors
favors mild human disturbance for plant species richness. For example- Mishra et al. (2004) in their study found
mild disturbance favors species richness but tree density was higher in low disturbed area. Likewise, Mishra et
al. (2008) found higher shrub and herb in high disturb area. Whereas, Tousignant (2010) found plant species
richness is negatively associated with disturbance variables. Similarly in the present study, richness of plant
species was lower in areas facing disturbance. With the increase in the elevation, human disturbance increased
148
Tahoor et al. (2016) 3(1): 142152

.
with a simultaneous decrease in tree population and mean tree height. Several studies conclude that at lower
elevation species richness is higher as compared to higher elevated areas (Rawal et al. 1991, Singh et al. 1994,
Sharma et al. 2009). The population of shrub was negatively associated with canopy cover because dense
canopy suppress the growth of lower strata by hindering sunlight (Sharma & Raghubanshi 2005). In the present
study, livestock grazing individually affects trees; sapling population and composition (density, diversity,
richness and evenness). But positive association with diversity, richness and evenness of grass is because due to
the fact that livestock helps in immigration of new species with tangling seeds in their hooves. But impact of
livestock grazing is a debate issue. Because some authors favors grazing for improvement of vegetation
composition in terms of plant species richness, diversity. For example- Olff & Ritchie (1992) reported increased
species diversity due to livestock grazing in grassland; Cooper (1960), Facelli (1994), Pearson (1934) and
Madany & Neil (1983) concluded that grazing improves the establishment of recruiting vegetation composition;
whereas Leopold (1924) found that livestock grazing would decrease the ground layer making the area naturally
as fire line or breaks.
Table 6. Rotated Principal Component matrix of vegetation variables in Kaimur wildlife sanctuary.
Variables
Tree density
Tree richness
Tree diversity
Tree evenness
Tree height
Sapling diversity
Sapling richness
Sapling evenness
Seedling diversity
Seedling richness
Seedling evenness
Distance from human habitation
Herb density
Herb cover
Grass cover
Canopy cover
Herb diversity
Herb richness
Weed density
Lopping density
Cow dung density
Component 1
0.540
0.852
0.829
0.89
0.515
0.556
0.777
0.716
0.631
0.640
0.629
0.548
0.510
-0.117
-0.243
Component 2
0.149
0.249
0.272
0.294
0.193
0.815
0.805
0.802
0.702
0.727
0.710
0.144
0.114
0.146
-0.110
-0.252
Weed presence was negatively associated with sapling and seedling because dense canopy created by
vertically could reduce the receiving amount of sunlight and so suppress the growth of regenerating species
(Sharma & Raghubanshi 2006). Weed was negatively associated with herbaceous vegetation because weed
presence would add woody debris and more litter to ground making it less favorable for growth of herbaceous
vegetation (Sharma & Raghubanshi, 2010).
Those sampling plots faced high level of tree cuts had canopy opening which may favored the growth of
sapling and seedling. This is so because of canopy opening or canopy gap, species received maximum amount
of sunlight and water which are important elements for growth and development of plants. Seng et al. (2004)
favored logging for regenerating species. Similarly, in the present study sapling and seedling density was
calculated highest in high disturbed area. Few literatures are also available on similar results favoring
regeneration composition due to human disturbance (Pandey & Shukla 2001, Buffum et al 2009, Muhanguzi
2009, Sharma & Raghubanshi 2010, Tripathi et al. 2010). Whereas Tripathi et al. (2008) found lower
regeneration population in disturbed area and reasoned human disturbance.
In the present study, presence of fire signs in the plot along with seedling indicates its post effects. Because
some of the seeds are dormant in nature, post adverse when received favorable conditions they germinate. These
disturbance variables alter the habitat and make it less favorable for growth of plant species. During the study
period, some of the species with only one individual was sampled which shows vulnerability on the basis of its
occurrence. And when there is very small number of population then it would have high chances of extinction
locally. In the present study very low density of some of the seedling illustrates poor generation of that
149
Tahoor et al. (2016) 3(1): 142152

.
respective species. Some of the species occurred as only seedling and sapling which shows that these species are
newly immigrants to that area. But some species were present as only woody species lacking sapling and
seedling which shows nil regeneration. Bhuyan et al. (2003) conclude that species lacking regenerative layer are
expected to face extinction locally in the coming future. In a given population, an equal or equivalent proportion
of tree, sapling and seedling may help in predicting its possible future status (Saxena & Singh 1984). Whereas in
the present study, none of the species was fit to the criteria given by Saxena & Singh (1984). On the basis of
present findings I believe that the reason behind instability of plants species of Kaimur wildlife sanctuary is
illegal biomass extraction done on larger scale by nearby villagers. Additionally, the Sorensons Index of
similarity indicates that dissimilarity of plant species increased with degree of disturbance i.e. from low to high
disturbed area. This suggests that human disturbance alters the vegetation community in the study area.
Similarly, the outcome of PCA in the present study conclude that livestock grazing and illegal tree lopping for
fuelwood collection are the main disturbing factors responsible for changes in vegetation community structure.
For example tree density, tree diversity, tree height, tree GBH, canopy cover, shrub density, shrub diversity, and
sapling diversity showed negative association to disturbance gradients.
Many of the plant species dont have minimum viable populations which in the coming years may face
extinction locally. Immediate actions are required in the KWS to conserve plant species facing biomass
extraction. Plantation of fast growing fuelwood and fodder plant species should be encouraged. This would
cover and compensate the needs of local people for forest resources. Land should be provided to these villagers
where agro-forestry or agro-farming like activities can be carried out. Crop-rotation method should be implied
for agricultural practices. Participation of local people along with large scale stakeholders are needed in the area.
Central and State government should launch various schemes to absorb local people in small scale industries.
State Forest Department and local NGOs should start awareness programs related to biodiversity conservation in
the sanctuary. Animal husbandry programs should be started for best and lesser number of cattle. For livestock,
stall feeding should be encouraged instead of illegal grazing inside the Sanctuary.
ACKNOWLEDGEMENT
This work was supported by grants provided by University Grants Commission, New Delhi. Authors are
thankful to the Forest Department, Uttar Pradesh for providing logistic support. Authors are thankful to Siraj
Mazumdar for giving valuable comments on the manuscript.
REFERENCES
Arjunan M, Puyravaud JP & Davidar P (2005) The impact of resource collection by local communities in dry
forest of Kalakud-Mundanthurai Tiger Reserve. Tropical Ecology 46(2): 135143.
Bajpai O, Kumar A, Mishra AK, Sahu N, Pandey J, Behera SK & Chaudhary LB (2012) Recongregation of tree
species of Katerniaghat Wildlife Sanctuary, Uttar Pradesh, India. Journal of Biodiversity and Environmental
Sciences 2: 2440.
Bajpai O, Kushwaha AK, Srivastava AK, Pandey J & Chaudhary LB (2015) Phytosociological status of a
monotypic genus Indopiptadenia: A near threatened tree from the Terai-Bhabar region of Central Himalaya.
Research Journal of Forestry 9(2): 3547.
Bhuyan P, Khan ML & Tripathi RS (2003) Tree diversity and population structure in undisturbed and human
impacted stands of tropical wet evergreen forest in Arunachal Pradesh, Eastern Himalayas, India.
Biodiversity and Conservation 12: 17531773.
Buffum B, Gratzer G & Tenzin Y (2009) Forest grazing and natural regeneration in a late successional broad
leaved community forest in Bhutan. Mountain Research and Development 29: 3035.
Champion HG & Seth SK (1968) A revised survey of the forest types of India. Government of India
Publications, Delhi, 200 p.
Chandra S (2010) Management Plan for Kaimur Wildlife Sanctuary. Wildlife Preservation Organization Forest
Department, Lucknow 98 p.
Cooper CF (1960) Changes in vegetation structure and growth of South Western Pine Forests since White
settlement. Ecological Monograph 30: 129164.
Curtis JT & McIntosh RP (1950) The interrelations of certain analytic and synthetic phytosociological
characters. Ecology 31: 434455.
150
Tahoor et al. (2016) 3(1): 142152

.
Damsereau (1960) The origin and growth of plant communities. In: Zarrow MX (ed) Growth in living systems:
Proceedings of International Symposium on growth. Purdue University, Indiana. Basic Books, New York
109 p.
Facelli JM (1994) Multiple indirect effects of plant litter affect establishment of woody seedlings on Old Field
Ecology. Ecology 75: 17271735.
Hall A (2005) The environ-mental gradients and plant communities of Bergen swamp, M.Sc. Thesis. Rochester
Institute of Technology, New York, USA.
Hammer O, Harper DAT & Ryan PD (2001) PAST: Plaentological staticitcs software Package for education and
Data Analysis. Palaentologia Electronica 4: 9 p.
Hoang VS, Baas P, Kebler PJA, Slik JWF, Steege HT & Raes (2011) Human and environmental influences on
plant diversity and composition in Ben En National park, Veitnam. Journal of Tropical Forest Science 23:
328337.
Jaccard P (1912) The distribution of the flora of Alpine zone. New Phytologist 11: 3750.
Karkee K (2004) Effects of deforestation on tree diversity and livelihoods of local community: a case study from
Nepal, M.Sc. Thesis. Tribhuwan University, Nepal.
Keel SA, Gentry H & Spinzi L (1993) Using vegetation analysis to facilitate the selection of conservation sites
in Eastern Paraguay. Conservation Biology 7: 6673.
Kothari A, Singh N & Suri S (1995) Conservation in India: a new direction. Economic and Political Weekly
30(43): 27552766.
Krebs CJ (1989) Ecological Methodology 2nd Edition. Harper and Row, New York, USA, 654 p.
Kumar R & Shahabuddin G (2005) Effects of biomass extraction on vegetation structure, diversity and
composition of forests in Sariska Tiger Reserve, India. Environmental Conservation 32: 248259.
Lalfakwma U, Sahoo K, Roy S, Vanlalhriatpuia K & Vanlalhluna P C (2009) Community composition and tree
population structure in undisturbed and disturbed tropical semi-evergreen forest stands of North-east India.
Applied Ecology and Environmental Research 7: 303318.
Leopold A (1924) Grass, bush, timber and fire in Northern Arizona. Journal of Forestry 22: 110.
Madany MH & Neil EW (1983) Livestock grazing- fire regime interactions within Montane Forest of Zion
National Park, Utah. Ecology 64: 661667.
Mishra AK, Upadhyaya VP & Mohanty RC (2008). Vegetation ecology of Simlipal Biosphere Reserve, Orissa,
India. Applied Ecology and Environmental Research 6(2): 8999.
Mishra BP, Tripathi OP, Tripathi RS & Pandey HN (2004) Effects of anthropogenic disturbance on plant
diversity and community structure of a sacred grove in Meghalaya, North-east India. Biodiversity and
Mueller-Dombois D & Ellenbergh H (1974) Aims and Methods of Vegetation Ecology. Wiley, New York Wiley
and Sons 135 p.
Muhanguzi DRH, Obua J, Oreym-Origa H & Vetaas OR (2005) Forest sites disturbance and seedling emergence
in Kalinzu forest, Uganda. Tropical Ecology 46: 9198.
Olff H & Ritchie ME (1998) Effects of herbivore on grassland plant diversity. Tree 13: 261265.
Pandey SK & Shukla SP (2001) Regeneration strategy and plant diversity status in degraded Sal forest. Current
Science 81: 95102.
Pearson GA (1934) Grass, pine seedlings and grazing. Journal of Forestry 32: 545555.
Rabha D (2014) Species composition and structure of Sal (Shorea robusta Gaertn. f.) forests along distribution
gradients of Western Assam, Northeast India. Tropical Plant Research 1(3): 1621.
Raghubanshi AS & Tripathi A (2009) Effect of disturbance, habitat fragmentation and alien invasive plants on
floral diversity in dry tropical forest of Vindhyan highlands: a review. Tropical Ecology 50: 5769.
Rawal RS, Bankoti NS, Samant SS & Pangtey YPS (1991) Phenology of tree layer species from the timberline
around Kumaun in Central Himalaya, India. Vegetatio 93: 109118.
Sagar R & Singh JS (2006) Local plant species depletion in a tropical dry deciduous forest of northern India.
Environmental Conservation 33: 256262.
Sagar R, Raghubanshi AS & Singh JS (2003) Tree species composition, dispersion and diversity along
disturbance gradient in a dry tropical forest of India. Forest Ecology and Management. 186: 6171.
151
Tahoor et al. (2016) 3(1): 142152

.
Sahoo S & Davidar P (2013) Effect of harvesting pressure on plant diversity and vegetation structure of Sal
forests in Simlipal Tiger Reserve. Tropical Ecology 54: 97107.
Sarkar M & Devi A (2014) Assessment of diversity, population structure and regeneration status of tree species
in Hollongapar Gibbon Wildlife Sanctuary, Assam, Northeast India. Tropical Plant Research 1(2): 2636.
Saxena AK & Singh JS (1984) Tree population structure in certain Himalayan forest associations and
implication concerning their future composition. Vegetatio 51: 6169.
Schwartz MW & Caro TM (2003) Effect of selective looging on tree and understorey regeneration in miombo
woodland in Western Tanzania. African Journal of Ecology 41: 7582.
Seng HW, Ratnam W, Noor SM & Clyde MM (2004) The effects of timing and method of logging on forest
structure in Peninsular Malaysia. Forest Ecology and Management 203: 209228.
Shahabuddin G & Kumar R (2006) Influence on anthropogenic disturbance on bird communities in a Tropical
Dry Forest: role of vegetation structure. Animal Conservation 9: 403414.
Shahabuddin G & Prasad S (2004) Assessing ecological sustainability of non-timber forest produce extraction:
the Indian scenario. Conservation and Society 2: 235250.
Sharma CM, Mishra AK, Prakash O, Dimri S & Baluni P (2014) Assessment of forest structure and woody
plant regeneration on ridge tops at upper Bhagirathi basin in Garhwal Himalaya. Tropical Plant Research
1(3): 6271.
Sharma CM, Suyal S, Gairola S & Gildhiyal SK (2009) Species richness and diversity along an altitudinal
gradient in moist temperate forest of Gharwal Himalaya. Journal of American Science 5: 119128.
Sharma G & Raghubanshi AS (2010) How Lantana invades dry deciduous forest: a case study from Vindhyan
highlands, India. Tropical Ecology 51: 305316.
Sharma GP & Raghubanshi AS (2009) Tree population structure, regeneration and expected future composition
at different levels of Lantana camara L. invasion in the Vindhyan tropical dry deciduous forest of India.
Lyonia 11: 2739.
Singh HB, Sundariyal RC & Sharma E (2003) Livestock grazing in the Khangchendzonga Biosphere Reserve of
Sikkim Himalaya, India: implications for management. The Indian Forester 4: 612623.
Singh SP, Adhikari BS & Zobel DB (1994) Biomass productivity, leaf longevity and forest structure in Central
Himalaya. Ecological Monograph 64: 401421.
Sundarapandian SM & Subbiah S (2015) Diversity and tree population structure of tropical dry evergreen
forests in Sivagangai district of Tamil Nadu, India. Tropical Plant Research 2(1): 3646.
Sutomo, Hobbs RJ & Cramer VA (2015) Plant Community Structure and Composition in Secondary Succession
Following Wildfire from Nues Ardentes of mount Merapi, Indonesia. Tropical Plant Research 2(3): 204
214.
Tousignant M, Pellerin S & Brisson J (2010) The relative impact of human disturbances on the vegetation of a
large Wetland complex. Wetland 30: 333344.
Tripathi OP, Pandey HN & Tripathi RS (2008) Effects of human activities on structure and composition of
woody species of the Nokrek Biosphere Reserve of Meghalaya, North-East India. Journal of Plant Ecology
32: 7379.
Tripathi OP, Upadhyaya K, Tripathi RS & Pandey HN (2010) Diversity, dominance and population structure of
tree species along fragment size gradients of a subtropical humid forest of North-east India. Research
Journal of Environmental and Earth Sciences 2: 97105.
152
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 153156, 2016
Short communication
New addition to lichen flora of Uttar Pradesh, India

Namita Gupta1*, Vartika Gupta2 and S. K. Dwivedi1
1
Department of Environmental Science, Babasaheb Bhimrao Ambedkar (A Central) University, Lucknow (U.P.)
2
Department of Environmental Science, Dr. R. M. L. Avadh University, Faizabad (U.P.)
*Corresponding Author: namitag09@gmail.com
[Cite as: Gupta N, Gupta V & Dwivedi SK (2016) New addition to lichen flora of Uttar Pradesh, India.
The lichen taxa collected from Uttar Pradesh are documented in different checklist, floristic, monographic
and revisionary studies (Awasthi, 1980, 1988, 1991, 2000, 2007, Srivastava, 2004, Dubey et al. 2007, Singh &
Sinha 2010, Karakoti et al. 2014, Gupta et al. 2015). Recently Nayaka & Upreti (2013) analyzed the status of
lichen diversity in Uttar Pradesh which revealed the occurrence of 135 species belonging to 46 genera and 25
families.
This state represented three distinct phytogeographical regions. The transitional belt running along the entire
length of the state of Uttarakhand and country of Nepal is called the Terai and Bhabhar area and have thick
forest cover, swamps and marshes. The Gangetic plain elongates the area from east to west is the most fertile as
well as agricultural land. The southern fringe of the Gangetic Plains is demarcated by the Vindhya Hills and
Plateau exhibit strong ground and low hills. Most of the central region of the state of Uttar Pradesh is most
fertile and utilized for agriculture from the ancient time. The region is devoid of forest, however, mango
orchards are quite common and provide suitable habitat for many lichen taxa to colonize.
The present investigation is carried out with an aim to document the lichen diversity pattern in mango
orchards of Gangetic plain. Three districts of this phytogeographical zone viz. Faizabad, Ambedkar Nagar and
Raebareli have been selected to conduct the present study. The identification of the lichen samples collected
revealed occurrence of five species as new addition to the lichen flora of the state.
The mango orchards in and around Tanda Thermal Power plant, Ambedkar Nagar (lies between coordinates
263300 N and 823900 E); Feroz Gandhi Unchahar National Thermal Power Plant Corporation, Raebareli
(between coordinates 2549 to 2636 N and 10041 to 8134 E and Faizabad district (situated at the latitude
2647N and longitude 8212 E) was surveyed for collection of lichens.
The collected specimens were identified by their morphological, anatomical and chemical characters and
specimens were preserved in the herbarium of CSIR-National Botanical Research Institute, Lucknow (LWG).
The LABOMED dissecting microscope was used for external morphology study while LEICA ATC 2000
compound microscope was used for microscopic anatomical details. The samples were mounted in water, 10%
KOH and Lugols solution. Measurements of asci and ascospores were made on material examined in KOH.
The colour test and Thin layer chromatography (TLC) of acetone extracts was performed using solvent system
A and C, followed by Orange et al. (2001), Culberson (1972) and Walker and James (1980). The microscopic
measurements were based on mature ascomata and are recorded for their minimum and maximum values.
Species description
1. Anisomeridium subnexum (Nyl.) R.C. Harris, More Florida Lich. (New York):150. 1995.
(Fig. 1A)
Arthopyrenia subnexa (Nyl.) Mll. Arg. Hedwigia. 30: 188. 1891. (Monoblastiaceae)
Thallus corticolous, crustose, yellow-grey, smooth, shining sometimes powdery, endophloeodal. Ascomata
solitary, 0.250.45 (0.50) mm diam., (0.10) 0.150.20 mm high, convex- hemispherical, globose completely
covered with thallus or naked around ostiole and black; ostiole indistinct, plane or sometimes slightly depressed;
centrum I-; pseudoparaphyses branched, anastomosed; asci cylindrical clavate,8-spored (90) 100161522
m, uniseriate or rarely biseriate; ascospore colourless, 1-septate, 2327(7) 911 m, oblong-ellipsoid,both
cells equal in size, slightly constricted at septum, epispore to 1 m thick. Pycnidia not seen. Thallus K-, C-, KC, P-, no lichen substance upon TLC.
153
Gupta et al. (2016) 3(1): 153156

.
Remarks: Hue (1892) reported this species from Andaman and Nicobar Islands and later recorded from
Madhya Pradesh and Karnataka (Upreti and Pant, 1993). The species is rare in the area, as it is known from a
single locality from the outskirts of the district Faizabad.
Specimen Examined: Faizabad district: Azamgarh road, Purabazar, Barauli, on tree trunk of Mangifera indica,
19th March, 2014, V. Gupta. 014-022614 (LWG).
2. Arthothelium chiodectoides (Nyl.) Zahlbr., Cat. Lich. Univ. 2: 122. 1922.
(Fig. 1B)
Arthonia chiodectoides Nyl. Flora 52: 72. 1869. (Arthoniaceae)
Thallus corticolous, crustose, most of the part endophloeodal, 88-166 m thick; ascomata yellowish brown
to dark blackish-brown, K-, punctuate, aggregated, covered with effuse thalline layer or naked; epithecium dark
blackish-brown; hymenium pale brown 100185 m tall, I+ blue; hypothecium dark blackish-brown; asci
butinicate, obovate to pyriform; paraphysoids profusely branched and anatomosed, strongly coherent; ascospore
8/ascus, hyaline, muriform, ovate to oblong, transversely 7 to 9-septate, vertically 1 to 3-septate, upper most cell
larger, undivided, 283635 m.
Chemistry: No chemical tested. Triterpenes detected upon TLC.
Remarks: This species is reported from Arunachal Pradesh, Goa, Himachal Pradesh, Karnataka, Maharashtra,
Sikkim and West Bengal. This species colonize on the bark of Mangifera indica, Azadirachta indica and Litchi
chinensis. Now, for the first time it is reported from Uttar Pradesh.
Specimens Examined: Faizabad district: Azamgarh road: Rajepur, on tree trunk of Mangifera indica, 19th
March, 2014, V. Gupta. 014-022610 (LWG); Purabazar, Jillu ka Purwa, on tree trunk of Mangifera indica, 19th
March, 2014, V. Gupta. 014-022615 (LWG). Sultanpur road: Khanpur, 6 km. away from the city, on bark of
Litchi chinensis, 20th March, 2014, V. Gupta. 014-022656 (LWG); Bikapur, Sugan rai ka purwa, on bark of
Mangifera indica, 20th March, 2014, V. Gupta. 014-022659 (LWG). Gonda road: Nawabganj, Ghuse ka purwa,
on tree trunk of Mangifera indica, 21st March, 2014, V. Gupta. 014-022707 (LWG); Birapur, on bark of
Azadirachta indica, 21st March, 2014, V. Gupta. 014-022708 (LWG). Raebareli road: Ranibazar, Roadside, on
tree trunk of Mangifera indica, 22nd March, 2014, V. Gupta. 014-022731 (LWG); Barun bazaar, Mahaveer
Mishra ka purwa, on tree trunk of Mangifera indica, 22nd March, 2014, V. Gupta. 014-022740 (LWG).
3. Bacidia medialis (Tuck. ex Nyl.) Zahlbr., Denskchr. Kaiserl. Akad. Wiss., Wien, Math.- Naturwiss. Kl. 83:
127. 1909.
(Fig. 1C)
Lecidea medialis Tuck. Zahlbruckner's Cat. Lich. Univ. 4: 221. (Ramalinaceae)
Thallus corticolous, crustose effuse, rough, cracked, granuolose-furfaraceous, greyish brown to grey, 5070
m thick, Apothecia constricted at base, 0.20.5 mm in diam., over mature apothecia sometimes split up into
lobes, sometime glomerulose aggregation of 34 apothecia, disc yellow brown, brown to red brown, plane to
convex, epruinose, margin entire, distinct, pale yellow to pale brown and later excluded exciple colourless to
pale, 3670 m thick at margin, K- epithecium colourless to pale brown, 1012 m thick, K-; hymenium 4070
m thick, I+ blue than vinose red; hypothecium colourless to pale yellow, 16-30 m thick, K; spores rod shaped
with both end rounded, rarely one ends narrower than the other, transversely (1-)35 septate, 16322.43.2
m; paraphyses simple to branched, thickened at apices. Thallus K-, C-, KC-, P-, no lichen substance upon
TLC.
Remarks: This species is also known from Himanchal Pradesh, Kerala, Lakshadweep, Orissa, Tamil Nadu and
West Bengal plains. The lichen species grows on the bark of Mangifera indica and Artocarpus heterophyllus.
Specimens Examined: Faizabad district: Azamgarh road: Bhikhapur, Shankargarh bazaar, on bark of
Mangifera indica, 19th March, 2014, V. Gupta. 014-022604 (LWG); Darshan nagar, Sirsanda, on tree trunk of
Mangifera indica, 19th March, 2014, V. Gupta. 014-022611 (LWG); Purabazar, Madna, on bark of Artocarpus
heterophyllus, 19th March, 2014, V. Gupta. 014-022631 (LWG); Sarai Rasi, on bark of Artocarpus
heterophyllus, 19th March, 2014, V. Gupta. 014-022632 (LWG). Sultanpur road: Near J.N.V., Dhabha Semar,
on bark of Mangifera indica, 20th March, 2014, V. Gupta. 014-022651 (LWG); Bikapur, Burma, on bark of
Ficus racemosa, 20th March, 2014, V. Gupta. 014-022665 (LWG); Bikapur, Sugan rai ka purwa, on bark of
Artocarpus heterophyllus, 20th March, 2014, V. Gupta. 014-022666 (LWG). Gonda road: Birapur, on bark of
Mangifera indica, 21st March, 2014, V. Gupta. 014-022710 (LWG). Raebareli road: Usroo, on bark of
Mangifera indica, 22nd March, 2014, V. Gupta. 014-022728 (LWG); Barun, Vill- Khiharan, on tree trunk of
Mangifera indica, 22nd March, 2014, V. Gupta. 014-022744 (LWG); Barun, Roadside, on bark of Madhuca
longifolia, 21st March, 2014, V. Gupta. 014-022745 (LWG). Lucknow road: Near Sohawal, Masoomganj, on
154
Gupta et al. (2016) 3(1): 153156

.
bark of Mangifera indica, 23rd March, 2014, V. Gupta. 014-022790 (LWG); Raunahi, on tree trunk of
Mangifera indica, 23rd March, 2014, V. Gupta. 014-022791 (LWG).
Figure 1. Lichen thallus of different species: A, Anisomeridium subnexum (Nyl.) Zahlbr.; B, Arthothelium chiodectoides
(Nyl.) Zahlbr.; C, Bacidia medialis (Tuck.) Zahlbr.; D, Pertusaria granulata (Ach.) Mll. Arg.; E, Pyxine sorediata (Ach.)
Mont.
4. Pertusaria granulata (Ach.) Mll. Arg., Flora, Regensburg. 68 (12): 259. 1885.
(Fig. 1D)
Porina granulata Ach., Syn. Meth. Lich.: 112. 1814. (Pertusariaceae)
Thallus corticolous, verrucose whitish grey to greenish grey, fertile verrucae with perithiceiod apothecia,
fertile verrucae; constricted at base; verrucose on surface, asci and spores not seen as the ascomata are
immature. Thallus K+ yellow, C-, KC-, P-; Atranorin and Perlatolic acid detected upon TLC.
Remarks: The species was previously reported from Karnataka, Kerela and Tamil Nadu. It is rare in the area, as
it is collected from a single locality in the outskirts of the district growing on Mangifera indica.
Specimens Examined: Faizabad district: Raebareli road: Masodha, Kadipur.on bark of Mangifera indica,
22nd March, 2014, V. Gupta. 014-022756 (LWG).
5. Pyxine sorediata (Ach.) Mont. in Sagra, Hist. Phys. Cuba, Bot. Pl. Cell. 2: 188. 1842.
Lecidea sorediata Ach., Syn. Meth. Lich. : 54. 1814. (Physciaceae)
(Fig. 1E)
155
Gupta et al. (2016) 3(1): 153156

.
Thallus corticolous, foliose; lobes 1.02.0 mm broad, pearl- white to light grey or dull yellow, branching sub
dichotomous, tightly or loosely adnate to the substrate; pseudo-cyphellae well developed along the margins but
rare on the lamina; pruina restricted to the lobe tips; soredia coarse, grey. Medulla yellow or light yellow, the
soralia which may be on marginal isidia- like lobules. Apothecia very rare, internal stipe colourless to pale
brown, K-; ascospores 121768 m. Thallus
K+ yellow; medulla, K-, Pd-, triterpenes at 4-5 detected upon TLC.
Remarks: The species is known from Arunachal Pradesh, Himachal Pradesh, Karnataka, Kerala, Madhya
Pradesh, Manipur, Nagaland, Sikkim, Tamil Nadu, Uttarakhand and West Bengal. The species is rare as it is
known from two different localities in the outskirts of Tanda and near to Unchahar thermal power plants
growing on Mangifera indica.
Specimens Examined: Ambedkar Nagar district, Tanda thermal power plant, Rajesultanpur road, Ismailpur
beldaha, Heerapur, on bark of Mangifera indica, 02nd April, 2015, N. Gupta 015-031723 (LWG); Raebareli
district, Unchahar thermal power plant, Manirampur, on tree trunk of Mangifera indica, 13th August, 2013, N.
Gupta. 013-023711 (LWG).
ACKNOWLEDGEMENTS
The authors are thankful to the Head, Department of Environmental Science, BBAU, Lucknow, Director,
CSIR-NBRI, Lucknow for providing laboratory facilities and Dr. D. K. Upreti, Head, Lichenology Laboratory,
CSIR-National Botanical Research Institute, Lucknow for the identification of lichen taxa. One of the authors
(Namita Gupta) is grateful to University Grant Commission, New Delhi for financial support by providing
Junior Research Fellowship.
REFERENCES
Awasthi DD (1980) Pyxine in India. Phytomorphology 30: 359379.
Awasthi DD (1988) A key to the macrolichens of India and Nepal. Journal of Hattori Botanical Laboratory
65:207302.
Awasthi DD (1991) A key to the Microlichens of India, Nepal and Sri Lanka. Biblioth Lichenol Bd. 40,
J.Cramer, Berlin, Stuttgart.
Awasthi DD (2000) Lichenology in Indian Subcontinent- A Supplement to A Handbook of Lichens. Bishen
Singh Mahendra Pal Singh, Dehradun.
Awasthi DD (2007) A Compendium of the Macrolichens from India, Nepal and Sri Lanka. Bishen Singh
Mahendra Pal Singh, Dehradun.
Culberson CF (1972) Improved conditions and new data for the identification of lichen products by a
standardized thin- layer Chromatography method. Journal of Chromatography 72: 113125.
Dubey U, Upreti DK & Rout J (2007) Lichen flora of Along Town, West Siang district, Arunanchal Pradesh.
Phytaxonomy 7: 2126.
Gupta N, Gupta V, Dwivedi SK &Upreti DK (2015) Comparative bioaccumulation potential of Pyxine cocoes
and Bacidia submedialis in and around Faizabad city, Uttar Pradesh, India. G- Journal of Environmental
Science and Technology 2 (6): 8692.
Hue AM (1892) Lichenes Exotici a professore W. Nylander discripti vel recognite. Paris.
Karakoti N, Bajpai R, Upreti DK, Mishra SK, Srivastava A & Nayaka S (2014) Effect of metal content on
chlorophyll fluorescence and chlorophyll degradation in lichen Pyxine cocoes (Sw.) Nyl.: A case study from
Uttar Pradesh, India. Environmental Earth Sciences 71(50): 21772183.
Nayaka S & Upreti DK (2013) Lichens of Uttar Pradesh. Uttar Pradesh State Biodiversity Board, Lucknow,
India.
Orange A, James PW & White FJ (2001) Micro chemical methods for the identification of lichens. British
Lichen Society, U. K.
Singh KP & Sinha GP (2010) Indian Lichnens of an Annotated Checklist. Botanical Survey of India, Kolkata.
Srivastava SK (2004) Floristic diversity in Uttar Pradesh- an overview. Journal of Economic & Taxonomic
Botany 28(2): 292334.
Upreti DK & Pant G (1993) Notes on Arthopyrenia Species from India. The Bryologist 96 (2): 226232.
Walker FJ & James PW (1980) A revised guide to the microchemical techniques for the identification of lichen
products. Bulletin of British Lichenology Society 46: 1329.
156
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 157161, 2016
Research article
New additions to the lichen flora of Jammu and Kashmir state

(India)
Mamta Bhat1, Reema Goni2*, Susheel Verma1 and D. K. Upreti3
1
Centre for Biodiversity Studies, School of Biosciences and Biotechnology,

BGSB University, Rajouri, Jammu & Kashmir, India
2
Department of Botany, Jammu University, Jammu & Kashmir, India
3
National Botanical Research Institute, Rana Partap Marg, Lucknow, Uttar Pradesh, India
*Corresponding Author: reemagoni@gmail.com
Abstract: Jammu & Kashmir is a Himalayan state of India which exhibits large altitudinal variation and thus
houses good lichen diversity in it. Rajouri is one of the border districts of state, situated in the lap of Pir Panjal
mountain range and was not well explored in the point of view of lichen diversity. Thus the present study was
conducted to explore the lichen diversity from this remote district. The study revealed addition of 12 new
records of lichen species for the state of Jammu and Kashmir. The reported species belongs to 11 genera of nine
families.
Keywords: Lichen - Rajouri - Western Himalaya - Jammu and Kashmir.
[Cite as: Bhat M, Goni R, Verma S & Upreti DK (2016) New additions to the lichen flora of Jammu and
Kashmir state (India). Tropical Plant Research 3(1): 157161]
INTRODUCTION
Jammu and Kashmir is one of the lichen rich regions of Himalaya and often called as Hot Spot of lichen
diversity in India (Sheikh et al. 2006). The state of Jammu & Kashmir exhibits large altitudinal variation
ranging from 300 - 6500 m amsl. The climate of Jammu and Kashmir thus varies from tropical to alpine. The
state falls in the lichenogeographic zone consisting of mountainous to semi mountainous plains, Shiwalik
ranges, mountains of Kashmir valley, Pir Panjal range, Trans-Himalayan range of Ladakh and Kargil. The
literature scanned revealed that Jammu and Kashmir State is represented by the occurrence of only 413 species
(Singh & Sinha, 2010, Rai et al. 2014, Goni et al. 2015, Goni & Sharma 2015).
Rajouri, one of the border districts of Jammu and Kashmir (J&K) state is situated in the lap of Pir Panjal
mountain range. It is located between 70744 East longitude and 32583335 North latitude. The total
geographical area of the district is 2630 Km2. It lies between elevations of 4006000 m asl. The district
experiences hot summers and moderately cold winters. The average temperature varies from 7C to 37C. The
climate varies from semi-tropical in the southern part to temperate in the mountainous northern part. Its
boundaries are connected with district Jammu and Reasi on the eastern side, district Poonch on the west,
Pulwama on the north and the famous Red Cliff Line (L.O.C) passes at the south end of district.
The unique topography of the district along with the climatic conditions supports a wide range of vegetation
i.e. from subtropical to alpine. Pinus roxburghii dominates the subtropical part of the region, covering 60% of
the total area along with Phyllanthus emblica, Quercus leucotricophora, Buxus wallichiana, Zanthoxylum
armatum, Dalbergia sissoo, Mallotus philippensis, Olea ferruginea, Cassia fistula, Acacia catechu, Syzygium
cumini, Ulmus wallichiana, Bauhinia variegate, Albizzia lebbeck, Ziziphus mauritiana, Celtis australis, Populus
ciliata, Pyrus pashia and Punica granatum. The temperate region is rich in Pinus wallichiana, Rhododendron
arboreum, Quercus semicarpifolia, Picea smithiana, Abies pindrow, Salix babylonica and Juniper communis.
Although some studies have been undertaken to document the information on higher plants of the district
(Rashid et al. 2008, Pant & Verma 2009, Sarver et al. 2009, Pant 2011), lower groups have been ignored. As
such scanty information is available on lower organisms from this district including lichens. Survey for the
exploration of lichens from the area resulted in the addition of 12 species of 11 genera belonging to 09 families
to the lichen flora of J&K state.
157
Bhat et al. (2016) 3(1): 157161

.
The frequent field visits were carried out from 20132015 for the collection of lichens from Rajouri district
of Jammu and Kashmir state. The specimens were collected from all the available substrata and the collected
specimens were dried and placed in separate herbarium packets along with details of locality, substratum, habit
and date. The identification of the specimens was done by using morphological, anatomical and chemical
details. Morphological details were studied using a Digi Zoom stereomicroscope and anatomical details under a
Nikon compound eclipse 400 microscope. Secondary metabolites were identified with the use of colour spot
tests which were performed using 10% KOH solution (K), calcium hypochlorite (C) and para-phenylene
diammine (P). Thin layer chromatography was done using solvent system A comprising of Toluene: 1, 4dioxane:acetic acid; 130:60:20 ml. The silica gel plate spotted with the lichen chemicals extracted in acetone
was placed in TLC jar, lined internally by a filter paper and containing solvent. The level of solvent was 1.0 cm
below the lichen spots. Solvent gradually rose up in the precoated silica plate and was allowed to rise up to 14
cm mark. Then plate was taken out. 10% aqueous solution of sulphuric acid was sprayed and the plate was
placed in hot air oven at 110C till the different colour spots appeared. The plate was observed under UV light
at 350 nm wavelength and finally Rf value was calculated. Identification was made of lichen substances on the
basis of position and colour of spots by comparing them with the published charts (Culberson 1972, Walker &
James 1980). The recent literature of (Awasthi 1991, 2000, 2007, Divakar & Upreti 2005) was also used for the
authentic identification of the specimens. After the complete identification and labelling the specimens were
deposited in the herbarium of Centre for Biodiversity Studies of BGSB University and National Botanical
Research Institute, Lucknow.
New Additions
A. Acarospora oxytona (Ach.) Massal., Ricerch.Auton.Lich.Crost.: 28. 1852. - Lecanora oxytona Ach. Lich.
Univ.: 436.1810.
(Fig. 1A)
Thallus saxicolous, marginally lobate, yellow, effigurate, areolate, forming continuous often circular
patches, circumference distinctly radiate; marginal lobes of thallus rough, subconvex and 1.52 mm long.
Apothecia plane, solitary to 0.11.0 mm in diameter, immersed in areolae; disc plane brown; margin thick,
persistent; ascospores simple, hyaline, ellipsoid, 451.72 m.
Chemistry: Thallus K-, Pd-, C-, KC-. No chemical present.
Specimen examined: Thannamandi, 1500 m, on rock, 25/10/2011, Mamta Bhat Acc. No. 034442 (LWG).
B. Anema decipiens (A. Massal.) Forss., Forssell, Nova Acta Reg. Soc. Sci. Upsal., ser. 3, 13: 92, 1885. (Fig. 1B)
Thallus saxicolous, minutely lobed, lobes upto 2 mm wide, bluish-grey to black. Apothecia red-brown to
black, to 0.5 mm in diameter; ascospores ellipsoidal, 915 (18)59(12) m.
Specimens examined: Dassal, Rajouri, 1060 m, on rock, 14/08/2013, Mamta Bhat Acc. No. 034483 (LWG).
C. Bacidia rubella (Hoffm.) Massal., Ricerch.Auton.Lich.Crost.: 118. 1852. - Verrucaria rubella Hoffm.
Deutschl. Fl. 2: 174. 1796.
(Fig. 1C)
Thallus corticolous, crustose, granular isidiate, grey green to yellow green, thin. Apothecia rare (0.4)
0.71(1.3) mm diameter, distinctly constricted below, flat sometimes convex, pale to dark red-brown,
sometimes white pruinose; ascospores acicular, (35)4070(75)2.53(4) m.
Specimens examined: Koteranka, 1667 m, on bark of Baxus wallichiana, 15/10/2012, Mamta Bhat Acc. No.
034543 (LWG).
D. Bulbothrix setschwanensis (Zahlbr.) Hale, Phytologia 28: 481. 1974. - Parmelia setschwanensis Zahlbr. In
Handel Mazzeti, Symbol.Sinic. 3.
(Fig. 1D)
Thallus saxicolous, foliose, adnate; lobes upto 6 mm wide, bulbate cilia along margins, often only basal bulb
distinct; isidia and soredia absent; lower side pale brown, densely rhizinate; medulla white. Apothecia upto 5
mm in diameter; ascospores 1219(22)610 m.
Chemistry: Medulla K+ yellow turning red, Pd+ orange - red, C-, KC-. Salizinic acid present.
Specimens examined: Bakori 1667 m, on bark of Quercus leucotrichophora, 09/10/2012, Mamta Bhat Acc.
No. 10195 (LWG).
158
Bhat et al. (2016) 3(1): 157161

.
Figure 1. A, Acarospora oxytona (Ach.) Massal.; B, Anema decipiens (A. Massal.) Forss.; C, Bacidia rubella (Hoffm.)
Massal.; D, Bulbothrix setschwanensis (Zahlbr.) Hale; E, Caloplaca ahmadiana Poelt & Hinteregger; F, Caloplaca
parviloba Wetmore; G, Diploschistes euganeus (Massal.) Steiner; H, Hyperphyscia granulata (Poelt) Moberg; I, Lecanora
pseudistera Nyl.; J, Peltula obscurans (Nyl.) Gyelink; K, Rinodina sophodes (Ach.) Massal.; L, Xanthoparmelia congensis
(B. Stein) Hale.
E. Caloplaca ahmadiana Poelt & Hinteregger, Bib. Lich. 50: 7273 (1993).
(Fig. 1E)
Thallus crustose, saxicolous, thick, 0.510.0 mm in diameter, often coalescing with other thalli to cover
large areas, orange to brownish-orange, uniformly squamulose, flat to slightly subconvex, margins of squamules
lifted from the substrate, primary squamules 0.81.5 mm wide, small lobules/squamules budding out from the
primary squamules and later on spreading entirely over whole of the surface, secondary squamules (lobules)
0.10.2 mm wide. Cortex paraplectenchymatous, 14.020.0 m thick; algal layer continuous.Medulla white,
prothallus absent.Apothecia and pycnidia not seen.
Chemistry: Thallus K+ purple, C -, Pd -. Medulla K -, C -, Pd -.Parietin present.
Specimens examined: Dhanore, 1100 m, on rock, 15/10/2012, Mamta Bhat Acc. No. 034553 (LWG).
F. Caloplaca parviloba Wetmore, Bryologist 106 (1): 148149 (2003).
(Fig. 1F)
Thallus crustose, saxicolous, areolate to sub squamulose, continuous with short narrow elongated lobes, 0.2
0.7 mm, areoles/squamules appressed to the substratum, margins slightly uplifted, flat to somewhat convex, 1.0
1.5 mm wide, margins of areoles with numerous short lobules 0.1 mm wide and 0.20.3 mm long, yellowwww.tropicalplantresearch.com
159
Bhat et al. (2016) 3(1): 157161

