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Research Report
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Abstract Dendrocalamus stocksii is an economically important strong solid and thorn less bamboo species which is endemic to
Westerns Ghats of India. Increase in utilization of this species has embarked the importance of conservation, diversity study,
propagation and plantation aspects.
Out study aims to analyse the morphological and genetic diversity among the ex-situ conserved 14 Candidate Plus Clumps (CPCs)
established at Bamboo Germplasm Bank of Institute of Wood science and Technology (IWST), Bangalore. This is the first report on
diversity studies of D. stocksii. The morphological diversity data among the 14 Candidate Plus Clumps (CPCs) originated from
different regions revealed variability in terms of culm height, diameter, internodal length, no. of culms/clump and solid and
hollowness of the culms. Highest culm diameter and number of culms/clump were recorded in PS-27.
In this study, the genetic diversity existing in the fourteen CPCs was estimated using ISSRPCR. Eight ISSR primers amplified fifty
three amplicons in the size ranging from 225 to 1480. The total number of polymorphic bands varied from three to nine with 71.26 %
polymorphic banding profiles. Unweighted Pair Group Method with arithmetic mean (UPGMA) revealed two major clusters. Dice
similarity coefficient ranges from 0.48 to 1.00. Genetic diversity studies based on location divided all the 14 genotypes in to three
clusters Sirsi, Dandeli and Ponda. Existence of 60-70% genetic diversity in the species dominated with vegetative multiplication and
sporadic flowering habit is a noteworthy for a germplasm bank and its contribution for future conservation programmes.
Keywords Ex situ conservation; Candidate plus clump; Sporadic flowering
1 Introduction
International Journal of Mol. Ecol. and Conserv. 2015, Vol.5, No. 3, 1-8
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batatas, Eleusine
coracana and Curcuma
longa
(Viswanath et al., 2014). In recent times due to the
scarcity of cane/rattan this species is increasingly been
seen as a substitute in furniture industry due to its
typical anatomical characteristics like the presence of
non-predominant nodes, solid nature of culms and
good culm wall thickness (Chandramouli et al., 2014).
Because of its multifarious uses this species was
considered as one among 15 industrially important
species by National Bamboo Mission (NBM) and it is
the most preferred species by the farmers in
Peninsular India.
International Journal of Mol. Ecol. and Conserv. 2015, Vol.5, No. 3, 1-8
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Morphological variation was recorded in terms of
clump characters (clump height) and culm characters
(culm diameter at 5th internode, wall thickness, culm
wall thickness to culm diameter ratio, no. of
culms/clump and new culm growth and culm
internodal length) for all 14 CPCs.
International Journal of Mol. Ecol. and Conserv. 2015, Vol.5, No. 3, 1-8
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Table1 Details of morphological variation in the 14 CPCs at 6 years in Germplasm Bank at Gottipura, Bangalore
Sl.
No.
CPCs
No.
Source
(loca.)
1
2
3
4
5
6
7
8
9
10
11
12
13
PS-14
PS-15
PS-17
PS-18
PS-24
PS-27
PS-28
PS-31
PS-32
PS-34
PS-50
PS-57
PS-58
14
PS-102
Sirsi
Sirsi
Sirsi
Sirsi
Sirsi
Sirsi
Dandeli
Dandeli
Dandeli
Dandeli
Sirsi
Sirsi
Dandeli
Ponda
(Goa)
Mean
SD
Coeffic.
of
variat.