.
orange to orange. Cortex paraplectenchymatous, 15.040.0 m thick. Algal layer continuous to uneven. Medulla
white. Prothallus not seen.
Chemistry: Thallus, apothecial disc and epihymenium K+ purple, C -, Pd -; Medulla K -, C -, Pd -. Parietin
present.
Specimens examined: Darhal, 1800 m, on rock, 07/09/2012, Mamta Bhat Acc. No. 034572 (LWG).
G. Diploschistes euganeus (Massal.) Steiner, Verhandl. Zool. Bot. Ges. Wein 69: 96. 1919. - Limboria euganea
Massal., Ric. Lich.Grost.: 152. 1852.
(Fig. 1G)
Thallus saxicolous, crustose, verrucose, areolate, ecorticate or with a corticiform layer, with a green algae,
apothecia perithicoid disc pale yellow to brown, disc narrowed and opening by a pore, 0.51 m diameter;
paraphysis simple, thallus whitish grey in colour, proper exciple, blackish 3040 m thick, hymenium hyaline,
80100 m high, spores 8 per ascus, biseriately arranged 24361518 m.
Chemistry: No chemicals present.
H. Hyperphyscia granulata (Poelt) Moberg, Moberg.Nord. J. Bot. 7: 721. 1987. - Physciopsis granulate Poelt,
Khumbu Himal 6(2): 91. 1974.
(Fig. 1H)
Thallus corticolous, to 5 cm across, branched; lobes to 3 mm wide, widest at tips; upper side grey brown to
brown, isidiate; isidia granular to globular; medulla orange red in lower part. India and Nepal specimens
sterile.
Chemistry: Skyrin present.
Specimens examined: Shadra Sharief, 1400 m, on bark of Salix babylonica, 17/11/2012, Mamta Bhat Acc. No.
034530 (LWG).
I. Lecanora pseudistera Nyl., Flora 55: 354. 1872.
(Fig. 1I)
Thallus crustose, dispersed-verrucose to areolate or subsquamulose, bulbate, whitish to greenish grey,
yellowish white to yellowish grey, whitish grey, epruinose; isidia and soredia absent; prothallus absent or
invisible. Apothecia numerous, immersed when young, becoming sessile to slightly constricted at the base, 0.3
1.2 mm in diameter; disc reddish orange to dark red brown, epruinose; margin thin, entire, smooth to
verruculose, entire to crenulate, concolorous with the thallus, ascus clavate, 38601015m; ascospores 8 per
ascus, ellipsoidal, 81558m.
Chemistry: Thallus and apothecial margin K+ yellow, C -, PD + yellow. Atranorin and 2 - O methylhyperlatolic acid present.
J. Peltula obscurans (Nyl.) Gyelink, Rep. Spec. Nov. Regn. Veg. 38: 308, 1935 - Endocarpiscum obsurans
Nyl. Bull. Soc. Linn. Normand. 2(6): 309, 1872.
(Fig. 1J)
Thallus saxicolous, squamulose, squamules greenish grey to olive, 0.752.0 mm in diameter, rounded to
angular, plain to convex, sometimes deeply lobed, rosette-shaped and attached by umbilicus. Thallus 100250
m thick, algal layer 40100 m thick, medulla of loose hyphae, 1030 m thick, lower cortex 35 cell layered,
1540 m thick. Apothecia 12 (rarely 3) per squamule, disc brown-orange, up to 0.4 mm in diameter, with
thalloid rim, hymenium 120150 m high, asci clavate, with gelatinous sheath, 72801520 m, multispored;
spores hyaline, simple, oval, 2312 m.
Chemistry: Thallus K-, I- , (hymenium I+ vinose - red).
Specimens examined: Nadian, Darhal, 1496 m, on rock, 07/09/2012, Mamta Bhat Acc. No. 034518 (LWG).
K. Rinodina sophodes (Ach.) Massal., Ricerch.Auton.Lich.Crost.14: 1852. - Lichen sophodesAch., Lich.
Suc.Prodrom.67:1798.
(Fig. 1K)
Thallus corticolous, crustose, grey to dark brown, in small patches, irregularly cracked, flat, determinate,
verrucose-areolate; prothallus dark, thin, entire. Apothecia 0.51.0 mm diameter immersed, sometimes
becoming sessile, frequent; ascospores 131678 m.
Chemistry: K-, Pd-, C -, KC -. No chemical present.
Specimens examined: Shadra Sharief, 1490 m, on bark of Pyruspashia, 17/11/2012. Mamta Bhat Acc. No.
034499 (LWG).
L. Xanthoparmelia congensis (B. Stein) Hale, Hale. Phytologia 28: 486. 1974. - Parmelia congensis B. Stein,
Jahr.Schles.Ges.Vaterl. Cultur 66: 140. 1889.
(Fig. 1L)
160
Bhat et al. (2016) 3(1): 157161

.
Thallus saxicolous, 24 cm across, centrally becoming subcrustose; lobes 0.31 mm wide; upper side
greenish yellow, isidiate; isidia globose, often bursting open at top, not forming soredia; lower side black,
sparsely rhizinate; medulla white. Apothecia not known.
Chemistry: Medulla K+ yellow, C-, KC-, P+ orange. Stictic, constictic, norstictic acids present.
ACKNOWLEDGEMENTS
We are grateful to the Director, Centre for Biodiversity Studies, BGSB University, Rajouri and Director,
National Botanical Research Institute, Lucknow for providing necessary facilities to undertake this work.
REFERENCES
Awasthi DD (1991) A key to Microlichens of India, Nepal and Srilanka. Biblioth. Lichenol 40: 1337.
Awasthi DD (2000) Lichenology in Indian Sub-continent. Bishen Singh Mahendra Pal Singh, Dehradun, India.
Awasthi DD (2007) A Compendium of the Macrolichens from India, Nepal and Sri Lanka. Bishen Singh
Mahendra Pal Singh, Dehradun, India.
Culberson CF (1972) Improved conditions and new data for the identification of lichen products by a
standardized thin- layer Chromatographic method. Journal of Chromatography 72: 113125.
Divakar PK & Upreti DK (2005) Parmeloid Lichens in India (A Revisionary study). Bishen Singh Mahendra Pal
Singh, Dehradun, India.
Goni R, Raina AKP, Magotra R & Sharma N (2015) Lichen flora of Jammu and Kashmir State, India: An
updated checklist. Tropical Plant Research 2(1): 6471.
Goni R & Sharma N (2015) Additions to lichen flora of Jammu and Kashmir, India. Tropical Plant Research
2(2): 7881.
Pant S (2011) Baxus wallichiana L.: A multipurpose Himalayan tree in peril. International Journal of
Biodiversity and Conservation 3: 175177.
Pant S & Verma S (2009) Diversity and economic importance of agroforestry species in Dhanore region of
Rajouri district, Jammu and Kashmir. Indian Journal of Forestry 32: 401405.
Rashid A, Anand VK & Serwar J (2008) Less known wild edible plants used by the Gujjar tribe of district
Rajouri, Jammu and Kashmir state. International Journal of Botany 4: 219224.
Rai H, Khare R, Upreti DK & Ahti T (2014) Terricolous Lichens of India: Taxonomic keys and description. In:
Rai H & Upreti DK (eds) Terricolous lichens in India, Vol. 2: Morphotaxonomic studies. Springer, New
York, pp. 17294.
Sarver J, Kumar S, Khan M, Ara M & Anand VK (2009) Diversity, distribution and utilization pattern of
economically important woody plants associated with agro-forestry in district Rajouri, Jammu and Kashmir
(Northwest Himalaya). Ethnobotanical Leaflets 13: 801809.
Sheikh MA, Upreti DK & Raina AK (2006) An enumeration of Lichens from three Districts of Jammu and
Kashmir, India. Journal of Applied Biosciences 32(2): 189191.
Singh KP & Sinha GP (2010) Indian Lichens: Annotated Checklist. Botanical Survey of India, Kolkata, India.
Walker FJ & James PW (1980) A revised guide to the microchemical techniques for the identification of lichen
products. Bulletin of British Lichenology Society 46: 1329.
161
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 162171, 2016
Research article
In vitro regeneration protocol development via callus formation

from leaf explants of tomato (Solanum lycopersicon Mill.)
Meherunnesa Papry1, S. M. Ahsan1*, Sayeed Shahriyar2, Maria Akter Sathi1, Prianka
Howlader1, Mahbub Robbani1, Soleh Akram3 and Md. Jamil Hossain Biswas4
1
Department of Horticulture, Patuakhali Science and Technology University, Bangladesh

Department of Biotechnology, Bangladesh Agricultural University, Mymensingh Bangladesh
3
Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh Bangladesh
4
Department of Entomology, Faculty of Agriculture, Bangladesh Agricultural University, Mymensingh
Bangladesh
*Corresponding Author: smvahsan@gmail.com
[Accepted: 02 April 2016]
2
Abstract: The study was conducted to develop an efficient regeneration protocol in tomato
through callus induction for subsequent plantlet regeneration. Seeds were inoculated on MS
medium where the germination rate was 78.4%. The leaves were used as explants. Different
concentration and combination of plant growth regulators (PGRs) were added with MS medium to
observe their efficacy on callus induction, shoot initiation and root formation. Leaf explants
cultured on MS medium fortified with 3 mg/L BAP gave the highest number of shoots (3.5) at 45
DAC. Among the concentrations of PGRs, 0.25 mg/L IAA produced the highest length (5.149 cm)
of plantlets, number (5.5) of leaves and fresh weight (0.781 g) of plantlets with the leaf explants at
45 DAC. The concentration of 0.5 mg/L IAA produced the highest number (25.25) of
roots/plantlet, length (8.785 cm) of roots at 45 DAC, from the same explants. The highest survival
rate of in vitro regenerated plantlets in the pot was 70.00 % with the leaf explants.
Keywords: Regeneration - Tomato - Callus - Explants - Plant growth regulator.
[Cite as: Papry M, Ahsan SM, Shahriyar S, Sathi MA, Howlader P, Robbani M, Akram S & Biswas MJH
(2016) In vitro regeneration protocol development via callus formation from leaf explants of tomato (Solanum
lycopersicon Mill.). Tropical Plant Research 3(1): 162171]
INTRODUCTION
Tomato (Solanum lycopersicon Mill.) belonging to the family Solanaceae, is one of the most popular,
important and nutritious vegetables in the world. Tomato is considered as the second most popular and highly
nutritive vegetable crop after potato (Mamidala & Nanna 2011) and is a model species for introduction of
agronomically important genes into dicotyledonous crop plants (Wing et al. 1994). Hundred grams of edible
parts of tomato contains 0.9 g protein, 0.1 g fat, 0.7 g fibre, 3.5 g carbohydrates, 1520 calorie energy, 500
1500 IU vitamin A, 0.1 mg thiamine, 0.02 mg riboflavin, 0.6 mg niacin, 2025 mg vitamin C, 69 mg calcium
and 0.10.3 mg iron (Uddin et al. 2004). Tomato is also an excellent source of lycopene (approximately 2050
mg/100g of fruit weight), a powerful antioxidant in the carotenoid family which protects human body from free
radicals which are responsible for the destruction of many body parts; lycopene is also known to prevent cancer
(Rao & Agarwal 2000). Tomato is cultivated all over Bangladesh due to its adaptability to wide range of soil
and climate (Ahamed et al. 1995).
To meet the increasing demand, it is necessary to develop good varieties with nutritional quality, higher
yield potential and wide adaptability. Conventional techniques of crop improvement are lengthy processes. The
technique of plant tissue culture has been emerged as a new and powerful tool for crop improvement like potato
(Shahriyar et al. 2015) and many other crops and vegetables (Kader et al. 2015).
The plant regeneration in tomato is genotype, explant, growth regulator and medium dependent. Many kinds
of plant growth regulators are used with varying concentration for tomato regeneration. The hormonal balance
between auxins and cytokinins can regulate the formation of roots, shoots and callus tissue in vitro. There are,
162
Papry et al. (2016) 3(1): 162171

.
however, considerable difficulties in predicting the effects of plant growth regulators. This is because of the
great differences in culture response between species, cultivars and even on the type of tissue in which the
interaction occurs.
Efficient plantlet regeneration in tomato was reported from meristems (Mirghis et al. 1995), leaf
(Ajenifujah-Solebo et al. 2012), stems (Liu et al. 2003), anthers, root (Singh & Bezei 2002), shoot tip (Selvi &
Khader 1993) and hypocotyls (Mamidala & Nanna 2011) in other countries, but very little studies are attempted
in Bangladesh on protocol development for high frequency plant regeneration of tomato.
Tomato seed production program in Bangladesh is practiced with imported virus free seeds which are
expensive. It is possible to bring down the cost of production by developing virus free seeds through tissue
culture. Moreover, maintenance of valuable germplasm in disease free conditions may be obtained by meristem
culture. But a standard tissue culture technique for tomato with suitable explants and plant growth regulators in
Bangladesh is yet to be established. On the above mentioned perspective, the present study was undertaken to
develop a suitable protocol for in vitro regeneration of tomato plantlets via callus formation of leaf explants.
The present research work was conducted at the Plant Biotechnology Laboratory, Department of
Horticulture, Patuakhali Science and Technology University. The seeds were collected from the Regional
Horticulture Research Station (RHRS), Lebukhali, Patuakhali. Winter variety of BARI tomato-14 was used as
the plant material. Leaves from in vitro grown tomato plants were cultured on MS medium and leaves as
explants. PH of the medium was adjusted to 5.8 with 0.1 N NaOH or 0.1 N HCI. All the media were autoclaved
for 20 minutes with 15 psi at 121C.
Collected tomato seeds were washed in tap water and surface sterilized with 70% ethanol for five minutes
with vigorous shaking followed by washing with sterile distilled water, surface disinfected with Sodium
hypochlorite (5.25%) for 10 minutes and rinsed 45 times with sterile water. Then they were washed with tween
20 for 23 minutes and rinsed with sterilized water till the foam was completely removed. The surface sterilized
seeds were then allowed to soak overnight to break dormancy. Then the seeds were placed in test tubes
containing MS medium and later transferred to growth room at 251C temperature under 16 hours
photoperiod with a light intensity (1500 lux) and relative humidity (6070%). Sterilized seeds were placed onto
seed germination medium in test tubes. In each test tube, 4 seeds were inoculated. The culture was then
incubated in incubation room till the germination of seeds. It was noticed that seeds started growing in dark and
later they were transferred to light. Thirty days old seedlings were used as the source of explants.
Callus proliferation
The seedlings raised in vitro culture were used as the source of leaf explants. Leaf discs were placed on the
sterile culture medium with various concentrations and combinations of BAP (1, 2, 3 mg/L) and NAA (0.25, 0.5
mg/L) and subsequent fresh weight and dry weight of the callus and changes in colours were recorded visually
after 15, 30 and 45 DAC.
Dry weight of the callus
The calli were kept in an oven (Model no.: NIIVE FN400) for drying for 72 hours at 50C after taking fresh
weights. After 72 hours, dried calli were weighed and the means were calculated.
Subculture of the callus for shoot regeneration
When the calli turned into green to yellow colour, those were removed aseptically. The pieces were again
cultured on freshly prepared medium supplemented with 0, 0.5, 1, 2 and 3 mg/L BAP for shoot induction from
callus and subsequent fresh weight and number of shoot were recorded after 15, 30 and 45 DAC.
Number of shoots /plantlet
The number of shoots emerged in each cultured bottle was calculated by counting the number of shoots
emerged. The data were recorded at 15 days of interval up to 45 days of culture.
In vitro plantlet regeneration with leaves
Initially 1.5 cm of plantlets were transferred to strength MS media containing 0.0, 0.1, 0.25, 0.50, 1.0
mg/L IAA and average length of plantlet and number of leaves per plantlet were counted at 15, 30 and 45 DAC.
163
Papry et al. (2016) 3(1): 162171

.
Subculture of the shoots for root induction
The sub cultured calli continued to proliferate and differentiated into shoots. When these shoots grew about
23 cm in length, those were rescued aseptically from the vial and separated from each other and again cultured
on freshly prepared half strength MS medium containing 0.0, 0.1, 0.25, 0.50, 1.0 mg/L IAA supplements for
root induction.
Number of roots/plantlet was recorded at 15 days interval up to 45 days of culture. The length of root was
also measured at 45 days of culture using a scale. After 45 days of inoculation, the fresh weight of plantlet was
taken with the electric balance.
Plantlets of the 57 cm length with well-developed roots were removed from culture vessel with the forceps
and transferred into pots containing garden soil, sand and well rotten cowdung at the ratio of 1:2:1. The plantlets
established within 5 to 7 days and the polythene bags were removed.
Statistical analyses
Data collected on different parameters under study were statistically analyzed to ascertain the significance of
the experimental results. The Analysis of Variance was performed and means were compared by Least
Significant Difference (LSD) test for interpretation of results. The significance of the difference between the
pair of means was evaluated using MSTAT-C computer package programs.
RESULTS
The experiment was conducted to assess the performance of the leaf explants of tomato for callus induction
and plantlet regeneration.
Seed germination
The seed germination rate on MS media was 78.4% wherein 2.4% seeds were contaminated and remaining
was unable to grow.
Callus proliferation from explants
The effects of different concentration and combination of PGRs in MS medium for leaf explants of tomato
(var. BARI tomato-14) for callus proliferation was observed.
Fresh weight of callus
The fresh weights of calli were recorded at 15, 30 and 45 days after culture (DAC) of leaf explants. The
maximum fresh weights of calli was 0.6100, 1.304 and 1.938 g produced by leaf explants at 15, 30 and 45 DAC
at 3 mg/L BAP + 0.25 mg/L NAA (Table 1)The minimum fresh weights of calli (0.3800, 0.956 and 1.097 g)
were produced in control (hormone free medium) at 15, 30 and 45 DAC respectively.
Table 1. Effect of plant growth regulators on the callus proliferation of leaf explants at different DAC.
Fresh weight (g) of callus at

different DAC
15
30
45
1 mg/L BAP + 0.25 mg/L NAA
0.4527 c
1.086 c
1.582 cd
0.4637 bc
1.110 bc
1.626 c
0.5427 a
1.243 a
1.822 b
0.4823 b
1.136 b
1.633 c
0.5607 a
1.264 a
1.902 a
0.4616 bc
1.081 c
1.560 d
Control
0.3543 d
0.849 d
1.144 e
LSD0.01 value
0.02612
0.04664
0.05234
CV (%)
5.08
3.88
3.00
Level of significance
**
**
**
Note: In a column, values having different letter (s) differ significantly at the 1% level
according to LSD.
** denotes significant at the 1% level of probability.
Concentrations and
combinations of PGRs
Fresh weight
(g) of explants
inoculated
0.005
0.005
0.005
0.005
0.005
0.005
0.005
Dry weight
(g) of callus
at 45 DAC
0.1600 c
0.1663 bc
0.1927 a
0.1730 b
0.1957 a
0.1607 c
0.1063 d
0.008258
4.52
**
of probability
Dry weight of callus

It varied significantly due to different concentration and combination of PGRs. The leaf explants cultured on
the MS medium containing 3 mg/L BAP + 0.25 mg/L NAA produced the maximum dry weight (0.1957 g) of
callus and the minimum dry weight (0.1040 g) of callus was in control.
164
Papry et al. (2016) 3(1): 162171

.
Changes in colours of explants
The change in colour was recorded at l5, 30 and 45 DAC (Fig. 1). The leaf explants became light yellow
after 15 and 30 days of inoculation. After 45 days of inoculation, the leaf explants became green at all
treatments except for 2 mg/L BAP + 0.25 mg/L NAA (Table 2).
Table 2. Relative colour change of callus from leaf explants of tomato at different
concentrations and combinations of PGRs.
Explants source and colour

at different DAC
Concentrations and combinations of PGRs
Leaf
15
30
45
Lye
Lye
Gre
Lye
Lye
Gre
Lye
Lye
Ye
Lye
Lye
Gre
Lye
Lye
Gre
Lye
Lye
Gre
Control
Lye
Lye
Gre
Note: Ye = Yellow, Lye=Light yellow, Gre= Green, Br= Brown.
Figure 1. Effect of NAA and BAP on callus formation from leaf explants of tomato at 45 DAC: A, 1 mg/L BAP + 0.25
mg/L NAA; B, 1 mg/L BAP + 0.50 mg/L NAA; C, 2 mg/L BAP + 0.25 mg/L NAA; D, 2 mg/L BAP + 0.50 mg/L NAA; E,
3 mg/L BAP + 0.25 mg/L NAA; F, 3 mg/L BAP + 0.50 mg/L NAA; G, Control.
165
Papry et al. (2016) 3(1): 162171

.
Shoot induction from callus
The effects of different concentrations and combinations of BAP on leaf explants of tomato for shoot
induction were observed.
Fresh weight of callus with shoots
The highest fresh weights of calli with shoots were 0.5920, 1.341 and 2.137 g obtained from leaf explants at
15, 30 and 45 DAC, respectively at the 2 mg/L BAP (Table 3).
There was significant difference among the different concentration and combination of PGRs in MS medium
in respect of fresh weight of callus with shoots at all sampling dates. The minimum fresh weights of calli with
shoots 0.3520, 0.744 and 1.224 g were produced from control at 15, 30 and 45 DAC, respectively.
Table 3. Interaction effects of leaf and PGRs on the fresh weight of callus with shoot and average number of
shoots at different DAC.
Fresh weight (g) of callus

Average no. of shoots at
with shoot at different DAC
different DAC
15
30
45
15
30
45
0.5 mg/L BAP
0.5225 de 1.143 ef
1.642 de
2.00 bc 2.250
1.0 mg/L BAP
0.5707 bc 1.264 bc
1.900 c
2.50 ab 3.000
2.0 mg/L BAP
0.5920 ab 1.341 ab
2.137 a
3.00 a
3.500
3.0 mg/L BAP
0.4980 e
1.115 f
1.701 d
1.25 de 2.000
Control
0.3520 f
0.744 h
1.224 f
LSD0.01 value
0.03481
0.08024
0.09988
0.4959 0.5642
CV (%)
3.64
3.80
3.00
18.18
15.50
**
*
**
**
ns
Note: In a column, values having different letter(s) differ significantly at the 1% and 5% levels of
probabilities according to LSD.
**, *, ns denotes significant at the 1%, 5% level and non-significant, respectively.
Concentrations
of PGRs
Fresh weight
(g) of explants
inoculated
0.25
0.25
0.25
0.25
0.25
Number of shoots
The maximum number of shoot was 3 and 3.5 produced by leaf explants at 30 and 45 DAC, respectively at
2.0 mg/L BAP (Fig. 2). In the present work, the number of shoots gradually increased with the advancement of
culture duration in all hormonal treatments. The increasing of BAP concentration up to 2 mg/L caused the
number of shoots to increase, but it fell down in presence of BAP (3 mg/L) that indicates the toxic effect of
growth regulators due to their accumulation.
Root development
The effects of different concentration of IAA in MS medium on root formation were observed.
Length of the plantlets
The lengths of the plantlet were recorded at 15, 30 and 45 DAC. The tallest plantlet 0.6317, 1.7830 and
5.149 cm from leaf explants at 15, 30 and 45 DAC, respectively at 0.25 mg/L IAA (Table 4). The smallest
plantlets were recorded at control.
Table 4. Interaction effects of leaf and plant growth regulators in MS medium on the average length of plantlets and average no. of
leaves/plantlet at different DAC.
Initial length of
plantlet (cm)
inoculated
1.5
1.5
1.5
1.5
1.5
Average length of plantlets (cm) at

Average no. of leaves/plantlet
different
DAC
at different DAC
Concentration of PGRs
15
30
45
15
30
45
MS + 0.10 mg/L IAA
0.3968 d
1.3620 cd
3.655 f
3.250 cd
3.500 cd
4.250 ef
MS + 0.25 mg/L IAA
0.6317 a
1.7830 a
5.149 a
4.250 a
4.500 a
5.500 a
MS + 0.50 mg/L IAA
0.5145 b
1.5700 b
4.111 c
3.750 b
4.250 b
5.250 ab
MS + 1.00 mg/L IAA
0.4800 bc
1.4270 c
3.873 e
3.250 cd
3.750 c
4.500 de
Control
0.2740 f
0.7470 f
1.598 h
2.333 f
2.500 e
3.250 gh
LSD0.01 value
0.04770
0. 07415
0.1119
0.3007
0.2480
0.4079
CV (%)
5.84
3.15
1.70
5.22
3.66
5.01
**
**
**
**
**
**
Note: In a column, values having different letter(s) differ significantly at the 1% level of probability according to LSD.
**denotes significant at the 1% level of probability.
166
Papry et al. (2016) 3(1): 162171

.
Figure 2. Effect of BAP on callus with shoot production from leaf explants at 45 DAC: A, 0.5 mg/L BAP; B, 1 mg/L BAP;
C, 2 mg/L BAP; D, 3 mg/L BAP; E, Control.
Number of leaves/plantlet
The highest numbers of leaves (4.25, 4.5 and 5.5) were produced from leaf explants at 15, 30 and 45 DAC,
respectively on the half strength medium supplemented with 0.25 mg/L IAA and the lowest number of leaves
2.333, 2.50 and 3.25 was for the hormone free medium at 15, 30 and 45 DAC, respectively.
Number of roots/plantlet
The highest numbers of roots (16.0, 21.00 and 25.25) were produced from leaf explants at 15, 30 and 45
DAC, respectively on MS medium supplemented with 0.5 mg/L IAA (Table 5). The lowest numbers of roots
were observed on MS medium supplemented with 0.1 mg/L IAA. No root formation was observed in control
treatment.
167
Papry et al. (2016) 3(1): 162171

.
Table 5. Interaction effects of leaf and PGRs in MS medium on the average no. of roots/plantlet, length
of roots and fresh weight of plantlets at different DAC.
Average no. of roots/plantlet at

Length of
Fresh weight
different DAC
roots (cm)
(g) of plantlets
at 45 DAC
at 45 DAC
15
30
45
MS + 0.10 mg/L IAA 11.75 ef
14.83 e
17.00 f
5.106 g
0.4420 fg
MS + 0.25 mg/L IAA 16.25 ab
19.08 b
21.75 c
6.122 de
0.7810 a
MS + 0.50 mg/L IAA 16.50 a
21.00 a
25.25 a
8.785 a
0.6220 d
MS + 1.00 mg/L IAA 12.17 de
16.00 d
19.58 d
6.266 d
0.4430 fg
Control
0.2930 i
LSD0.01 value
1.140
1.047
0.9414
0.2250
0.03327
CV (%)
6.06
4.32
3.26
2.32
3.57
**
**
**
**
**
Note: In a column, values having different letter(s) differ significantly at the 1% level of
probability according to LSD.
** denotes significant at the 1% level of probability.
Concentrations
of PGRs
Length of roots
Leaf explants produced the longest root (8.785 cm) at 45 DAC on 0.5 mg/L in MS medium. The shortest
root (5.106 cm) was observed on MS medium supplemented with 0.1 mg/L IAA. No root was formed in
control (Fig. 3A).
Fresh weight of plantlets
The highest fresh weight of plantlet was 0.7810 g produced from leaf explants at 45 DAC on 0.25 mg/L IAA
in MS medium. The fresh weights of plantlets were significantly influenced by the application of IAA.
Oppositely, the lowest weight 0.2930 g was produced by control at 45 DAC. The survival rate of regenerated
plants from leaf explants was 70% (Fig. 3B).
Figure 3. A, Root initiation from leaf explants of tomato in MS + 0.25 mg/L IAA; B, Establisment of tomato plantlets in
pots containing a mixture of garden soil, sand and cow dung at the ratio of 1:2:1 from leaf explants.
DISCUSSION
Callus proliferation from explants
The effects of different concentration and combination of plant growth regulators (PGRs) in MS medium for
leaf explants of tomato (var. BARI tomato-14) for callus proliferation was observed.
Fresh weight of callus
As the maximum and the minimum fresh weight of calli of leaf explants were 1.938 g and 1.097 g at 45
DAC at 2 mg/L BAP + 0.25 mg/L NAA and control respectively. Liu et al. (2003) reported similar results while
working with leaf and stem explants with 2.5 mg/L BAP + 0.2 mg/L NAA. The fresh weights of calli varied
significantly due to different concentrations and combinations of PGR at all observing dates. The minimum
168
Papry et al. (2016) 3(1): 162171

.
fresh weights of calli (0.3800, 0.956 and 1.097 g) were produced in control (hormone free medium) at 15, 30
and 45 DAC, respectively. Kayum (2004) observed the best callus formation of tomato with the same
concentrations and combinations of PGRs. These findings also support the results of Harish et al. (2010) while
working with leaf disc, stem and hypocotyl of six tomato cultivars (Sindhu, Shalimar, CO3, PKM, Vaishnavi
and Ruchikar) with 0.5 mg/L NAA + 2 mg/L BAP.
Dry weight of callus
The dry weights of calli varied significantly due to different concentrations and combinations of PGRs. The
leaf explants cultured on the MS medium containing 3 mg/L BAP + 0.25 mg/L NAA produced the maximum
dry weight of callus. Papry et al. (2015) also found similar results in case of callus formation from stem explants
of tomato. Capote et al. (2000) also reported similar results while working with leaf tissue and stem segments of
different cultivars with BAP + NAA combinations of PGRs.
Changes of colour in explants
After inoculation of explants to culture media, the leaf segments showed light yellow appearance at the first
sight and gradually became green. The colour changes were observed gradually with the advancement of culture
period. The results appeared that the colour change of inoculated explants also showed clear variation due to
different PGRs treatments. Harish et al. (2010) also observed the colour change of tomato explants while they
worked with tomato for regeneration. The findings of his results support the present experiment.
Shoot induction from callus
A. Fresh weight of callus with shoots
The highest fresh weight of calli with shoots was 2.137 g obtained from leaf explants at 45 DAC at the
hormonal concentration of 2 mg/L BAP. There was significant difference among the different concentrations
and combinations of PGRs in MS medium in respect of fresh weight of callus with shoots at all sampling dates.
The minimum fresh weight of calli with shoots was 1.224 g produced from control at 45 DAC. Ugandhar et al.
(2012) reported similar results in MS medium supplemented with 2 mg/L BAP.
B. Number of shoots
The maximum number of shoot was 3.5 produced by leaf explants at 45 DAC at 2.0 mg/L BAP. In the
present work, the number of shoots gradually increased with the advancement of culture duration in all
hormonal treatments. The increasing of BAP concentration up to 2 mg/L caused the number of shoots to
continue developing, but it fell down in presence of BAP (3 mg/L) that indicates the toxic effect of growth
regulators due to their accumulation.
These findings support the results of Janani et al. (2013), Shah et al. (2013), Otroshy et al. (2013) and Khan
et al. (2006); they found the highest shoot regeneration response from leaf explants. These results are also
similar with the findings of Mohamed et al. (2010) who had reported the highest number of shoots in MS
medium supplemented with BAP (2 mg/L) and no adventitious shoots was noticed in the control (hormone free
medium). The cytokinin (BAP), if added at high dosage to plants induces programmed cell death (PCD) by
accelerating senescence (Carimia et al. 2004). That event was observed in presence of BAP (3 mg/L).
Length of the plantlets
The tallest plantlet was found as 5.149 cm from leaf explants at 45 DAC at 0.25 mg/L IAA. The smallest
plantlets were recorded at control (hormone free medium).
Number of leaves/plantlet
The highest number (5.5) of leaves per explant was produced from leaf explants at 45 DAC on the half
strength medium supplemented with 0.25 mg/L IAA and the lowest number of leaves was 3.25 for the hormone
free medium (control) at 45 DAC.
Number of roots/plantlet
The highest number of roots 25.25 was produced from leaf explants at 45 DAC, on MS medium
supplemented with 0.5 mg/L IAA. Leaf explants showed the most important organogenesis capacity in
comparison to cotyledon explants (Majoul et al. 2007).The lowest number of roots was observed on MS
medium supplemented with 0.1 mg/L IAA. No root formation was observed in control treatment. Liu et al.
(2003) reported that tomato initiated high rooting at the same concentration and produced thick and strong roots.
Similarly the highest number of roots/shoot (22.1) was observed on MS medium supplemented with IAA 0.5
169
Papry et al. (2016) 3(1): 162171

.
mg/L (Osman et al. 2010). Devi et al. (2008) also reported that the best rooting in tomato was obtained on
MS medium.
Length of roots
The results revealed that the explants differed significantly in respect of root length. Leaf explants produced
the longest root (8.785 cm) at 45 DAC on 0.5 mg/L in MS medium. The shortest root (5.106 cm) was
observed on MS medium supplemented with 0.1 mg/L IAA. No root was formed in control. This result is
similar to the findings of Ishag et al. (2009), Osman et al. (2010) and Parmar et al. (2012) who had also
observed the longest roots on MS medium supplemented with IAA at 0.5 mg/L.
Fresh weight of plantlets
The highest fresh weight of plantlet was 0.7810 g produced from leaf explants at 45 DAC on 0.25 mg/L IAA
in MS medium. The fresh weights of plantlets were significantly influenced by the application of IAA.
Oppositely, the lowest weight 0.2930 g was produced in control at 45 DAC.
ACKNOWLEDGEMENT
Authors are grateful to Department of Horticulture, Patuakhali Science and Technology University,
Bangladesh.
REFERENCES
Ahmed H, Chaudhry Z., Abbas S, Yasmin A & Rashid H (1995) Tissue culture studies in tomato
(Lycopersiconesculentum) var. Moneymaker. Pakistan Journal of Botany 42(1): 155163.
Ajenifujah-Solebo SOA, Isu NA, Olorode O, Ingelbrecht I & Abiade OO (2012) Tissue culture regeneration of
three Nigerian cultivars of tomatoes. African Journal of Plant Science 6(14): 370375.
Capote RA, Fundora MZ & Perez DO (2000) Effect of different factors on the in vitro plant regeneration from
leaflets of five genotypes of tomato (Lycopersicon esculentum Mill.). Revista del-Jardin-Botanico-Ncional
21(1): 7176.
Carimia F, Terzi M, De Michele R, Zottini M & Lo Schiavo F (2004) High levels of the cytokinin BAP induced
PCD by accelerating senescence. Plant Science 166(4): 963969.
Harish MC, Rajeev kumar S & Sathish kumar R (2010) Efficient in vitro callus induction and regeneration of
different tomato cultivars of India. Asian Journal Biotechnology 2: 178184.
Ishag S, Osman MG & Khalafalla MM (2009) Effects of growth regulators, explant and genotype on shoot
regeneration in tomato (Lycopersicon esculentum cv. omdurman). International journal of sustainable crop
production 4(6): 713.
Janani C, Girija S & Kumari BDR (2013) In vitro culture and agrobacterium mediated transformation in high
altitude tomato (Lycopersicon esculentum Mill.) cultivar Shalimar. International Journal of Pharmacy
Teaching & Practices 4(1): 483488.
Kader A, Sinha SN & Ghosh P (2015) Contribution of environmental factors on in vitro culture of an
endangered and endemic mangroves HeritierafomesBuch.-Ham. and Bruguieragymnorhiza (L.) Lam.
Tropical Plant Research 2(3): 192203.
Kayum MA (2004) In vitro plantlets regeneration system for protocol development in tomato. MS Thesis, Dept.
of Crop Botany, Bangladesh Agricultural University, Memensingh.
Khan M, Usman M & Lilla MI (2006) Facile plant regeneration from tomato leaves induced with
spectinomycin. Pakistan Journal of Botany 38(4): 947952.
Liu Q, Zhao Y, Caj N, Liu QB, Zhao Y & Cai N (2003) Studies on the rapid propagation of peral tomato.
Journal of Hunan Agricultural University 29(2): 128126.
Majoul H, Lengliz R, Gharsallah-Chouchane S, Gorsane F, Fakhfakh H & Marrakchi M (2007) Factors
affecting tomato (Lycopersicon esculentum Mill.) transformation frequency. Acta Horticulturae 758: 4351.
Mamidala P&Nanna RS (2011) Effect of genotype, explant source and medium on in vitro regeneration of
tomato. International Journal of Genetics and Molecular Biology 3: 4550.
Mirghis E, Mirghis R & Lacatus V (1995) Analysis of tomato cultivars and hybrids for in vitro callus formation
and regeneration. Acta Horticulturae 412: 111116.
Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures.
Plant Physiology 15: 473497.
170
Papry et al. (2016) 3(1): 162171

.
Osman MG, Elhadi EA & Khalafalla MM (2010) Callus formation and organogenesis of tomato (Lycopersicon
esculentum Mill. cv. Omdurman) induced by thidiazuron. African Journal of Biotechnology 9(28): 4407
4413.
Otroshy M, Khalili Z, Ebrahimi MA, Nekoui MK & Moradi K (2013) Effect of growth regulators and explant
on plant regeneration of Solanum lycopersicum L. var. cerasiforme. Russian Agricultural Resources 39(3):
226235.
Papry M, Ahsan SM & Shahriyar S (2015) In vitro regeneration protocol development via callus formation from
stem explant of tomato. Asian Journal of Medical and Biological Research 1 (3): 589599.
Parmar P, Subramanian RB & Achakzai AKK (2012) Optimization of growth regulators for shoot Induction and
regeneration of tomato (Lycopersicum esculentum Mill.). Academic Journal of Plant Sciences 5(4): 114
118.
Rao A & Agarwal S (2000) Role of antioxidant lycopene in cancer and heart disease. Journal of the American
College of Nutrition 19: 56356.
Selvi DT & Khader MA (1993) In vitro morphogenetic capacity of tomato (Lycopersicon esculentum Mill.) var.
PKM.1.South Indian Horticulture 41(5): 251258.
Shah SH, Ali S & Ali GM (2013) A novel approach for rapid in vitro morphogenesis in tomato (Solanum
lycopersicum Mill.) with the application of cobalt chloride. European Academic Research 1(9): 27022721.
Shahriyar S, Akram S, Khan K, Miya MF & Sarkar MAR (2015) In vitro plant regeneration of potato (Solanum
tuberosum L.) at the rate of different hormonal concentration. Asian Journal of Medical and Biological
Research 1(2): 297303.
Singh R & Bezei TS (2002) Effects of PGR on high rate of callus proliferation of tomato cv. Roma. VP.
Canadian Journal of Botany 80: 928931.
Uddin MDF, Islam SMA, Sultana N & Shiragi MHK (2004) Effect of variety and plant growth regulators in MS
medium on callus proliferation from virus infected tomato plant. Asian network for scientific information
Ugandhar T, Venkateshwarlu M & Sammaiah D (2012) In vitro multiple shoot induction in Solanum
lycoperiscon Mill.through nodal culture. Plant Sciences Feed 2(11): 163169.
Wing RA, Zhang HB & Tanksley SD (1994) Map based cloning in crop plants: tomato as a model system. I.
Genetic and physical mapping of jointless. Molecular General Genetics 242: 681688.
171
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 172175, 2016
Research article
Assessment of antibacterial activity of Amorphophallus

paeoniifolius tuber and its peel extracts
Venkata Narasimha Kadali1*, Tadi Ramesh2, Sudhakara Rao Pola1 and B. V. Sandeep1
1
Department of Biotechnology, Andhra University, Visakhapatnam, Andhra Pradesh, India

Department of Biotechnology, SVKP & Dr. K.S. Raju Arts and Science College, Penugonda, Andhra Pradesh,
India
*Corresponding Author: vnsimhakadali@gmail.com
2
Abstract: The aim of the present study was to assess Amorphophallus paeoniifolius tuber and its
peel for its antibacterial activity. The tuber and peel extracts of the selected plant were tested
against five pathogenic bacteria. The ethanolic tuber extract of the plant shown inhibitory effect on
four bacterial species such as Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli
and Streptococcus mutans. The ethanolic tuber extract displayed no effect on Bacillus subtilis. The
ethanolic extract of tuber showed diameter of inhibition zones ranging from 6 mm-18 mm. The
ethanolic extract of peel showed inhibitory effect only on two bacterial species such as
Staphylococcus aureus and Pseudomonas aeruginosa. The ethanolic extract of peel exhibited
inhibition zone ranging from 7 mm to 16 mm. Water extracts of tuber (inhibition zone ranging
from 7 mm to 9mm) and peel (inhibition zone ranging from 6 mm to 9 mm) inhibited only one
bacterial species such as Staphylococcus aureus and Streptococcus mutans respectively.
Keywords: Suran - Tuber - Peel - Ethanolic extract - Pathogenic bacteria.
[Cite as: Kadali VN, Pola SR, Ramesh T & Sandeep BV (2016) Assessment of antibacterial activity of
Amorphophallus paeoniifolius tuber and its peel extracts. Tropical Plant Research 3(1): 172175]
INTRODUCTION
Medicinal plants gaining lot of importance now days because of their efficacy in healing different disorders
traditionally (Kadali & Sandeep 2015). The best source of drugs without lethal effects to human systems could
be the plant source and this has been proved by the traditional healing system and the recent studies conducted
on the experimental animals (Kadali et al. 2015). Herbs are the source of magnificent inhibitors that could act
on wide variety of diseases. One of the great aspect of herbs is they show 100% results when comes to the
healing. Herbs have all sorts of answers aginst various diseases (Kadali et al. 2016).
Amorphophallus paeoniifolius (Dennst) Nicolson belongs to family Araceae known as Suran is a wellknown plant in the Indian traditional system of medicine and distributed throughout India (Pramod et al. 2012).
It is known to have Cytotoxic activity (Angayarkanni et al. 2007), CNS depressants activity (Das et al. 2009). It
also possesses hepatoprotective effect against paracetamol-induced liver damage in rats (Pramod et al. 2012).
Methanolic extract of Amorphophallus paeoniifolius tuber proved to be antihelmenthic (Dey & Ghosh. 2010).
Methanolic extract of Amorphophallus has the gastro protective ability against pylorus ligation induced
gastotoxicity in albino rats (Nataraj et al. 2011). Petroleum ether extracts of Amorphophallus showed dosedependent activity regarding onset of convulsion (De et al. 2012). Ethanolic extract of A. paeoniifolius leaves
exhibited a statistically significant reduction in the severity and frequency of diarrhoea produced by castor oil
(Purwal et al. 2011). The ethanolic extract of Amorphophallus paeonifolius has shown significant antitumor and
antioxidant effect in animals and tuber stimulates both cellular and humoral immunity (Jagadheesh et al. 2010).
In this study an attempt has been made to assess the anti-bacterial activity using ethanol and water extracts of
tuber and peel of Amorphophallus paeonifolius against five pathogenic bacteria.
Collection of Plant material
Tubers of Amorphophallus paeoniifolius (Dennst) Nicolson were collected from village Seshammachruvu,
172
Kadali et al. (2016) 3(1): 172175