Aver.
culms
height
(m)
No. of
nodes/
culm
Internode
length
(cm)
Culm wall
thickness
(mm)
Culm
diameter
(mm)
Culm wall
thick.:
culm
diameter
Total
(culms/
clump)
7.0
6.5
6.0
7.2
7.1
6.3
6.2
8.1
6.3
6.5
6.3
7.0
7.5
24
20
22
22
19
23
21
22
20
22
21
21
24
39.1
39.16
34.74
30.9
37.94
38.58
32
38.16
35
20.0
36.08
34.83
35.83
35.65 (S)
11.6
34.74 (S)
12.9
10.72
37.15 (S)
34.35 (S)
8.2
26.9
36.2 (S)
10.3
29.97
33.7
35.65
32.6
34.74
34.3
36.82
37.15
34.35
34.2
26.9
36.2
32.7
29.97
33.7
1:1
1:2.81
1:1
1:2.66
1:3.44
1:1
1:1
1:4.17
1:1
1:1
1:3.17
1:1
1:1
52
77
61
72
68
65
8
69
74
12
71
51
52
No. of
new
shoots
emerged
in 6th year
8
4
3
5
7
7
9
6
3
0
11
8.0
22
36.35
7.55
29.8
1:3.95
52
6.86
0.67
21.64
1.45
34.91
4.96
23.57
12.33
33.51
2.92
0.02
0.02
56.00
21.44
5.67
2.99
9.77
6.69
14.21
52.32
8.72
87.18
38.29
52.85
International Journal of Mol. Ecol. and Conserv. 2015, Vol.5, No. 3, 1-8
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Table 2 Details of ISSR primers used for the genetic analysis of Dendrocalamus stocksii (Munro.)
Primer
code
864
814
826
852
849
818
835
843
Sl. No.
1
2
3
4
5
6
7
8
Primer sequence
GGC GGCGGCGGCGGCGGC
CTC TCT CTC TCT CTC TA
ACA CAC ACA CAC ACA CC
TCT CTC TCT CTC TCT CRA
GTGTGTGTGTGTGTGTYA
CACACACACACACACAG
AGA GAG AGA GAG AGA GYC
CTC TCT CTC TCT CTC TRA
Total no.
of bands
8
3
9
6
7
6
7
7
No. of polymorphic
bands
8
2
7
2
4
3
6
6
Percent
polymorphism
100
66.67
77.78
83.33
57.14
50.00
85.71
85.71
Molecular
Weight (bp)
359-1305
416-1545
314-1580
506-1411
420-1307
574-1532
225-1029
266-1584
Table 3 Similarity matrix of D. stocksii generated from Dice estimate similarity based on the number of shared fragments
PS-14
PS-15
PS-17
PS-18
PS-24
PS-27
PS-28
PS-31
PS-32
PS-34
PS-50
PS-57
PS-58
PS-102
1
1.00
0.99
0.99
0.95
0.99
0.97
0.83
0.97
0.95
0.84
0.77
0.95
0.54
0.95
10
11
12
13
14
1.00
1.00
0.96
1.00
0.98
0.81
0.98
0.93
0.83
0.76
0.97
0.53
0.93
1.00
0.96
1.00
0.98
0.82
0.98
0.93
0.83
0.76
0.97
0.53
0.93
1.00
0.96
0.93
0.81
0.93
0.89
0.82
0.80
0.93
0.50
0.94
1.00
0.98
0.82
0.98
0.98
0.83
0.76
0.97
0.53
0.94
1.00
0.79
0.98
0.93
0.80
0.73
0.95
0.52
0.93
1.00
0.79
0.83
0.99
0.95
0.81
0.66
0.78
1.00
0.93
0.80
0.73
0.95
0.48
0.93
1.00
0.84
0.77
0.90
0.54
0.91
1.00
0.93
0.82
0.64
0.80
1.00
0.75
0.63
0.72
1.00
0.55
0.93
1.00
0.52
1.00
PS-58
PS-50
PS-28
PS-34
PS-32
PS-102
PS-18
PS-57
PS-14
PS-24
PS-15
PS-17
PS-27
PS-31
joining tree.