.
Achanta mandalam, West Godavari district. The tubers were authenticated by Dr. N. Suryanayana raju,
Department of Botany, SVKP & Dr. K S Raju Arts and Science College, Penugonda. Tubers were cut in to
small pieces and dried in sunlight for a week and then powdered using blender to get coarse powder.
Test Microorganisms
The Tuber and Peel extracts of Amorphophallus paeoniifolius were tested against five pathogenic bacteria.
The test organisms include Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Bacillus subtilis
and Streptococcus mutans.
Extraction process
Preparation of extracts was done according to the procedure done by (Sharmila & Gomathi. 2011). 25g of
tuber powder was packed in soxhlet extraction unit and exhaustively extracted using 100 ml of ethanol and
water at 60C for 12 hours .The extract was completely dried in water bath at 40C and subsequent stored at 4C.
Peel extracts were also prepared by using above procedure.
Determination of antibacterial activity
Antibacterial activity was measured by well diffusion method (Perez et al. 1990). Nutrient agar (Hi media)
was prepared and poured in to the petriplates. After solidification of media, overnight bacterial cultures were
inoculated on the surface of media. By using a sterile gel puncher 4 mm of wells were made in each petri plates.
Then 15 l, 20 l, 25 l of ethanol, and water extracts of tuber and peel of Amorphophallus paeoniifolius were
added in to the three wells respectively. The plates were incubated in the incubator at 37C for optimum
bacterial growth. In the next day, diameter of the zone of inhibition was measured.
RESULTS
The ethanolic tuber extract of Amorphophallus paeoniifolius (Dennst) Nicolson showed inhibitory effect on
four bacterial species such as Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and
Streptococcus mutans. The ethanolic tuber extract exhibited no effect on Bacillus subtilis. The inhibition zones
for S. aureus were 8 mm, 10 mm & 13 mm; P. aeruginosa were 13 mm, 14 mm & 18 mm; E. coli were 6 mm, 7
mm & 8 mm and S. mutans were 12 mm, 13 mm & 15 mm (including well 4 mm) at concentrations of 15 l, 20
l and 25 l respectively (Fig. 1). The ethanolic extract of peel displayed inhibitory effect only on two bacterial
species such as Staphylococcus aureus and Pseudomonas aeruginosa with the inhibition zones 7 mm, 11 mm &
12 mm and 11 mm, 14 mm & 16 mm respectively at concentrations of 15 l, 20 l and 25 l (Fig. 2). The water
extract of tuber inhibited only one bacterial species S. aureus (7 mm, 8 mm & 9 mm). On the other hand the
water extract of peel inhibited only Streptococcus mutans (6 mm, 7 mm & 9 mm) (Table 1).
Figure 1. Inhihition zones of ethanolic extract of Amorphophallus paeoniifolius tuber on different bacterial species.
173
Kadali et al. (2016) 3(1): 172175

.
Figure 2. Inhihition zones of ethanolic extract of Amorphophallus paeoniifolius peel on different bacterial species.
Table 1. Inhibition zones of water extract of Amorphophallus paeoniifolius tuber and peel against test pathogenic bacteria.
Test bacterial species
15 l/Tuber
Staphylococcus aureus
7 mm
Pseudomonas aeruginosa
Bacillus subtilis
Escherichia coli
Streptococcus mutans
Note: - indicates no zone of inhibition.
Inhibition zones in mm (including well 4mm)

20 l/Tuber 25 l/Tuber 15 l/Peel 20 l/Peel
8 mm
9 mm
6 mm
7 mm
25 l/Peel
9 mm
DISCUSSION
As the modern antibiotics have innumerable anarchic toxic effects, plant extracts could assist as alternative
antibacterial agents. Researchers centring on the traditional healers in order to find plant based drugs (Kadali et
al. 2015). This study showed that the Amorphophallus paeoniifolius exhibited significant antibacterial activity
against five pathogenic bacteria. The ethanolic extract of tuber showed diameter inhibition zones ranging from
618 mm. The water extract of tuber showed ranging from 7 mm to 9 mm. The ethanolic extract of peel
exhibited inhibition zone ranging from 7 mm to 16 mm. The water extract of tuber and peel exhibited inhibition
zones ranging from 7 mm to 9mm and 6 mm to 9 mm respectively. The ethanolic extract has exhibited
significant anti-bacterial activity than the water extract may be due to the release of bio active compounds which
are responsible for the anti-bacterial activity in to the ethanol than water.
CONCLUSION
In this present study it can be concluded that the Amorphophallus paeoniifolius has anti-bacterial activity in
its tuber and peel extracts. This tuber accounts for several pharmacological effects. Hence essentially, effective
work should be done to isolate the compounds liable for its various medicinal activities.
ACKNOWLEDGEMENT
The authors wish to thank the secretary and correspondent of S.V.K.P & Dr. K. S. Raju Arts & Science
College, Penugonda for providing the research laboratory and also chemicals which enabled us to complete this
work. The authors are greatful to expertise available in the campus that helped a lot in species identification.
REFERENCES
Angayarkanni J, Ramkumar KM, Poornima T & Priyadarshini U (2007) Cytotoxic activity of Amorphophallus
174
Kadali et al. (2016) 3(1): 172175

.
paeniifolius tuber extracts in vitro. American-Euresian Journal of Agricultural & Environmental Sciences 2:
395398.
Das S. S, Sen M, Dey Y. N., De S & Ghosh AK (2009) Effects of Petroleum Ether Extract of Amorphophallus
paeoniifolius Tuber on Central Nervous System in Mice. Indian Journal Pharmaceutical Science 71 (6):
651655.
De S, Dey YN, Gaidhani S & Ota S (2012) Effects of the petroleum ether extract of Amorphophallus
paeoniifolius on experimentally induced convulsion in mice. International Journal of Nutrition
Pharmacology Neurological Disorder 2: 132134.
Dey YN & Ghosh AK (2010) Evaluation of anthelmintic activity of the methanolic extract of Amorphophallus
paeoniifolius Tuber. International Journal of Pharmaceutical Sciences and Research 1 (11): 117121.
Jagadheesh K, Arumugam V, Elangovan N & Kumar PP (2010) Evaluation of Anti-tumour and Antioxidant
Activity of Amorphophallus paeoniifolius on DMBA Induced Mammary Carcinoma. International Journal
of Chemical and Pharmaceutical Sciences 1(2): 4050.
Kadali VN, Kindangi KR, Rao PS & Sandeep BV (2016) Wonder Herbs Having Anti Asthmatic Activity
Present in West Godavari District, Andhra Pradesh, India- A Mini Review. Advances in Biology,
Biotechnology and Genetics 03(01): 0106.
Kadali VN & Sandeep BV (2015) Anti-hyperglycemic plants used by the traditional healer of west Godavari
District, Andhra Pradesh, India. International Journal of Pharmacognosy 2(9): 47377.
Kadali VN, Kindangi KR, Peter AE, Rao PS, Bindiya P & Sandeep BV (2015) Hepato-Protective HerbsPresent In West Godavari District Of Andhra Pradesh, India- A Mini Review. International Journal of
Medical and Health Research 1(1): 1518.
Nataraj HN, Murthy RLN & Setty SR (2011) In vitro evaluation of gastro-protective activity of suran- a
possible explanation through HPTLC analysis. International Research Journal of Pharmacy 2 (9): 103106.
Perez C, Pauli M & Bazerque P (1990) An antibiotic assay by agar-well diffusion method. Acta Biologiae et
Medecine Experimentaalis 15: 113115.
Pramod JH, Pournima AS, Siddhalingesh GP, Yuvaraj DM & Khedkar AS (2012) Hepatoprotective activity of
Amorphophallus paeoniifolius tubers against paracetamol-induced liver damage in rats. Asian Pacific
Journal of Tropical Biomedicine 2 (1): S238S242.
Purwal L, Shrivastava V & Jain UK (2011) Studies on Anti-Diarrhoeal Activity of Leaves of Amorphophallus
paeoniifolius in Experimental Animals. International Journal of Pharmaceutical Sciences Research 2(2):
468471.
Sharmila N & Gomathi N (2011) Antibacterial, Antioxidant activity and Phytochemical studies of Crossandra
infundibuliformis leaf extracts. International Journal of Phytomedicine 3: 151156.
175
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 176181, 2016
Research article
Uses of wild edible macro fungi by Bodo community of Kokrajhar

district, Assam, India
Miniswrang Basumatary* and Mohesh Gogoi
Biotech Hub, Department of Botany, Science College, Kokrajhar, BTAD, Assam, India
*Corresponding Author: miniswrangb@gmail.com
Abstract: The study deals with ethnomycological knowledge of Bodo community of Kokrajhar
district, BTAD, Assam. The community has extensive mycological knowledge on which they can
easily differentiate the edibility of wild macro fungi and consume nearly 1315 varieties that grow
in wild mostly during the rainy season from May to September. Among different edible macro
fungi the commonly occurring 5 species viz. Volveriella volvacea, Agaricus semotus, Lentinus
polychrous, Stropharia semiglobata and Termitomyces eurrhizus belonging to family Plutaceae,
Agaracaceae, Lentinaceae, Strophariaceae and Tricholomataceae were identified and their
morphological characters were discussed along with the traditional method of recipe preparation
by the said ethnic community.
Keywords: Ethnomycology - Macro fungi - Bodo community - Traditional recipe.
[Cite as: Basumatary M & Gogoi M (2016) Uses of wild edible macro fungi by Bodo community of Kokrajhar
district, Assam, India. Tropical Plant Research 3(1): 176181]
INTRODUCTION
Mushrooms or macro-fungi are the fleshy, spore-bearing fruiting body of higher fungi (Mitra et al. 2013),
typically produced above ground on soil or on their food source. They are edible as well as poisonous (Sharma
1989). The diversity of macro fungi occupies important place both in terms of their ecological and economic
value. It has a very close association with the food habit of different ethnic tribe of Assam. Indigenous
knowledge of edible macro-fungi and their utilization by tribal is an important component of ethno mycology
(Das et al. 2014). Bodos are major ethnic tribal community of Assam most dominantly residing in the BTAD
region (Lower Assam) which lies roughly in between (8950' E to 9610' E and 2430' N to 2810' N). It is one
of the richest biodiversity zones in NE region of India. Since pre-historic times the wild macro fungi have been
consumed by the indigenous people and it has been prized as an important source of natural dietary product and
delicious food supplement (Sarma et al. 2010). They have acquired this traditional mycological knowledge from
elders through oral transmission from generation to generation.
The availability of wild edible mushrooms and their ethno-mycological usage have been reported by many
workers from different states of North-East India (Sing et al. 2002, Boruah et al. 1997). Paul et al. (2015)
recently identified 13 species of macro fungi some of which are edible and ethnomycologically important from
Ultapani Reserve Forest under Manas Biosphere Reserve. Wild macro-fungi have been used extensively in
traditional systems of health and subsistence throughout the history. However, their actual food values and
macro nutrients contents, edibility and medicinal properties have not been properly dealt (Jonathan & Fasidi
2003). Although many work on the availability of wild edible macro-fungi have been done from Assam (Sharma
et al. 2010, Baruah et al. 1971) but report on ethno mycological knowledge of Bodo community and their
traditional knowledge of using wild macro fungi is very rare. It is, therefore, an urgent need to document this
traditional knowledge of using wild edible macro-fungi before it comes to an end among young generation. In
view of the above an effort has been made to explore the number of commonly occurring wild edible macro
fungi and document the ethnomycological knowledge of Bodo community.
The field survey was carried out in different Bodo inhabited villages and forest areas of Kokrajhar district,
BTAD, Assam. Local market survey was done to gather information from the local mushroom vendors and
176
Basumatary & Gogoi (2016) 3(1): 176181

.
study on their natural habitat has been carried out. The macrofungi encountered were collected in polythene
bags and brought to the laboratory for identification. Specimens were preserved according to the method
followed by Paul et al. (2015). The information on ethnic uses and methods of preparation of different cuisine
from wild macro-fungi were recorded from elderly people of the said community. Identification of the species
was done on the basis of macro morphological characters viz. size, shape, colour, texture, structure of the gills
etc. and verified by comparing standard literatures. The classification and details identification of the macro
fungi observed is labeled according to the classification proposed by Ainsworth (1973) given in the book An
Introduction to Fungi by Dube (2005). Similarly, the plant substratum where the common wild edible
mushrooms grow has been identified with the help of taxonomic literature (Baruah & Ahmed 2014, Kanjilal &
Bor 2005).
Preliminary investigation revealed that the Bodo community consumes nearly 1315 varieties of wild macro
fungi. During the survey we could identify five popularly used species of macro fungi belonging to family
Lentinaceae, Agaracaceae, Plutaceae, Strophariaceae and Tricholomataceae which are saprophytic in nature and
specific to their habitat (Table 1). Their diversity greatly depends on specific environmental condition of the
habitat where they grows. The parameters like soil or substratum, temperature, moisture, light condition and
rainfall plays a very important role in their growth and development. The wild macro fungi have been used as a
delicious food supplement since prehistoric time by Bodo people. The harvesting of wild edible macro fungi is
purely on the basis of the ethnomycological knowledge that has been passed from one generation to the next.
The community people primarily gather this natural product for their own consumption as well as for earning
livelihood by selling in local markets. The gatherers who usually choose mushrooms as food have good
knowledge about the morphological appearance of the edible macro fungi and can easily differentiate the edible
and poisonous macro fungi.
Table 1. Identified macro fungi varieties.
Mushroom Species
Lentinus polychrous
Volvariella volvacea
Agaricus semotus
Stropharia semiglobata
Class
Basidiomycetes
Basidiomycetes
Basidiomycetes
Basidiomycetes
Family
Lentinaceae
Plutaceae
Agaricaceae
Strophariaceae
Local Name
Salni mwikhun
Jigabni mwikhun
Mwikhun Ghai
Mwikhun Jujai
Termitomyces eurrhizus
Basidiomycetes
Tricholomataceae
Mwikhun Hapaw
Habitat/ Substrate
Dead Sal wood, wild
Rotten Paddy straw
Manure rich Soil
Grassy areas inhabited
by sheep and cows
Abandoned termite
nest infested soil
Although many wild varieties of macro fungi availably grow in the forest patches of the region during the
rainy season, but it has been learnt that only 34 species are extensively used for consumption and sold in the
market. The most commonly preferred macro fungi are Volveriella volvacea, Agaricus semotus and
Termitomyces eurrhizus which are commonly found to grow in wild during the month of May to September.
Notwithstanding to grow a number of varieties, Volvereilla volvacea is considered to be most delicious and it is
highly priced and sold at around Rs. 7080 per 250 gm. The wild macro-fungi are mainly collected by the
villagers who are living adjacent to a forest patches and sell it in the local market. The consumption of wild
macro fungi varies from region to region and the knowledge on macro fungi is extensive due to which the report
on mushroom poisoning is very rare. The report of accidental consumption of wild poisonous mushrooms is
occasional (34 cases yearly) and those who accidentally consumed poisonous mushrooms are reported to face
problems such as nausea, vomiting, diarrhea, jaundice and hepatic or renal failure etc. but no causalities have
been reported in the recent year which shows that the community people is well acquainted in differentiating the
edibility of wild macro fungi.
Ethno mycological knowledge of Bodos in identification of edibility of macro fungi:
People from ethnic tribal societies have close association with and have good knowledge about forest
resources (Das et al. 2014). Among different inhabitants of the region, mostly the Bodos collect the wild macro
fungi and they are well aware of the existence of poisonous mushrooms and can differentiate them very easily
from edible variety through their ethnomycological knowledge. The rare cases of mushroom poisoning among
Bodos reflect the extensive mycological knowledge of Bodos in precise identification of edible macro fungi.
However, they do not follow any standardized method to differentiate the edibility; they identify through visual
177

.
method and smelling the fruiting body. The knowledge not only provided an extensive idea to collect
mushrooms as delicious food but also provided economic subsistence to the local people. Thus, documentation
of such knowledge prevailing among Bodos may be helpful to identify edible mushrooms and avoid mushroom
poisoning. The identifying features and knowledge that are used by Bodos to differentiate the edible and
poisonous macro fungi is as follows (Table 2).
Table 2. Identifying feature between edible and poisonous fungi.
Edible
1. Macro fungi which have a clear distinct ring on
the stalk at maturity.
Poisonous
1. Macro fungi directly grow on partially decomposed
cow dung without ring and black gills.
2. Macro fungi with peculiar and unpleasant odour.
2. Macro fungi having familiar and pleasant odour.

3. Fruiting body or gills becomes red brown when
harvested.
4. Stipe turns brown on breakage and when kept in
water it releases brown colour in the water.
5. Macro fungi grow in familiar substratum or
death tree trunk viz. Gambari (Gmelina arborea
L.), Sal (Shorea robusta Gaertn.), Taijou
(Mangifera indica L.), Khwdwm (Anthocephalus
cadamba Miq.), Taighir (Dillenia indica L.),
Thalir (Musa balbiciana Colla.), Kantal
(Artocarpus heterophyllus Lamk.), Jolpi
(Elaeocarpus floribendus Blume.), Jiya (Lannea
grandis A. Rich.), Sumli (Bombax malabaricum
DC.), Kharo Khandai (Oroxyllum indicum Vent.)
and Sefang (Stercospermum chelonoides DC.)
3. Colorful fruiting body whose gills bear black spots

and turn black on breakage.
4. Stipe becomes black on breakage or when picked
up from the soil.
5. Net Mushrooms known as Jeymwikhun (Bodo);
the macro fungi having net on its fruiting body are
considered highly poisonous.
6. Macro fungi on breakage release mucilage
substance from stalk.
7. Macro fungi when kept in a mixture of salt water
along with lemon juice turns black or blue.
Different recipe preparation from wild macro fungi:

The wild macro fungi are used for preparing different delicious recipe by Bodos rather than using as
medicine. During the rainy season when mushrooms flourish in wild, they are collected from the field and are
eaten as a delicious food supplement with dinner and lunch. They have acquired their own technique of
preparation of different recipe from wild macro fungi since time immemorial. Different types of local recipe are
prepared according to their choice from wild macro fungi. The most common delicious and favorite local
recipes are Mwikhun Paja (Mushroom Fry), Ondlajwng Mwkhunjwng (Mushroom with Rice Gravy).
i. Mwikhun Paja (Mushroom Fry) is prepared from the macro fungi with larger fruiting body as side dish. The
macro fungi are collected and washed thoroughly with water and then sliced off and again kept in water to
reduce the level of contaminant. It is prepared in a continuous heated metal pan called Sarai in Bodo with
mustard oil and finely chopped onion and chilies. First, the oil is heated in Sarai and then finely chopped
onion and the chilli are poured and stirred for 12 mins. After that sliced mushroom is poured and stirred
frequently until the mushrooms starts to release their moisture. When all the moisture content is removed a
little amount of water is added and allowed to boil. There after ingredients like salt, garlic paste, jheera
powder and turmeric powder are added at different concentration to make its texture attractive and boiled for
a few minutes until thick gravy is formed.
ii. Ondlajwng Mwikhunjwng (Mushroom with Rice Gravy) is another popular curry of Bodos which is being
prepared with rice powder and some specific plants, edible roots or flowers and macro fungi. The macro
fungi growing in tree trunk of Gmelina arborea Linn., Shorea robusta Gaertn. etc. and small sized
saprophytic macro fungi such as Mwikhun Jujai Stropharia semiglobata are used for preparing the same.
At first mushroom is washed and sliced off and half fried with mustard oil, onion and green chilies and kept
in a container. After that rice gravy is prepared with mustard oil, salt, chilies and garlic in hot water with
powdered rice grain and Kharwi a traditionally prepared alkali material from the burnt ashes of the bark
and other parts of local variety of banana, mustard plant, stem of sesame. Later the half fried mushrooms are
178

.
poured into the boiled rice gravy and allowed to cook. In order to make it more delicious sometimes meat
like-duck, pork, chicken etc. and small fishes are being added.
Apart from this the macro fungi is also eaten with potato, poneer, chicken, pork etc. to enhance the flavor.
Sometimes it is eaten by roasting which is known as Menanwi Janai in Bodo. It is prepared by exposing the
mixture of macro fungi in ember. The macro fungi were collected first and then wash thoroughly and sliced off.
After that the fungi are mixed well with mustard oil, salt, garlic paste and turmeric powder and wrapped with
banana leaf and then put into the ember for cook. They prepared different local cuisine according to their
individual choice. It has got a very important place in the food habit of Bodo community, so further
development towards large scale cultivation mushroom based on wild edible varieties can promote the economic
growth of the people of the region. It has also been recommended as food item contributing significantly to the
protein nutrition of the developing countries like India (FAO) which can be used as an alternative against
starvation because of its high protein and vitamin content. Therefore, cultivation and preservation of traditional
mycological knowledge can also contribute to food security as it is easily available, affordable and usable for
the poor from the wild (Das et al. 2014).
Figure 1. Some edible macro fungi: A, Lentinus polychrous; B, Lentinus polychrous growing on dead Sal wood; C,
Stropharia semiglobata; D, Volvareilla volvacea; E, Agaricus semotus; F, Termitomyces eurrhizus.
Identification and morphological description of the characterized wild edible macro fungi:
Lentinus polychrous : Commonly known as Sal mwikhun (Bodo) grows singly or in clump on death Sal wood
or decayed tree trunk rich in mosses and algal growth. The fruiting body is soft when young becoming tough
at old, whitish brown in colour with the edge bent downward, finely dispersed brown scales is present on the
surface of the fruiting body, stalk short measuring 23 cm, attached with the cap at the side (Fig. 1A). Pileus
38 cm, surface smooth, margin incurved, gills brown becomes black when dried and the fruiting body
becomes leathery when dried. Fruiting body is used for consumption; good when eaten at young and mature
one is tough. Spore print white.
Stropharia semiglobata : Common, grows gregariously in the field and other grassy areas inhabited by cows
and sheep during rainy season and early winter. Commonly known as Mwikhun jujai (Bodo). It is often
eaten as an additive with different curry of Bodos. The fungus is small (Fig. 1C). Pileus 0.52 cm,
semiglobate; soft, round, conical becoming nearly convex when matured, brownish to yellow, viscid, shiny,
glabrous, smoth; mergin regular, not splitting, non-striate; cuticle separable; context thin, pale unchanging;
gills free brown and odor not distinctive. Stipe cylindrical, long 26 cm slightly bulbous at the base, hollow,
surface brownish yellow, shredding at maturity; annulus absent, a narrow dark zone on the stipe is present
near the apex representing the presence of evanescent veil. Spore print golden yellow.
179

.
Volvareilla volvacea : It is called paddy straw mushroom (Fig. 1D) commonly known as Jigabni
mwikhun(Bodo), which grows in solitary or gregarious in decayed paddy straw preserved for cattle feed
slightly attached with the humus rich soil. The fruiting body is dark gray colour, brown to black on the top,
gills crowded, distinctly formed, thin, flesh colored becomes brown when harvested. Stipe central,
cylindrical, hollow with cottony presence, attenuated upward, 58 cm long, whitish, ending below with a
solid bulbous base, volva well developed and membranous with margin free (Fig. 1D). Pileus usually 6.0
12.0 cm in diameter, soft and smooth, margin sometimes split when attains maturity. Spore print salmon
pink colored on white paper.
Agaricus semotus : Common throughout Assam, grows singly or in group on soil where households cattle
dung is disposed off in a specific spot for organic manure production. Commonly known as Mwikhun ghai
(Bodo). The fruiting body is round, convex when young becoming nearly flat when matured measuring 713
cm, white in colour with finely dispersed brown scales on the surface, centre of the cap darker (Fig. 1E),
gills crowded free, smooth creamy white in appearance, stalk long nearly 613 cm with a distinct ring in the
upper portion of the stalk, base somewhat thickened and bulbous.
Termitomyces eurrhizus : Commonly found in Assam and other parts of India. Grows solitary on termite soil or
near the termite nests, usually appears just after the pre monsoon showers. Fruiting body fleshy, 3- 7 cm in
diameter; pileus glabrous, fleshy, with obtusely rounded perforations surface brown and off-white at
margins, globose towards the centre; hymenophore lamellate; lamellae free to adnexed, cylindrical; Gills
crowded, distinctly formed, free to sub adnate and white (Fig. 1F). Stipe central, usually long up to 20.0 cm,
firm and sometimes with a bulbous base, surface white above and brownish below, spores sub hyaline,
ellipsoid and smooth. Spore print salmon pink.
CONCLUSION
The identified macro fungi species grow in wild mostly during May to September. Diverse form of macro
fungi has been reported to be present in different localities of Kokrajhar district and the inhabitants of the region
consume nearly 1315 varieties. Selective mushroom varieties are consumed by preparing delicious recipe. The
knowledge about the edibility of wild edible macro-fungi is mainly restricted to only few elderly people.
Therefore, proper documentation of this ethnomycogical knowledge of using wild macro fungi is very important
along with creation of data base of wild edible macro fungi of the region. And many varieties of the wild edible
macro fungi which can serve as additional source of nutrition that are found to grow in the region are not
reported earlier. Further study will reveal more information related to ethnic use and distribution pattern of wild
macro fungi varieties. Therefore an extensive study is recommended for detail characterization of these
economically important natural food products growing in different localities of the region.
ACKNOWLEDGMENTS
Authors are thankful to the Department of Biotechnology, Govt. of India for providing financial assistance
by sanctioning Institutional Level Biotech Hub, Science College, Kokrajhar. Thanks are due to the Principal,
Science College, Kokrajhar for his constant support. We also express our thanks to S. Narzary, Forest Guard,
Ultapani Forest Range, Kokrajhar and the local Bodo people who have extended their cooperation during the
study.
REFERENCES
Ainsworth GC (1973) Introduction and keys to higher taxa. In: Ainsworth GC, Sparrow FK & Sussman AS
(eds) The Fungi. An Advance Treatise,Vol. IV. Academic Press, New York, London.
Bano Z & Rajarathanum S (1988) Pleurotus mushroom part II. Chemical composition nutritional value, post
harvest physiology, preservation and role as human food. Critical Reviews in Food Science and Nutrition 27:
87158.
Baruah C & Ahmed I (2014) Plant Diversity of Assam: A checklist of Angiosperm and Gymnosperms. Assam
Science Technology and Environment Council, BigyanBhawan, G.S. Road, Guwahati, Assam, India.
Baruah HK, Sing DK & Islam M (1971) On the distribution of higher Basidiomycetes in the Sibsagar district,
Assam. Bulletin of the Botanical Survey of India 13(3&4): 285289.
Boruah P, Kailta P, Bordoloi D, Gogoi P & Adhikary RK (1997) Some fleshy fungi of ethnobotanic use from
north east India. Advances in Forestry Research in India 16:165171.
180

.
Chang ST (1980) Mushroom as human food. Bio Science 30: 339401.
Das K, Lamo A, Paul D & Jha LK (2014) Ethnomycological Knowledge on Wild Edible Mushroom of Khasi
Tribes of Meghalaya, North- Eastern India. European Academic Research 2(3): 34333443.
Dube HC (2005) An Introduction to Fungi, 3rd Eds. Vikas Publishing House Pvt. Ltd., New Delhi, pp. 306351.
Grangeia C, Heleno SA, Barros L, Martins A & Ferreira ICFR (2011) Effects of tropism on nutritional and
nutraceutical potential of wild edible mushrooms. Food Research International 44: 10291035.
Jonathan SG & Fasidi IO (2003) Antimicrobial activities of two Nigerian edible macro-fungi Lycoperdon
pusilum (Bat.Ex) and L. giganteum. African Journal of Biomedical Research 6: 85 90.
Kanjilal UN & Bor NL (2005) Flora of Assam, Vol. I-V. Omson Publication, New Delhi.
Mitra JN, Mitra D & Chowdhuri SK (2013) Studies in Botany, Vol- I (Revised Edition). Moulik Library,
Kolkata, pp. 825830.
Ozturk C, Kasik G, Dogan HH & Aktas S (2003) Macrofungi of Alanya district. Turkish Journal of Botany 27:
303312.
Paul M, Sarma TC & Sarma GC (2015) Occurrence of Some Economically Important Macrofungi in Ultapani
Reserve Forest under Manas Biosphere Reserve, Assam. International Journal of Advanced Research 3(9):
319325.
Purkayastha RP & Chandra A (1985) Manual of Indian Edible Mushrooms, Jagendra Book Agency, New Delhi,
India.
Sarma TC, Sarma I & Patiri BN (2010) Wild edible mushrooms used by some ethnic tribes of Western Assam.
The Bioscan 3: 613625.
Shrama OP (1989) Text Book of Fungi. Tata McGraw- Hill Publishing Company Ltd., New Delhi, pp. 208255.
Sing NI, Sing SM & Th C (2002) Fleshy Fungi of Manipur. In: Plant Genetic Diversity: Exploration,
Evaluation, Conservation. Affiliated East West Press Pvt. Ltd., New Delhi, pp. 913.
181
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 182185, 2016
Research article
A study of effect of induced mutation on flowering of plant in M2

& M3 generations in chickpea (Cicer arietinum L.)
Navnath G. Kashid1* & Subhash B. More2
1
Department of Botany, Vasant Mahavidyalaya Kaij, Dist: Beed. Maharashtra. India

2
Department of Biology, Champawati College Beed. Maharashtra. India
*Corresponding Author: ngkashid@gmail.com
Abstract: In the present investigation of induced mutation in chickpea (Cicer arietinum L.) both
the chemical mutagens (EMS and SA) succeeded in inducing variability in days to flowering of
plants in both the cultivars. In case of SA treatments, an increasing trend with increasing
concentrations was observable in both the cultivars of chickpea in M2 and M3 generations as
regards days to flowering. The frequency of viable mutant showed the highest value at the 0.10%
EMS and 0.02% SA concentration in both the cultivars of chickpea. Most of the viable mutants
have been observed as true breeding in the subsequent M3 generation. They can be very well
utilized on a commercial scale in view of the varied positive attributes carried by them.
Keywords: Induced mutation - Flowering - Chickpea.
[Cite as: Kashid NG & More SB (2016) A study of effect of induced mutation on flowering of plant in M2 &
M3 generations in chickpea (Cicer arietinum L.). Tropical Plant Research 3(1): 182185]
INTRODUCTION
Chickpea belongs to family Leguminosae a view accepted by the majority of biologist (Verdcourt 1970).
However some taxonomist follow Hutchinsons classification and according to this classification, chickpea
belong to the family Fabaceae (Papilionaceae) of the order leguminales and class dicotyledonous. The binomial
nomenclature of chickpea is Cicer arietinum L., where cicer is the genus and arietinum is the species.
The chickpea flower is typically papilionaceous and zygomorphic. The calyx forms a tube which is
persistent and green. The corolla is papilionaceous and consists of standard; the wings and keel. The colour of
flower varies from pinkish, purplish, redish-blue to white. The stamens are ten diadelphous, arranged 9+1; the
anthers are bicelled, basifixed and orange in colour. The ovary is superior, sessile and oval with terminal bent
style and a blunt knob like stigma.
Mutation breeding can constitute a valuable tool to the conventional breeding methods in widening the
genetic base of cultivated germplasm in crops through creation of some useful mutants, henceforth, mutation
breeding finds a prominent place in the augmentation and recreation of genetic variability and has played a
significant role in the development of many crop varieties (Micke 1988, Maluszynski et al. 2001).
It is now the known fact that the availability of the large genetic variability within the species is prerequisite
for the improvement of the cultivated plants and the mutagenesis has proved to be a handy tool to enhance the
mutation rate and thereby enlarging the genetic variability and increasing the scope for obtaining the desired
selections.
In recent years a lot of work has been undertaken on induced mutagenesis through physical and chemical
mutagens with keen interest to know its impact on food security and malnutrition conditions. It has been clearly
shown in a number of plant species that the effect induced, varies with the varying mutagens and with the
variation in mutagen doses. Thus selecting a mutagen and its optimum dose for a genotype in any plant species
is an important step in mutation breeding programme.
The experimental seeds of chickpea (Cicer arietinum L.), cultivar BDN 9-3 were procured from Agricultural
Research Station Badnapur, Dist: Jalna (Maharashtra) and PG-5 from Mahatma Phule Krishi Vidyapeeth,
Rahuri, Dist: A. Nagar (Maharashtra) India, ethyl methane sulphonate (EMS) and sodium azide (SA) were
182
Kashid & More (2016) 3(1): 182185

.
employed in present study for the treatments of seeds. Ethyl methane suphonate (CH3SO2O2C2H5) a
monofunctional alkylating agent, with molecular weight of 124.16 manufactured by sisco research laboratory,
Mumbai and Sodium Azide (NaN3), with molecular weight of 65.01, manufactured by Spectrochem pvt. Ltd.
Mumbai was used in the present work.
Healthy and dry uniform seeds of chickpea cultivars with moisture content of 10-12 % were treated with
0.05, 0.10 & 0.15 % Ethyl methane sulphonate, while 0.01, 0.02 % 0.03 % concentration of sodium azide. The
treated seeds (675) from each treatment were used for raising M1 generation in field. All the experiments were
carried out in triplicate following RBD design. The seeds of individually harvested M1 plants were sown in the
experimental field to raise M2 generation. M2 plant population was screened for scoring viable mutations in the
field. The spectrum and frequency of viable mutations was calculated.
A thorough statistical analysis was carried out by computing the mean, slandered error and coefficient of
variation using slandered formulae (Mungikar 1997). Differences in means between controls and treated
populations were estimated to study the amount of variability induced by two mutagenic treatments. Days to
flowering was recorded as the number of days from the date of sowing to the opening of first flower on the
plant.
In the present study, the days to flowering were observed to be delayed in many treatments of all the
mutagens in case of both the cultivars (Table 12). An early flowering feature could be well correlated with
early maturing characters possessed by the concerned plant types. Early flowering mutants were observed by
several researchers in many plant systems after different mutagenic treatments. Delay in flowering has been
attributed to delay in germination (Bianchi et al. 1963) or slowness in growth of the plant (Iqbal 1972).
Table 1. Effect of mutagens on days to flowering in M2 generation of chickpea.
Variety: PG-5
Variety: BDN 9-3
Treatment
Concentration (%)
Control
0.05
0.10
EMS
0.15
0.01
0.02
SA
0.03
Control
EMS
SA
0.05
0.10
0.15
0.01
0.02
0.03
Mean
47.80
49.10
45.40
46.20
46.10
47.40
48.60
Standard error
0.34
0.63
0.60
0.72
0.54
0.69
0.60
47.80
49.10
45.40
46.20
46.10
47.40
48.60
0.34
0.63
0.60
0.72
0.54
0.69
0.60
Shift in mean Coefficient of variation

1.25
1.30
2.24
-2.40
2.31
-1.60
2.70
-1.70
2.06
-0.40
2.53
0.80
2.16
1.30
-2.40
-1.60
-1.70
-0.40
0.80
1.25
2.24
2.31
2.70
2.06
2.53
2.16
Table 2. Effect of mutagens on days to flowering in M3 generation of chickpea.
Variety: PG-5
Variety: BDN 9-3
Treatment
Concentration (%)
Control
0.05
0.10
EMS
0.15
0.01
0.02
SA
0.03
Control
EMS
SA
0.05
0.10
0.15
0.01
0.02
0.03
Mean
47.10
48.80
45.80
46.60
47.00
48.10
48.80
48.60
50.10
47.20
46.10
49.00
50.10
51.40
Standard error Shift in mean

0.46
0.66
1.70
0.72
-1.30
0.75
-0.50
0.60
-0.10
0.57
1.00
0.63
1.70
0.49
0.60
0.66
0.63
0.72
0.77
0.69
1.50
-1.40
-2.50
0.40
1.50
2.80
Coefficient of variation
1.69
2.35
2.72
2.78
2.23
2.07
2.25
1.74
2.09
2.43
2.38
2.55
2.69
2.33
183
Kashid & More (2016) 3(1): 182185

.
It is an important character which plays a significant role in altering the life cycle of any plant. Both the
mutagens succeeded in inducing variability in days to flowering of plants in both the cultivars of chickpea.
Some of the mutants showed an early flowering in control were 47.80 and 49.80 in BDN 9-3 and PG-5,
respectively. In case of EMS treatments, the values for days to flowering varied with the varied concentrations
in BDN 9-3, where as in PG-5 the values were in inducing order with increasing concentrations, in both M2 and
M3 populations of chickpea. In case of SA treatments, an increasing trend with increasing concentrations was
observable in both the cultivars of chickpea in M2 and M3 generations as regards days to flowering.
The early flower mutants were characterized by development of flowers as early as control. They attained
flowering in 30.20 and 31.60 days as against 47.80 and 49.80 days in control of BDN 9-3 and PG-5,
respectively. They mature as early as control in both the cultivars in chickpea.
Kaul (1980b) suggested that the mutation of two dominant genes to their recessive forms makes for an early
flowering in peas. Higher doses of both the mutagens induced late flowering while lower doses induced early
flowering. Similar results were also observed by Chopde (1976), Khan & Veeraswamy (1974), Brij & Pandya
(1986), Micke et al. (1990) and Bhatia et al. (1991). EMS treatment was found to be most effective in inducing
early flowering followed by gamma rays. Result obtained was in confirmation with results of Rao et al. (1984),
Biradar (2004), Shinde (2007) in pigeon pea, Manjaya & Nandanvar (2007) and Tambe (2009) in soybean.
The reports of delayed flowering with increasing concentrations of mutagenic treatments have been stated by
Gregory (1956), Doly (1961), Das & Chowdhary (1962), Chowta & Dnyansagar (1974), Chary (1983),
Kothekar (1987), Vandana & Dubey (1990), Padmavathi (1993), Satpute (1994), Rayyan (1995), Panchbhaye
(1997).
ACKNOWLEDGMENTS
The authors were thankful to Prof. Dr. Vijay Kothekar for their valuable guidance in this work. We also
thankful Head Department of Botany, Dr. Babasaheb Ambedkar Marathwada University Aurangabad,(
Maharashtra) for their laboratory and field facilities.
REFERENCES
Bianachi A, Marchesis & Soressi GP (1963) Some results in radiogenetical experiments with tomato varieties.
Radiation Botany 3: 333343.
Biradar AB (2004) Gamma Ray and Ethyl Methane Sulphonate (EMS) Induced Mutation Studies in Pigeonpea
[Cajanus Cajan (L.) Millsp.]. Ph.D Thesis. Mahatma Phule Krishi Vidyapeeth, Rahuri, India.
Bhatia CR, Thakare RG, Pawar SE, Kale DM & Kitto PH (ed) (1991) Induced mutations for yield and yield
components showing altered partitioning of dry matter. In: Plant mutation breeding for crop improvement.
Organized by IAEA and FAO, U.N. (Vienna), 18-22 June 1990, 2: 4353.
Brij VS & Pandya BP (1986) Ultra-early semidwarf variant in pigeonpea. Current Science 55(9): 466467.
Chary SN (1983) Mutagenic studies in pigeonpea (Cajanus cajan L. Millsp.) Ph.D. Thesis, Osmania University,
Hyderabad, India.
Chopde PR (1976) Chemical mutagenesis in pigeonpea [Cajanus cajan (L.) Millsp.]. Journal of Marathwada
Agriculture University 1976: 1720.
Chowta CD & Dnyansagar VR (1974) Abnormalities induced in flowering and floral parts by gamma rays and
EMS in Chlorophytum tuberosum Baker. M.V.R.Patrika 9: 7176.
Das A & Choudhary AK (1962) Effect of radio- active isotopes on the flowering behavior of jute. Transactions
of the Bose Research Institute 25: 2123.
Doly K (1961) The effect of fast neutrons on quantitative variability in Arabidopsis thaliana. Genetica 46: 861.
Gregory WC (1956) Induction of useful mutations in the peanut. In: Genetics in Plant breeding. Proceeding of
Brookhavean Symposium in Biology 9: 177190.
Iqbal J (1972) Effects of acute gamma radiation on the survival growth and radiosensitivity of apical meristem
of capsicum annum at different stages of seedling development. Radiation Botany12: 197204.
Kaul MLH (1980) Seed protein variability in rice. Z. Pflanzenzucht 84: 302312.
Khan WMA & Veeraswamy R (1974) Mutations Induced in Red gram [Cajanus cajan (L.) Millsp.] By Gamma
radiation and EMS. Radiation Botany 14: 237242.
Kothekar VS (1987) Differential mutagenic sensitivity in coriandrum sativum Linn. Current Science 56: 491
492.
184
Kashid & More (2016) 3(1): 182185

.
Maluszynski M, Szarejko I, Barriga P & Balcerzyk A (2001) Heterosis in crop mutant crosses and production of
high yielding lines using doubled haploid systems. Euphytica 120: 387398.
Manjaya JG & Nandanwar RS (2007) Genetic improvement of soybean variety JS 80-21 through induced
mutations. Plant Mutation Reports 1(3): 3640.
Micke AK (1988) Genetic improvement of grain legumes using induced mutation.; Proc. FAO/IAEA
Workshop on Improvement of grain legume production using induced mutation, 1-5 July, 1986 Pullman,
USA, IAEA, Vienna, pp.151.
Micke A., Domini B & Maluszyski M (1990) Induced mutations for crop improvement; Mutation breeding
Review 7: 41.
Mungikar AM (1997) An introduction to biometry. Saraswati printing press. Motikaranja, Aurangabad M.S.
India.
Padmawati T (1993) Mutagenic studies for the improvement of sunflower (Helianthus annuus) Ph.D. Thesis.
Osmania University, Hyderabad.
Panchabhaye PM (1997) Mutational breeding of sunflower (Helianthus annus) Ph.D. Thesis. Dr. B.A.M.
University, Aurangabad, Maharashtra.
Rayyan A (1995) Mutagenesis and tissue culture studies in vigna mungo L. Hepper. Ph.D. Thesis. Osmania
University, Hyderabad.
Rao DM, Reddy TP & Manohar RD (1984) Induction of mutations in pigeonpea (Cajanus cajan L.). Mutation
Breeding Newsletter 24: 8.
Satpute RA (1994) Mutational studies in safflower (Carthamus tictorious L.) Ph. D. Thesis. Dr. B. A.
Marathwada University, Aurangabad, Maharashtra, India.
Shinde MD (2007) Assessment of variability in M3 lines of pigeonpea [Cajanus cajan (L.) Millisp.], M.Sc.
(Agri.) Thesis. M.P.K.V. Rahuri, India.
Tambe AB (2009) Induction of genetic variability in soybean [Glycine max L.) Merrill.] for yield contributing
traits. Ph.D. Thesis. University of Pune, India.
Verdcourt B (1970) Studies in the leguminosae, papilionoideae for the flora of Tropical East Africa.II. Kew
Bulletin 24: 235307.
Vandana & Dubey DK (1990) Effect of EMS and DES on germination, growth, fertility and yield of lentil var.
K-85 (Macrosperma). Lens Newsletter 17(2): 812.
185
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 186190, 2016
Research article
An update on biological activities of medicinal plant

Dipak Paul and Sankar Narayan Sinha*
Environmental Microbiology Research Laboratory, Department of Botany, University of Kalyani, Kalyani
741235, West Bengal, India
*Corresponding Author: sinhasn62@yahoo.co.in
Abstract: Ipomoea quamoclit belonging to Convolvulaceae family is an annual, herbaceous plant,
commonly known as mayil manikkam, akasamulla, kunjalata, tarulata, kamalata, getphul in India
and distributed throughout the tropical areas of the world. Ipomoea quamoclit is used as folk
medicine around the world for various illnesses. This paper reviews the important biological
activities of I. quamoclit reported over the last few decades. These include antioxidant activity,
antimicrobial activity, anticancer activity, antidiabetic activity as well as insecticidal activity.
These studies reveal that I. quamoclit have various biological activities; hence, it is encouraging to
find its new therapeutic uses.
Keywords: Ipomoea quamoclit - Folk medicine - Biological activities - Therapeutic uses.
[Cite as: Paul D & Sinha SN (2016) An update on biological activities of medicinal plant Ipomoea quamoclit L.
INTRODUCTION
Since the initiation of human civilization plants play a major role for the survival and development of human
beings. Plants have been used for several years as a prime natural source of traditional medicine and alternative
medicine all over the world to treat various diseases (Razali et al. 2008, Huang et al. 2014, Mehra et al. 2014,
Truyen et al. 2015). This practice has been in existence since prehistoric times (Mittal et al. 2014). Traditional
Chinese medicines of China; Ayurvedic, Siddha, Unani medicine system of Indian subcontinent and so many
other systems are present in other countries of the globe (Ishtiaq et al. 2012, Kaur 2015). Atharvaveda (~1000
BC), Charak Samhita (~700 BC) and Sushruta Samhita (~200 BC) are the major manuscripts which give
elaborate description of about 1200 herbs (Jain et al. 2014, Bajpai et al. 2016).
In a report by the World Health Organization (WHO), approximately 80% of the worlds population for their
primary health care currently depends on traditional herbal medicine (Verma & Singh 2008, Hiremath &
Taranath 2013). Search of complementary and alternative plant derived medicine has gained gradually
importance in the recent decade due to the less or no side effects and health hazards of the chemically
synthesized drugs (Sibanda & Okoh 2007, Bajpai et al. 2016). The therapeutic properties of these medicinal
plants are due to the presence of bioactive substances such as flavonoids, alkaloids, glycosides, tannins,
coumarins, vitamins and other phenolic compounds (Meng et al. 1999, Sinha & Paul 2014, Kumaran &
Citarasu, 2015, Marcus 2016, Sen et al. 2016). Although, number of studies have been performed to know the
bioactive substances responsible for any medicinal property of a plant, but still there are several plants and their
medicinal properties for which the bioactive substances are unknown (Kumaran & Citarasu 2015). Therefore, a
thorough knowledge about the traditionally utilized medicinal plants is of utmost importance for exploration of
its elite bioactive components (Ghosh et al. 2015). The present study reviews the important biological activities
of Ipomoea quamoclit L. for their therapeutic properties.
PLANT MATERIAL
Ipomoea quamoclit L. is a less studied medicinal plant, which is an annual, herbaceous; twining vine belongs
to the family Convolvulaceae. It is commonly known as cypress vine, star glory, cardinal vine, cardinal creeper,
Indian pink, hummingbird vine and cupids flower (Chetty et al. 2008, Haque & Ghosh 2013). Ipomoea
quamoclit is one of the most commonly seen plant throughout the tropics in and around of the living area from
186
Paul & Sinha (2016) 3(1): 186190

.
northern South America to Mexico. In India, it is called mayil manikkam, akasamulla, kunjalata, tarulata,
kamalata, getphul etc.
Plant annual or perennial herb, twining vine which grows upto a height of 13 m. Leaves are long (29 cm),
pinnately lobed, each side of the leaf containing 919 lobes. The flowers are long (34 cm) with a diameter of
2 cm, five points trumpet-shaped, and can be red, pink or white. It flowers in summer and fall (Lowell &
Lucansky 1990) (Fig. 1).
Figure 1. Different parts of Ipomoea quamoclit: A, Flowers; B, Leaf; C, Fruits.
ETHNOMEDICINAL USES
Ipomoea quamoclit is used as folk medicine around the world for various illnesses (Rajendran et al. 2007,
Sajem et al. 2008) (Table 1). The plant is considered cooling and purgative; used in chest pain. Pounded leaves
are used as remedy for bleeding piles and carbuncles (Pullaiah et al. 1997, Yusuf et al. 2009). Leaves are also
used as poultices for bleeding haemorrhoids (Kumar & Akhtar 2013, Sorathia 2014). Plant found its significant
use in Siddha medicine where the decoction of leaves and stems are used to treat fever, diabetes and in Thailand,
it is used for snake bites and bloody cough (Khare 2007, Hasan et al. 2009).
Table 1. Ethnomedicinal uses of Ipomoea quamoclit L.
Plant species
Family
Convolvulaceae
Part used
Leaves,
Root,
Seeds
Ethnomedicinal uses
Treatment of physical weakness,
abnormal behaviour, sinking of voice,
bleeding from cuts and wounds, piles,
snakebites. Used as purgative.
BIOLOGICAL ACTIVITIES
Various studies have confirmed that Ipomoea quamoclit exhibit a vast range of bioactivities like antioxidant
activity, antimicrobial activity, anticancer activity, antidiabetic activity as well as insecticidal activity.
187
Paul & Sinha (2016) 3(1): 186190

.
Anticancer Activity
The cytotoxic studies indicate that Ipomoea quamoclit inhibits Caco-2 (colon cancer) cell viability which is
dose dependent. The IC50 values estimated were >100g/ml for ethanolic plant extract. Their findings revealed
that the ethanolic extract of Ipomoea quamoclit has significant cytotoxic activity (Renuka & Ravishankar 2014).
Ho et al. (2015) studied the anti-proliferative effect of the dichloromethane, methanol, hexane and ethyl acetate
extracts of leaves of Ipomoea quamoclit on HeLa (cervix adenocarcinoma), MCF-7 (breast adenocarcinoma),
CNE-1 (nasopharyngeal carcinoma), 3T3 (normal mouse fibroblast) and HT-29 (colorectal adenocarcinoma)
cell lines. Among different solvent, the methanol extract of Ipomoea quamoclit leaf against the tested cell lines
was shown to possess the highest anti-proliferative activity. The crude aqueous extracts of leaves of Ipomoea
quamoclit studied had significant cytotoxic property on a cell line (HEP G2) and exhibited remarkable
inhibitory effect on A549 cell line (Rane & Patel 2015).
Antioxidant Activity
Hydromethanol extracts of Ipomoea quamoclit, were evaluated for antioxidant activity using radical
scavenging assay with 1,1-diphenyl-2-picrylhydrazyl (DPPH) (Hasan et al. 2009). Their study suggests that
Ipomoea quamoclit extract have moderate antioxidant activity. Clarke et al. (2013) also studied antioxidant
activity of ethanol and ethyl acetate extract of Ipomoea quamoclit whole plant with the 2,2-diphenyl-1picrylhydrazyl (DPPH) and ferric reduction activity potential (FRAP). Uddin et al. (2013) evaluated antioxidant
activity of hydro-methanol extract of Ipomoea quamoclit aerial part by DPPH scavenging assay and reported
that Ipomoea quamoclit extract possess significant antioxidant activity. Renuka & Ravishankar (2014)
determined antioxidant activity of ethanol extract Ipomoea quamoclit aerial part by four DPPH free radical
scavenging activity method, reducing power method, phosphomolybdenum method and nitric oxide scavenging
method. The tested Ipomoea quamoclit extract have strong antioxidant activity against various oxidative system
in-vitro. They also found that the tested extract indicated the highest radical scavenging activity with the greatest
amount of phenolic content. According to them antioxidative nature of the extracts might depend on their
phenolics. Kumar et al. (2015) performed in vitro antioxidant activity of aqueous ethanolic extract of Ipomoea
quamoclit whole plant by DPPH assay, superoxide anion scavenging activity assay, hydrogen peroxide
scavenging assay, nitric oxide scavenging activity assay and metal chelating activity and reported that Ipomoea
quamoclit extract exhibit significant antioxidant activity.
Antidiabetic Activity
Reddy et al. (2015) evaluated antidiabetic activity of the hydroalcoholic extract of whole plant by
streptozotocin induced diabetic rats. Thirty days administration of hydroalcoholic extract of Ipomoea quamoclit
whole plant exhibited a significant depletion in blood glucose levels.
Antimicrobial Activity
Ipomoea quamoclit possess antimicrobial properties. The hexane extract of Ipomoea quamoclit stems
inhibited the bacterial growth of Salmonella enteritidis, Bacillus cereus, Escherichia coli, and Staphylococcus
aureus, whereas their methanolic extract only affected the growth of Escherichia coli. The hexane extract of
Ipomoea quamoclit leaves inhibited the growth of Bacillus cereus. The hexane extract of Ipomoea quamoclit
flowers arrested the growth of Staphylococcus aureus; their methanolic extract inhibited the growth of Bacillus
cereus, Staphylococcus aureus and Salmonella enteritidis. Their results cast Ipomoea quamoclit as a likely
source of bioactive molecules capable of inhibit the growth of pathogenic bacteria (Moreno et al. 2007). Moin et
al. (2012) evaluated antimicrobial activity of cationic protein from mature seeds of Ipomoea quamoclit against
several pathogenic bacterial and fungal strains. Crude protein fraction from Ipomoea quamoclit seed exhibited
significant antibacterial activity and potent minimum inhibitory concentration against tested bacterium Bacillus
subtilis. In vitro screening of ethanolic extract of Ipomoea quamoclit exhibited species specific activity in
inhibiting bacterial and fungal growth. The ethanol extract of the plant showed good activity against selected
Gram positive bacteria (Bacillus subtilis, Staphylococcus warneri, Staphylococcus aureus) and Gram negative
bacteria (Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Pseudomonas putida, Proteus
mirabilis) and fungal strain (Candida albicans) (Renuka & Ravishankar 2014).
Insecticidal Activity
Plant extracts are one of the best alternatives of chemically synthesized insecticides and can protect the flora
as well as the global environments. Prodhan et al. (2012) investigated effect of chloroform extract of Ipomoea
188
Paul & Sinha (2016) 3(1): 186190

.
quamoclit whole plant on salivary gland chromosomes of house fly (Musca domestica L.). Dose mortality test
result showed the intensity of activity of I. quamoclit was 911.83 ppm. The test results showed significant
effects on salivary gland chromosomes.
CONCLUSION
In the present study biological activities Ipomoea quamoclit L., a medicinal plant found in different tropical
areas of the world, has been critically reviewed. It exhibits different bio activities like antimicrobial activity,
antioxidant activity, anticancer activity, antidiabetic activity as well as insecticidal activity. From this study it is
concluded that Ipomoea quamoclit is a source of biologically active compounds and have various biological
activities, hence, it is encouraging to find its new therapeutic. Due to various promising biological activities and
ethnomedicinal importance, further studied should be carried out on drug development from plant extracts and
constituents of the Ipomoea quamoclit.
ACKNOWLEDGMENTS
The authors were thankful to Head Department of Botany, University of Kalyani, Kalyani, West Bengal for
their laboratory and field facilities.
REFERENCES
Bajpai O, Pandey J & Chaudhary LB (2016) Ethnomedicinal uses of tree species by Tharu tribes in the
Himalayan Terai region of India. Research Journal of Medicinal Plant 10: 1941.
Chetty KM, Sivaji K & Rao KT (2008) Flowering plants of Chittoor district, Andhra Pradesh, India. 1st eds.
Student Offset Printers, Tirupati, India.
Clarke G, Ting KN, Wiart C & Fry J (2013) High correlation of 2, 2-diphenyl-1 picrylhydrazyl (DPPH) radical
scavenging, ferric reducing activity potential and total phenolics content indicates redundancy in use of all
three assays to screen for antioxidant activity of extracts of plants from the Malaysian
rainforest. Antioxidants 2: 110.
Ghosh S, Parihar VS, Dhavale DD & Chopade BA (2015) Commentary on therapeutic potential of Gnidia
glauca: a novel medicinal plant. Medicinal Chemistry 5: 351353.
Haque SM & Ghosh B (2013) In vitro completion of sexual life cycle: production of R1 plants of Ipomoea
quamoclit L. Propagation of Ornamental Plants 13: 1924.
Hasan SR, Hossain MM, Akter R, Jamila M, Mazumder MEH & Rahman S (2009) DPPH free radical
scavenging activity of some Bangladeshi medicinal plants. Journal of Medicinal Plants Research 3: 875
879.
Hiremath VT & Taranath TC (2013) Indigenous traditional knowledge on tree species survey of Jogimatti
forest, Chitradurga district, Karnataka, India. The Journal of Ethnobiology and Traditional Medicine 118:
222227.
Ho KL, Chung WE, Choong KE, Cheah YL, Phua EY & Srinivasan R (2015) Anti-proliferative activity and
preliminary phytochemical screening of Ipomoea quamoclit leaf extracts. Research Journal of Medicinal
Plant 9: 127134.
Huang SH, Agrawal DC, Wu FS & Tsay HS (2014) In vitro propagation of Gentiana scabra Bunge an
important medicinal plant in the Chinese system of medicines. Botanical Studies 55: 56.
Ishtiaq M, Mumtaz AS, Hussain T & Ghani A (2012) Medicinal plant diversity in the flora of Leepa Valley,
Muzaffarabad (AJK), Pakistan. African Journal of Biotechnology 11: 30873098.
Jain P, Sharma HP, Basri F, Baraik B, Kumari S & Pathak C (2014) Pharmacological profiles of ethnomedicinal plant: Plumbago zeylanica L.- a review. International Journal of Pharmaceuticals Science Review
and Research 24: 157163.
Kaur R (2015) Ethnobotanical studies of some of the traditionally important medicinal plants of Punjab
(India). International Journal of Current Research and Academic Review 3: 262271.
Khare CP (2007) Indian medicinal plants. 1st eds. Springer, New Delhi, India.
Kumar AS, Reddy JR & Gupta VRM (2015) In vitro antioxidant activity of Porana paniculata and Ipomoea
quamoclit two ethnomedicinally important plants of Convolvulaceae family. British Journal of
Pharmaceutical Research 5: 286293.
Kumar V & Akhtar M (2013) Medicinal Convolvulaceous plants of eastern Uttar Pradesh. Indian Journal of
Life Sciences 2: 6365.
189
Paul & Sinha (2016) 3(1): 186190

.
Kumaran T & Citarasu (2015) Ethnopharmacological investigation and antibacterial evaluation of the
methanolic extract of Asparagus racemosus (Shatavari). Tropical Plant Research 2: 175179.
Lowell C & Lucansky TW (1990) Vegetative anatomy and morphology of Ipomoea quamoclit
(Convolvulaceae). Bulletin of Torrey Botanical Club 117: 232246.
Marcus AC (2016) Preliminary investigation of some phytochemicals and organic functions in n-hexane extracts
of Chromolaena odorata leaf and stem. Journal of Chemical, Biological and Physical Sciences 6: 91100.
Mehra A, Bajpai O & Joshi H (2014) Diversity, utilization and sacred values of ethno-medicinal plants of
Kumaun Himalaya. Tropical Plant Research 1: 8086.
Meng JC, Lu H, Li H, Yang L & Tan RX (1999) A new antibacterial sesquiterpenes glycoside and other
bioactive compounds from Biebersteinia heterostemon. Spectroscopy Letters 32: 10051012.
Mittal J, Sharma MM & Batra A (2014) Tinospora cordifolia: a multipurpose medicinal plant- a review. Journal
of Medicinal Plants Studies 2: 3247.
Moin S, Shibu BS, Wesley PS & Devi BC (2012) In vitro screening of antibacterial protein activity from
medicinal and economically important plants seed. Drug Invention Today 4: 533536.
Moreno S, Herrera H, Crescente O, Vallera H, Moreno M, Mundaray R & Materano Y (2007) Phytochemical
evaluation and antibacterial activity of extracts of Ipomoea quamoclit L. (Convolvulaceae). Saber,
Universidad de Oriente, Venezuela 19: 205209.
Prodhan ZH, Biswas M, Rahman M, Islam N & Golam F (2012) Effects of plant extracts on salivary gland
chromosomes of house fly (Musca domestica L.). Life Science Journal 9: 19301935.
Pullaiah T & Chennaiah E (1997) Flora of Andhra Pradesh. 1st ed. Scientific Publishers, Jodhpur, India.
Rajendran K, Srinivasan KK & Shirwaikar A (2007) Pharmacognostical identification of stem and root of
Ipomoea quamoclit (Linn.). Natural Product Sciences 13: 273278.
Rane VA & Patel BB (2015) In-vitro cytotoxic activity of leaf extracts of Ipomoea Jacq. species against A 549
and HEP-G2 cell lines. International Journal of Pharmaceutical Sciences and Research 6: 294299.
Razali N, Razab R, Junit SM & Aziz AA (2008) Radical scavenging and reducing properties of extracts of
cashew shoots (Anacardium occidentale). Food Chemistry 111: 3844.
Reddy RJ, Kumar SA & Gupta RMV (2015) Anti-diabetic activity of Ipomoea quamoclit in streptozotocin
induced diabetic rats. Journal of Pharmacognosy and Phytochemistry 4: 6871.
Renuka K & Ravishankar K (2014) Preliminary phytochemical investigation and in vitro study of antioxidant
antimicrobial and anticancer activities of ethanolic extract of Ipomoea quamoclit. World Journal of
Pharmaceutical Research 3: 612626.
Sajem AL, Rout J & Nath M (2008) Traditional tribal knowledge and status of some rare and endemic medicinal
plants of North Cachar Hills district of Assam, Northeast India. Ethnobotanical Leaflets 12: 261275.
Sen N, Paul D & Sinha SN (2016) In vitro antibacterial potential and phytochemical analysis of three species of
chilli plant. Journal of Chemical and Pharmaceutical Research 8: 443447.
Sibanda T & Okoh AI (2007) The challenges of overcoming antibiotic resistance: plant extracts as potential
sources of antimicrobial and resistance modifying agents. African Journal of Biotechnology 6: 28862896.
Sinha SN & Paul D (2014) Antioxidant potentials of Parthenium hysterophorus L. leaf extracts. Scientific
Research Journal of India 3: 8086.
Sorathia KD (2014) Ethnomedicinal explorations for certain Convolvulaceae members of Anjar Taluka. Life
Sciences Leaflets 49: 2833.
Truyen DM, Mansor M & Ruddin AS (2015) A note on Aroids Ethnobotany in Hau River, Vietnam. Tropical
Plant Research 2: 5863.
Uddin N, Islam R, Hasan N, Hossain MS, Roy A, Hossain MM & Rana MS (2013) DPPH scavenging assay of
eighty four Bangladeshi medicinal plants. IOSR Journal of Pharmacy and Biological Sciences 6: 6673.
Verma S & Singh SP (2008) Current and future status of herbal medicines. Veterinary World 1: 347350.
Yusuf M, Chowdhury JU, Haque MN & Begum J (2009) Medicinal plants of Bangladesh. Bangladesh Council
of Scientific and Industrial Research, Chittagong, Bangladesh.
190
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 191198, 2016
Research article
Microsatellite markers based heterozygosity assessment in

Jatropha curcas L.: A potential bioenergy crop
Ramanuj Maurya1* and Hemant Kumar Yadav1, 2
1
CSIR- National Botanical Research Institute, Rana Pratap Marg, Lucknow-226001, India
2
Academy of Scientic and Innovative Research (AcSIR), New Delhi, India
*Corresponding Author: ramanuj_1985@rediffmail.com
Abstract: The tree breeding is more difficult by the changes that occur during the transition from
juvenility to maturity. A correlation between individual heterozygosities of parents and their
offspring arises from the fact that, at most allelic frequencies, heterozygous parents produce higher
proportion of heterozygous progeny than do homozygous parents. Microsatellite markers are an
efficient tool for the assessment of heterozygosity and homozygosity. In order to assess the level
of heterozygosity of Jatropha curcas, 56 SSRs markers were used for genotyping 48 progeny
derived from selfed seeds of a single J. curcas plant. Out of 56, 7 SSRs could not produce
sufficient and significant data as they failed to amplify in more than 70% genotypes and thus not
considered for further analysis. Therefore, genotypic data of 49 SSRs were used for heterozygosity
assessment. Out of 49 SSRs, 31 SSRs were found to be monomorphic and 18 polymorphic
indicating homozygosity and heterozygosity on plants, respectively. The polymorphic SSRs
showed allele variation from 2 to 9 with an average of 3.56 alleles per SSRs. The SSR
JGM_CD232 showed maximum of 9 alleles followed by SSR JGM_CD348, JGM_CD421, and
JGM_CD092 with 5 alleles. The heterozygosity, calculated as proportion of heterozygous
individuals in population, varied from 0.00 to 1.00 with an average of 0.37. However, majority of
the markers (61%, 11 out of 18) showed heterozygosity variation from 0.00 to 0.22 indicating low
level of heterozygosity at these loci. The rest 7 SSRs showed heterozygosity from 0.6 to 1.0 (mean
0.84) indicating higher proportion of heterozygosity at these loci. In the present investigation, the
heterozygosity assessment in J. curcas indicating low level of heterozygosity showed there is need
to create genetic variability in J. curcas for genetic improvement.
Keywords: Jatropha curcas - SSR marker - Heterozygosity - Bioenergy crop.
[Cite as: Maurya R & Yadav HK (2016) Microsatellite markers based heterozygosity assessment in Jatropha
curcas L.: A potential bioenergy crop. Tropical Plant Research 3(1): 191198]
INTRODUCTION
Climate change is one of the biggest threats to the earths biodiversity loss. Due to high energy demand
humans are continuously deployed the natural resources like coal, gas and oil which continuously increase the
level of carbon dioxide in the environment and contribute to the global climate change. Over exploitation of
fossils fuel reserves and the increasing incidences of environmental pollution demand the search for alternate
and renewable sources of biofuels, including biodiesel. In order to fight the climate change biodiesel production
from Jatropha oil has potential effect to reduce the hunger of carbon dioxide and to fight against the global
climate change. Biofuels are renewable in nature and less polluting than petroleum based fuels (Openshaw
2000, Mandpe et al. 2005, Ong et al. 2011a). Among the various biodiesel sources, Jatropha curcas attracted
more attention as a potential source of biofuels due to its non-food nature, oil-rich and widely adaptable
properties (Heller 1996, Openshaw 2000, Sujatha et al. 2008, Makkar et al. 2009, King et al. 2009, Devappa et
al. 2010, Johnson et al. 2011). The biodiesel production from J. curcas has been considered as an environment
friendly renewable fuel alternative to alleviate the energy crisis (Fairless 2007, Ghosh et al. 2007). J. curcas L.
is belonging to the family Euphorbiaceae having chromosome number 2n=22 (Dehgan 1984) with relatively
smaller genome size of ~416 Mb (Carvalho et al. 2008, Sato et al. 2011). J. curcas L. is a tropical species native
191
Maurya & Yadav (2016) 3(1): 191198

.
to Mexico and Central America, but widely distributed in other tropical and sub-tropical areas of the world,
especially in Africa, India and South-East Asia (Heller 1996, Sujatha & Prabhakaran 1997, Openshaw 2000).
All part of Jatropha can be used for a wide variety of purposes rising from traditional medicine for common
human and animal ailments, protection against land erosion and as a boundary fence or live hedge to newly
found high economic potential as a fossil fuel replacement (Openshaw 2000, Sirisomboon et al. 2007, Rao et al.
2008).
Considering its global importance as promising biofuel plant genetic studies in J. curcas have been
undertaken towards its genetic improvement through various means including traditional as well as
biotechnological methods. A traditional approach of genetic improvement for polygenic traits is time and labor
consuming. The molecular markers technique is most widely exploited with conventional breeding program to
enhance the more accuracy and save the time. Number of molecular markers have been developed and used for
the study of genetic diversity in J. curcas such as RAPD (Rosado et al. 2010, Machua et al. 2011, Rafii et al.
2012, Kumar et al. 2013, Pamidimarri & Reddy 2014), ISSR (Kumar 2011, Grativol et al. 2011, Camellia et al.
2012, Soonthornyatara et al. 2015), AFLP (Quintero et al. 2011, Shen et al. 2012, Sinha & Tripathi 2013,
Osorio et al. 2014, Avendano et al. 2015), SSRs (Yadav et al. 2011, Ricci et al. 2012, Kumari et al. 2013,
Osorio et al. 2014, Maurya et al. 2013, 2015) and SNPs markers (Gupta et al. 2012, Montes et al. 2014, Trebbi
et al. 2015).
Knowledge of genetic nature of J. curcas such as homozygosity and heterozygosity is one of the important
aspects for the genetic improvement. Traditionally, biochemical markers were used for the detection of
heterozygosity but facing problem for detecting polymorphic loci and heterozygosity (David 1998). Over recent
years microsatellite have been applied extensively as a genetic markers for the detection of homozygosity and
heterozygosity which replacing other techniques, such as self-pollination with subsequent progeny testing and
morphological markers (Murovec et al. 2007). The most frequently used molecular markers for homozygosity
and heterozygosity testing used to be randomly amplified polymorphic DNAs (Eimert et al. 2003, Yahata et al.
2005), but microsatellite markers are used much more nowadays because of their co-dominant nature and
unambiguous result. Genetic enhancement of this important plant is now a major target for its sustainable
productivity. Genetic improvements of J. curcas with the implementation of molecular markers with
conventional breeding could be increased the productivity. Recently, some efforts have been done by some
researcher for the development of improved better yielding genotypes through interspecific hybridization J.
curcas with other related Jatropha species (Basha et al. 2009, Dhillon et al. 2009, Popluechai et al. 2009,
Muakrong et al. 2013, Laosatit et al. 2014, Aruna et al. 2015).
The previous literature showed that the number of researchers have worked on the genetic diversity
assessment in J. curcas using different molecular markers. However, there are no studies conducted in J. curcas
for the heterozygosity assessment using SSR markers. Therefore, present investigation was undertaken for the
assessment of heterozygosity in J. curacs which could be utilized in future for the genetic improvement of J.
curcas to develop high yielding genotype.
Plant materials and genomic DNA isolation
For the assessment of heterozygosity level in Jatropha curcas, one accessions i.e. NBJC147 was selfed to
check cross pollination. The mature seeds were collected to raise seedling for heterozygosity assessment. Total
genomic DNA was extracted from fresh young leaves of 48 seedling of selfed Hansraj and Chhatrapati using a
modified CTAB (Cetyl Trimethyl Ammonium Bromide) method. The quality of DNA was checked on 0.8%
agarose gel, and DNA concentration was determined using a Nanodrop spectrophotometer ND1000 (Nanodrop
Technologies, DE, USA).
Polymerase chain reaction and fragment analysis
PCR amplification was carried out in 10l reaction mixtures that contained 10 ng genomic DNA, 1X PCR
master mix (AmpliTaq Gold, Applied Biosystems, USA), 0.1 l (5pmol/l ) of forward primer (tailed with
M13 tag), 0.3l (5pmol/l) each of both reverse primer and M13 tag (labeled with either 6- FAM, VIC, NED
and PET). PCR was performed on Verti Thermal Cycler (Applied Biosystems, USA) using the following
cycling condition: initial denaturation at 95C for 5 min followed by 36 cycle of 94C for 30 s, 50-55C (primer
specific) for 45 s and 72C for 1 min. Subsequently, 10 cycles of denaturation for 30 s at 94C, annealing for 45
192
Maurya & Yadav (2016) 3(1): 191198

.
s at 53C, extension for 45 s at 72C followed by final extension for 15 min at 72C was performed. The
amplified PCR products from the parents and hybrids were resolved by TBE agarose gel electrophoresis using
1.5% Agarose (Geni) and then post PCR multiplex sets was prepared based on fluorescence labeled primers. For
post PCR multiplex set, 1 l FAM and 2 l of each VIC, NED, and PET labeled PCR product were combined
with 13 l of water. 1 l of this mixture was then added to 10 l Hi-Di formamide containing 0.25 l GeneScan
TM
600 LIZ as internal size standard. This was then denatured for 5 min at 950C, quick chilled on ice for 10 min
and run on a capillary-based 3730xl DNA analyzer (ABI, USA). Microsatellite loci repeats were assayed on the
basis of their observed heterozygosity and number of alleles detected with PCR amplification profile. Fragment
analysis was performed using GeneMapper software ver.4.0 and data were scored as allele size (bp).
Data scoring and Statistical analysis
SSR bands were scored as single independent locus. Well-resolved fragments were scored as present (1) or
absent (0) for each marker locus. The allelic data of polymorphic SSRs were subjected to statistical analysis
using Power Markers (Liu & Muse 2005) to calculate observed heterozysity (Ho), gene diversity or expected
heterozygosity (He), major allele number and polymorphic information content (PIC) value. The PIC value was
calculated following Botstein et al. (1980) as follows:
[
Where, Pi and Pj are the frequencies of ith and jth allele.

RESULTS
Identification of polymorphic markers
Initially, 56 polymorphic SSR markers were selected from previously developed SSR markers from four
SSR enriched genomic libraries (Maurya et al. 2013, 2015). These 56 SSR polymorphic markers were selected
on the basis of PIC value which polymorphism was detected with a panel of 7 accessions of Jatropha curcas
and 1 another species of J. integgerima (Maurya et al. 2013, 2015).
Heterozygosity Assessments
Table1. Details of 56 SSRs for used for the heterozygosity detection.
Total SSRs screened

56
No. of polymorphic SSRs

18
No. of monomorphic SSRs No. of failed SSRs

31
7
These selected 56 SSR markers were used for the heterozygosity assessment with 48 selfed J. curcas
genotype. Among these, 49 showed clear PCR products across all the genotype, while only 9 were found nonspecific amplifications (Table 1). Out of 49 SSRs, 31 SSRs were found monomorphic indicating that all the
plants were homozygous at these loci and the rest, 18 SSRs were found polymorphic producing more than one
allele and thus indicating heterozygous condition on these loci (Table 2). The heterozygosity, calculated as
proportion of heterozygous individuals in population, varied from 0.00 to 1.00 with an average of 0.37.
Table 2. Polymorphism feature of 18 polymorphic SSRs among 48 progenies of single plants used for
heterozygosity assessment.
Marker
JGM_A439
JGM_A464
JGM_B034
JGM_B038
JGM_B041
JGM_B054
JGM_B062
JGM_B190
JGM_B479
JGM_B586
JGM_CD005
JGM_CD092
JGM_CD106
JGM_CD232
Allele No.
3.00
2.00
4.00
4.00
2.00
3.00
3.00
2.00
3.00
3.00
3.00
5.00
3.00
9.00
Gene Diversity
0.06
0.49
0.48
0.12
0.17
0.10
0.25
0.50
0.50
0.54
0.06
0.29
0.52
0.25
Heterozygosity
0.02
0.63
0.22
0.04
0.06
0.02
0.13
0.96
0.84
1.00
0.02
0.00
0.77
0.08
PIC
0.06
0.37
0.40
0.12
0.16
0.10
0.23
0.37
0.39
0.44
0.06
0.28
0.41
0.25
193
JGM_CD421
JGM_CD469
JGM_CD128
JGM_CD348
Range
MeanSD
5.00
3.00
2.00
5.00
2.00-9.00
3.561.69
0.53
0.50
0.04
0.55
0.04-0.55
0.330.20
Maurya & Yadav (2016) 3(1): 191198

.
1.00
0.42
0.09
0.40
0.00
0.04
0.73
0.48
0.00-1.00
0.04-0.48
0.370.41
0.280.45
However, majority of the markers 11 (61%) showed heterozygosity variation from 0.00 to 0.22 with an average
of 0.22 indication low level of heterozygosity at these loci. The rest 7 SSR showed heterozygosity from 0.6 to
1.0 with an average 0.84 indicating higher proportion of heterozygosity at these loci. The JGM_CD232 was
showed maximum seven alleles followed by JGM_CD170, JGM_CD348 produced six allele while
JGM_CD421, JGM_CD092 and JGM_B034 produced five alleles. The di-nucleotide SSR markers (JGM_B)
showed less amplification with 48 genotype. However, tri-nucleotide SSR markers (JGM_CD) showed
maximum amplification with 48 genotypes. A representative snap shot from GeneMapper ver. 4.0 showing the
polymorphic and monomorphic allele in heterozygosity assessment (Fig. 1).
Figure 1. Snap shot showing A, Polymorphic (heterozygous) B, monomorphic (homozygous) allele.
DISCUSSIONS
Genetic variability is one of the essential requirements for crop improvement through plant breeding. The
prime step in this process involves germplasm screening for establishment of genetic diversity. With the
increasing knowledge on genome, molecular markers have been assisting the other marker system like
morphological and quantitative data trait for genetic characterization studies. There are number of molecular
markers have been developed in J. curcas and used for the genetic diversity assessment of Jatropha from
different growing regions like China (Sun et al. 2008), Brazil (Rosado et al. 2010, Grativol et al. 2011), Mexico
(Quintero et al. 2011), Thailand (Tanya et al. 2011), India (Bhasha et al. 2009, Maurya et al. 2013, 2015), but
there is no any study on the screening of heterozygosity on the basis of molecular markers. The number of
molecular markers like RAPD and AFLP were very quickly used for the genetic diversity analysis in J. curcas
but these markers has its own disadvantages lacking of reproducibility (Karp et al. 1997, Hansen et al. 1998,
Virk et al. 2000). However, microsatellite markers are codominant and highly polymorphic which can
discriminate the homozygosity and heterozygosity of an individual. The selfed 48 genotypes were assessed with
56 SSR markers showed the 18 polymorphic which indicate the heterozygosity of these markers at different loci.
The understanding the genetic basis genotypes will be quite useful to select suitable parental lines for
hybridization programmes for the genetic improvement of J. curcas. Some preliminary research of
homozygosity and heterozygosity have been done in different crops like rice (Liang et al. 2011), Mimulus
aurantiacus (Murovec et al. 2007). The tree breeding is more difficult by the changes that occur during the
transition from juvenility to maturity. Breeding populations can be characterized by quantifying the levels and
organization of genetic variation within and between different breeding groups. Under the appropriate
conditions, markers can replace phenotypic selection, thereby removing the need for growing or rearing of
individuals (Chen et al. 2010). Markers- based systems have been used to study and compare the levels of
194
Maurya & Yadav (2016) 3(1): 191198

.
random genetic variation throughout the different cycles of a breeding programme, thus allowing much greater
flexibility and control over the rate of reduction of genetic variability (Lia & Wua 2007). A correlation between
individual heterozygosities of parents and their offspring arises from the fact that, at most allelic frequencies,
heterozygous parents produce higher proportion of heterozygous progeny than do homozygous parents (Mitton
et al. 1993). Microsatellite markers are an efficient tool for the assessment of heterozygosity and homozygosity.
In the present investigation, the heterozygosity was assessed in J. curcas using SSR markers. Majority of the
SSRs (64%) used to assess the heterozygosity were found to be monomorphic and 36% polymorphic. Majority
of the markers (61%, 11 out of 18) showed heterozygosity variation from 0.00 to 0.22 indicating low level of
heterozygosity at these loci. According to Lerner (1954) the high levels of heterozygosity enhanced the
developmental homeostasis among individuals within populations and highly heterozygous individuals have
lower level of phenotypic variation than predominantly homozygous individuals.
CONCLUSION
In conclusion, the present study was the heterozygosity assessment using SSR markers of selfed Jatropha
curcas conducted at juvenility stage. The heterozygosity assessment in J. curcas indicating low level of
heterozygosity which warrants the urgent need to create genetic variability in J. curcas for genetic improvement.
ACKNOWLEDGEMENT
The authors would like to thanks the Dr. C.S. Nautiyal, Director, CSIR-NBRI, for providing the necessary
facilities during the investigation.
REFERENCES
Aruna R, Singh PS, Prakash CR, Ghosh A & Agarwal PK (2015) Development of Jatropha hybrids with
enhanced growth, yield and oil attributes suitable for semi-arid wastelands. Agroforestry System 89: 113.
Avendano R, Diaz EG, Melara MV, Solano NC, Villalobos AM, Cascante FA, Benavides BW & Solis-Ramos
LY (2015) Genetic diversity analysis of Jatropha species from Costa Rica using AFLP markers. American
Journal of Plant Sciences 6: 24262438.
Basha SD, Francis G, Makkar HPS, Becker K & Sujatha M (2009) A comparative study of biochemical
traits and molecular markers for assessment of genetic relationships between Jatropha curcas L.
germplasm from different countries. Plant Science 176: 812823.
Botstein D, White RL, Skolnick M & Davis RW (1980) Construction of genetic linkage map in man using
restriction fragment length polymorphisms. American Journal of Human Genetics 32: 314331.
Camellia NA, Thohirah LA & Abdullah NAP (2012) Genetic relationships and diversity of Jatropha curcas
accessions in Malaysia. African Journal of Biotechnology 11: 30483054.
Carvalho CR, Clarindo WR, Prac MM, Araujo FS & Carels N (2008) Genome size, base composition and
karyotype of Jatropha curcas L.: an important biofuel plant. Plant Science 174: 613617.
Chen SY, Lin YT, Lin CW, Chen WY, Yang CH & Ku HM (2010) Transferability of rice SSR markers to
bamboo. Euphytica 175: 2333.
David P (1998) Heterozygosity fitness correlations: new perspectives on old problems. Heredity 80: 531537.
Dehgan B (1984) Phylogenetic significance of interspecific hybridization in Jatropha (Euphorbiaceae).
Systematic Botany 9: 467478.
Devappa RK, Makkar HPS & Becker K (2010) Quality of biodiesel prepared from phorbol ester extracted
Jatropha curcas oil. Journal of the American Oil Chemist Society 87: 697704.
Dhillon RS, Hooda MS, Jattan M, Chawla V, Bhardwaj M & Goyal SC (2009) Development and molecular
characterization of interspecific hybrids of Jatropha curcas X J. integerrima. Indian Journal of
Biotechnology 8: 384390.
Eimert K, Reutter G & Strolka B (2003) Fast and reliable detection of doubled-haploids in Asparagus officinalis
by stringent RAPD-PCR. Journal of Agricultural Science 141: 7378.
Fairless D (2007) Biofuel: the little shrub that couldmaybe. Nature 449: 652655.
Ghosh A, Chaudhary DR, Reddy MP, Rao SN, Chikara J, Pandya JB, Patolia JS, Gandhi MR, Adimurthy S,
Veghela N, Mishra S, Rathod MR, Prakash AR, Shethia BD, Upadhyay SC, Balakrishna V, Praksh CR &
Ghosh PK (2007) Prospects for Jatropha methylester (biodiesel) in India. International Journal of
Environmental Studied 64: 659674.
195
Maurya & Yadav (2016) 3(1): 191198

.
Grativol C, Fonseca D, Lira MC, Hemerly AS & Ferreira PC (2011) High efficiency and reliability of intersimple sequence repeats (ISSR) markers for evaluation of genetic diversity in Brazilian cultivated Jatropha
curcas L. accessions. Molecular Biology Report 38: 245256.
Gupta P, Idris A, Mantri S, Asif MH, Yadav HK, Roy JK, Tuli R, Mohanty CS & Sawant SV (2012) Discovery
and use of single nucleotide polymorphic (SNP) markers in Jatropha curcas L. Molecular Breeding 30:
13251335.
Hansen M, Hallden C & Sall T (1998) Error rates and polymorphism frequencies for three RAPD protocols.
Plant Molecular Biology Report 16: 139146.
Heller J (1996) Physic NutJatropha curcas L. Promoting the Conservation and Use of Underutilized and
Neglected Crops, PhD dissertation. Institute of Plant Genetic and Crop Plant Research,
Gatersleben/International Plant Genetic Resource Institute, Germany/Rome, Italy.
Johnson TS, Eswaran N & Sujatha M (2011) Molecular approaches to improvement of Jatropha curcas L. as a
sustainable energy crop. Plant Cell Report 30: 15731591.
Karp A, Kresovich S, Bhat KV, Ayada WG & Hodgkin T (1997) Molecular tools in plant genetic resources
conservation: a guide to the technologies. International Plant Genetic Resources Institute (IPGRI) Technical
Bulletin no. 2.
King AJ, Montes LR, Clarke JG, Affleck J, Li Y, Witsenboer H, van der Vossen E, van der Linde P, Tripathi Y,
Tavares E, Shukla P, Rajasekaran T, van Loo EN & Graham IA (2013) Linkage mapping in the oilseed crop
Jatropha curcas L. reveals a locus controlling the biosynthesis of phorbol esters which cause seed toxicity.
Plant Biotechnology Journal 11: 986996.
Kumar S, Kumaria S & Tandon P (2013) SPAR methods coupled with seed-oil content revealed intra-specic
natural variation in Jatropha curcas L. from northeast India. Biomass and Bioenergy 54: 100106.
Kumar S, Kumaria S, Sharma SK, Rao SR & Tandon P (2011) Genetic diversity assessment of Jatropha curcas
L. germplasm from northeast India. Biomass and Bioenergy 35: 30633070.
Kumari M, Grover A, Yadav PV, Arif M & Ahmed Z (2013) Development of EST-SSR markers through data
mining and their use for genetic diversity study in Indian accessions of Jatropha curcas L.: a potential
energy crop. Genes Genome 35: 661670.
Laosatit K, Tanya P, Muakrong N & Srinives P (2014) Development of interspecic and intergeneric hybrids
among Jatropha-related species and verication of the hybrids using ESTSSR markers. Plant Genetic
Resources: Characterization and Utilization 12: S58S61.
Lerner IM (1954) Genetic homeostasis. Oliver and Boyd, Edinburgh and London, 134 p.
Lia L & Wua LC (2007) Genetic diversity and molecular markers of five snapper species. Chinese Journal of
Agricultural Biotechnology 4: 3946.
Liang YS, Zhang QJ, Wang SQ, Cao LY, Gao ZQ, Li P & Cheng SH (2011) Microsatellite analysis of
homozygosity progression of heterozygous genotypes segregating in the rice subspecies cross
Peiai64s/Nipponbare. Biochemical Genetics 49: 611624.
Liu K & Muse SV (2005) Power marker: integrated analysis environment for genetic marker data.
Bioinformatics 21: 21282129.
Machua J, Muturi G, Omondi SF & Gicheru J (2011) Genetic diversity of Jatropha curcas L. populations in
Kenya using RAPD molecular markers: implication to plantation establishment. African Journal of
Biotechnology 10: 30623069.
Makkar HPS, Maes J, De Greyt W & Becker K (2009) Removal and degradation of phorbol esters during pretreatment and transesterification of Jatropha curcas oil. Journal of the American Oil Chemist Society 86:
173181.
Mandpe S, Kadlaskar S, Degen W & Keppeler S (2005) On road testing of advanced common rail diesel
vehicles with biodiesel from the Jatropha curcas plant. Society of Automotive Engineers of Japan 26:
356364.
Maurya R, Gupta A, Singh SK, Rai KM, Chandrawati, Sawant SV & Yadav HK (2013) Microsatellite
polymorphism in Jatropha curcas L.-A biodiesel plant. Industrial Crops and Products 49: 136142.
Maurya R, Gupta A, Singh SK, Rai KM, Chandrawati, Katiyar R, Sawant SV & Yadav HK (2015) Genomicderived microsatellite markers for diversity analysis in Jatropha curcas. Tree Structure and Functions 29:
849858.
196
Maurya & Yadav (2016) 3(1): 191198

.
Mitton JB (1993) Theory and data pertinent to the relationship between heterozygosity and fitness. In: Thornhill
NW (ed) The Natural History of Inbreeding and Outbreeding: Theoretical and Empirical Perspectives.
University of Chicago Press, Chicago, IL, pp. 352374.
Mitton JB, Schuster WSF, Cothran EG & De Fries JC (1993) The correlation between the individual
heterozygosity of parents and their offspring. Heredity 71: 5963.
Montes JM, Technow F, Martin M & Becker K (2014) Genetic diversity in Jatropha curcas l. assessed with
SSR and SNP markers. Diversity 6: 551566.
Muakrong N, One KT, Tanya P & Srinives P (2013) Interspecic Jatropha hybrid as a new promising source of
woody biomass. Plant Genetic Resources: Characterization and Utilization 12: S17S20.
Murovec J, Stajner N, Jakse J & Javornik B (2007) Microsatellite marker for homozygosity testing of putative
doubled haploids and characterization of Mimulus species derived by a cross-genera approach. Journal of
American Society for Horticultural Science 132: 659663.
Ong HC, Mahlia TMI, Masjuki HH & Norhasyima RS (2011) Comparison of palm oil, Jatropha curcas and
Calophyllum inophyllum for biodiesel: a review. Renewable Sustainable Energy Review 15: 35013515.
Openshaw K (2000) A review of Jatropha curcas: an oil plant of unfulfilled promise. Biomass and
Bioenergy19: 115.
Osorio LRM, Salvador AFT, Jongschaap REE, Perez CAA, Sandoval JEB, Trindade LM, Visser RGF & van
Loo EN (2014) High level of molecular and phenotypic biodiversity in Jatropha curcas from Central
America compared to Africa, Asia and South America. BMC Plant Biology 14: 77.
Pamidimarri DVNS & Reddy MP (2014) Phylogeography and molecular diversity analysis of Jatropha curcas
L. and the dispersal route revealed by RAPD, AFLP and nrDNA-ITS analysis. Molecular Biology Report
41: 32253234.
Pecina-Quintero V, Anaya-Lopez JL, Colmenero AZ, Garcia NM, Colin CAN, Bonilla JLS, Agilar-Rangel MR,
Langarica JLS & Bustamante DJM (2011) Molecular characterization of Jatropha curcas L. genetic
resources from Chiapas, Mexico through AFLP markers. Biomass and Bioenergy 35: 18971905.
Pecina-Quintero V, Anaya-Lopez JL, Zamarripa-Colmenero A, Nunez-Coln CA, Montes-Garca N, SolsBonilla JL & Jimenez-Becerril MF (2014) Genetic structure of Jatropha curcas L. in Mexico and probable
centre of origin. Biomass and Bioenergy 60: 147155.
Popluechai S, Breviario D, Sujatha M, Makkar HPS, Raorane M, Reddy AR, Palchetti E, Gatehouse AMR,
Syers JK, Donnell AGO & Kohli A (2009) Narrow genetic and apparent phenetic diversity in Jatropha
curcas: initial success with generating low phorbol ester interspecific hybrids. Nature Precedings
hdl:10101/npre.2009.2782.1.
Rafii MY, Shabanimofrad M, Edaroyati PMW & Latif MA (2012) Analysis of the genetic diversity of physic
nut, Jatropha curcas L. accessions using RAPD markers. Molecular Biology Report 39: 65056511.
Rao GR, Korwar GR, Shanker AK & Ramakrishna YS (2008) Genetic associations, variability and diversity in
seed characters, growth, reproductive phenology and yield in Jatropha curcas (L.) accessions. Trees 22:
697709.
Ricci A, Chekhovskiy K, Azhaguvel P, Albertini E, Falcinelli M & Saha M (2012) Molecular characterization
of Jatropha curcas resources and identification of population-specific markers. Bioenergy Research 5: 215
224.
Rosado TB, Laviola BG, Faria DA, Pappas MR, Bhering LL, Quirino B & Grattapaglia D (2010) Molecular
marker reveal limited genetic diversity in large germplasm collection of the biofuels crop Jatropha curcas L.
in Brazil. Crop Science 50: 23722382.
Sato S, Hirakawa H & Isobe S (2011) Sequence analysis of the genome of an oil-bearing tree, Jatropha curcas
L. DNA Research 18: 6576.
Shen J, Khongsak P, David B & Xiaoyang C (2012) AFLP-based molecular characterization of 63 populations
of Jatropha curcas L. grown in provenance trials in China and Vietnam. Biomass and Bioenergy 37: 265
274.
Sinha P & Tripathi SB (2013) Molecular characterization of Jatropha genetic resources through amplified
fragment length polymorphism (AFLP) markers. International Journal of Chemical and Technical Research
5: 735740.
197
Maurya & Yadav (2016) 3(1): 191198

.
Sirisomboon P, Kitchaiya P, Pholpho T & Mahuttanyavanitch W (2007) Physical and mechanical properties of
Jatropha curcas L. fruits, nuts and kernels. Biosystem Engineeing 97: 201207.
Soonthornyatara S, Sripichitt P, Kaveeta R & Hongtrakul V (2015) Assessment of genetic diversity of Jatropha
curcas L. using AFLP and ISSR markers. Chiang Mai Journal of Science 42: 614625.
Sujatha M & Prabakaran AJ (1997) Characterization and utilization of Indian Jatropha curcas. Indian Journal
of Plant Genetic Resources 10: 12328.
Sujatha M, Reddy TP & Mahasi MJ (2008) Role of biotechnological interventions in the improvement of
castor (Ricinus communis L.) and Jatropha curcas L. . Biotechnology Advance 26: 42435.
Sun QB, Li LF, Li Y, Wu GJ & Ge XJ (2008) SSR and AFLP markers reveal low genetic diversity in the
biofuel plant Jatropha curcas in China. Crop Science 48:1865-1871.
Tanya P, Taeprayoon P, Hadkam Y & Srinives P (2011) Genetic diversity among Jatropha and Jatropha related
species based on ISSR markers. Plant Molecular Biology Reports 29: 252264.
Trebbi D, Papazoglou EG, Saadaoui E, Vischi M, Baldini M, Stevanato P, Cettul E, Sanzone AP, Gualdi L &
Fabbri A (2015) Assessment of genetic diversity in different accessions of Jatropha curcas. Industrial Crops
and Products 75: 3539.
Virk PS, Zhu J, Newbury HJ, Bryan GJ, Kackson MT & Ford-Lloyd BV (2000) Effectiveness of different
classes of molecular markers for classifying and revealing variations in rice (Oryza sativa) germplasm.
Euphytca 112: 275284.
Yadav HK, Ranjan A, Asif MH, Mantri S, Sawant SV & Tuli R (2011) EST-derived SSR markers in Jatropha
curcas L.: development, characterization, polymorphism, and transferability across the species/genera. Tree
Genetics and Genomes 7: 207219.
Yahata M, Harusaki S, Komatsu H, Takami K, Kunitake H, Yabuya T, Yamashita K & Toolapong P (2005)
Morphological characterization and molecular verification of a fertile haploid pummelo (Citrus grandis
Osbeck). Journal of American Society for Horticultural Science 130: 3440.
198
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 199212, 2016
Research article
Leaf life-span dynamics of woody species in tropical dry forests of

India
R. K. Chaturvedi1* and A. S. Raghubanshi2
1
Centre for Integrative Conservation, Xishuangbanna Tropical Botanical Garden,

Chinese Academy of Sciences, Menglun, Mengla, Yunnan 666303, China
2
Institute of Environment and Sustainable Development, Banaras Hindu University, Varanasi 221005, India
*Corresponding Author: ravikantchaturvedi10@gmail.com
Abstract: We analysed the diversity of leaf life-span in the woody species of Vindhyan highlands
and grouped the plant species according to their leaf flushing period, leaf life span and leaf fall
period. We also studied the factors influencing the foliar phenology of the woody species. The
study was conducted on five sites (Hathinala, Gaighat, Harnakachar and Ranitali) within the
tropical dry deciduous forest in northern India. Leaf life-spans of the woody species were highly
variable. About 67% species had peak of their leaf flush initiation during summer period (pre-rain
leaf flushing) and rest species had their peak leaf initiation during rainy season (post-rain leaf
flushing). The peak period of leaf flushing initiation at all the sites was May when 52% of the
species initiated their leaf formation and the peak period of leaf flushing completion was August
when 38% of the plant species completed their leaf formation. The peak period of leaf fall
initiation in maximum species (54%) at all sites was November and the peak period of leaf fall
completion at all the sites was February when 63% of the species shed their leaves. The peak
period of leaf flush initiation as well as leaf fall initiation in most of the species at dry sites was
one month before as compared to that of moist sites.
Keywords: Leaf phenology - Tropical dry forest - Soil moisture content - Deciduousness - Woody
species.
[Cite as: Chaturvedi RK & Raghubanshi AS (2016) Leaf life-span dynamics of woody species in tropical dry
forests of India. Tropical Plant Research 3(1): 199212]
INTRODUCTION
The three most important factors influencing the phenological changes in the tropical dry forests (TDFs) are:
(i) rainfall (Daubenmire 1972, Bullock & Solis-Magallanes 1990, Borchert 1994a, Eamus & Prior 2001, Bajpai
et al. 2016); (ii) photoperiod (Bullock & Solis Magallanes 1990, Rivera et al. 2002, Borchert et al. 2004, Elliot
et al. 2006, Bajpai et al. 2012, 2016); and (iii) stem water status (Borchert 1994a, b, 1998). Growth and
reproduction of plant species in the TDFs are significantly influenced by the spatial and temporal variations in
the foliar phenology (Suresh & Sukumar 2011, Nanda et al. 2014, Borah & Devi 2014). Many tree species in
these forests flush their leaves during the dry season, before the onset of the rains (Bullock & Solis-Magallanes
1990, Mooney et al. 1995). For this mechanism, two principal reasons have been suggested: (1) new leaves may
be able to make maximum use of the higher radiation during the dry season (Wright & van Schaik 1994); and
(2) new leaves avoid predation when herbivores are at their least abundant in the dry season (Murali & Sukumar
1993). The variations in foliar phenology in the TDFs, could be an adaptation to the pressures exerted by
animals, plants and may rely on the changes in environmental conditions to trigger the event (van Schaik et al.
1993). Phenology is sometimes concurrently or at different times of the year, controlled by multiple factors
(Nilsen & Muller 1981, White et al. 1997, Jolly et al. 2005). Therefore, it is important to understand the factors
influencing the foliar phenology in the woody species of TDFs.
Water stored in the tree stem, or remaining in the subsoil, buffers the impact of low water availability and
allows the production of new leaves during the dry season (Borchert 1980, 1983, 1994b, c, Reich & Borchert
1984). This indicates that tree water status, rather than a climatic factor directly, is probably the principal
determinant of tree phenology in the TDFs. Borchert & Rivera (2001) have shown that leaf buds remain
Received: 02 February 2016
199
Chaturvedi & Raghubanshi (2016) 3(1): 199212

.
dormant during the dry season in many tree species of semi-deciduous tropical forests, and bud-break is induced
by an increasing photoperiod after the spring equinox. Bud-break is highly synchronous in conspecifics of these
spring-flushing trees, although some within species differences occurred. One likely explanation for the latter
is the amount of stem-, soil- or rain-water available to the tree. Whether leaf flush is triggered by photoperiod or
other factors, sufficient water supply is a prerequisite. Bud-break and leaf expansion during the dry season occur
only when the trees are fully rehydrated (Borchert 1994b, c, Borchert et al. 2002), and the rate of shoot
development and the duration of leaf expansion varies strongly with water availability (Borchert 1994c,
Borchert & Rivera 2001).
Intraspecific variations in leaf lifespan are reported in plant species growing in the tropics (Ackerly &
Bazzaz 1995, Reich et al. 2004, Vincent 2006) and in temperate zones (Jurik & Chabot 1986, Oikawa et al.
2004). It is a general belief that plants adjust leaf life-span so as to maximize whole-plant photosynthesis
(Franklin & Agren 2002, Hikosaka 2003, 2005, Kikuzawa & Lechowicz 2006, Oikawa et al. 2006, 2008).
Models predict that when whole-plant photosynthesis is high, leaves are more frequently shed as the plant
prioritizes investment in new leaves (Kikuzawa 1991, Hikosaka 2003), and this pattern is consistently observed
across different biomes and taxa (Reich et al. 1999, Wright et al. 2004, Karst & Lechowicz 2007). Hence,
intraspecific variation in leaf life-span should be considered as an adaptation for carbon gain (Oikawa et al.
2004).
In general, deciduous species have a higher photosynthetic rate and shorter leaf life span than evergreens,
while evergreens are more shade-tolerant (Chabot & Hicks 1982). Similarly, within a single species, an
individual in a well-lit place has a higher photosynthetic rate (Noda et al. 2004) than a shaded individual.
Therefore, cost-benefit analysis also shows that having a higher photosynthetic rate leads to a high leaf-turnover
rate, which in turn leads to a shorter leaf life span. Accordingly, a shorter mean leaf life span has been reported
for individuals in well-lit environments relative to shaded conspecifics (Seiwa & Kikuzawa 1991, Ackerly &
Bazzaz 1995, Hikosaka 2005, Vincent 2006). Apart from high rates of photosynthesis, species with short leaf
life span also have high respiration rate, leaf nutrient concentrations and SLA (Wright et al. 2004).
In the present study, we analyse the diversity of leaf life-span in the woody species of Vindhyan highlands
and group the plant species according to their leaf flushing period, leaf life span and leaf fall period. We also
study the factor influencing the foliar phenology of the woody species.
Study area
We selected five study sites in the forests of Vindhyan highlands (241807250017 N, 823738
832305 E). Among the five sites, Hathinala, Gaighat, Harnakachar and Ranitali sites are situated in
Sonebhadra district and Kotwa in Mirzapur district of Uttar Pradesh (Fig. 1). They occupy land area of 2555,
394, 1507, 2118 and 199 hectares, respectively. The selected sites represented a range in soil water availability.
The area experiences tropical monsoon climate with three seasons in a year, viz., summer (Aprilmid June),
rainy (mid JuneSeptember) and winter (NovemberFebruary). The months of March and October constitute
transition periods, respectively between winter and summer, and between rainy and winter seasons. The mean
maximum monthly temperature varies from 20C in January to 46C in June, and the mean minimum monthly
temperature reaches 12C in January and 31C in May. According to the data collected from the meteorological
stations of the state forest department for 19802010, the mean annual rainfall ranges from 865 to 1196 mm
(Chaturvedi et al. 2011a). About 85 % of the annual rainfall occurs during the monsoon (rainy) season from the
south-west monsoon, and the remaining from the few showers in December and in MayJune. There is an
extended dry period of about 9 months (Octobermid June) in the annual cycle (Jha & Singh 1990). The
monthly rainfall varies from 6 mm in April to 334 mm in August (Chaturvedi et al. 2012). Soils of the study
area are residual, ultisols, sandy-loam in texture, reddish to dark gray in colour and extremely poor in nutrients
(Chaturvedi & Raghubanshi 2011). Recently estimated physico-chemical properties of soils of the study region
have been described in Chaturvedi & Raghubanshi (2015). The forest region exhibit patchiness in the species
composition due to small variations in the environmental variables (Chaturvedi et al. 2011b). Species
composition, distribution and diversity (Chaturvedi & Raghubanshi 2014), seasonal growth (Chaturvedi et al.
2011c, 2013, 2014), and functional traits (Chaturvedi & Raghubanshi 2013) of woody species are highly
influenced by soil moisture content. The density-DBH distributions in the forest region exhibited low DBHwww.tropicalplantresearch.com
200

.
biased structure, where the average density of seedlings, saplings and adults were 9,2611,511, 799154 and
29762, respectively (Chaturvedi & Raghubanshi 2014).
Figure 1. Map showing the location of study areas.
Study Design
For the purpose of this study, three plots, each of 4 ha (200 m 200 m), were established randomly at each
site. Fifteen individuals each, of the species were randomly selected from the three 4 ha plots at each site and
were marked. Plant species which were not present at all the five sites were selected from only those sites where
they were present. On each marked individual, one twig (currently growing shoots of last-order branches) on
each of four major branches (one in each direction was marked with metal tags). On these twigs monthly count
of leaf number was made from January 2005 to December 2006.
Soil moisture content was measured at 10 locations, randomly in each plot, at each site, as percentage by
volume every month, at a depth of 10 cm at 1-month intervals for 2 years (i.e. January 2005 to December 2006)
using a theta probe instrument (type ML 1, Delta-T Devices, Cambridge, UK). The following phenological
events were derived from the monthly leaf counts: initiation of leaf flush, completion of leaf flush, leaf-fall
initiation, and completion of leaf fall. Leaf-flush period of a species is the duration (days) from the first leaf
flush to the last flush amongst its individuals. Leaf fall period of a species represents the time duration from the
estimated first leaf fall to the last amongst individuals. The leaf life-span period for each species was calculated
as the mean leaf life-span of all individuals of the species. Species were classified as Group I and Group II on
the basis of pre- and post-rainfall leaf flushing, respectively. Also, on the basis of leaf life-span, species were
categorized into four groups as (i) 1012 mo life-span, (ii) 810 mo life-span, (iii) 68 mo life-span and (iv) 46
mo life-span, excluding the extreme values in each case. Species with 812 mo leaf life-span belong to Group I
and with 48 mo leaf life-span to Group II. Site wise variations in the leaf flush and leaf fall was analysed for
these two groups.
201

.
Data analysis
Soil moisture content measured at 10 locations in each plot was averaged to get three values for each site, for
each month. Site wise variations of soil moisture content in each month was analysed by multivariate ANOVA
using the SPSS (v. 16) package.
RESULTS
Soil moisture content
Across the five study sites, monthly measured soil moisture content was generally maximum at Hathinala
and minimum at Kotwa (Fig. 2). Across the 12 months in a year, July and August showed higher soil moisture
content. Lower soil moisture content was observed in November and December. Across the 12 months, site wise
variations in soil moisture content were not significant only in July, August and September (Table 1).
Figure 2. Monthly soil moisture content at the five study sites from January 2005 to December 2006.
Table 1. ANOVA on soil moisture content measured monthly across the five study sites. The residual df is 10.
Month
Jan
Feb
Mar
Apr
May
Jun
Jul
Aug
Sep
Oct
Nov
Dec
d.f.
4
4
4
4
4
4
4
4
4
4
4
4
F
13.375
8.393
67.031
7.576
16.701
14.708
2.157
1.508
2.426
115.35
51.659
41.954
P
< 0.010
< 0.010
< 0.001
< 0.010
< 0.001
< 0.001
> 0.050
> 0.050
> 0.050
< 0.001
< 0.001
< 0.001
Diversity of Leaf Life span

Leaf life-spans of the woody species were also highly variable (Fig. 3). Leaves of Hardwickia binata and
Shorea robusta had life-span of 10 to 12 months. Life-span of 8 to 10 months was observed in the leaves of
seven species (Acacia auriculiformis, Albizia odoratissima, Azadirachta indica, Bauhinia racemosa, Carissa
202

.
spinarum, Diospyros melanoxylon and Ficus racemosa). In 31 species (Acacia catechu, Anogeissus latifolia,
Bridelia retusa, Buchanania lanzan, Cassia fistula, Cassia siamea, Chloroxylon swietenia, Dendrocalamus
strictus, Elaeodendron glaucum, Flacourtia indica, Gardenia turgida, Grewia hirsuta, Grewia serrulata,
Hollarrhena antidysenterica, Holoptelia integrifolia, Hymenodictyon excelsum, Indigofera cassioides,
Lagerstroemia parviflora, Madhuca longifolia, Miliusa tomentosa, Mitragyna parvifolia, Nyctanthes
arbortristis, Ougeinia oogenesis, Pterocarpus marsupium, Schleichera oleosa, Schrebera swietenioides,
Soymida febrifuga, Terminalia chebula, Terminalia tomentosa, Zizyphus nummularia and Zizyphus oenoplea),
life-span of 6 to 8 months was detected, whereas, in rest of the 12 species (Abrus precatorius, Adina cordifolia,
Boswellia serrata, Emblica officinalis, Eriolaena quinquelocularis, Gardenia latifolia, Lannea coromandelica,
Lantana camara, Semecarpus anacardium, Sterculia urens, Woodfordia fruticosa and Zizyphus glaberrima), the
life-span was of 4 to 6 months (Fig. 3). In our study, about 67% species had peak of their leaf flush initiation
during summer period (Group I species, pre-rain leaf flushing) and rest species had their peak leaf initiation
during rainy season (Group II, post-rain leaf flushing).
Figure 3. Periods of major (solid black horizontal bars) and minor (hatched horizontal bars) leaf flush in woody species of Vindhyan
highlands. Periods of substantial leaf drop is indicated by downward arrows. 0 indicates when most individuals were leafless (Ls = Lifespan in months).
Diversity of Leaf Flushing

There occurred wide diversity among species in terms of duration of leaf flush (Fig. 3). F. racemosa, L.
camara and S. robusta produced new leaves for 6-8 months. Seven species (A. catechu, A. odoratissima, A.
latifolia, C. spinarum, G. serrulata, H. binata and L. parviflora) produced new leaves through 5-6 months, 18
species (A. auriculiformis, B. retusa, B. lanzan, C. siamea, D. melanoxylon, E. glaucum, F. indica, G. latifolia,
H. antidysenterica, M. tomentosa, M. parvifolia, N. arbortristis, S. oleosa, Soymeda febrifuga, T. chebula, T.
tomentosa, W. fruticosa and Z. nummularia) flushed through 4-5 months and 24 species (A. precatorius, A.
cordifolia, A. indica, B. racemosa, B. serrata, C. fistula, C. swietenia, D. strictus, E. officinalis, E.
quinquelocularis, G. turgida, G. hirsuta, H. integrifolia, H. excelsum, I. cassioides, L. coromandelica, M.
longifolia, O. oogenesis, P. marsupium, S. swietenioides, S. anacardium, S. urens, Z. glaberrima and Z.
oenoplea) flushed through 3-4 months (Fig. 3).
203

.
Figure 4. Initiation and completion of leaf flushing. Data for Hathinala are represented by circles, for Gaighat by squares,
for Harnakachar by diamond, for Ranitali by up triangle and for Kotwa by down triangle. Solid symbols are for initiation and
open symbols are for completion of phenological event. The smooth solid curves represent initiation and dash dotted curves
represent completion of phenological event.
In general, the peak period of leaf flushing initiation at all the sites was May when 52% of the species
initiated their leaf formation and the peak period of leaf flushing completion was August when 38% of the plant
species completed their leaf formation (Fig. 4). At the community level, May constituted the peak period of leaf
flushing initiation at Hathinala (63%), Gaighat (48%) and Harnakachar (51%), whereas, the peak period leaf
flushing initiation at Ranitali (42%) and Kotwa (43%) was April (Fig. 5). Similar to the peak period leaf
flushing initiation, the peak period of completion of leaf flushing was same for Hathinala, Gaighat and
Harnakachar where 46%, 55% and 48% of the species have shown completion of their leaf formation
respectively. In the drier sites, the completion of leaf flushing was August and September (both 42%) for
Ranitali and August (50%) for Kotwa (Fig. 5).
Figure 5. Initiation and completion of leaf flush of species in the study sites. HN, Hathinala; GG, Gaighat; HK,
Harnakachar; RT, Ranitali; KT, Kotwa.
Leaf Flush Initiation

Peak period of leaf flush initiation in majority of species in group I (pre-rainfall group) occurred in March
(67%), however, species in group II (post-rainfall group) were highly deciduous and most of them (63%)
showed their peak period of leaf flush initiation in May (Fig. 6). In group I, 80% species at Hathinala, 50%
204

.
species at Gaighat and Harnakachar showed their peak period of leaf flush initiation in April (Fig. 6). At
Ranitali, S. robusta showed peak period of leaf flush initiation in February, H. binata in March and D.
melanoxylon in April. Only two species at Kotwa were in group I, in which the peak period of leaf flush
initiation was in April (for D. melanoxylon) and March (for F. racemosa). Most of the species at Hathinala
(72%), Gaighat (63%) and Harnakachar (59%), belonging to group II showed their peak period of leaf flush
initiation in May (Fig. 6). At Ranitali, most of the species (43%) showed the peak period of leaf flush initiation
equally in April and May, whereas, at Kotwa, majority of the species (42%) showed the peak period of leaf
flush initiation in April (Fig. 6).
Figure 6. Initiation and completion of leaf flushing of species having 8-12 months (Group I) and 4-8 (Group II) months of
leaf life span. Data for Hathinala are represented by circles; Gaighat by squares; Harnakachar by diamond; Ranitali by up
triangle; Kotwa by down triangle. Solid and solid cross haired symbols are for initiation; open and open cross haired
symbols are for completion of phonological event. The smooth solid curves represent initiation in Group I species; broken
curves represent initiation in Group II species; dash dotted curves represent completion in Group I species; dash double
dotted curves represent completion in Group II species.
Leaf Flush Completion

Phenological event of leaf flush completion was much variable among species, the peak period in group I for
A. indica and B. racemosa was June, for A. odoratissima, it was July, for C. spinarum and D. melanoxylon it
was August, for H. binata and A. auriculiformis it was September, for F. racemosa it was October and for S.
robusta it was November. The peak period of leaf flush completion in species of group II was also variable but
majority of species showed their peak period in August for 42% and September for 40% of the species (Fig. 6).
The species of group II were highly deciduous and leaf flush completion ended in October, however, in L.
camara, the peak period of leaf flush completion was November. The peak period of leaf flush completion at
Hathinala was much variable among the species where, B. racemosa showed the peak in June, A. odoratissima
in July, D. melanoxylon in August, H. binata in October and S. robusta in November.
At Gaighat, A. indica showed maximum peak in June, D. melanoxylon in July (similar at Harnakachar,
Ranitali and Kotwa), C. spinarum in August (similar at Harnakachar), H. binata (similar at Harnakachar and
Ranitali) and A. auriculiformis in September and S. robusta in October (similar at Harnakachar and Ranitali). At
Kotwa, the site specific species F. racemosa showed the peak in October. In group II most of the species at
Hathinala (53%), Gaighat (63%) and Harnakachar (52%) showed their peak of leaf flush completion in
September, however, at Harnakachar and Kotwa the peak for most of the species was observed in August (48%
for Ranitali and 58% for Kotwa) (Fig. 6).
Duration of Leaf Fall
Generally, the duration of leaf fall for most of the species ranged through 34 months (DecMar), however,
in F. racemosa and S. robusta minor leaf fall was also observed in September and October. Duration of leafless
period ranged from one to three months, however, individuals of A. auriculiformis, C. siamea, F. racemosa and
205

.
S. robusta were never found completely leafless in their annual cycle. The only species leafless for 4 months
was S. urens (Fig. 3).
Leaf Fall Initiation
Figure 7. Initiation and completion of leaf fall. Data for Hathinala are represented by circles, for Gaighat by squares, for
Harnakachar by diamond, for Ranitali by up triangle and for Kotwa by down triangle. Solid symbols are for initiation and
open symbols are for completion of phenological event. The smooth solid curves represent initiation and dash dotted curves
represent completion of phenological event.
The peak period of leaf fall initiation in maximum species (54%) at all sites was November and the peak
period of leaf fall completion at all the sites was February when 63% of the species shed their leaves (Fig. 7).
When the leaf fall in plant species was studied at different sites, it was observed that the peak period of leaf fall
initiation for majority of species at Hathinala (53%), Gaighat (65%) and Harnakachar (52%) was December,
whereas at drier sites, i.e. Ranitali and Kotwa, the peak period of leaf fall initiation for most of the species was
November (59% at Ranitali and 64% at Kotwa) which is one month before than that of the moist sites (Fig. 8).
The peak period of leaf fall completion at the three comparatively moist sites was February when 73% of the
species at Hathinala, 58% at Gaighat and 55% at Harnakachar shed their leaves. At the drier sites, the peak
period of leaf fall completion was January when 52% of species at Ranitali and 36% of species at Kotwa shed
their leaves (Fig. 8).
Figure 8. Initiation and completion of leaf fall of species in the study sites. HN, Hathinala; GG, Gaighat; HK, Harnakachar;
RT, Ranitali; KT, Kotwa.
206

.
Similar to the peak period of leaf flush initiation, the peak period of leaf fall initiation for most of the species
in group I was one month after that of group II species. In group I, the peak reached in December whereas in
group II, the peak was in November (Fig. 9). At Hathinala, the peak period of H. binata, A. odoratissima and B.
racemosa was November, whereas, for the other two species (S. robusta and D. melanoxylon), it was
September.
Figure 9. Initiation and completion of leaf fall of species having 8-12 mo (Group I) and 4-8 mo (Group II) of leaf life span.
Data for Hathinala are represented by circles; Gaighat by squares; Harnakachar by diamond; Ranitali by up triangle; Kotwa
by down triangle. Solid and solid cross haired symbols are for initiation; open and open cross haired symbols are for
completion of phonological event. The smooth solid curves represent initiation in (Group II) species; broken curves
represent initiation in (Group I) species; dash dotted curves represent completion in (Group II) species; dash double dotted
curves represent completion in (Group I) species.
At Gaighat, the peak for S. robusta reached in September (similar for Harnakachar and Ranitali), for C.
spinarum in October (similar for Harnakachar), for H. binata and D. melanoxylon in November (similar for
Harnakachar, Ranitali and Kotwa) and for A. auriculiformis in December. F. racemosa, the site specific species
at Kotwa showed its peak of leaf fall initiation in September. Species in group II at all the five sites had same
peak period of leaf fall initiation i.e. November (Fig. 9), where 56% species at Hathinala, 71% at Gaighat, 69%
at Harnakachar, 57% at Ranitali and 75% species at Kotwa started their leaf fall.
Leaf Fall Completion
Peak period of leaf fall completion for most of the species in group I (67%) and group II (84%) was
February (Fig. 9). In the species of group I at Hathinala, A. odoratissima and B. racemosa showed the peak of
leaf fall in February, whereas, H. binata, S. robusta and D. melanoxylon attained their peak in March. At
Gaighat, A. auriculiformis reached the peak in June, however, the other five species showed the peak in
February. All the species at Harnakachar and Ranitali reached the peak of leaf fall completion in February,
whereas, at Kotwa, D. melanoxylon showed its peak in February and F. racemosa in March. Similar to the leaf
fall initiation, the peak period of leaf fall completion for most of the species in group II was same at all the five
sites (Fig. 9), where 81% species at Hathinala, 88% at Gaighat, 93% at Harnakachar, 87% at Ranitali and 50%
at Kotwa were leafless in February.
In deciduous woody species of TDFs, both spring flushing and summer flushing generally precede the first
rains by 12 months, suggesting their timing has been selected for by the rainfall pattern. The rates of leaf flush
and leaf fall in different plants vary with available soil moisture, leaf structure, depth of root system, and other
variables and, therefore, leaf phenology is asynchronous within a population. Our study showed variations in
leaf phenological events of woody species in moist and dry sites. Within site variation of leaf flushing and leaf
fall events were also very prominent.
207

.
The characteristic phenology of dry period flushing species is also controlled by non-climatic environmental
variables, such as water storage in deep soils and photoperiodic induction of leafing, which also determine tree
phenology in TDF species around the globe (Borchert 1994d, Rivera et al. 2002). These variations may be due
to patchy distribution of microhabitats in the TDF soils (Roy & Singh 1994). To exploit the resources present in
these microhabitats, species evolve differently which causes variations in their leaf phenology.
In our study, most of the species had peak of their leaf flush intiation during dry period (Group I species,
pre-rain leaf flushing) while rest species had their peak leaf initiation during rainy season (Group II, post-rain
leaf flushing). Species belonging to Group I are often capable of physiological adjustment in low soil moisture
condition when atmospheric humidity is also low, resulting in greater photosynthetic carbon gain and lower
transpirational water loss than otherwise would be possible (Mulkey et al. 1992). In contrast, the species
belonging to Group II are not able to adjust to dry season conditions because they are produced during period of
high water availability.
Besides the adjustment of plant species to low soil water availability, early leaf flushing has been reported to
protect the young leaves from the herbivore damage which occurs when a leaf is young (Coley & Aide 1991).
This damage affects growth, survivorship, and reproduction of the plant species (Crawley 1985, Dirzo 1984,
Krischik & Denno 1983, Louda 1984, Marquis 1984). Chemical and physical defenses, particularly leaf
toughness, are effective herbivore defense of mature leaves (Coley 1983), but these defenses are not as well
developed in young leaves. However, because young leaves are an ephemeral stage in the life of a leaf,
characteristics of leaf phenology also may influence the degree of vulnerability of young leaves to herbivores
(Aide 1988, Clark & Clark 1991). Therefore, in TDFs, some species "anticipate" the wet season and produce
leaves under drought conditions during the dry season (Frankie et al. 1974, Rockwood 1973; Shukla &
Ramakrishnan 1982). Herbivore abundances are usually low during the dry season (Wolda 1978, but see
Boinski & Fowler 1989), and thus leaves produced at this time should temporally escape herbivores (Aide
1988).
Although, there was wide variation in the leaf flush initiation and leaf fall initiation in the woody species
across the study sites, the peak period of leaf flush initiation as well as leaf fall initiation in most of the species
at dry sites was one month before as compared to that of moist sites. Plant species growing at the dry sites
tolerate water stress to a greater extent as compared to moist site. Early leaf flushing helps them to extend the
wet growing season which is shorter than the moist site due to early leaf fall in the dry season.
During rainy season, when soil moisture becomes uniform across the landscape, the period of leaf flush
initiation and completion at different sites for most of the species form similar pattern. This similarity of leaf
flushing event was clearly seen in Fig. 5 where similar curves having sharp boundary was observed for all the
five study sites. This synchrony in leaf flushing event is due to similarity in soil moisture availability for the
plant species at different sites. Therefore, site variability is not the determining factor for leaf flush initiation in
the rainy season. However, during the phenological event of leaf fall, there occurs much difference in the
phenological pattern which is seen in Fig. 8, where the peak period of leaf fall for most of the species at
different study sites form variable curves. Here, the asynchrony is due to site-wise variation in soil moisture
availability. Plants acquire water according to their rooting depth and the water holding capacity of the site.
Woody species in the Vindhyan region exhibit a broad range of deciduousness, and four classes can be
recognized; semi-evergreen, <2-month deciduous, 24-mo-deciduous and >4-month deciduous. This study
detected two species in semi-evergreen category, seven species in 2-4-months deciduous and 43 species in >4month deciduous category. Leafless period in woody species, generally occurring in response to water stress,
represents the time period during the annual cycle when resources (light, water, nutrients etc.) are not being
exploited or are being used at a low intensity. The duration of the tolerance of water stress in different
leaflessness categories should decrease in the order: semi-evergreen, <2-mo-deciduous, 24-mo-deciduous, >4mo-deciduous. Prolonged leaflessness may strongly affect the water relations of woody species, leading to
selection for low-density wood with greater water storage (e.g. >4-mo-deciduous B. serrata and L.
coromandelica which show characteristics equivalent to stem succulents of Borchert 2000).
Semi-evergreen and short-deciduous woody species show adaptations such as deep root systems with access
to subsoil water. Skarpe (1996) reported that evergreen species have deep root systems, deciduous fine leaved
trees have deep to moderately shallow roots, and deciduous broad-leaved trees have moderately deep to
moderately shallow roots.
208

.
Our study area experience a long dry season, the woody species are adapted in such a way that they can
tolerate water stress for the maximum possible duration to maintain relative growth rate by retaining leaves for
longest duration. When water stress reaches the threshold, they shed their leaves rapidly to avoid water stress.
Site wise variation in the foliar phenology reveals that the soil moisture content is an important factor in TDFs,
influencing foliar phenology of the woody species. However, many other factors still needs to be studied to
completely understand the phenological event.
ACKNOWLEDGMENTS
We thank the Divisional Forest Officer, Renukoot, Sonebhadra, Uttar Pradesh, India, for granting permission
to work in the forest. The study was financially supported by Research Associate scheme of Council of
Scientific and Industrial Research (award no. 09/13(452)/2012-EMR-I; to R.K.C.).
REFERENCES
Ackerly DD & Bazzaz FA (1995) Leaf dynamics, self-shading and carbon gain in seedlings of a tropical pioneer
tree. Oecologia 101: 289298.
Aide TM (1988) Herbivory as a selective agent on the timing of leaf production in a tropical understory
community. Nature 336: 574575.
Bajpai O, Kumar A, Mishra AK, Sahu N, Behera SK & Chaudhury LB (2012) Phenological study of two
dominant tree species in tropical moist deciduous forest from the northern India. International Journal of
Botany 8(2): 6672.
Bajpai O, Pandey J & Chaudhury LB (2016) Periodicity of different phenophases in selected trees from
Himalayan Terai of India. Agroforestry Systems [DOI: 10.1007/s10457-016-9936-9]
Boinski S & Fowler NL (1989) Seasonal patterns in a tropical lowland forest. Biotropica 21: 223233.
Borah M & Devi A (2014) Phenology, growth and survival of Vatica lanceaefolia Bl.: A critically endangered
tree species in moist tropical forest of Northeast India. Tropical Plant Research 1(3): 112.
Borchert R & Rivera G (2001) Photoperiodic control of seasonal development and dormancy in tropical stemsucculent trees. Tree Physiology 21: 213221.
Borchert R (1980) Phenology and ecophysiology of tropical trees: Erythrina poeppigiana O. F. Cook. Ecology
61: 10651074.
Borchert R (1983) Phenology and control of flowering in tropical trees. Biotropica 15: 8189.
Borchert R (1994a) Induction of rehydration and budbreak by irrigation or rain in deciduous trees of a tropical
dry forest in Costa Rica. Trees 8: 198204.
Borchert R (1994b) Soil and stem water storage determine phenology and distribution of tropical dry forest
trees. Biotropica 15: 8189.
Borchert R (1994c) Water status and development of tropical trees during seasonal drought. Trees-Structructure
and Function 8: 115125.
Borchert R (1994d) Water storage in soil or tree stems determines phenology and distribution of tropical dry
forest trees. Ecology 75: 14371449.
Borchert R (1998) Response of tropical trees to rainfall seasonality and its long-term changes. Climate Change
39: 381393.
Borchert R (2000) Organismic and environmental controls of bud growth in tropical trees. In: Viemont JD &
Crabbe J (eds) Dormancy in plants: from whole plant behavior to cellular control. Wallingford: CAB
International, pp. 87107.
Borchert R, Meyer SA, Felger RS & Porter-Bolland L (2004) Environmental control of flowering periodicity in
Costa Rican and Mexican tropical dry forests. Global Ecology and Biogeography 13: 409425.
Borchert R, Rivera G & Hagnauer W (2002) Modification of vegetative phenology in a tropical semi-deciduous
forest by abnormal drought and rain. Biotropica 34: 2739.
Bullock SH & Solis-Magallanes JA (1990) Phenology of canopy trees of a tropical deciduous forest in Mexico.
Biotropica 22: 2235.
Chabot BF & Hicks DJ (1982) The ecology of leaf life spans. Annual Review of Ecology and Systematics 13:
229259.
Chaturvedi RK & Raghubanshi AS (2011) Plant Functional Traits in a Tropical Deciduous Forest: An
Analysis. Berlin, Germany: Lambert Academic Publishing GmbH and Co. KG.
209

.
Chaturvedi RK & Raghubanshi AS (2013) Phenotypic plasticity in functional traits of woody species in tropical
dry forest. In: Valentino JB & Harrelson PC (eds) Phenotypic Plasticity: Molecular Mechanisms,
Evolutionary Significance and Impact on Speciation. New York: Nova Science Publishers, Inc, pp. 3566.
Chaturvedi RK & Raghubanshi AS (2014) Species composition, distribution and diversity of woody species in
tropical dry forest of India. Journal of Sustainable Forestry 33: 729756.
Chaturvedi RK & Raghubanshi AS (2015) Assessment of carbon density and accumulation in mono- and multispecific stands in Teak and Sal forests of a tropical dry region in India. Forest Ecology and Management
339: 1121.
Chaturvedi RK, Raghubanshi AS & Singh JS (2011a) Carbon density and accumulation in woody species of
tropical dry forest in India. Forest Ecology and Management 262: 15761588.
Chaturvedi RK, Raghubanshi AS & Singh JS (2011b) Effect of small scale variations in environmental factors
on the distribution of woody species in tropical deciduous forests of Vindhyan Highlands, India. Journal of
Botany 2011: Article ID 297097. [doi:10.1155/2011/297097]
Chaturvedi RK, Raghubanshi AS & Singh JS (2011c) Leaf attributes and tree growth in a tropical dry forest.
Journal of Vegetation Science 22: 917931.
Chaturvedi RK, Raghubanshi AS & Singh JS (2012) Effect of grazing and harvesting on diversity, recruitment
and carbon accumulation of juvenile trees in tropical dry forests. Forest Ecology and Management 284: 152
162.
Chaturvedi RK, Raghubanshi AS & Singh JS (2013) Growth of tree seedlings in a dry tropical forest in relation
to soil moisture and leaf traits. Journal of Plant Ecology 6: 158170.
Chaturvedi RK, Raghubanshi AS & Singh JS (2014) Relative effects of different leaf attributes on sapling
growth in tropical dry forest. Journal of Plant Ecology 7: 544558.
Clark DB & Clark DA (1991) Herbivores, herbivory, and plant phenology: patterns and consequences in a
tropical rain-forest cycad. In: Price PW, Lewinsohn TM, Fernandes GW & Benson W (eds) Plant-animal
interactions: evolutionary ecology in tropical and temperate regions. New York: John Wiley & Sons, Inc,
pp. 209225.
Coley PD & Aide TM (1991) Comparison of herbivory and plant defenses in temperate and tropical broadleaved forests. In: Price PW, Lewinsohn TM, Fernandes GW & Benson W (eds) Plant-animal interactions:
evolutionary ecology in tropical and temperate regions. New York: John Wiley & Sons, Inc, pp. 2549.
Coley PD (1983) Herbivory and defensive characteristics of tree species in a lowland tropical forest. Ecological
Monographs 53: 209233.
Crawley MJ (1985) Reduction of oak fecundity by low density herbivore populations. Nature 314: 163164.
Daubenmire R (1972) Phenology and other characteristics of tropical semi-deciduous forest in northeastern
Costa Rica. Journal of Ecology 60: 147170.
Dirzo R (1984) Insect-plant interactions: some ecophysiological consequences of herbivory. In: Medina E,
Mooney HA & Vazquez-Yanes C (eds) Physiological ecology of plants in the wet tropics. The Hague,
Netherlands: Dr. Junk W, pp. 209224.
Eamus D & Prior L (2001) Ecophysiology of trees of seasonally dry tropics: Comparisons among phenologies.
Advances in Ecological Research 32: 113197.
Elliot S, Baker PJ & Borchert R (2006) Leaf-flushing during the dry season: The paradox of Asian monsoon
forests. Global Ecology and Biogeography 15: 248257.
Frankie GW, Baker HG & Opler PA (1974) Comparative phenological studies of trees in tropical wet and dry
forest in the lowlands of Costa Rica. Journal of Ecology 62: 881913.
Franklin O & Agren GI (2002) Leaf senescence and resorption as mechanisms of maximizing photosynthetic
production during canopy development at N limitation. Functional Ecology 16: 727733.
Hikosaka K (2003) A model of dynamics of leaves and nitrogen in a plant canopy: An integration of canopy
photosynthesis, leaf life span, and nitrogen use efficiency. American Naturalist 162: 149164.
Hikosaka K (2005) Leaf canopy as a dynamic system: ecophysiology and optimality in leaf turnover. Annals of
Botany 95: 521533.
Jha CS & Singh JS (1990) Composition and dynamics of dry tropical forest in relation to soil texture. Journal of
Vegetation Science 1: 609614.
210

.
Jolly WM, Nemani R & Running SW (2005) A generalized, bioclimatic index to predict foliar phenology in
response to climate. Global Change Biology 11: 619632.
Jurik TW & Chabot BF (1986) Leaf dynamics and profitability in wild strawberries. Oecologia 69: 296304.
Karst AL & Lechowicz MJ (2007) Are correlations among foliar traits in ferns consistent with those in the seed
plants? New Phytologist 173: 306312.
Kikuzawa K & Lechowicz MJ (2006) Toward synthesis of relationships among leaf longevity, instantaneous
photosynthetic rate, lifetime leaf carbon gain, and the gross primary production of forests. American
Naturalist 168: 373383.
Kikuzawa K (1991) A cost-benefit analysis of leaf habit and leaf longevity of trees and their geographical
pattern. American Naturalist 138: 12501263.
Krischik VA & Denno RR (1983) Individual, population, and geographic patterns in plant defense. In: Denno
RF & McClure MS (eds) Variable plants and herbivores in natural and managed systems. New York:
Academic Press, pp. 463512.
Louda S (1984) Herbivory effects on stature, fruiting, and leaf dynamics of a native crucifer. Ecology 65: 1379
1386.
Marquis RJ (1984) Leaf herbivores decrease fitness of a tropical plant. Science 226: 537539.
Mooney HA, Bullock SH & Medina E (1995) Introduction. In: Mooney HA, Bullock SH & Medina E (eds)
Seasonally dry tropical forests. Cambridge: Cambridge University Press, pp. 18.
Mulkey SS, Smith AP, Wright SJ, Machado JL & Dudley R (1992) Contrasting leaf phenotypes control
seasonal variation in water loss in a tropical forest shrub. Proceedings of the National Academy of Sciences
USA 89: 90849088.
Murali KS & Sukumar R (1993) Reproductive phenology of a tropical dry forest in Mudumalai, southern India.
Journal of Ecology 82: 759767.
Nanda A, Suresh HS & Krishnamurthy YL (2014) Phenology of a tropical dry deciduous forest of Bhadra
wildlife sanctuary, southern India. Ecological Processes 3: 112.
Nilsen ET & Muller WH (1981) Phenology of the droughtdeciduous shrub Lotus scoparius: climatic controls
and adaptive significance. Ecological Monographs 51: 323341.
Noda H, Muraoka H & Washitani I (2004) Morphological and physiological acclimation responses to
contrasting light and water regimes in Primula sieboldii. Ecological Research 19: 331340.
Oikawa S, Hikosaka K & Hirose T (2006) Leaf lifespan and lifetime carbon balance of individual leaves in a
stand of an annual herb, Xanthium canadense. New Phytologist 172: 104116.
Oikawa S, Hikosaka K & Hirose T (2008) Does leaf shedding increase the whole-plant carbon gain despite
some nitrogen being lost with shedding? New Phytologist 178: 617624.
Oikawa S, Hikosaka K, Hirose T, Shiyomi M, Takahashi S & Hori Y (2004) Cost-benefit relationships in leaves
emerging at different times in a deciduous fern Pteridium aquilinum. Canadian Journal of Botany 82: 521
527.
Reich PB & Borchert R (1984) Water stress and tree phenology in a tropical dry forest in the lowlands of Costa
Rica. Journal of Ecology 72: 6174.
Reich PB, Ellsworth DS, Walters MB, Vose JM, Gresham C, Volin JC & Bowman WD (1999) Generality of
leaf trait relationships: a test across six biomes. Ecology 80: 19551969.
Reich PB, Uhl C, Walters MB, Prugh L & Ellsworth DS (2004) Leaf demography and phenology in Amazonian
rain forest: a census of 40,000 leaves of 23 tree species. Ecological Monographs 74: 323.
Rivera G, Elliott S, Caldas LS, Nicolossi G, Coradin VTR & Borchert R (2002) Increasing day-length induces
spring flushing of tropical dry forest trees in the absence of rain. Trees 16: 445456.
Rockwood LL (1973) The effect of defoliation on seed production of six Costa Rican tree species. Ecology 54:
13631369.
Roy S & Singh JS (1994) Consequences of habitat heterogeneity for availability of nutrients in a dry tropical
forest. Journal of Ecology 82: 503509.
Seiwa K & Kikuzawa K (1991) Phenology of tree seedlings in relation to seed size. Canadian Journal of Botany
69: 532538.
Shukla R & Ramakrishnan P (1982) Phenology of trees in a sub-tropical humid forest in north-eastern India.
Vegetatio 49: 103109.
211

.
Skarpe C (1996) Plant functional types and climate in a southern African savanna. Journal of Vegetation
Science 7: 397404.
Suresh HS & Sukumar R (2011) Vegetative phenology of tropical montane forests in the Nilgiris, South India.
Journal of the National Science Foundation of Sri Lanka 39: 337347.
van Schaik CP, Terborgh JW & Wright SJ (1993) The phenology of tropical forests: adaptive significance and
consequences for primary consumers. Annual Review of Ecology and Systematics 24: 353377.
Vincent G (2006) Leaf life span plasticity in tropical seedlings grown under contrasting light regimes. Annals of
Botany 97: 245255.
White MA, Thornton PE & Running SW (1997) A continental phenology model for monitoring vegetation
response to interannual climatic variability. Global Biogeochemical Cycles 11: 217234.
Wolda H (1978) Seasonal fluctuations in rainfall, food and abundance of tropical insects. Journal of Animal
Ecology 47: 369381.
Wright IJ, Reich PB, Westoby M, Ackerly DD, Baruch Z, Bongers F, Cavender-Bares J, Chapin FS,
Cornelissen JHC, Diemer M, Flexas J, Garnier E, Groom PK, Gulias J, Hikosaka K, Lamont BB, Lee T, Lee
W, Lusk C, Midgley JJ, Navas ML, Niinemets , Oleksyn J, Osada N, Poorter H, Poot P, Prior L, Pyankov
VI, Roumet C, Thomas SC, Tjoelker MG, Veneklaas EJ & Villar R (2004) The worldwide leaf economics
spectrum. Nature 428: 821827.
Wright SJ & van Schaik CP (1994) Light and the phenology of tropical trees. American Naturalist 143: 192
199.
212
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 213220, 2016
Research article
Characterization and validation of teak plus trees ramets of

national teak germplasm bank through microsatellites
Shashank Mahesh1, Vivek Vaishnav1*, Pramod Kumar1,
Naseer Mohammad1 and S. A. Ansari2
1
Genetics and Plant Propagation Division, Tropical Forest Research Institute, Jabalpur, Madhya Pradesh, India
2
Institute of Forest Productivity, Ranchi, Jharkhand, India
*Corresponding Author: vivekvaishnaw@live.in
Abstract: National Teak Germplasm Bank, Chandrapur (Maharashtra) holds one of the largest
collections of teak plus tree ramets collected in the form of bud grafts from 12 natural teak
populations of India. These grafts were planted with three ramets of each tree. The objective of the
present investigation was to validate 118 ramets of 48 plus trees and also to characterize the
germplasm bank using microsatellite markers. 12 microsatellite primers amplified 27 loci with
estimates of expected and observed heterozygosity (0.33 and 0.2). Moderate values of resolving
power (1.05) and polymorphic information content (0.27) confirmed its stringency for analysis.
The genetic structure of the germplasm was determined through the FST (0.075), Shannons
information index (0.59), Neis heterozygosity (0.41) and Chi-square tests to confirm its HWE
state. 7.54% variation existed among populations and 92.46% within populations. Ramets were
validated through UPGMA dendrograph. Majority of the plus trees grouped as per their sampling
location. 35 ramets belonging to 14 plus trees (APT-22, MHSC-A1, MHSC-A2, MHSC-A3,
ORPUB-6, UP-M, UP-A, AI-K, MHALP-8, ST-26, ST-35, ORANP-3, ORPUB-23 and ORPUB0) clustered strictly as per their parental origin (ortet). For eight plus trees (APKEA-23, APKEA24, APKEN-1, APNLP-1, MHSC-J1, MYHV, ORPB-12 and ORPUB-10) with three ramets, two
ramets grouped with each other; whereas the third ramet could not be validated.
Keywords: Ortet - HWE - PIC - RP - Gene diversity.
[Cite as: Mahesh S, Vaishnav V, Kumar P, Mohammad N & Ansari SA (2016) Characterization and validation
of teak plus trees ramets of national teak germplasm bank, Chandrapur, Maharashtra through microsatellites.
INTRODUCTION
Teak (Tectona grandis L. f., 2n=36) is one of the premier timber tree species of the world belonging to
family Lamiaceae, planted widely in the tropics (Gill et al. 1983). It is native to India, Thailand, Myanmar and
Laos. Molecular level characterization of the species has confined India and Laos as the centers of the genetic
origin (Verhaegan et al. 2010). In India, teak improvement program started in 1960s and intended mostly on the
selection of phenotypically superior plus trees, establishment of provenance/progeny trials and seed orchards
and creation of seed production areas. In teak the traditional breeding methods to find out good genotype is a
long process. Therefore, germplasm of the plus trees are considered fit to be used for raising seed orchards. The
scientific program related to molecular breeding and tree genomics also needs a structured population of the
species.
To fulfill the need, national teak germplasm bank (NTGB) at Chandrapur (Maharashtra) was established in
1978. It is having collection of teak plus tree clones raised from the selected plus trees bud grafts of 12 natural
populations of India. 260 plus tree clones with three ramets of each had been planted in row/column design with
8m X 8m spacing (Kumar et al. 1998). It is the only collection of its kind in the country. The main object of the
germplasm bank is to conserve a broad spectrum of genetic variability to serve as a reservoir for different
conservation and management needs.
213
Mahesh et al. (2016) 3(1): 213220

.
Assessment of genetic variability employing molecular markers has proved to be a keystone to
understanding the genomic constitution, categorizing the genes responsible for important traits, the classification
and conservation of genetic variation in plant germplasm and developing selective proliferation approaches for
breeding and ex-situ conservation. The neutral genetic diversity of teak from the natural area and introduced
populations has been studied with molecular markers (Vaishnaw et al. 2014). Studies based on highly stringent
dominant (RAPD, ISSR and AFLP) and co-dominant marker (SSR) technique state that the teak populations
have genetic variability from 57% to 99.6% within population and from 0.04% to 43% between populations
(Ansari 2014). Teak in India exhibits high variability not only at gene level (Verhaegan et al. 2010, Vaishnaw
et al. 2014, Ansari 2014, Fofana et al. 2009, Fofana et al. 2013) but varies greatly in timber characteristics
(mainly central and southern Indian teak) also. The germplasm bank must have collection covering varied
characteristic of teak and it must represent the entire genetic variability found in natural population as well.
Therefore, present investigation was performed aiming, (1) validation of plus trees ramets, (2) assessment of
the genetic variability and (3) molecular level characterization of national teak germplasm bank using
microsatellite markers.
Plant material and DNA extraction
Branch cuttings were collected from total 118 ramets representing 48 plus trees maintained at NTGB,
Chandrapur (Maharashtra). Three ramets from each of 22 plus trees and two ramets from each of 26 plus trees
were sampled which were the collection from nine states of the country (Table 1). The surface sterilized cuttings
were administered 200 ppm IAA + 200 ppm thiamine for 4 hours followed by sealing of the top cut end with
paraffin wax. The treated cuttings were planted in polybags filled with potting mixture. After one month of the
planting, the cuttings produced leaf sprouts, which were harvested for the extraction of genomic DNA.
Genomic DNA was extracted following the modified Doyle and Doyle method (Narayanan et al. 2006). To
remove large amounts of polyphenols and polysaccharides, 3% PVP was added to the extraction buffer. An
additional washing step with cold ethanol was also included for removal of remaining impurities. Purity and
quantity of the extracted genomic DNA were estimated through spectrophotometer and the quality of DNA was
visually verified on 1% agarose gel electrophoresis.
Table 1. Teak (Tectona grandis L. f.) plus tree clones selected for the characterization of germplasm bank through
microsatellite markers.
Locations
All India
Andhra Pradesh
Maharashtra
Karnataka
Orissa
Madhya Pradesh
Tamilnadu
Uttar Pradesh
West Bengal
TOTAL
Accessions
With three Ramets
APKEA-23, APKEA-24, APKEN-1,
APNLP-1, APNPL-0, APT-22
MHAL-P2, MHALP-3, MHALP-8,
MHSC-A1, MHSC-A2, MHSC-A3,
MHSC-J1
MYHD-1, MYHD-3, ST-26, ST-27, ST-35,
MYHV, ST-43,
ST-44, ST-45
ORANP-3, ORANP-7, ORPUB-23, ORPUB-0, ORANR-4, ORPB-12, ORPUB-10,
ORPUB-00
ORPUB-11, ORPUB-2, ORPUB-6,
PT-3
TNT-10,
TNT-8
UP-G,
UP-M, UP-A,
WB-4
26 genotypes X 2 ramets each = 52 trees
22 genotypes X 3 ramets each = 66 trees
With two Ramets
AI, AI-8, AI-C, AI-D, AI-I, AI-K, AI-N
APT-20,
Microsatellite markers assay

In prior investigation, 35 microsatellite primers were screened in teak genome. Out of them twelve primers
could amplify the genome and were selected for final analysis with selected genotypes (Table 2). PCR
amplification was carried out in 15 l of reaction volume containing 15 ng genomic DNA, 1 unit Taq
polymerase (Promega GoTaq M3001), 0.2 mM of each dNTPs, 1X Taq polymerase buffer and 0.66 M each of
forward and reverse primers. Ampli cation cycle consisted of an initial 5 min denaturation at 94oC, 34 cycles
for 30 sec at 94oC, 30 sec at 55oC (Ta), 1 min at 72oC, and nal extension step for 8 min at 72oC. The PCR
214
Mahesh et al. (2016) 3(1): 213220

.
products were mixed with 3L gel loading dye and were separated through electrophoresis on a 3.5% (Agarose
SFR) gel at 100 V in 1X TBE buffer with 0.5 g/ml ethidium bromide.
Data analysis
Banding profiles generated by the microsatellites compiled into a data matrix based on the molecular weight
of amplicons. The informativeness of primers was determined through genetic parameters; expected and
observed heterozygosity (He and Ho) and polymorphic information content (PIC) value were calculated by
software POWERMARKER Version 3.25 (Liu & Muse 2005). The stringency of the primers to discriminate the
genotypes was confirmed through calculation of resolving power (RP) as described by Prevost and Wilkinson
(1999).
To evaluate the genetic variability and structure of the germplasm bank, the analysis of molecular variance
(AMOVA) and FST was conducted using software Arlequin Version 3 (Excoffier et al. 1980). Shannons
information index (I), Neis heterozygosity (H) and Hardy-Weinberg equilibrium (HWE) through chi-square test
confirmation was determined applying software POPGENE Version 1.32 (Yeh et al. 1999). The ramets were
validated through their pairing with identical ramet in a neighbor joining dendrograph based on the Neis
genetic distance (1983) method implemented in software POWERMARKER Version 3.25 (Liu & Muse 2005).
Molecular markers are widely used in plant genetics, breeding, biological diversity analysis, and cultivar
identification since they can directly manifest genetic differences at the DNA level. SSR motifs are
polymorphic, abundant, and randomly distributed in eukaryotic genomes (Tautz 1989). Compared to other
biomarkers, such as RAPDs and AFLPs, SSR markers are stable, co-dominant, and low cost. Therefore, they
have been widely used in genetic analysis and genomic linkage mapping of tree population. A total of 118
ramets belonging to 48 teak plus trees were sampled (Table 1) from NTGB located at Chandrapur, Maharashtra.
However many accessions in the assemblage of NTGB were initially acquired from different states of India and
the morphological data of the accessions have been maintained but their original geographic origin and
parentage is unknown due to slipshod management of the tags in each bud graft. This study is first and only
report for molecular characterization of the National teak germplasm bank.
Informativeness of primers
27 markers were amplified by 12 microsatellite primers and the number of alleles detected among the 27
markers studied ranged from 2 to 7 (Table 2). The expected heterozygosity (also called gene diversity) of the
primers ranged from 0.22 (by Tg-7) to 0.47 (Tg-11) with average value of 0.33. The observed heterozygosity of
primers was found with average value of 0.2 (Table 2). These values of heterozygosity are comparably low to
the SSR primers resulted (0.576) for population of Quercus suber (Gomez et al. 2001) and closely similar to the
values of SSR primers resulted (0.22 to 0.54) for Birch (Hao et al. 2015). Low value of observed heterozygosity
than expected indicates low variability covered by the microsatellites. With average value of 1.05, RP ranged
from 0.2 (by Tg-13) to 5.42 (by Tg-11). The PIC is determined by both, allele numbers and allele frequency
distribution and can be used to evaluate the variation of microsatellites (Botstein et al. 1980). In this study, the
PIC values of the primers ranged from 0.2 (by Tg-7) to 0.36 (by Tg-11) with average 0.27 (Table 2). PIC and
RP have been used in several studies (Prevost & Wilkinson 1999, Smith et al. 1997, Korkovelos et al. 2008) to
analyze markers for their informativeness in genotyping, genetic diversity assessment, and discriminatory
power. In the current research, markers with higher RP values were more informative in other genetic
parameters also. These results are in agreement with Prevost & Wilkinson (1999) who observed a strong linear
relationship between resolving power and discriminatory power of a marker. However, these values are dynamic
and changeable, depending upon the number and nature of the genetic material involved. The low to moderate
values of gene diversity and PIC indicated that the microsatellites applied for the investigation were stringent
but covered lower genetic variability of entire germplasm bank. Primer Tg-11 showed highest values not only
for gene diversity and heterozygosity but for resolving power and PIC also (Table 2). Therefore, the amplicons
of primer Tg-11 can be used further for linkage disequilibrium or association mapping of teak.
Genetic structure of germplasm bank
AMOVA resulted moderate values of genetic variation. 7.54 % variation was found among populations and
92.46% variation within populations of germplasm bank. In earlier reports, teak populations have been
215
Mahesh et al. (2016) 3(1): 213220

.
characterized through dominant (RAPD, AFLP and ISSR) and co-dominant (SSR) marker systems (Ansari
2014) and among populations variation was revealed minimum 0.04% (Kumar 2011) to maximum 43%
(Shreshta et al. 2005). On the other hand, within population variation was found minimum 57% (Shreshta et al.
2005) to maximum 99.6% (Kumar 2011). The earlier report based on microsatellites based characterization of
teak populations revealed that only 0.17% variation exhibited among six natural teak populations of India and,
within population variation was found non-significant (Verhaegan et al. 2010). The FST value of the germplasm
bank was found 0.075 which is lower than the estimates (0.15 to 0.34) revealed through dominant marker
systems (Ansari, 2014) but it was found higher than the estimate (0.02) revealed by microsatellite systems
(Verhaegan et al. 2010). Since the germplasm bank has a large coverage of genotypes collected from north,
central and south states of India therefore, higher value of FST was obtained in present investigation compared to
earlier report (Verhaegan et al. 2010) in which genotypes only from south Indian states were sampled. The
Shannons information index and Neis heterozygosity was found 0.59 and 0.41 respectively. These values
strengthen the fact that the germplasm bank exhibits maximum variability of the natural teak population. Chisquare values for different loci indicate that 89% loci deviated from HWE at significant level (p<0.05). Since
the germplasm bank had been established by the selection of phenotypically superior trees from several natural
populations, it deviated from the equilibrium state. Estimates of parameters depicting genetic variability of the
germplasm bank confirm that the bank maintains high genetic diversity comparing to the earlier reported
variability through microsatellites and it represents the genetic structure and variability of natural population of
teak in India. Of course its mode of establishment through selection of superior trees causes its deviation from
the HWE state but the available resources can be managed for breeding and mapping population.
Table 2 Genetic parameters showing informativeness of 12 microsatellite primer sets on teak genotypes.
Primer
ID
Tg-2
Tg-3
Tg-7
Tg-8
Tg-9
Tg-11
Tg-12
Tg-13
Tg-15
Tg-16
Tg-18
Tg-21
Forward (F) and Reverse (R) sequences (5-3)

F GCTTTAGTGATTCTCGCCTA
R CTCAATAATTCCAAACCGAC
F TCTCTCATTCTCTICGGTTC
R TTTCTAGAGGCCCATAATGA
F
TTATTGCTCTTTGGGTTTGT
R TATTCTCGCTTCCATGACTT
F AAGAAAGACGACAACCTTG
R GCTTTAGTGATTCTCGCCTA
F CAACTGGAAATCCACAATTT
R GGCCTATATTTCTTTCCTCC
F TCATGCACACATGTAACACA
R ACCGCAAATAATCATAATGG
F GCTATCAAATTTGCTGCTTT
R ACTGATTGCTATAAAGGCCA
F ATCGTATTGCAGCTTTGTCT
R GGAACTCCTTTCTCGTCTTT
F TTATCAACTTCTGCAACCCT
R GAGTATGTCACCTGCCTGTT
F AACCATGACAGAAACGAATC
R GACATTCAGCCTGCTACTTC
F TCTTGATGGTTTGCCTATTT
R TATCTTCATGGTTGCCTTCT
F GCAGTAATGAAAGGATTTC
R ACATTACTTCTCACATGCCC
Amplicon
size (bp)
622 -644
He
Ho
RP
PIC
0.36
0.25
1.02
0.29
651-693
0.30
0.11
0.42
0.25
522-559
0.22
0.07
0.27
0.20
722-749
0.31
0.21
0.85
0.26
478-577
0.45
0.28
1.10
0.35
797-849
0.47
0.68
5.42
0.36
687-760
0.38
0.16
0.64
0.31
708-769
0.25
0.05
0.20
0.22
610-739
0.24
0.12
0.69
0.21
520-596
0.42
0.23
0.89
0.33
820-862
0.34
0.15
0.61
0.27
806-846
0.24
0.11
0.45
0.21
Average
0.33 0.20 1.05 0.27
Note: He= expected heterozygosity, Ho= observed heterozygosity, RP= resolving power, PIC= polymorphic
information content.
Validation of ramets
The NJ dendrograph revealed that out of 118 ramets belonging to 48 PTs, 35 ramets exactly grouped as per
their parental origin/ortet (Fig. 1). 100% ramets from the ortets APT-22, MHSC-A1, MHSC-A2, MHSC-A3,
ORPUB-6, UP-M, UP-A, AI-K, MHALP-8, ST-26, ST-35, ORANP-3, ORPUB-23 and ORPUB-0 were
validated (Table 3). 66% ramets could be validated from the ortets APKEA-23, APKEA-24, APKEN-1,
216
Mahesh et al. (2016) 3(1): 213220

.
APNLP-1, MHSC-J1, MYHV, ORPB-12 and ORPUB-10. Only two out of three ramets could be validated of
these ortets (Table 3). Unlike the earlier characterization of these 48 genotypes based on dominant marker
Table 3 Validated ramets of teak assembled in National teak germplasm bank Chandrapur,
Maharashtra through microsatellite markers.
S. N.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
Overall
Accessions/ Ortet
AI
AI-8
AI-C
AI-D
AI-I
AI-K
AI-N
APKEA-23
APKEA-24
APKEN-1
APNLP-1
APNPL-0
APT-20
APT-22
MHAL-P2
MHALP-3
MHALP-8
MHSC-A1
MHSC-A2
MHSC-A3
MHSC-J1
MYHD-1
MYHD-3
MYHV
ORANP-3
ORANP-7
ORANR-4
ORPB-12
ORPUB-10
ORPUB-11
ORPUB-2
ORPUB-23
ORPUB-6
ORPUB-0
ORPUB-00
PT-3
ST-26
ST-27
ST-35
ST-43
ST-44
ST-45
TNT-10
TNT-8
UP-A1
UP-G
UP-M
WB-4
Ramets
2
2
2
2
2
2
2
3
3
3
3
3
2
3
2
2
2
3
3
3
3
2
2
3
2
2
3
3
3
3
3
2
3
2
2
3
2
2
2
3
2
2
2
3
3
2
3
2
118
Validated Ramets
0
2
0
0
0
0
0
3
2
2
3
0
0
3
0
0
0
3
3
3
2
0
0
0
2
0
0
2
2
0
0
2
3
2
0
0
0
0
0
0
0
0
0
0
3
0
3
2
47
Validation (%)
0
100
0
0
0
0
0
100
66.6
66.6
100
0
0
100
0
0
0
100
100
100
66.6
0
0
0
100
0
0
66.6
66.6
0
0
100
100
100
0
0
0
0
0
0
0
0
0
0
100
0
100
100
39.8
(Narayanan et al. 2007), the majority of PTs were grouped together as per their geographical location of
sampling (Fig. 1). Few of the ramets could not be validated through applied microsatellites and characterized
distinct from the labeled ortet. There could be chances of mislabeling or failure of bud graft vis--vis rootstock
overtaking the growth. The genetic fidelity of these invalidated ramets needs should be further checked using
217
Mahesh et al. (2016) 3(1): 213220

.
multiple marker system supported by the authentic phenotyping of these ramets. However, microsatellites
markers efficiently revealed the status of genetic diversity in these collections and also clustered most of
geographically related PTs clones together.
100
23
96
ST-45-1
ORANR-4-1
MHAL-P2-2
MYHD-1-1
3
ST-44-2
26 ST-35-1
35
ST-35-2
MYHD-3-1
10
TNT-10-1
ORPUB-2-3
20 ORPUB-11-3
61
ORPUB-2-2
2
ORPUB-11-1
25
1
ORPUB-2-1
MYHV-2
18
MYHV-3
APKEA-23-2
40
APKEA-23-3
AI-C-1
11 AI-I-1
44
MHAL-P2-1
APT-20-2
12
ORPUB-00-2
APT-22-3
57 APT-22-1
60
APT-22-2
APNPL-0-1
MHSC-A2-1
14
71 MHSC-A2-2
69
MHSC-A2-3
27
UP-M-3
95 UP-M-1
41
UP-M-2
W B-4-1
9 MHALP-3-1
38
W B-4-2
AI-8-1
AI-K-2
AI-K-1
1
5 AI-1
11
ST-27-1
3
2
AI-8-2
23
ORANP-7-1
MHALP-3-2
17
MHALP-8-2
MHALP-8-1
8 ORPUB-0-1
69
ORPUB-0-2
MHSC-J1-1
PT-3-1
APKEA-24-2
9 ORANR-4-2
23
PT-3-2
1
APKEA-24-1
46
APKEA-24-3
MHSC-A3-3
56 MHSC-A3-1
39
MHSC-A3-2
APNLP-1-2
31
MHSC-J1-2
1
MHSC-J1-3
27
ORANR-4-3
MYHD-3-2
6
APNLP-1-3
15 APNLP-1-1
36
APNPL-0-3
TNT-8-2
ST-43-1
44 APKEN-1-2
88
APKEN-1-3
5
ST-43-2
15
TNT-8-1
APNPL-0-2
27
TNT-8-3
2
ORPB-12-1
40
ORPB-12-2
ST-43-3
9
MHSC-A1-1
69 MHSC-A1-2
69
MHSC-A1-3
ORPB-12-3
APKEN-1-1
20
31 ORPUB-10-2
75
ORPUB-10-3
16
UP-A-3
51 UP-A-1
69
UP-A-2
UP-G-2
AI-D-2
2
11 ST-26-1
20
ST-26-2
2
ORPUB-00-1
52 ORPUB-23-1
98
ORPUB-23-2
AI-C-2
6
ORANP-3-1
23 ORANP-3-2
24
ST-45-2
ORPUB-6-3
6 ORPUB-6-1
24
ORPUB-6-2
1
ORPUB-10-1
24
ORPUB-11-2
7 MYHD-1-2
TNT-10-2
AI-I-2
51
ST-44-1
1
AI-N-2
47
MYHV-1
ORANP-7-2
AI-2
4
35
11 ST-27-2
AI-D-1
40
APT-20-1
PT-3-3
36 AI-N-1
76
UP-G-1
Figure. 1 Neighborjoining (NJ) dendrograph of 118 ramets of plus trees of teak (Tectona grandis L. f.) based on Neis
(1983) genetic distance method.
CONCLUSION
Gene diversity is a result of gene evolution in plant species (Guo et al. 2012) and becomes a foundation of
genetic improvement of species. The results of the present study indicated that the National teak germplasm
bank located at Chandrapur, Maharashtra was a good initiative to assemble clones of plus trees collected from
all India. Prior information related to its genetic variability and validation of the ramets with their ortets will be
helpful to manage mapping and breeding population for next generation breeding program. Our study confirms
that the germplasm bank represents the actual variability of teak population that has been reported by earlier
findings. Mismatch of the ramets with their ortet can be considered as a limit which can be overcome by the
218
Mahesh et al. (2016) 3(1): 213220

.
establishment of bud grafts from same ortet. The investigation recommends for increasing number of selected
PTs from natural teak populations to widen the genetic base of National Teak Germplasm Bank, Chandrapur
because a large proportion of genetic variation in teak occurs among individuals within populations. It is also
recommended to maintain the equal number and size of germplasm representing various geographical locations
of teak natural population of the country. Further, this study highlights the effectiveness of microsatellite
markers to distinguish the ramets with respect to genetic diversity estimates of assembled germplasm, promoting
efficient conservation of genetic resources of teak in India.
ACKNOWLEDGEMENT
We are thankful to Department of Biotechnology, Government of India, New Delhi for funding the project
and to the Maharashtra Van Sanshodhan Sansthan (Maharashtra State Forest Department) for permitting access
to the National Teak Germplasm Bank, Chandrapur. There is no conflict of interest among authors.
REFERENCES
Ansari SA (2014) A molecular diversity of teak (Tectona grandis L. f). In: Silviculture Conference 2014, held in
ICFRE Dehradun, India.
Botstein D, White RL, Skolnick M & Davis RW (1980) Construction of a genetic linkage map in man using
restriction fragment length polymorphism. American Journal of Human Genetics 32: 314331.
Excoffier L, Laval G & Schneider S (2005) Arlequin ver. 3.0: An integrated software package for population
genetics data analysis. Evolionary Bioinformatics Online 1: 4750.
Fofana IJ, Ofori D, Poitel M & Verhaegen D (2009) Diversity and genetic structure of teak (Tectona grandis L.
f.) in its natural range using DNA microsatellite markers. New Forest 37: 175195.
Fofana IJ, Silue S, Diarrassouba N, Kadio AA & Sangare A (2013) Comparative analyses of amplified fragment
length polymorphism (AFLP) and simple sequence repeat (SSR) in genetic diversity of teak (Tectona
grandis L. f.). International Journal of Advance Agriculture Research 1: 114123.
Gill BS, Bedi YS & Bir SS (1983) Cytopalynological studies in woody members of family Verbenaceae from
north-west and central India. Journal of Indian Botanical Society 62: 235244.
Gomez A, Pintos B, Aguiriano E, Manzanera A & Bueno MA (2001) SSR Markers for Quercus suber tree
Identification and Embryo Analysis. Journal of Heredity 92 (3): 292295.
Guo Y, Li YL, Huang Y, Jarvis D, Sato K, Kato K, Tsuyuzaki H, Chen L & Long C (2012) Genetic diversity
analysis of hulless barley from Shangari-la region revealed by SSR and AFLP markers. Genetic Resources
and Crop Evolution 59: 15431552.
Hao W, Wang S, Huajing L, Zhou B, Wang X & Jiang T (2015) Development of SSR markers and genetic
diversity in White Birch (Betula platyphylla). PLoS ONE 10(4): e0125235 [DOI:
10.1371/journal.pone.0125235]
Korkovelos AE, Mavromatis AG, Huang WG, Hagidimitriou M, Giakoundis A & Goulas CK (2008)
Effectiveness of SSR molecular markers in evaluating the phylogenetic relationships among eight Actinidia
species. Scientia Horticulturae 116: 305310.
Kumar R (2011) Studies on genetic diversity analysis of teak (Tectona grandis L.F.) of central India using
RAPD marker. Ph.D. Thesis (Forest Biotechnology), FRI Deemed University, Dehra Dun.
Kumar V, Kotrange HR & Dhotekar UP (1998) Genetic improvement of teak. Indian Forester 124: 687695.
Liu K & Muse SV (2005) Power Marker: Integrated analysis environment for genetic marker data.
Bioinformatics 21: 21282129.
Narayanan C, Dubey S, Wali SA, Shukla N, Kumar R, Mandal AK & Ansari SA (2006) Optimization of DNA
extraction for ISSR studies in Tectona grandis L. f. - an important forest tree species. African Journal of
Narayanan C, Wali SA, Shukla N, Kumar R, Mandal AK & Ansari SA (2007) RAPD and ISSR markers for
molecular characterization of teak (Tectona grandis) plus trees. Journal of Tropical Forest Science 19 (4):
218225.
Prevost A & Wilkinson MJ (1999) A new system of comparing PCR primers applied to ISSR fingerprinting of
potato cultivars. Theoretical and Applied Genetics 98: 107112.
Shreshta MK, Volkaert H, & Straeten DVD (2005) Assessment of genetic diversity in Tectona grandis using
amplified fragment length polymorphism markers. Canadian Journal of Forest Research 35(4): 10171022.
219
Mahesh et al. (2016) 3(1): 213220

.
Smith JSC, Chin ECL, Shu H, Smith OS, Wall SJ, Senior ML, Mitchell E, Kresovich S & Ziegle J (1997) An
evaluation of the utility of SSR loci as molecular markers in maize (Zea mays L.): comparisons with data
from RFLPs and pedigree. Theoretical and Applied Genetics 95: 163173.
Tautz D (1989) Hypervariabflity of simple sequences as a general source for polymorphic DNA markers.
Nucleic Acid Research 17: 64636471.
Vaishnaw V, Mohammad N, Wali SA, Kumar R, Tripathi SB, Negi MS & Ansari SA (2014) AFLP markers for
analysis of genetic diversity and structure of teak (Tectona grandis) in India. Canadian Journal of Forest
Research 45(3): 297-306.
Verhaegan D, Fofana IJ, Logossa ZA & Ofori D (2010) What is the genetic origin of teak (Tectona grandis L.)
introduced in Africa and Indonesia? Tree Genetics and Genomics 6(5): 717733.
Yeh FC, Yang RC & Boyle TJ (1999) POPGENE, Version 1.3. Microsoft Windows-based freeware for
population genetic analysis. Distributed by the author (http://www.ualberta.ca/~fyeh/fyeh/).
220
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 221223, 2016
Short communication
Wild food plants of Mishing tribe- An ethnobotanical survey

Gitamani Dutta1*, Gautam Baruah1 and Ashalata Devi2
1
Balipara Tract & Frontier Foundation, Nameri Field Station, Sonitpur, Assam, India
2
Department of Environmental Science, Tezpur University, Assam, India
*Corresponding Author: gitadtt@gmail.com
[Cite as: Dutta G, Baruah G & Devi A (2016) Wild food plants of Mishing tribe- An ethnobotanical survey.
The wild edible plants are important in the livelihood strategies of tribal people. The value of wild edible
vegetables in food security has not been given sufficient attention in India (Reddy et al. 2007). Mishing is a
tribal community belonged to Mongoloid group a multitude of people that followed Austro-Asiatic races to
India (Singh et al. 1996). Mishing or Miri tribe inhabiting the districts of Dhemaji, North Lakhimpur, Sonitpur,
Tinsukia, Dibrugarh, Sibsagar, Jorhat and Golaghat of Assam, Northeast India. The Mishings are known to use a
good number of wild plants as traditional food and they are also known to be highly passionate for cooking
traditionally unique food items (Barua et al. 2007). The present study highlights some of the important wild
food plants of Mishing tribe of Assam.
The study was conducted in the Bokagaon of Sonitpur district of Assam, Northeast India. The information
was accrued after discussions with the village head and the missing inhabitant of the village following a semi
structured questionnaire (Fig. 1). Plant specimens were collected and identified with the help of Flora of Assam
(Kanjilal & Bor 1934).
Figure 1. Interviewing the Mishing people during the study (A & B).
A total of 41 plant species belonging to 34 families has been recorded in the present study that used as food
by the Mishing tribe. The live photographs of some of them have also been provided (Fig. 2). Table 1 enlists the
wild plant species commonly used as food by the Mishing tribe of the study area. The preparation of rice bear
locally known as Apong is one of the common activity of the Mishing house hold. Leaves of few species like
Artocarpous heterophyllus, Clerodendrum cloebrookianum, Scoparia dulcis, Solanum indium also used in the
preparation of Apong. Apong is not only an alcoholic refreshing drink but an integral part of the social,
cultural and religious life of the Mishing community of North East India (Pegu et al. 2013).
Table 1. Wild plant species used as food by the Mishing tribes of Sonitpur district of Assam
S. N.
1
2
3
Scientific name
Adhatoda vasica Nee
Albizzia procera (Roxb.) Benth.
Alocasiaa cuminata Schott
Mishing/Assamese name
Bahaka (As; M)
Koroi (As); Tantari-asing (M)
Kochu (As.); Ange (M)
Family
Acanthaceae
Mimosaceae
Araceae
Plant part used

Leaves, Flowers
Leaves
Shoots, leaves,
tubers
221
Alpinia alughas (Retz) Rose
Amaranthus spinosus L.
Artocarpous heterophyllus Lamk
7
8
9
10
11
Azadiracta indica A. Juss.

Baccaurea sapida L.
Bambusa balcooa Roxb.
Cassia tora L.
Centella asiatica Urb.
12
Ceratopteris thalictroides (L.) Ad.

Brongn.
Clerodendrum cloebrookianum L.
Colocasia esculenta (L.) Schott.
13
14
Tora (As); Talayangakhan

(M)
Hati-Khutora (As); Geang
(M)
Kothal (As); Bilangaai (M)
Moha Neem (As; M)
Leteku (As; M)
Bhalookaabaah (As; M)
Horumedelua (As)
Bormanimuni (As); Bortan
Manimuni (M)
Pani dhekia (As); Okangoing
(M)
Nefafu (As); Pakcoom (M)
Kochu (As); Ange (M)
Dutta et al. (2016) 3(1): 221223

.
Zingiberaceae
Leaves, Young
shoot
Amaranthaceae
Leaves stem
Moraceae
Meliaceae
Euphorbiaceae
Poaceae
Caesalpinaceae
Apiaceae
Stem, leaves,
fruit
leaves
Fruit
Shoot
Young leaves
Whole plant
Parkeriaceae
Fronds
Verbenaceae
Araceae
Leaves
Tender leaves,
tubers
15
Corchorus capsularis L.
Tita Morapat (As); Mura (M) Tiliaceae
Young plant
16
Dillenia indica L.
Outenga (As); Champa (M) Dilleniaceae
Fruit
17
Dioscorea alata L.
Kathalu (As); Ali (M)
Dioscoreaceae
Tuber
18
Diplazium esculentum (Retz.)SW
Dhekia (As) Okang (M)
Athyriaceae
Tender leaf
19
Drymaria cordata (L.) Willd ex Roem. Laijabori (As; M)
Carryophyllaceae Tender leaves
shoots
20
Ficus glomerata Roxb
Dimoru (As); Takpiyang (M) Moraceae
Leaves
21
Flacourita cataphracta L.
Ponniyal (As; M)
Flacortiaceae
Fruit
22
Garcinia cowa L
Kujithekera (As; M)
Cluciaceae
Fruit
23
Hibiscus subdarifa L.
Tenga Mora (As; M)
Malvaceae
Leaves, fruits
24
Houttuynia cordata Thunb
Mosundori (As; M)
Saururaceae
Leaves
25
Hydrocotyle sibthopioides L
Harumanimuni (As);
Apiaceae
Whole plant
26
Leucas aspera Link.
Doron (As); Durun (M)
Lamiaceae
Leaves
27
Meliosma pinnata (Roxb.) Maxim.
Bon pachala (As);
Sabiaceae
Young leaves
Dermiesing (M)
28
Meliosma simplicifoila (Roxb.) Walp. Dhapapatia (As); Nitak (M) Sabiaceae
Young leaves
29
Mikania micrantha
Kolialota (As)
Asteraceae
Young leaves
30
Moringa pterygosperma Gaetern
Sajina (As; M)
Moringaceae
Leaves, flower
31
Nycthanthus arbor-tristis L.
Shewali (As; M)
Oleaceae
Leaves, flower
32
Oxalis corniculata L.
Horutengesi (As)
Oxalidaceae
Whole plant
33
Paederia foetida L
Bhedailota (As);
Rubiaceae
Stem, leaves
Bungkirupug (M)
34
Sarcochlamys pulcherrima Gaud.
Mesaki (As)
Urticaceae
Young leaves
35
Scoparia dulcis L.
Bondhonia (As); Tirsirkosa Scrophulariaceae Leaves
(M)
36
Solanum indicum L.
Tit-bhekuri (As); Bangko
Solanaceae
Leaves, Fruit
(M)
37
Spilanthes paniculata L
Swoni (As); Malsa (M)
Asteraceae
Leaves
38
Stenochlaena palustris (Burm. f.)
Dhekialota (As);Tarong (M) Baleachnaceae
Young frond
Bodd.
39
Teteli (As; M)
Caesalpinaceae
Seed, young
leaves
40
Vitex neguno L
Pochotiya (As; M)
Verbenaceae
Leaves
41
Zanthoxylam oxyphyllum Edgn
Mezenga (As); Onger(M)
Rutaceae
Tender shoots
As means Assamese, M means Mishing
ACKNOWLEDGEMENTS
Authors are thankful to Shri Kamison Mili, the village head of Bokagaon for his help and support during the
field visit. The help received from the Budhesor Mili, Bineshory Mili, Ganak Pau and Bimol Mili during
fieldwork is duly acknowledged. Authors are duly acknowledged the financial support received from Globally
Managed Services (GMS).
222
Dutta et al. (2016) 3(1): 221223

.
Figure 2. Some ethnobotanically used food plants: A, Sarcochlamys pulcherrima; B, Spilanthes paniculata; C, Nycthanthus
arbor-tristis; D, Amaranthus spinosus; E, Leucas aspera; F, Houttuynia cordata; G, Hibiscus subdarifa; H, Cassia tora.
REFERENCES
Reddy KN, Pattanaik C, Reddy CS & Raju VS (2007) Traditional knowledge on wild food plants in Andhra
Pradesh. Indian Journal of Traditional Knowledge 6: 223229.
Singh J, Bhuyan TC & Ahmed A (1996) Ethnobotanical studies on the Mishing tribes of Assam with special
reference to food and medicinal plant. Journal Economic Taxonomy Botany (Additional Series) 12: 350
356.
Barua U, Hore DK & Sarma R (2007) Wild edible plants of Majuli island and Darang districts of Assam. Indian
Journal of Traditional Knowledge 6: 191194.
Pegu R, Gogoi J, Tamuli AK & Teron R (2013) Apong, an alcoholic beverage of cultural significance of the
Mishing community of Northeast India. Global Journal of Interdisciplinary Social Sciences 2: 1217.
Kanjilal UN & Bor NL (2005) Flora of Assam. Omsons Publication, New Delhi
223
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 224229, 2016
Research article
New addition to the Lichen flora of Uttarakhand, India

Sugam Gupta1, 2*, Himanshu Rai2, 3, Dalip Kumar Upreti3,
Pradeep Kumar Sharma1 and Rajan Kumar Gupta2
1
Department of Environmental Science, Graphic Era University, Dehradun, Uttarakhand, India

Department of Botany Pt. L.M.S. Government Post Graduate College, Rishikesh, Uttarakhand, India
3
Lichenology laboratory; Plant Diversity, Systematic and Herbarium Division; CSIR-National Botanical
research Institute, Lucknow, Uttar Pradesh, India
*Corresponding Author: sugam2606@gmail.com
2
Abstract: The present paper revealed the occurrence of nine lichen species from Uttarakhand for
the first time. The species belong to six families (Cladoniaceae, Lecanoraceae, Parmeliaceae,
Peltigeraceae, Physciaceae, Verrucariaceae) and represents four growth forms of lichens found
growing on soil, rock and soil over rock. Brief moropho-taxonomic details of all the nine species
have been provided with their ecology and distribution.
Keywords: Lichens - Uttarakhand - New addition - Ecology - Distribution.
[Cite as: Gupta S, Himanshu Rai H, Upreti DK, Sharma PK & Gupta RK (2016) New addition to the Lichen
flora of Uttarakhand, India. Tropical Plant Research 3(1): 224229]
INTRODUCTION
Indian Himalayas has been known for its floral composition and considered as one of the main hotspot
consisting of different plant groups including lichens. The lichens are slow growing organism that retains
uniform morphology and dependent on the atmosphere for their water and nutritional demand. The varied
climatic conditions and altitudinal ranges in the Himalayas provide different substrate for the colonization and
growth of lichens (Vetaas & Grytnes 2002, Grau et al. 2007, Kumar et al. 2014). Other than vascular plants,
lichens are considered as most significant indicator of ecosystem fluctuations as they are more sensitive towards
habitat and climatic alternation (Herk et al. 2002, Saipunkaew et al. 2007, Rai et al. 2011). Approximately
20,000 species of lichens are so far known from the world and it is estimated that Indian lichen flora comprises
of 2319 species under 305 genera and 74 families (Singh & Sinha 2010).
India has vast geographical area, different phytogeographical regions and varied climate conditions which
exhibit variation in the diversity of lichens. Among eight lichenogeographical regions in India, the Western
Ghats and the Himalayas shows higher lichen diversity. The Eastern Himalayas archives higher lichen diversity
with 1162 species, followed by Western Ghats and Western Himalayas with 1157 and 812 species respectively
(Rai et al. 2014). The lichen distribution as compared to other cryptograms is highly influenced by
microclimatic factors of a particular region.
The Western Himalayas occupies the extreme northwestern margins of India including Jammu & Kashmir,
Himachal Pradesh and Uttarakhand and sustains significant assemblage of lichen flora. The varied climatic
conditions and altitudinal ranges provide varied habitats for colonization and growth of lichens. Among the
different state of India, Uttarakhand represents more than 600 species of lichens followed by Himachal Pradesh
and Jammu & Kashmir with 503 and 386 species respectively (Upreti & Negi 1998, Sheikh et al. 2006, Nayaka
et al. 2010, Singh & Sinha 2010, Goni et al. 2015, Goni & Sharma 2015, Mishra 2015).
The topography of the state provide a wide altitudinal range from plain foothills to higher alpine region,
hence the state exhibit tropical type of climate in the lower Himalayan region and temperate to alpine type of
climate condition in higher Himalayas. About 95% of the total geographical area of Uttarakhand comprises of
Himalayan Mountains which can be further divided into Garhwal and Kumaon regions in the West and East
correspondingly.
224
Gupta et al. (2016) 3(1): 224229

.
MATERIALS & METHODS
Study Area
During the study the lichen samples were collected from in and around Badrinath Valley including Mana,
Bhimpul and Vasudhara area (Fig. 1). The region characterized by an average maximum temperature 18C to a
minimum of -22C. Precipitation occurs in the form of snow and heavy rainfall. Snowfall occurs in the month of
DecemberMarch with the maximum rainfall in month of July and October. The area represents typical alpine
habitats, extreme temperature and vegetation of valley comprises of alpine grasslands or alpine herbs and trees
like Betula spp., Salix spp. and Rhododendron spp. The lichen growth flourish on rock and soil and this plant
group make one of most eye catching vegetation in the area.
Figure 1. Location of study sites in Uttarakhand, India.
Lichen Identification
The specimens were collected from all available substrates and the sample segregated identified and
preserved in the lichenology laboratory of CSIR-National Botanical Research Institute, Lucknow. The
morphological characters were studied with Leica E24 binocular and anatomical structures were studied by
Nikon Eclipse E 400 compound microscope. Secondary metabolites in specimens were determined using thin
layer chromatography (TLC) in solvent system A (180 Toluene: 60 dioxane: 8 acetic acid) and spot test
performed by (Elix & Ernst-Russel 1993, Organge et al. 2001). The species identified on the basis of
morphological, anatomical and chemical chararactertics using relevant keys for various lichen taxa (Divakar &
Upreti 2005, Awasthi 2007, Upreti 2008). The voucher specimens with details of locality, date of collection and
substratum were deposited at the Lichen herbarium (LWG), National Botanical Research Institute, Lucknow.
RESULT AND DISCUSSION
Lichen Flora
The present study enumerates the addition of nine lichen species as new additions to the lichen flora of
Uttarakhand. Normandina pulchella is reported first time from the Himalayas, previously this species was
reported only from Tamil Nadu (Upreti et al. 2008). Similarly Rinodina megaspora has been reported for the
first time from Western Himalayas as earlier it has restricted distribution in the Eastern Himalayas only. Out of
the nine new addition recorded, Lecidella alaiensis and Rinodina megaspora are only two, microlichen genera
(crustose), Remototracgyna incognita, Melaniella disjuncta, Melanohalea infumata, Parmelia squarrosa,
Peltigera collina and Normandina pulchella have foliose growth form, while Cladonia subsquamosa have a
fruticose growth.
Since the study area located at higher elevation of more than 3000m, usually devoid of tress therefore all the
species reported found growing on rocks and soil over rocks and on mosses. Normandina pulchella having
squamulose to subfoliose thallus forms small colonies with other lichens and sometimes difficult to locate.
Rinodina megaspora a crustose lichen also exhibit interesting habit forms brown powdery patches over moss
tuft. The brief species description of each species with their ecology and distribution is mentioned below.
225
Gupta et al. (2016) 3(1): 224229

.
Enumeration of species
1. Cladonia subsquamosa Kremp. in Warming., Vidensk. Meddel. Dansk. Naturhist. Foren. Kjobenhavn 5:
366.1873 (1874).
(Fig. 2A)
Thallus terricolous, fructicose, dimorphic, primary squamules small, fragile, persistent. Secondary thallus
podetiate, podetia grey-green to brownish, 1020 mm tall, 12 mm thick at base, Podetial surface sorediate
throughout along with microsquamules. Podetia K-, or K + yellow, P+ yellow-red; fumarprotocetraric acid,
present in TLC.
Ecology and distribution: This species found growing on soil over rocks in moist places. In India the
species is reported from Tamil Nadu-Nilgiri and Palni Hills. Outside India from Pantropical Asia, Africa, N.
and S. America and Australasia.
Specimen examined: Badrinath, alt. 3175 m, on Soil, 12.10.2013, Rai H, Khare R & Gupta S 13021127(LWG).
2. Remototracgyna incognita (Kurok.) Divakar & A. Crespo., Am. J. Bot. 97(4): 586 (2010).
(Fig. 2B)
Thallus saxicolous, foliose loosely adnate to the substratum, lobes sub irregular, imbricate, thick, margin
crenate, upper surface mineral grey to grey, smooth, densely isidiate; rhizines black, short, dichotomously
branched. Apothecia lecanorine. Spore 8 in ascus, colourless, oval 101546 m. Thallus K+ yellow,
Medulla K-, C+ rose, KC + red, P-; protolichesterinic acid present in TLC.
Ecology and distribution: The species grows on exposed rocks or soil over rock both in open exposed dry
and moist habitats. In India the species is known from Meghalaya, Sikkim and West Bengal-hills. Outside
India from Japan and Nepal.
Specimen examined: Between Bhimpul to Vasudhara, on rock, alt. 3229 m, 13.10.2013, Rai H, Khare R &
Gupta S 13-021103 (LWG).
3. Lecidella alaiensis (Vain.) Hertel., Herzogia 2(4): 501.1973.
(Fig. 2C)
Thallus saxicolous, crustose, white to sooty yellow 0.51.8 mm thick, smooth to chinky, verru-culose.
Apothecia to 0.51.5 mm broad, andante, shining, disc black, margin smooth persistent lecedeine or
biatorine. Spores 8 in ascus, ellipsoid to broadly ellipsoid 10.517.06.010.5m. Thallus K-,C-,KC-,P-,
medulla K+ yellow; atranorin and zeorin is present in TLC.
Ecology and distribution: The species grows on exposed rocks and boulders. In India the species is known
from Himachal Pradesh and Jammu & Kashmir. Outside India the species is reported from Afghanistan,
China, Iran, Mongolia and Russia.
Specimen examined: Badrinath, East of temple, on way to Devdarshani, on boulders, alt. 3150 m,
28.09.1976, Dange K 76.799 (LWG-LWU).
4. Melanelia disjuncta (Erichsen). Essl., Mycotaxon 7(1): 46. 1978.
(Fig. 2D)
Thallus saxicolous, foliose, adnate, lobes to 1.5(-3) mm wide, upper side dark olive brown to blackish,
lower side dark brown to black, rhizate. Apothecia to 3 mm in diam, lecedeine. Spore 8 in ascus oval
ellipsoid, colourless,91257m.Thallus K-,C-,KC- or KC+ faint rose, P-.; perlatolic and stenosporic acids
present in TLC.
Ecology and distribution: The species is known to be saxicolous but occasionally it is found on tree trunk.
In India the species exhibit its restricted distribution in Jammu & Kashmir. Outside India it is known from
Europe, North America.
Specimen examined: Between Bhimpul to Vasudhara, on rock, alt. 3229 m, 13.10.2013, Rai H, Khare R &
Gupta S 13-021112 (LWG).
5. Melanohalea infumata (Nyl.) O. Blanco, A. Crespo, Divakar, Essl., D. Hawksw. & Lumbsch, Mycol. Res.
108(8): 882. 2004.
(Fig. 2E)
Thallus saxicolous, loosely adnate to 7 cm across, lobes flat, shot and rounded, 1.04.0mm wide, upper
side olive-green to reddish brown, without pseudocyphellae, isidiate, lower side dark brown to black,
sparsely rhizinate. Apothecia not known. Thallus K-,C-,KC-,P-; No substance present in TLC.
Ecology and distribution: This species grows on exposed rocks or on soil over rock. The species is
reported from Himachal Pradesh and Jammu & Kashmir. Outside India it is reported known from
Karakorum, Europe and North America.
226
Gupta et al. (2016) 3(1): 224229

.
Specimen examined: Badrinath, on soil, alt. 3173 m, 13.10.2013, Rai H, Khare R & Gupta S 13-021136
(LWG).
Figure 2. Microscopic image of new addition to Uttarakhand: A, Cladonia subsquamosa Kremp.; B, Remototrachyna
incognita (Kurok.) Hale.; C, Lecidella alaiensis (Vain.) Hertel.; D, Melanelia disjuncta (Erichsen). Essl.; E, Melanohalea
infumata (Nyl.) O. Blanco & al.; F, Normandina pulchella (Borrer) Nyl.; G, Parmelia squarrosa Hale.; H, Peltigera collina
(Ach.) Schrad.; I, Rinodina megaspore (D.D. Awasthi & M.R. Agarwal) D.D. Awasthi.
6. Normandina pulchella (Borrer) Nyl., Ann.Sci.Nat.Bot.Ser.4,15:382 (1861).
(Fig. 2F)
Thallus corticolous, squamulose, scattered or partly contiguous forming dense colonies in irregular
patches, of small cochleate to rounded squamules; squamules plane to concave, concentrically ridged,
undivided or distinct with lobes ,12 mm wide, upper surface pale grey to greenish-grey, soredia on the
surface and along the margins, medulla indistinct, photobiont layer distinct. Apothecia absent. Thallus K-,
KC-, C- and P-; zeroin is present in TLC.
Ecology and distribution: The species exhibit special habit as found growing in small colonies in
association with of on lichen or mosses or humus. In India the species is only known from Tamil Nadu.
Outside India it is reported from all the continents except Antarctica.
Specimen examined: Between Bhimpul to Vasudhara, on soil over rock, alt. 3229 m, 13.10.2013, Rai H,
Khare R & Gupta S 13-023569 (LWG).
7. Parmelia squarrosa Hale., Phytologia 22(1): 29, 1971.
(Fig. 2G)
Thallus corticolous rarely saxicolous, foliose, adnate; lobes sublinear, upper side pale to mineral grey,
shiny, pseudocyhellae forming reticulate network, isidia along the ridges of pseudocyhellae, lower side black
and shiny, densely rhizinate. Rhizines richly branched. Apothecia rare substipitate. Spores 8 in ascus, 30
351016 m. Thallus K+ yellow, medulla K+ yellow turning red C-, KC-, P+ orange-red; atranorin and
salazinic acid ias present in TLC.
Ecology and distribution: The species is found growing on trunks of small shurbs or exposed rock. In India
the species is distributed in Himachal Pradesh and Sikkim. Outside India it is known from Bhutan, China,
Japan, Nepal, Europe and North America.
Specimen examined: Badrinath, on rock, alt. 3199 m, 12.10.2013, Rai H, Khare R & Gupta S 13-021359
(LWG).
227
8. Peltigera collina (Ach.) Schrad., J. Bot. (Schrader) 3: 78. 1801.
Gupta et al. (2016) 3(1): 224229

.
(Fig. 2H)
Thallus terricolous, foliose, adnate, upto 3 cm across; lobes 46 mm wide; upper surface yellowish
brown, etomentose, marginally soraliate with granular soredia, lower surface with diffused,indistinct;brown
vein; rhizines simple to confluent; photobiont a nostoc; medulla pale brown. Apothecia not present.
Gyrophoric acid, zeorin, tenuiorin and unknown substances present in TLC.
Ecology and distribution: The species grows on soil or soil over rock or over mosses and both in moist
shady and exposed areas. In India the species exhibit restricted distribution to Sikkim and Tamil Nadu.
Outside India the species is reported from China; Central Europe and North America.
Specimen examined: Badrinath, on soil, alt. 3144 m, 12.10.2013, Rai H, Khare R & Gupta S 13-021138
(LWG).
9. Rinodina megaspora (D.D. Awasthi & M.R. Agarwal) D.D. Awasthi., Biblioth. Lichenol. 40: 4. 1991. (Fig. 2I)
Thallus crustose, granular leprose, grey to dark grey; photobiont a green alga (Trebouxia). Apothecia
0.81.5 mm diam., disc brown-black to black, margin thalline. Spores 48 in ascus, brown, 3 septate,
smooth, lumina rounded, 31391318m. Thallus K-, C, KC-, P-; no secondary compounds present in in
TLC.
Ecology and distribution: The species grows on exposed rocks over soil and exhibit its restricted
distribution in Eastern Himalayas and reported only from West Bengal hills. Outside India the species has
wide distribution from Australia, Bhutan, New Gueinea, New Zealand, temperate region of Central and
Southern Europe and North America.
Specimen examined: Badrinath, on rock, alt. 3198 m, 12.10.2013, Rai H, Khare R & Gupta S 13-021547
(LWG).
ACKNOWLEDGEMENTS
The authors are grateful to Department of Environment Science, Graphic Era University, Dehradun and
Director, CSIR-National Botanical Research Institute for providing necessary laboratory facilities.
REFERENCES
Awasthi DD (2007) A compendium of the Macrolichens from India, Nepal and Sri Lanka. Bishen Singh &
Mahendra Pal Singh, Dehra Dun.
Elix JE & Ernst-Russel KD (1993) A catalogue of standardized thin layer chromatographic data and
biosynthetic relationships for lichen substances, 2nd edn. Australian National University, Canberra.
Goni R & Sharma N (2015) Additions to lichen flora of Jammu and Kashmir, India. Tropical Plant Research
2(2): 7881.
Goni R, Raina AKP, Magotra R & Sharma N (2015) Lichen flora of Jammu and Kashmir State, India: An
updated checklist. Tropical Plant Research 2(1): 6471.
Grau O, Grytnes JA & Birks HJB (2007) A comparison of altitudinal species richness patterns of bryophytes
with other plant groups in Nepal, Central Himalaya. Journal of Biogeography 34: 19071915.
Herk van CM, Aptroot A & Dobben van HF (2002) Longterm monitoring in the Netherlands suggests that
lichens respond to global warming. The Lichenologist 34: 141154.
Kumar J, Rai H, Khare R, Upreti DK, Dhar P, Tayade AB, Chaurasia OP & Srivastava RB (2014) Elevational
controls of lichen communities in Zanskar valley, Ladakh, a Trans Himalayan cold desert. Tropical Plant
Research 1(2): 4854.
Mishra GK & Upreti DK (2015) Lichen flora of Kumaun Himalaya. Lap Lambert Academic, Deutschland,
Germany.
Nayaka S, Upreti DK & Rai H (2011). An outline of lichen diversity in Uttarakhand, India. In: 6th Uttarakhand
State Science and Technology Congress, pp. 99.
Orange A, James PW & White FJ (2001) Microchemical methods for the identication of lichens. British
Lichen Society, London.
Rai H, Khare R, Gupta RK & Upreti DK (2011) Terricolous lichens as indicator of anthropogenic disturbances
in a high altitude grassland in Garhwal (Western Himalaya), India. Botanica Orientalis-Journal of Plant
Science 8: 1623.
228
Gupta et al. (2016) 3(1): 224229

.
Rai H, Khare R, Upreti DK & Ahti T (2014) Taxonomic Keys and Description. In: Rai H & Upreti DK (eds)
Terricolous Lichens of India Vol 2: Morphotaxonomic Studies. Springer, New York, USA.
Saipunkaew W, Wolseley PA, Chimonides PJ & Boonpragob K (2007) Epiphytic macrolichens as indicators of
environmental alteration in northern Thailand. Environmental Pollution 146: 366374.
Sheikh MA Upreti DK & Raina AK (2006) Lichen Diversity in Jammu and Kashmir, India. Geophytology
36(1&2): 6985.
Singh KP & Sinha GP (2010) Indian lichens: An annotated checklist. Government of India, Botanical Survey of
India, Ministry of Environment and Forest, New Delhi, India.
Upreti DK & Negi HR (1998) Lichen flora of Chopta-Tungnath, Garhwal Himalayas. Journal of Economic and
Taxonomic Botany 22: 273286.
Upreti DK, Joshi Y, Divakar PK, Lumbsch HT & Nayaka S (2008) Notes on some interesting lichens from
Western Ghats in India. Phytotaxonomy 8: 113116.
Vetaas OR & Grytnes JA (2002) Distribution of vascular plant species richness and endemic richness along the
Himalayan elevation gradient in Nepal. Global Ecology and Biogeography 11: 291301.
229
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 230242, 2016
Research article
Floristic diversities and medicinal importance of selected

sacred groves in Thrissur district, Kerala
Deepa MR*, Sheema Dharmapal P. and P. S. Udayan
P.G. and Research Department of Botany, Sree Krishna College, Guruvayur, Ariyannur (P.O),
Thrissur District, Kerala, India
*Corresponding Author: deepakrishna56@gmail.com
Abstract: Sacred groves are forest fragments of varying sizes, existing outside conventional forest
areas in and around human inhabitation. It acts as safe sites for reproduction of variety of floral
and faunal resources.An exploratory survey of different sacred groves of Thrissur District (10.52
N to 76.21 E), namely Adipparambukavu, Daivathinkavu, Kanisherykavu, Kottaichalippattukavu
and Kottarathkavulead to the collection of 119 species coming under 104 genera and 51 families,
representing 08vulnerable, 12 endemic and 03 near threatened species. It includes 17.64% herbs,
19.33% shrubs, 41.18% trees and 21.85% climbers. Maximum diversity is present in
Kanisherykavu (57 plant species and 36 families) and minimum diversity is present in
Adipparambukavu (27 plant species belonging to 18 families). Fabaceae and Moraceae were the
dominant families present in these sacred groves. Among these Kottarathkavu is well protected
and is under the observation of Kerala Forest Department. It also harbours several valuable and
medicinal plants beneficial for mankind. These groves serve as seed banks for future afforestation
and can further help in education and research. Since sacred groves are gradually declining
immediate attempts are needed to scientifically document and explore them to ensure their long
term conservation.
Keywords: Traditional lore - Climax vegetation - Conservation - Bioresources.
[Cite as: Deepa MR, Sheema Dharmapal P & Udayan PS (2016) Floristic diversities and medicinal importance
of selected sacred groves in Thrissur district, Kerala. Tropical Plant Research 3(1): 230242]
INTRODUCTION
Sacred groves have existed in India from time immemorial as patches of densely wooded areas, venerated on
religious grounds. In Kerala it is a common practice among Hindus to assign a part of their land near the
Tharavadu or house as the abode of goddess Durga or serpent god Naga or Shasta and the place is called
Kavu or Sarpakavu. These are one of the informal approaches of conserving the biological diversity of a
region and play an important role in preservation of depleting resource elements such as medicinal plants and
occur in India and in other parts of Asia and Africa (Bhandary & Chandrashekar 2003).Total number of sacred
groves in India varies between 100,000 and 150,000 (Malhotra 1998) and harbour good number of rare and
endemic plants, medicinal herbs and shrubs. These sacred plants have been using since time immemorial by
local tribals and traditional practitioners (Mehra et al. 2014, Bajpai et al. 2016). In sacred groves the number of
herbs and shrubs are more in the disturbed zone (Nair 1992). Due to urbanization and industrialization coupled
with rationalization, scarcity of land leading to the depletion of the cover and shrinkage of kavu as a result the
large chunk of the areas are diverted for other activities and only a small portion is left with especially adjacent
to the temple (Devaraj et al. 2005). Onyekwelu & Olusola (2014) reported sacred groves were preserved by fear
of deity, cultural importance and place of worship. Tree felling within groves was regarded as abomination and
sacrifices must be offered before any tree was felled. The rules and taboos used to protect the groves are
crumbling, which must be addressed if they are to continue playing important role in in-situ biodiversity
conservation. Sacred groves lose their prominence nowadays, but are still relevant in Indian rural landscapes
inhabited by traditional communities (Ray et al. 2014). Sacred groves act as the ancient method of water
harvesting and resource sharing system and are pockets of almost climax vegetation (Karunakran et al. 2005).
230
Deepa et al. (2016) 3(1): 230242

.
They are mostly available in remote tribal areas of the district and become threatened due to loss of traditional
lores and beliefs.
These groves have distinct floral characters making it unique ecosystem (Oommen et al. 2000). It also
harbours 100% valuable and medicinal plants beneficial for mankind. Groves help to maintain water table in
that areas. Filling up of ponds and removal of sacred groves, which used to help maintain the ecological balance
play a major role for the drop in ground water table (The Hindu Business Line 2004). Sacred groves in Kerala
preserve more than 800 species of angiosperms (20% of total flowering plants recorded from the state). Out of
which 150 plants are medicinal and 40% are rare and endangered (Chandrashekara & Sankar 2000). These
groves serve as seed banks for future afforestation and can further help in education and research. Hence it is
necessary to evolve strategies for effective conservation and management of sacred groves.
Sacred groves in Kerala are located mainly in Kasargod, Kannur, Kozhikode, Thrissur, Palakkad, Ernakulam
and Alappuzha districts. About 761 sacred groves have been reported so far from Kerala State
(Balasubramanyan & Induchoodan 1999), which bears many threatened species (Nair & Mohanan 1981). The
present study was conducted to know the status of plant diversity in the sacred groves of Thrissur district of the
State and uses of these plants by the local people.
Study Area
The study was conducted in the sacred groves of Thrissur district, Kerala state, located between 10 52' N
latitude and 76 21' E longitude with an area of 120.26 Km (Fig. 1). Study area includes Adipparambukavu,
Daivathinkavu, Kanisherykavu, Kottaichalippattukavu and Kottarathkavu (Fig. 2). The management of these
kavu is under the control of different families. The main deity is Nagam. Other deities are also present.
Protection of these kavu is mainly due to the presence of deities.
Figure 1. Location of sacred groves in Thrissur district, Kerala.
Data collection
Floristic composition of each grove was analysed during field visits conducted over different seasons
between April 2013 and September 2015, specimens were collected in each species and tagged. All the
Angiosperms including trees, shrubs, herbs and climbers were considered for the study. Important field
observation like habit, phenology of the plant, colour, texture and smell of leaves, local names and local uses
available were also noted. Each species in fresh condition was critically studied with the help of floras like,
Flora of Presidency of Madras (Gamble 19151936); Flowering plants of Thrissur district (Sasidharan &
231
Deepa et al. (2016) 3(1): 230242

.
Sivarajan 1996) and provisional determination was made. The plants were identified with the help of floras and
finally by comparing with the reference collections available in the herbarium of Kerala Forest Research
Institute (KFRI), Peechi. The species were often poisoned, processed and labeled, by standard herbarium
methods given by Jain & Rao (1977).The voucher specimens are deposited at Sree Krishna College, Guruvayur.
IUCN categories are used to evaluate the plants and arranged in to RET species (IUCN 2012).
Figure 2. Selected sacred groves in Thrissur district: A, Adipparambukavu; B, Daivathinkavu; C, Kanisserykavu; D,

Kottaichalippattukavu; E, Kottarathkavu.
232
Deepa et al. (2016) 3(1): 230242

.
RESULTS
Table 1. Location and Deities of Sacred groves studied.
Sacred groves
Taluk
Panchayath
Adipparambukavu
(ADP)
Daivathinkavu
(DAV)
Kanisherykavu
(KNS)
Kodungallur
Valappad
Area
in Ha.
0.741
Kodungallur
Kaippamangalam
0.494
Talappilly
Porkulam
1.359
Kottaichalippattukavu
Chavakkad
Vadanappilly
0.741
(KTC)
Kottarathdharmadai
vamkavu (KTR)
Kodungallur
Kaippamangalam
0.741
Latitude/
Longitude
10.3982 N,
76.0918 E
10.3167 N,
76.1333 E
10.65 N,
76.08 E
10.4667 N,
76.0833 E
10.3167 N,
76.1333 E
Deity
Nagayakshi, Sarpam
Rakshassu, Nagam
Nagam, Bhagavathy
Annapoorneswari,
Veerabhadran
Darmadaivam, Nagam,
Manikandabhootham
The present study conducted in the sacred groves of Adipparambukavu, Daivathinkavu, Kanisherykavu,
Kottaichalippattukavu and Kottarathkavu (Table 1). In these groves 119 species of angiosperms coming under
104 genera and 51 families representing 8 vulnerable, 12 endemic and 3 near threatened species were collected
(Table 2).
Table 2. Number of Angiosperms in Sacred groves.
Sacred groves
Adipparambukavu (ADP)
Daivathinkavu (DAV)
Kanisherykavu (KNS)
Kottaichalippattukavu (KTC)
Kottarathkavu (KTR)
Species
27
31
57
39
40
Genus
24
31
53
38
38
Family
18
20
36
29
26
Endemic Plants
3
4
5
6
9
Vulnerable Plants
3
2
2
3
5
Out of 119 species Hydnocarpus pentandra(Buch.-Ham.) Oken, Leea indica (Burm. f.) Merr. and Pothos
scandens L. are common in these five sacred groves. Caryota urens L. and Chassalia curviflora (Wall ex Kurz)
Thw. are common in Adipparambukavu, Daivathinkavu, Kanisherykavu and Kottarathkavu. Derris scandens
(Roxb.) Benth. and Holigarna arnottiana Hook. f. are common in Adipparambukavu, Daivathinkavu,
Kottaichalippattukavu and Kottarathkavu. Aphanamixis polystachya (Wall.) Parker, Artocarpus hirsutus Lam.,
Calophyllum calaba L., Dalbergia latifolia Roxb., Gloriosa superba L., Hydnocarpus pentandra (Buch.-Ham.)
Oken, Saraca asoca (Roxb.) de Wilde and Smilax zeylanica L. are Vulnerable, Artocarpus hirsutus Lam.,
Briedelia stipularis (L.) Blume, Calophyllum calaba L., Chionanthus mala-elengi (Dennst.) P.S. Green ssp.
mala-elengi, Holigarna arnottiana Hook. f., Hydnocarpus pentandra (Buch.-Ham.) Oken, Memecylon
talbotianumBrandis, Mussaenda frondosa L., Olea dioica Roxb., Pandanus kaidaKurz, Sida rhomboidea Roxb.
ex Fleming and Tabernaemontana alternifolia L. are endemic and Garcinia gummi-gutta (L.) Robs., Magnolia
champaca (L.) Baill. ex Pierre and Tinospora sinensis (Lour.) Merr. are near threatened species present in these
sacred groves (Sasidharan & Sivarajan 1996, Ravikumar et al. 2000). Aeginetia indica L. in Orobanchaceae is a
root parasite present in Kanisherykavu. It includes 17.64% herbs, 19.33% shrubs, 41.18% trees and 21.85%
climbers. All 119 species of plants are medicinal. Food plants of these groves includes Anacardium occidentale
L., Artocarpus heterophyllus Lam., Artocarpus hirsutus Lam., Chrysophyllum cainito L., Citrus medica L.,
Cocos nucifera L., Colocasia esculenta (L.) Schott , and Passiflora edulis Sims (Fig. 3).
Such kind unique plant wealth shows the importance of conservation of sacred groves. Fabaceae and
Moraceae were the dominant families present in these sacred groves (Table 3). Among these Kottarathkavu is
well protected and is under the observation of Kerala Forest Department. Maximum number of Endemic plants
present in Kottarathkavu (6.72%). Adipparambukavu and Kottarathkavu are protected with compound wall.
These groves have distinct floral characters making it unique ecosystem.
Table 3. Species recorded from sacred groves and medicinal uses.
S. N.
1
Botanical name
(Family)
Abrus precatorius L.
(Fabaceae)
Col. No.
1
Sacred
Habit
grove(s)
ADP, KTC, C
KTR
Plant part(s)
used
Leaves, Roots,
Seeds
Uses
Hair growth, fever, difficult breathing,
thirst, eye and skin disease.
233
Adenanthera pavonina L. 2
(Mimosaceae)
ADP,
KNS
Aeginetia indica L.
(Orobanchaceae)
153
KNS
Albizia odoratissima (L.

f.) Benth. (Mimosaceae)
ADP,
T
DAV, KNS
Albizia saman (Jacq.)

F.Muell.
(Mimosaceae)
155
KNS
Alstonia scholaris (L.) R.

Br.
(Apocynaceae)
13
KTC, KTR T
Alternanthera
bettzickiana (Regel) Voss
(Amaranthaceae)
Anacardium occidentale
L.
(Anacardiaceae)
Aphanamixis polystachya
(Wall.) Parker
(Meliaceae)
Areca catechu L.
(Arecaceae)
Artocarpus heterophyllus
Lam.
(Moraceae)
Artocarpus hirsutus Lam.
(Moraceae)
Asparagus racemosus,
Willd.
(Liliaceae)
203
KTR
152
DAV
14
Deepa et al. (2016) 3(1): 230242

.
Bark, Leaves,
Ulcers, pharyngopathy, burning
Seeds,
sensation, hyperdipsia, vomiting, fever,
Heartwood
giddiness, dysentery, pain in joints,
warts and emetic.
Whole plant
Renal cancer, diabetes, acute nephritis,
chronic liver diseases, cough, and
arthritis.
Bark
Insect bites, ulcers, leprosy, skin
diseases, cough, bronchitis, diabetes and
burning sensation.
Root, Seeds,
Stomach cancer, colds, diarrhoea,
headache, intestinal ailments and
stomach ache, sore throat,
Mycobacterium tuberculosis
Bark, Leaves,
Malaria, asthma, skin and respiratory
Milky exudate diseases, cardiac troubles, beri-beri,
fever, abdominal disorders, leprosy, foul
ulcers, bronchitis and congested liver.
Leaves, Stem
Given to anaemic children in order to
improve their health.
Fruits, Seeds,
Roots, Bark
Diabetes, poisoning, ulcers, corn and

aphrodisiac.
ADP, KTR T
Bark, Seeds
Liver enlargement, spleen and

abdominal complaints and tumors.
68
DAV, KNS T
60
DAV,
T
KNS, KTR
Roots, Leaves,
Nut
Roots, Seeds,
Leaves, Fruits
Sore lips, lumbago, urinary disorders and

anorexia. Nuts prevent decay of tooth.
Boils, wounds, skin diseases, fever,
ulcers, vata and pitta disorders.
5
156
ADP, KNS, T
KTR
DAV, KTR C
Fruits, Leaves,
Bark
Tubers
Azadirachta indica A.
Juss.
(Meliaceae)
201
KTC
15
Bambusa bambos (L.)

Voss (Poaceae )
244
KNS
16
Bambusa vulgaris Schrad 69

(Poaceae)
DAV
17
Breynia vitis-idaea
(Burm. f.) C.E.C. Fisch.
(Euphorbiaceae)
Briedelia retusa (L.) A.
Juss.
(Euphorbiaceae)
Briedelia stipularis (L.)
Blume
(Euphorbiaceae)
264
KTC
Anorexia, small pimples, cracks on the

skin and sores.
Urinary diseases, gynaecological
disorders, hyperacidity, gastritis,
improves memory power, increases
breast milk, piles, eye diseases, and
leucorrhoea.
Bark, Leaves,
Skin and eye diseases, rheumatism,
Flowers, Fruits, intestinal worms, diabetes, small pox,
Seeds
chiken pox, ulcers, ringworm, scabies ,
leprosy, liver disorders, cough, anorexia,
polyuria, wounds, fever and poisoning.
Roots, Fruits
Haemorrhoid, diarrhoea, wounds, fever,
Leaves,
cough, shortness of breath, vomiting,
Calcareous
cardiac and skin diseases.
deposits,
Resin
Infantile epilepsy, kidney troubles,
coughs, excess mucous, fever and
reduced risk of digestive disorders.
Bark, Leaves
Haemorrhage and tonsillitis.
238
KNS
Bark , Roots
Pain in lumbago and sciatica.
199
KTC, KTR S
Leaves, Bark
Jaundice, anaemia, cough, fever, asthma

and as gargle for sores in mouth.
10
11
12
13
18
19
234
20
Butea monosperma
(Lam.) Taub
(Fabaceae)
59
KNS
21
Calophyllum calaba L.
(Clusiaceae)
Calycopteris floribunda
Lam.
(Combretaceae)
158
KTC
Deepa et al. (2016) 3(1): 230242

.
Bark, Flowers, Piles, tumours, menstrual disorders,
Seeds, Resin
dysentery, intestinal worms,
anthelmintic, rectal diseases,
hepatopathy, diabetes and hydrocele.
Kernel oil
Healing properties
15
KNS
Leaves, Fruits
Capsicum frutescens L.
(Solanaceae)
Carallia brachiata
(Lour.) Merr.
(Rhizophoraceae)
Caryota urens L.
(Arecaceae)
159
KNS
Fruits
Colic, intestinal worms, leprosy, malaria,

dysentery, ulcers, vomiting, skin
diseases, snake-bite poisoning, thrist and
diarrhoea.
Carminative and rubefacient.
58
KTR
Bark, Fruits
Contagious ulcers and itches.
Shoot apex,
Toddy
Diarrhoea, migraine and scorpion-sting

poisoning.
26
Cassia fistula L.
(Caesalpiniaceae)
245
ADP,
T
DAV,
KNS, KTR
KNS
T
Root, Leaves,
Bark, Fruits,
Flowers
27
Cayra tiapedata (Lam.)

A. Juss. ex Gagnep.
(Vitaceae)
Cayra tiatrifolia (L.)
Domin
(Vitaceae)
Centrosema molle Benth.
(Fabaceae)
Chassaliacurviflora
(Wall. ex Kurz) Thw.
(Rubiaceae)
Chionanthus mala-elengi
(Dennst.) P. S. Green
(Oleaceae)
Chromolaena odorata
(L.) King & Robins.
(Asteraceae)
Chrysophyllum cainito L.
(Sapotaceae)
Cinnamomum verum Presl
(Lauraceae)
150
KTC, KTR C
whole plant
Skin and cardiac diseases, leprosy, fever,

promotes digestion, leucoderma,
eczema, diabetes, jaundice, polyuria, and
urticaria.
Uterine reflexes and applied on cracked
heels.
KNS
Roots
210
KNS
Seed
ADP,
S
DAV,
KNS, KTR
KTC
T
Roots
Leaves
Giddiness, epilepsy and similar

affections of the brain.
211
KNS, KTC, S
KTR
Leaves
Leaf juice is applied externally on cuts

and wounds to stop bleeding
ADP
Fruit
Diarrhoea
10
ADP, DAV T
Bark, Leaf oil
35
Citrus medica L.
(Rutaceae)
56
KNS
Fruits
36
Cleome burmannii Wight

& Arn.
(Capparaceae)
Clerodendrum
infortunatum L.
(Verbenaceae)
Coccinia grandis (L.)
Voight
(Cucurbitaceae)
197
KNS
Whole plant
Anorexia, bronchitis, asthma, diseases of

heart, mouth and teeth, chronic cold,
vomiting, diarrhoea, uropathy and
restoring normal skin colour.
Pain, piles, indigestion, vomiting,
constipation, flatulence, tumours,
helminthiasis, hiccough, hyperdipsia,
anorexia, hepatopathy and dysentery.
Anti-inflammatory
246
KNS
Leaves , Bark
Diabetes, leprosy, skin diseases,

inflammations and proctoptosis
212
KTC
Whole plant,
Rhizomes
Polyuria, cough, diabetes, skin and liver

diseases, fever, ulcers, anorexia,
bronchitis, rheumatism, dysentery,
vomiting, burning sensations, leprosy,
asthma, jaundice and helminthiasis.
22
23
24
25
28
29
30
31
32
33
34
37
38
57
160
Tumours, fever and splenopathy, ulcers,

hepatopathy, cardiac disorders, Wounds
dropsy and haemorrhoids.
Scorpion and snake bites. Antimicrobial,
Wound Healing
Cough and malaria.
235
Deepa et al. (2016) 3(1): 230242

.
Inflorescence,
Bronchitis, hepatopathy, uterine
Fruits, Roots,
disorders, helminthiasis, gastritis,
Seeds
haemorrhage, polyuria, leucorrhoea,
hyperdipsia, tumours, skin diseases,
dysentery, diarrhoea, dehydration and
diabetes.
Rhizomes
Internal haemorrhages, adenitis,
somatalgia, congestion of the portal
system, otalgia and general debility.
Whole plant
Haemorrhage, leprosy and diseases of
vata.
39
Cocos nucifera L.
(Arecaceae)
12
DAV, KNS T
40
Colocasia esculenta (L.)

Schott
(Araceae)
Commelina benghalensis
L.
(Commelinaceae)
Costus speciosus
(Koenig) J.E. Smith
(Costaceae)
Curculigoo rchioides
Gaertn.
(Hypoxidaceae)
Cyclea peltata (Lam.)
Hook. f. & Thoms.
(Menispermaceae)
162
KNS
235
KTR
149
KNS
Rhizomes
54
KNS
Tubers
18
ADP, KTC C
Roots
45
Dalbergia latifolia Roxb.

(Fabaceae)
53
KNS
Roots, Bark,
Leaves
46
Delonix regia (Boj. ex

Hook.) Rafin.
(Caesalpiniaceae)
Derriss candens ( Roxb.)
Benth.
(Fabaceae)
Dioscorea bulbifera L.
(Dioscoreaceae)
234
KTC
Leaves
11
195
ADP,
C
DAV,
KTC, KTR
KNS
C
Seeds, Leaves,
Whole plant,
Bark
Tubers
49
Elephantopus scaber L.
(Asteraceae)
76
DAV
Whole plant
50
Euphorbia thymifolia L.
(Euphorbiaceae)
168
KTC
Whole plant
51
(Moraceae)
247
ADP
Bark, Aerial
roots, Buds
52
Ficus hispida L. f.
(Moraceae)
Ficus racemosa L.
(Moraceae)
Ficus religiosa L.
(Moraceae)
Ficus tinctoria G. Forst.
(Moraceae)
Garcinia gummi-gutta
(L.) Robs.
(Clusiaceae)
169
KNS
Bark, Fruits
51
ADP
Bark
Haemorrhage, fever, cough and other

respiratory diseases, diabetes, blood and
skin diseases and leprosy.
Urinary and skin diseases, menorrhagia,
piles, jaundice, asthma, diarrhoea and
gonorrhea.
Purifies blood and beneficial in treating
skin diseases, poisonous affections, colic
pain, fever, vomiting, diarrhoea and
respiratory disorders.
Polyuria, sciatica, chronic ulcer, leprosy,
urinary bladder disorders, burning
sensation, oedema, brain tonic,
diarrhoea, obesity and worms.
Diseases of vata, constipation,
inflammations, arthritis, hemiplegia and
dysmenorrhoea.
Unripe beans loosen the bowels with
gripe. Leaves reduced to plasma are
good in erysipelas.
Ulcers, piles, leprosy, worm infestation,
cardiac diseases, polyuria, urinary
calculi, aphrodisiac, rejuvenator,
dysentery and syphilis.
Diarrhoea, hemorrhage, urinary calculi,
leprosy, retention of urine, bronchitis,
skin disease, intermittent fevers,
hepatopathy, ophthalmopathy, cough and
swellings.
Cough, asthma, respiratory and skin
diseases, worms, poisonous affections,
dyspnoea and purification of blood.
Skin diseases, dysentery, diarrhoea,
leucorrhoea, nervous disorders and
reduces blood sugar in diabetes.
Ulcers, leucoderma, psoriasis, anaemia,
jaundice, and inflmmations.
Skin and vaginal diseases and ulcers.
77
Bark
Skin and vaginal diseases and ulcers.
144
ADP,
T
DAV, KTC
ADP
T
Root, Leaves
193
ADP, DAV T
Leaves, Fruits,
Seed oil
Women during childbirth, relieve

swollen eyes.
Ulcers, inflammations, bleeding piles,
diarrhoea, cold, dysentery, indigestion,
hyperdipsia, antiobesity, dropsy and
worm cases.
41
42
43
44
47
48
53
54
55
56
236
57
Gliricidia sepium (Jacq.)

Kunth ex Walp.
(Fabaceae)
Gloriosa superba L.
(Liliaceae)
232
ADP, KTC, T
KTR
Leaves, bark,
Seeds
50
KTC, KTR C
Tubers
Grewia nervosa (Lour.)

Panigrahi
(Tiliaceae)
Grewia tiliifolia Vahl
(Tiliaceae)
217
KNS, KTC S
Whole plant
248
KTC
Bark, Leaves
61
Hemidesmus indicus (L.)

R.Br.
(Periplocaceae)
172
KNS, KTR C
Roots
62
Hibiscus hispidissimus
Griff.
(Malvaceae)
Hibiscus rosa-sinensis L.
(Malvaceae)
21
KNS
Leaves, Roots
218
KNS
Leaves,
Flowers, Roots
Holarrhena pubescens
143
(Buch.-Ham.) Wall. ex G.
Don
(Apocynaceae)
Holigarna arnottiana
174
Hook.f.
(Anacardiaceae )
Hydnocarpus pentandra 192
(Buch.-Ham.) Oken,
(Flacourtiaceae)
KTR
Bark, Seeds
58
59
60
63
64
65
66
67
68
69
70
71
72
Hyptis suaveolens (L.)

Poit.
(Lamiaceae)
Ichnocarpus frutescens
(L.) R.Br.
(Apocynaceae)
Indigofera cassioides
Rottl. ex. DC.
(Fabaceae)
Indigofera hirsuta L.
(Fabaceae)
Ipomoea staphylina
Roem. & Schult.
(Convolvulaceae)
Ixora coccinea L.
(Rubiaceae)
Deepa et al. (2016) 3(1): 230242

.
Headache, cold and cough.
Swelling, piles, oedema, leprosy, ulcers,

pain in the bladder, toxicosis, itching,
antidote against cobra poison; easy and
quick expulsion of the placenta after
delivery.
Indigestion, eczema and itch, typhoid,
dysentery and syphilitic ulceration of the
mouth.
Burning sensation, hyperdipsia,
pharyngopathy, cough, skin, blood and
cardiac diseases, wounds, ulcers,
diarrhoea, haemorrhages and seminal
weakness.
Dyspepsia, dysentery, cough, bronchitis,
gout, uterine haemorrhage, wounds,
leprosy, blood and skin diseases,
anaemia, jaundice, dysuria, fever, thirst,
vomiting and rheumatism.
Improves digestion, inflammations,
helminthiasis, dyspepsia and
opthalmopathy.
Skin diseases, diarrhoea, piles,
haemorrhage, polyuria, hair falling,
menorrhagia, cough, contraceptive,
fever, cystitis and irritable conditions of
genito urinary tract.
Diarrhoea, piles, haemorrhage, leprosy,
worm infestation, thrist, pain, erysipelas,
hepatopathy, gastropathy, chronic
bronchitis, boils, ulcers and dysentery.
Arthritis, beriberi, tumours, leucoderma,
ulcers, diabetes, leprosy and warts.
ADP,
T
DAV,
KTC, KTR
ADP,
T
DAV,
KNS, KTC,
KTR
DAV,
H
KTC, KTR
Fruits
175
KTC
Roots
249
KTC
Roots
Coughs, pains in the chest.
254
DAV
Leaves
stomach problems and yaws
176
KTC, KTR C
Stem latex
Skin disease.
80
DAV,
S
KNS, KTR
Roots, Leaves,
Flowers
Blood purifier, antiseptic, infantile skin

ailments, diarrhoea, dysentery, fever,
sores, ulcers, hemoptysis, catarrhal
bronchitis, eye troubles, scabies, cholera
and gonorrhoea.
154
Seeds, Seed oil
Whole plant
Leprosy, skin diseases, eczema,

dermatitis, tubercular laryngitis, chronic
ulcers, dyspepsia, flatulence and
verminosis.
Worm infestation, wounds and
inflammations of the navel of the
newborn and also emetic.
Dyspepsia, diabetes, fever, skin troubles
and stones in bladder.
237
73
74
Deepa et al. (2016) 3(1): 230242

.
Poisoning, herpes, leprosy,
opthalmopathy, and wounds.
Jasminum angustifolium
(L.) Willd.
(Oleaceae)
Leea indica (Burm.f.)
Merr.
(Leeaceae)
Macaranga peltata
(Roxb.) Muell.-Arg.
(Euphorbiaceae)
Magnolia champaca (L.)
Baill. ex Pierre
(Magnoliaceae)
49
KNS
Leaves
48
ADP,
Roots
Diarrhoea, dysentery, hyperdipsia, ulcer

and skin diseases.
Leaves, Bark,
Gum
Used as vulnerary. Gum used for

venereal sores.
DAV, KNS,
267
KTC, KTR
KNS
T
178
KNS
Bark, Flowers
Manihot carthaginensis
ssp. glaziovii (Muell.Arg.) Allem
(Euphorbiaceae)
Memecylon talbotianum
Brandis
(Melastomataceae)
Merremia vitifolia (Burm.
f.) Hall. f.
(Convolvulaceae)
Mikania micrantha Kunth
in HBK
(Asteraceae)
Mimosa pudica L.
(Mimosaceae)
251
DAV
Stem, Root
Chronic gastritis, fever, strangury,

cough, bronchitis, nausea, leprosy,
wounds, ulcers, anorexia, colic,
flatulence, helminthiasis, cephalalgia,
and ophthalmia.
Skin infections.
123
DAV,
T
KTC, KTR
Bark, Root,
Seeds, Leaf
Anti-diarrhoeal, Hypoglycemic,
Antimicrobial, Wound healing.
119
CHL, KNS C
Whole plant,
Roots
Strangury and urethral discharges. Root

eaten by tribals as a stomachic.
47
DAV, KNS C
Leaves
139
KTC, KTR, H
DAV
Whole plant,
Roots
82
Mimusops elengi L.
(Sapotaceae)
84
ADP, KNS T
Bark, Flowers,
Fruits
83
Morinda pubescens J. E.
Smith
(Rubiaceae)
Mussaenda frondosa L.
(Rubiaceae)
Naravelia zeylanica (L.)
DC.
(Ranunculaceae)
Ocimum tenuiiflorum L.
(Labiatae)
180
KNS
Bark, Roots,
Fruits
118
KNS, KTR S
46
KNS
Roots, Leaves,
Stem
Whole plant
86
KNS
Whole plant
Olea dioica Roxb.

(Oleaceae)
Oplismenus compositus
(L.) P. Beauv.
(Poaceae)
Pandanus kaida Kurz
(Pandanaceae)
252
KNS
Bark, Leaves
Snake bites, eliminating discomfort of

hornet, bee and ant stings antimicrobial
activity from the leaves
Urinary complaints, sores, piles,
diarrhoea, dyspnoea, leprosy, uterine
disorders, haemorrhage, wounds,
oedema, skin diseases and burning
sensation.
Urethrorrhoea, diarrhoea, dysentery,
cephalalgia, leprosy, constipation,
dental, cardiac and eye diseases,
burning sensation, thirst, uterine
disorders, fever, headache, poisoning
and aphrodisiac.
Eczema, fever, ulcers, glandular
swellings and digestive disorders
especially in children.
Leprosy and eye troubles, coughs and
against intestinal worms.
Helminthiasis, leprosy, dermatopathy,
rheumatalgia, odontalgia, wounds,
cephalalgia, inflammations, and ulcers.
Cough, cold, bronchitis, dysentery,
improves appetite, skin and ear diseases,
itches, ringworm, leprosy, intestinal
worms, ulcers, poisonous affections and
specific for all kinds of fevers.
Febrifuge and emetic.
116
KTC
Whole plant
Relieve pain of snakebite
270
DAV, KTR S
Stem, Sap,
Flower
Wounds, Fevers, Pains, Epilepsy, Skin

diseases, Ear diseases, Headaches, Back
pains, Rheumatoid arthritis, Diabetes
mellitus, Psychological disorders.
75
76
77
78
79
80
81
84
85
86
87
88
89
238
90
Passiflora edulis Sims

(Passifloraceae)
138
KNS
91
Pavetta indica L.
(Rubiaceae)
114
DAV, KTC S
92
Phyllanthus reticulatus
Poir.
(Euphorbiaceae)
183
KTR
93
Plumeria rubra L.
(Apocynaceae)
Polyalthia longifolia
(Sonner.) Thw.
(Annonaceae)
Pothos scandens L.
(Araceae)
44
KTC
188
KTR
137
Racosperma
auriculiforme
(Benth.) Pedley
(Mimosaceae)
Saraca asoca (Roxb.) de
Wilde
(Caesalpiniaceae)
42
ADP,
C
DAV,
KNS, KTC,
KTR
DAV
T
41
ADP
Bark, Flowers
Sarcostigma kleinii Wight 187

& Arn.
(Icacinaceae)
Sida acuta Burm. f.
108
(Malvaceae)
ADP
Bark, Leaves,
Seed oil
KTC
Roots
Sida fryxellii Sivar. &

Pradeep (Malvaceae)
Sida rhomboidea Roxb.
ex Fleming
(Malvaceae)
Smilax zeylanica L.
(Smilacaceae)
(Moraceae)
253
KTC
Whole plant
94
KTR
Roots, Leaves
Fever, heart diseases, burning sensations,

piles and inflammations.
31
Roots
128
DAV,
C
KTC, KTR
CHL, KNS T
104
Strychnos nux-vomica L.
(Loganiaceae)
39
KNS, KTC T
Bark, Seeds
105
Swietenia mahagoni (L.)

Jacq. (Meliaceae)
Tabernaemontana
alternifolia L.
(Apocynaceae)
Tabernaemontana
divaricata (L.) R. Br.
(Apocynaceae)
105
KNS
Bark
135
ADP, KNS, T
KTR
Roots, Bark
Venereal diseases, rheumatism,

urinarycomplaints and dysentery.
Sinusitis, inflammations, elephantiasis,
cough, bronchitis, ulcers, diarrhoea,
dysentery, fever, swellings,
hyperhidrosis, neuralgia and
haemorrhages.
Intermittent fevers, dyspepsia, dysentery,
paralytic and neuralgic affections,
chronic rheumatism, insomnia, colic,
impotence, spermatorrhoea and skin and
heart disease.
Anti-pyretic, tonic and astringent; used
as a substitute for Cinchona bark.
Toothaches, inflammations of cornea
and also as a vermicide.
225
KNS
Flowers, Roots
94
95
96
97
98
99
100
101
102
103
106
107
Deepa et al. (2016) 3(1): 230242

.
Flower
Nervous disorders, bronchial conditions,
arthritis, asthma, insomnia,
gastrointestinal disorders and
menopausal symptoms.
Roots, Leaves
Visceral obstructions, urinary diseases,
jaundice, dropsical affections, ulcerated
nose and for haemorrhoids.
Bark, Leaves,
Rheumatism, dysentery and venereal
Fruits
diseases, burning sensation, gastropathy,
obesity, ophthalmodynia, sores, burns,
and skin eruptions.
Roots, Bark,
Ulcers, herpes and scabies, itch,
Latex
rheumatism and gum troubles.
Bark
Rheumatism, constipation, worm
infestation, polyuria, skin diseases and
fever.
Whole plant
Skin diseases, boils, swellings, wounds,
ulcers, dropsy, menorrhagia, vomiting,
flatulence, strangury and burning
sensation.
Root, Bark
Aches and pains and sore eyes,
rheumatism.
Bark, Roots,
Seeds
Uterine disorders, cures enlargement of

cervical glands, burning sensation,
dyspepsia, worms and biliousness,
bleeding piles, scabies and other skin
diseases.
Cephalalgia, gastropathy, helminthiasis,
leprosy, skin diseases, epilepsy and
indolent ulcers.
Uropathy, arthritis, leucorrhoea,
gonorrhoea, diarrhoea and to promote
strength.
Antibacterial properties
Eye diseases, burning sensation, skin

diseases, toothache, and joint pains.
239
Deepa et al. (2016) 3(1): 230242

.
Tender leaves, Bronchitis, dysentery, skin diseases,
Bark, Flowers, inflammation, pruritus, ulcers,
Fruits, Seeds
haemorrhage, haemoptysis, vesical
calculi, stomatitis, dipsia, and strangury,
arthritis, neuralgia and dyspepsia.
Whole plant,
Inflammations, skin diseases,
Roots
elephantiasis, dyspepsia, stomachalgia,
flatulence, asthma, bronchitis, anaemia,
fever, boils, pimples, syphilis,
gonorrhoea and rat poisoning.
Roots
Boils and ulcers, traumatic bleeding,
snakebites
108
Tectona grandis L.f.

(Verbenaceae)
38
KNS
109
Tephrosia purpurea (L.)

Pers.
(Fabaceae)
104
KTR
110
Tetrastigma
leucostaphylum (Dennst.)
Alston ex Mabb.
(Vitaceae)
Tiliacora acuminata
(Poir.) Miers. ex Hook. f.
& Thoms.
(Menispermaceae)
Tinospora sinensis
(Lour.) Merr.
(Menispermaceae)
Triumfetta rhomboidea
Jacq.(Tiliaceae)
Urena lobata L.
(Malvaceae)
Vanda tessellata (Roxb.)
Hook. ex D. Don
(Orchidaceae)
Vernonia cinerea (L.)
Less.
(Asteraceae)
33
ADP
255
ADP, KTR C
Roots
Antidote to snake poison
37
KTC
Stems
134
KNS
Whole plant
227
KTR
Roots
Piles and ulcerated wounds, liver

complaints, Chronic rheumatism and
also as muscle relaxant.
Dysentery, intestinal ulcers, diarrhoea
and leprosy.
Flatulent colic, cough, and sore throat.
273
KTC
Roots
Dyspepsia, bronchitis, inflammations

and piles.
35
KNS
Whole plant
Vernonia elliptica DC.

(Asteraceae)
Zanthoxylum rhetsa
(Roxb.) DC.
(Rutaceae)
256
KTR
133
KNS
Stem, Leaf,
Flower
Bark, Fruits
Fever, leucorrhoea, excessive bleeding,

skin diseases, dysuria, bladder stones,
piles, worms and haematological
disorders.
Fever, body tonic, parasites.
111
112
113
114
115
116
117
118
Dyspepsia, asthma, bronchitis, heart

diseases, toothache, diseases of eye and
ear, worm infestation, leprosy, diseases
of head, rheumatism, cholera and
treating pimples.
119 Zingiber zerumbet (L.)
101
H
Rhizomes
Cardiac disorders, oedema, cures
J.E. Smith
KNS
vomiting, piles, filariasis, anaemia,
(Zingiberaceae)
cough, dyspnoea, anorexia, fever,
diarrhoea, dyspepsia, diseases, diabetes,
eye and neurological diseases.
Note: ADP- Adipparambukavu, DAV- Daivathinkavu, KNS- Kanisherykavu, KKTC- Kottaichalippattukavu, KTRKottarathdharmadaivamkavu, C- Climber, H- Herb, S- Shrub, T- Tree.
Sacred groves are considered as store house of rare, endemic and endangered plants because of floristic
wealth and biodiversity conservation. This study shows that natural vegetation is maintained inside the sacred
grove and all species are medicinal. Here the percentages of tree species are large compared to herbs, shrubs and
climbers. Compound walls were absent in Daivathinkavu (DAV), Kottaichalippattukavu (KTC) and
Kanisherykavu (KNS). Due to this increase the external interference of human beings and cattle grazing. At the
time of heavy rainfall soil erosion is common and therefore fertilized upper soil lost, these adversely affected the
plant growth. Invasive weeds like Chromolaena odorata (L.) King & Robins and Mikania micrantha Kunth
adversely affect the growth of other plants inside. These are major threat to conservation. Wastes including
plastics deposited in groves are other threats. Plants are considered as lungs of earth. Conservation of groves
means conservation of floras and faunas inside groves. Sacred groves in undisturbed state conserve biodiversity
240
Deepa et al. (2016) 3(1): 230242

.
and ecological balance. Clearing of vegetation for construction of temples and roads has resulted in shrinkage of
sacred groves. Protection of sacred ponds, RET species and keystone species like Ficus trees, nesting birds are
necessary for biodiversity conservation inside the grove. In this circumstance suitable management measures
and awareness programmes about medicinal plants inside the sacred groves are necessary for sustainable
utilization of the valuable bioresources.
Figure 3. Some important plant species with their plant part used as food present in sacred groves: A, Artocarpus
heterophyllus Lam.; B, Artocarpus hirsutus Lam.; C, Citrus medica L.; D, Cocos nucifera L.; E, Colocasia esculenta (L.)
Schott; F, Passiflora edulis Sims.
ACKNOWLEDGEMENTS
Authors are grateful to Sri. D. Jayaprasad, Principal and Dr. G. Jayakrishnan, Department of Botany, Sree
Krishna College, Guruvayur for providing valuable suggestions for the work. Authors acknowledge the family
members of these sacred groves for granting permission to conduct the study and providing information about
the groves.
REFERENCES
Bajpai O, Pandey J & Chaudhary LB (2016) Ethnomedicinal uses of tree species by Tharu tribes in the
Himalayan Terai region of India. Research Journal of Medicinal Plant 10(1): 1941.
Balasubramanyan K & Induchoodan NC (1999) Can the endemics of the Sacred Groves in Kerala withstand
human onslaught? In: Kumaravelu G & Chaudhuri KK (eds) Endemic and endangered plant and animal
241
Deepa et al. (2016) 3(1): 230242

.
species of Eastern and Western Ghats. Proceedings of the national seminar conducted by Research Wing,
Tamil Nadu Forest Department, Chennai, pp. 5964.
Bhandary MJ & Chandrashekar KR (2003) Sacred Groves of Dakshina Kannada and Udupi districts of
Karnataka. Current Science 85(12): 16551656.
Chandrashekara UM & Sankar S (2000) Structure and functions of sacred groves: Case studies in Kerala. In:
Ramakrishnan PS, Saxenaand KG & Chandrashekara UM (eds) Conserving the sacred for Biodiversity
Management. Oxford and IBH Publishing Co. Pvt. Ltd., pp. 323335.
Devaraj P, Ramanujam MP & Ganesan T (2005) Status report of Sacred groves of Pondicherry Region and
Strategies for Conservation. Institute of Forest Genetics and Tree Breeding PB 1061, R.S. Puram,
Coimbatore 641002, India, pp. 1621.
Gamble JS & Fischer CEC (19151936) The Flora of the Presidency of Madras. Parts 111 (parts 1-7 by
Gamble and 8-11 by Fischer), Vols. 13. Adlard & Sons Ltd., London.
Jain SK & Rao RR (1977) A Handbook of Field and Herbarium Methods. Today & Tomorrow, New Delhi.
Karunakran PV, Balasubramanian M & Ramesh BR (2005) Conservation and Management of Sacred Groves in
Kerala as Community Reserves. Institute of Forest Genetics and Tree Breeding, PB 1061, R.S.Puram,
Coimbatore 641002, India, pp. 233238.
Malhotra KC (1998) Anthropological dimensions of sacred groves in India: an overview. In: Ramakrishnan PS,
Saxena KG &Chandrashekara UM (eds) Conserving the Sacred for Biodiversity Management. Oxford and
IBH, New Delhi, pp. 423438.
Mehra A, Bajpai O & Joshi H (2014) Diversity, utilization and sacred values of Ethno-medicinal plants of
Kumaun Himalaya. Tropical Plant Research 1(3): 8086.
Nair HG (1992) Ecological studies of a Sacred Grove. Project report submitted to W.W.F, New Delhi, pp. 55.
Nair NC & Mohanan CN (1981) On the rediscovery of four threatened species from the sacred groves of Kerala.
Journal of Economic and Taxonomic Botany 2: 233235.
Onyekwelu JC & Olusola JA (2014) Role of sacred grove in in-situ biodiversity conservation in rainforest zone
of southwestern Nigeria. Journal of Tropical Forest Science 26(1): 515.
Oommen S, Ved DK & Krishnan R (2000) Tropical Indian Medicinal Plants Propagation methods. Foundation
for Revitalisation of Local Health Traditions (FRLHT) #50, MSH layout, 2nd stage, 3rd main Anandnagar,
Bangalore- 560 024, India. pp. 26, 326.
Ray R,Chandran MDS & Ramachandra TV (2014) Biodiversity and ecological assessments of Indian sacred
groves. Journal of Forestry Research 25 (1): 2128.
Ravikumar K, Ved DK Assisted by VijayaSankar R & Udayan PS (2000) 100 Red-Listed Medicinal Plants of
Conservation Concern in Southern India. Foundation for Revitalisation of Local Health Traditions
(FRLHT), 50, MSH Lay out, 2nd stage, 3rd main, Anand Nagar, Banglore, India.
Sasidharan N & Sivarajan VV (1996) Flowering Plants of Thrissur Forest(Western Ghats, Kerala, India).
Scientific Publishers, Jodhpur.
The Hindu Business Line (2004) Available from: http://www.thehindubusinessline.com/2004/03/18/
stories/2004031800421700.htm (accessed: 18 May 2004).
242
ISSN (E): 2349 1183

ISSN (P): 2349 9265
3(1): 243248, 2016
Research article
Variation in rock phosphate solubilization by three isolates of

Aspergillus niger van Tieghem grown in liquid media
supplemented with different carbon and nitrogen sources
Hruda Ranjan Sahoo and Nibha Gupta*
Division of Plant Pathology and Microbiology, Regional Plant Resource Centre, Bhubaneswar, Odisha, India
*Corresponding Author: nguc2003@yahoo.co.in
Abstract: In the present study, we worked out the phosphate solubilization potential of three
different isolates of Aspergillus niger from various sources such as leaf (L), root (R) and soil (S)
under different carbon and nitrogen supplementation in liquid culture. The fungal cultures were
inoculated in Czapek Dox Medium containing different carbon and nitrogen sources and
supplemented with Moroccan rock phosphate. The fungal strains exhibited good potential favoring
the solubilization of rock phosphate in laboratory conditions. On carbon sources modification,
Aspergillus niger (L) and (S) showed highest P solubilization activity 27.6% and 29.6%
respectively in presence of glucose whereas Aspergillus niger (R) showed highest P solubilization
of 27.8% in presence of inositol. Similarly on modification of nitrogen sources, Aspergillus niger
(L) and (S) showed maximum solubilization of 35.25% and 40.8% respectively, but Aspergillus
niger (R) showed maximum solubilization of 37% in presence of amino acid L-phenylalanine in
culture broth. Further pot experiment with different soil composition and host plants may exhibit
its exploitable potential.
Keywords: Rock phosphate - Phosphate solubilization - Carbon sources - Nitrogen sources.
[Cite as: Sahoo HR & Gupta N (2016) Variation in rock phosphate solubilization by three isolates of
Aspergillus niger van Tieghem grown in liquid media supplemented with different carbon and nitrogen sources.
INTRODUCTION
Phosphorus is an important nutritional element required indirectly for plant growth and development by
incorporating itself in several biomolecules such as phospholipids, nucleic acids and nucleotides (Ahemad et al.
2009). Besides, cellular organization, phosphorus plays key role in many physiological and biochemical role and
ultimately leads to crop growth and yield (Bagyaraj et al. 2000). However it is deficient in soil as it is fixed
resulting into low availability and demands more input of chemical fertilizer. Under such poor soil conditions,
phosphate solubilizing microbes can be helpful in mineralization of fixed phosphorus (Whitelaw 2000). A wide
range of fungi especially Aspergillus and Penicillium are reported to solubilize insoluble form of phosphorus
depending upon their ability to produce and release organic acids to their surrounding environment (Seshadri et
al. 2004, Wakelin et al. 2004). The organism metabolic activity is dependent upon the cultural, nutritional and
environmental factors (Nahas 1996, Kundu et al. 2002). Mostly carbon sources are the important factor behind
the microbial metabolism and proliferation (Yadav et al. 2010, Khan et al. 2013). Different nitrogenous
components in the growth media has also impact upon the microbial activity (Habte & Osorio 2012). Keeping
this view, a study was planned to evaluate the efficiency of three different isolates of Aspergillus niger van
Tieghem for phosphate solubilization under different nutritional conditions added with rock phosphate.
The Fungal cultures of Aspergillus niger van Tieghem Leaf, Root and Soil isolated from leaf, root and soil,
were grown on Sabouraud Dextrose Agar slant for 3 days at 30C for further study. The plate solubilization
index of the three isolates was calculated by the formula given by Gaur et al. (1990) and Premono et al. (1996).
Triplicate sets of Czapek Dox broth medium with rock phosphate as phosphate sources and separately with
243
Sahoo & Gupta (2016) 3(1): 243248

.
different carbon and nitrogen sources were used and inoculated with these fungal isolates and incubated at
282C for 10 days. P content was estimated by vanadophosphomolybdate method and represented in the form
of % solubilised (Jackson 1958).
Phosphate solubilizing potential of three isolates of Aspergillus niger in solid state and liquid culture
conditions is depicted in table 1. Solubilisation index evaluated through the plate test with PK medium added
with TCP showed good activity of these isolates of Aspergillus niger and ranged as 1.27 to 1.85. All the three
Aspergillus niger showed 23.7% to 25.83% whereas difference in solubilization of rock phosphate in liquid
culture have been observed among these fungal isolates. Aspergillus niger isolated from root showed highest
solubilization (36.2%) followed by other two isolated from soil (31.3%) and leaf (22.7%). Though fungi
contribute more to the soil biomass being the important constituents of soil microflora depending on the
conditions like soil depth and nutrient conditions, Aspergillus niger sourced from leaf showed better
solubilization efficiency in plate test.
Table 1. Phosphate solubilising potential of three isolates of Aspergillus niger in solid state and liquid culture conditions.
P solubilized
Solubilization Efficiency (%)
Plate test
(solid medium) Solubilization Index
Broth culture TCP Solubilization (%)
(liquid medium) RP solubilization (%)
Aspergillus niger (L) Aspergillus niger (R) Aspergillus niger (S)

87.5
50
27
1.875
1.5
1.27
25.836.54
23.92.27
23.74.3
22.70.46
36.25.6
31.31.99
Carbon and nitrogen sources greatly influence phosphate solubilization process. In the presence of various
carbon and nitrogen sources, micro-organisms have diverse levels of phosphate solubilization activity. Rock
phosphate solubilization under carbon and nitrogen supplementation in media separately has shown in table 23.
It is clearly evident that carbon sources affect the P solubilization capacity of fungi indicating that microorganisms utilizes different carbon sources as energy sources and most of the test sugars support phosphate
solubilization activity. Aspergillus niger (R) was able to solubilize rock phosphate more in different carbon
source used except lactose and sorbose (Table 2). The pH measured after 10 day incubation period of fungal
culture under such circumstances varies from 4.46 to 6.28 (Fig. 1). Similarly Aspergillus niger (S) performed
good for rock phosphate solubilization in liquid culture and exhibited solubilised P % ranged 24.329.6 except
for lactose. Presence of glucose and maltose in culture confirmed to be better carbon sources for this organism
as far as phosphate solubilization is concerned. Though, all carbon sources tested in the present experiment
showed good support for the metabolic activity pertaining to phosphate solubilization, lactose did not contribute
well in this regard. Data measured on the drift of pH after 10 days of incubation period of 10 days are presented
in figure 2. It is observed that carbon metabolism in presence of sugar fructose is more as compared to other
sugars used as pH of the final culture was drifted to 3.9 (Fig. 3). Aspergillus niger (L) did not show much
solubilization efficiency in presence of different carbon sources used except glucose (27.6 %). Though the fungi
showed decrease in pH in presence of fructose (3.9), it could not affect the solubilization as Aspergillus niger
(L) showed 14.95.6% phosphate solubilization only.
Table 2. Effect of Carbon sources on phosphate solubilization activity (% P solubilized) of different
isolates of Aspergillus niger.
Carbon sources
Aspergillus niger (L) Aspergillus niger (R)
Aspergillus niger (S)
14.95.6
22.92.1
25.21.45
Fructose
27.62.5
261.8
29.61.16
Glucose
3.40.49
27.81.15
24.34.16
Inositol
0.80
0.90.45
3.10.95
Lactose
0.750.07
22.22.25
28.90.71
Maltose
11.61.8
21.71.16
26.51.52
Mannose
8.80
21.90.51
26.21.8
Raffinose
30.42
13.91.46
24.44.7
Sorbose
3.40.49
24.50.7
27.32.48
Sucrose
All the fungi showed diverse levels of RP solubilization activity in presence of different carbon sources.
Glucose being the simplest sugar is favorable for growth of organism and acid production thereby enhances
solubilized P release in the medium. All the 3 fungi showed good P solubilization activity in presence of
glucose. The pH drift and P solubilization potential of organisms was estimated after 10 days of incubation.
244
Sahoo & Gupta (2016) 3(1): 243248

.
Acid production is commonly observed during P solubilization (Scervino et al. 2011). Hence, P solubilization
takes place due to release of organic acids by fungi resulting in reduction of pH (Yadav & Singh 1991, El-Komy
2005). The pH change from neutral to acidic condition is found in presence of most of the sugars at the end of
10 days. However, no correlation could be established between P solubilization in liquid broth and acidic pH
recorded for the same. Similar observations were also noticed in experiments conducted by Wani et al. (1979).
8
6.88
7.04
6.88
6.29
7
6
5
3.9
4.86
4.39
4.53
4.56
pH
4
3
2
1
0
Fructose Glucose Inositol Lactose Maltose Mannose Raffinose Sorbose Sucrose
Carbon sources
Figure 1. Effect of carbon sources on pH drift during P solubilisation in Aspergillus niger (L).
6.28
6
5
4.46
4.8
6.08
5.94
5.32
5.13
4.82
4.69
pH
4
3
2
1
0
Carbon sources
Figure 2. Effect of carbon sources on pH drift during P solubilisation in Aspergillus niger (R).
6.34
6.06
pH
4.21
4.33
4.28
4.31
4.39
4.24
3.9
4
3
2
1
0
Carbon sources
Figure 3. Effect of carbon sources on pH drift during P solubilisation in Aspergillus niger (S).
245
Sahoo & Gupta (2016) 3(1): 243248

.
Table 3. Effect of nitrogen sources on phosphate solubilisation activity (% P solubilized) of different isolates
of Aspergillus niger.
Nitrogen sources
Ammonium chloride
Ammonium Sulphate
L-Glutamine
L-Phenylalanine
L-Threonine
L-valine
Potassium nitrate
Urea
Sodium nitrate
Aspergillus niger (L)

23.44.81
27.453.75
30.89.05
29.86.4
35.250.07
25.711
32.62.12
24.857.7
34.20.14
Aspergillus niger (R)

29.41.41
34.90.7
22.41.6
372.3
22.72.76
34.13.6
34.92
6.84.1
32.751.34
Aspergillus niger (S)

39.31.1
39.71.35
38.70.36
390.75
40.81.67
35.20.51
30.23.1
22.81.46
28.91.3
All the tested Aspergillus niger L, R, S influenced RP solubilization process in presence of different nitrogen
sources (salts/amino acids). The results obtained during experiment have been presented in table 3. All the three
fungi have shown good solubilization potential in presence of different nitrogen sources in combination of
glucose as basal carbon source. Aspergillus niger (L) solubilised rock phosphate at 35.25 and 34.2% in presence
of L-threonine and sodium nitrate respectively whereas Aspergillus niger (R) preferred L-phenylalanine and
could be able to solubilize 37.2% rock phosphate in liquid culture conditions. Aspergillus niger (S) showed
good P solubilization activity in presence of most of the nitrogen sources which demonstrates that this organism
is well adapted and can grow and function in different Cultural conditions. However, no significant changes in
the pH of the culture filtrate was observed as an effect of different nitrogen sources (Fig. 4, 5). The decline in
the pH of the culture filtrate of Aspergillus niger (S) has been observed and presented in figure 6. Highest drift
in pH to highly acidic range i.e. 23 was measured in the presence of L-phenylalanine followed by potassium
nitrate (3.3) and sodium nitrate (3.45). However, it is known that nitrates were more efficient nitrogen source for
P solubilization activity due to presence of assimilatory enzymes for nitrate reduction in organisms (Dave &
Patel 2003). Moreover inorganic nitrogen sources proved to be better source for P solubilization activity as
compared to organic ones (Selvi et al. 2012).
7
6.22
6.38
4.96
5.06
5.08
5.23
4.98
5.56
5.22
5
pH
4
3
2
1
0
Nitrogen sources
Figure 4. Effect of nitrogen sources on pH drift during P solubilisation in Aspergillus niger (L).
The role of Aspergillus niger towards solubilization of phosphate sources is clearly evident from the present
study. Though many fungi obtained from the different sources other soil have been observed as potent candidate
to be exploited for the bioinoculant development, the role of soil fungi stand unbeatable as fungi present in the
soil convert unavailable forms of phosphorus to available phosphorus for plant to absorb by employing different
strategies. Soil P transformations are mediated by microbial activity and influenced by various factors such as
plant species, soil type and environmental factors (Chen et al. 2004). In the present study, the fungal strain
exhibited good potential favoring the solubilization of rock phosphate in laboratory conditions with
supplementation of different carbon and nitrogen sources but further pot experiment with different soil
composition and host plants may exhibit its exploitable potential.
246
7
6
6.19
5.01
5.06
5.65
5.77
Sahoo & Gupta (2016) 3(1): 243248

.
6.21
6.26
6.12
6.01
5
pH
4
3
2
1
0
Nitrogen sources
Figure 5. Effect of nitrogen sources on pH drift during P solubilisation in Aspergillus niger (R).
6.66
7
6
4.9
pH
4.7
5.5
5.26
4.82
3.3
4
3
3.45
2.3
2
1
0
Nitrogen sources
Figure 6. Effect of nitrogen sources on pH drift during P solubilisation in Aspergillus niger (S).
ACKNOWLEDGEMENT
The financial assistance obtained through INSPIRE programme, (No. DST/INSPIRE Fellowship/2013/506)
DST, Govt. of India is gratefully acknowledged.
REFERENCES
Ahemad M, Zaidi A, Khan MS & Oves M (2009) Biological importance of phosphorus and phosphate
solubilising micro-organisms- an overview, In: Khan MS & Zaidi A (eds) Phosphate solubilizing microbes
for crop improvement. Nova, New York, pp. 14.
Bagyaraj DJ, Krishnaraj PU & Khanuja SPS (2000) Mineral phosphate solubilization: agronomic implication,
mechanism and molecular genetics. Proceedings of the Indian National Science Academy 66: 6982.
Chen CR, Condron LM, Davis MR & Sherlock RR (2004) Effects of plant species on microbial biomass
phosphorus phosphatase activity in a range of grassland soils. Biology and Fertility of Soils 40: 313322.
Dave A & Patel HH (2003) Impact of different carbon and nitrogen sources on phosphate solubilization by
Pseudomonas fluorescens. Indian Journal of Microbiology 43(1): 3336.
El-Komy MAH (2005) Coimmobilization of Azospirillum lipoferum, Bacillus megaterium for Successful
Phosphorus, Nitrogen Nutrition of Wheat Plants. Food Technology and Biotechnology 43 (1): 1927.
Gaur A., Rana J, Jalali B & Chand H (1990) Role of VA mycorrhizae, phosphate solubilizing bacteria and their
interactions on growth and up-take of nutrients by wheat crops. In: The National Conference on Mycorrhizae
(1990: Hisar, India). Proceeding. Hisar, India. Trends in Mycorrhizal Research 105106.
247
Sahoo & Gupta (2016) 3(1): 243248

.
Habte M & Osorio NW (2012) Effect of nitrogen form on the effectiveness of a P-solubilizing fungus to
dissolve rock phosphate. Journal of Biofertiliser and Biopesticides 3: 14.
Jackson ML (1958) Soil Chemical Analysis. Prentice Hall Inc. Englowoodcliff, New Jersey, USA.
Khan MS, Ahemad E, Zaidi A & Oves M (2013) Functional Aspect of phosphate solubilizing bacteria:
Importance in crop production, In: Maheswari DK (ed) Bacteria in Agrobiology: crop productivity.
Springer, Berlin. pp. 237265.
Kundu BSR., Gera N, Sharma A & Bhatia R (2002) Host specificity of phosphate solubilizing bacteria. Indian
Journal of Microbiology 42: 1921.
Nahas E (1996) Factors determining rock phosphate solubilization by microorganisms isolated from soil. World
Journal of Microbiology and Biotechnology 12: 567572.
Premono EM, Moawad MA & Vleck PLG (1996) Effect of phosphate solubilizing Pseudmonas putida on the
growth of maize and its survival in the rhizosphere, Indonasian Journal of Crop Science 11: 1323.
Scervino JM, Papinutti VL, Godoy MS, Rodriguez MA, Monica ID, Recchi M, Pettinari MJ & Godeas AM
(2011) Medium pH, carbon and nitrogen concentrations modulate the phosphate solubilization efficiency of
Penicillium purpurogenum through organic acid production. Journal of Applied Microbiology 110(5): 1215
1223.
Selvi KB & Ravindran AD (2012) Influence of different carbon and nitrogen sources on insoluble inorganic
phosphate solubilization by Bacillus subtilis. International Journal of Advanced Biological Research 2(3):
441445.
Seshadri S, Ignacimuthu S & Lakshminarasimhan C (2004) Effect of nitrogen and carbon sources on the
inorganic phosphate solubilization by different Aspergillus niger strains. Chemical Engineering
Communications 191: 10431052.
Wakelin SA, Warren RA, Harvey PR & Ryder MH (2004) Phosphate solubilization by Penicillium spp. closely
associated with wheat roots. Biology and Fertility of Soils 40: 3643.
Wani PV, More BB & Patil PL (1979) Physiological studies on the activity of phosphorus solubilizing
microorganisms. Indian Journal of Microbiology 19(1): 2325.
Whitelaw MA (2000) Growth promotion of plants inoculated with phosphate solubilizing fungi. Advances in
Agronomy 69: 99151.
Yadav J, Verma JP, Yadav SK & Tiwari KN (2010) Effect of salt concentration and pH on soil inhabiting
fungus Penicillium citrinum Thom. For solubilisation of tricalcium phosphate. Microbiology Journal 72:
625630.
Yadav K & Singh T (1991) Phosphorus solubilization by microbial isolate from a calcifluvent. Journal of
Indian Society of Soil Science 39(7): 8993.
248

Volume 3, Issue 1 of Tropical Plant Research

Hochgeladen von

Dokumentinformationen

Copyright

Verfügbare Formate

Dieses Dokument teilen

Dokument teilen oder einbetten

Freigabeoptionen

Stufen Sie dieses Dokument als nützlich ein?

Sind diese Inhalte unangemessen?

Copyright:

Verfügbare Formate

Volume 3, Issue 1 of Tropical Plant Research

Hochgeladen von

Copyright:

Verfügbare Formate

ISSN (E): 2349 1183

ISSN (P): 2349 9265

Tree species diversity in tropical forests of Barak valley

School of Environmental Sciences, Jawaharlal Nehru University, New Delhi, India

Borah et al. (2016) 3(1): 0109

Figure 1. Map of Barak valley, Assam, India.

Borah et al. (2016) 3(1): 0109

RESULTS AND DISCUSSION

Borah et al. (2016) 3(1): 0109

Bridelia Montana (Roxb.) Willd.

Borah et al. (2016) 3(1): 0109

Garciniacowa Roxb. ex DC.

Borah et al. (2016) 3(1): 0109

Pajanelia longifolia (Willd.) Schum.

Borah et al. (2016) 3(1): 0109

Vatica lancaefolia (Roxb.) Bl.

Borah et al. (2016) 3(1): 0109

Borah et al. (2016) 3(1): 0109

ISSN (E): 2349 1183

Soil organic carbon stocks in different land uses in Pondicherry

Sundarapandian et al. (2016) 3(1): 1017

Figure 1. Aerial view of Pondicherry University Campus. (Source: Google Earth)

Sundarapandian et al. (2016) 3(1): 1017

Soil moisture (%)

Sundarapandian et al. (2016) 3(1): 1017

Soil depth (cm)

Figure 3. pH in different land uses in Pondicherry University campus, Puducherry, India.

Bulk density (g/m2)

Soil depth (cm)

Soil depth (cm)

Sundarapandian et al. (2016) 3(1): 1017

Soil depth (cm)

Total Organic Carbon (t/ha)

Sundarapandian et al. (2016) 3(1): 1017

Logarithm of SOM (%)

Logarithm of SOM (%)

Logarithm of SOC (%)

Sundarapandian et al. (2016) 3(1): 1017

Sundarapandian et al. (2016) 3(1): 1017

ISSN (E): 2349 1183

Predicting suitability of tree species in various climatic conditions

Institute of Forest Productivity, Lalgutwa, Ranchi, Jharkhand

Tiwari & Mishra (2016) 3(1): 1832

Table 2. Location details and climatic conditions.

Tiwari & Mishra (2016) 3(1): 1832

Data type & Width

Table 4. User master.

Data type & Width

Table 5. Location master.

Data type & Width

Tiwari & Mishra (2016) 3(1): 1832

Data type & Width

Tiwari & Mishra (2016) 3(1): 1832

FORM LEVEL VALIDATION :

Tiwari & Mishra (2016) 3(1): 1832

DATABASE LEVEL VALIDATION:

MESSAGE: RECORD ADDED.

Tiwari & Mishra (2016) 3(1): 1832

Edit User Privileges

Tiwari & Mishra (2016) 3(1): 1832

Search in Database table

Data not found

Figure 2. Flow chart for advance search.

Tiwari & Mishra (2016) 3(1): 1832