Ponda
Dandeli
Sirsi
International Journal of Mol. Ecol. and Conserv. 2015, Vol.5, No. 3, 1-8
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ISSR markers target a small segment between the two
microsatellites of the genome which possibly makes
few loci available for amplification by these primers
(Zietkiewicz et al., 1994). ISSR primers are able to
amplify highly variable but small segments (Mc
Gregor et al., 2000). Thats why few ISSR primers can
capture much more variability from genomic segment
than several RAPD primers with random coverage of
entire genome (Moreno et al., 1998).
4 Conclusions
There exists large morphological and genetic
variability in this the germplasm bank with fourteen
CPCs.
5 Recommendations
Due to poor seed setting and non-gregarious nature of
this bamboo, genetic diversity could be highly
restricted and continuous vegetative propagation from
a narrow genetic base could have serious implication
for conservation of the species. So, to increase the
diversity in future generations, planting clones of
different origin may be encouraged. More CPCs are to
be selected from natural growing areas of D. stocksii,
and widening of the existing gemplam in Germplasm
Bank is recommended.
6 Acknowledgements
The authors are grateful to the Director and the Group
Coordinator of Research of the Institute of Wood
Science and Technology for the encouragement and
for providing the facilities. Germplasm Bank,
Gottipura was established with the financial support of
International Journal of Mol. Ecol. and Conserv. 2015, Vol.5, No. 3, 1-8
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Moreno S., Martin J.P., and Ortiz J.M., 1998, Inter-simple sequence repeats
PCR for characterization of closely related grapevine germplasm,
Euphytica, 101: 117125
http://dx.doi.org/10.1023/A:1018379805873
Muyeed Ahmed S., 2012, Studies on genetic fidelity of in vitro propagated
plants of Bambusa bambos (L.) Voss. and Dendrocalamus stocksii
(Munro) M. Kumar, Remesh & Unnikrishnan using DNA markers,
PhD Thesis, Forest Research Institute University, Dehra Dun,
Uttaranchal, India
Nagaoka, and Ogihara Y., 1997, Applicability of inter simple sequence
repeat polymorphisms in wheat for use as DNA markers in comparison
to RFLP and RAPD markers, Theor. Appl. Genet., 94:597-602
NiM, and Li WH,1979, Mathematical model for studying genetic variation
in terms of restriction endonucleases, Proceedings of National
Academy of Sciences USA 76: 52695273
http://dx.doi.org/10.1073/pnas.76.10.5269
Panetsos K.P., Economou, and Scaltsoyiannes A., 1987, Propagation in vitro
of aspen hybrid Populus spartiatica x Populus tremula from mature
trees, Agricultural Research, 11: 449-459
Panse V.G., and Sukhatme P.V., 1985, Statistical Methods for Agricultural
Workers, 4th ed., ICAR, New Delhi, 347 p
Ramanayake S.M.S., Meemaduma V.N., and Weerawardene T.E., 2007,
Genetic diversity and relationships between nine species of bamboo in
Sri Lanka, using random amplified polymorphic DNA, Pl. SystEvol.,
269:5561
Rao A.N., 2013, Genetic diversity of woody bamboos-their conservation
and improvement
www.biodiversityinternational.ora/fileadmin/bioversity/publications/w
eb version/572/ch21. htm
Reddy Y., and Yekanthappa A.N., 1989, Propagation technique of
Oxytenanthera stocksii, Myforest, 25: 3032
Sanjaya., Rathore T.S., and Ravishankar Rai V., 2005, Micropropagation of
Pseudoxitenanthera stocksii, In Vitro Cellular and Developmental
Biology Plant, 41(3): 333-337
Seethalakshmi K.K., and Kumar M., 1998, Bamboo of India: A
compendium published by the KFRI, Peechi: 342p
Seethalakshmi K. K., Muktesh Kumar, M S., Sankara Pillai K. and Sarojam,
N., 1998, Bamboo of India - A compendium, Bamboo Information
Centre-India, INBAR, Technical Report, No. 17.
Singhal R.M., and Gangopadhyay P.B., 1999, Bamboos in India and data
base, Publication Division, Indian Council of Forestry Research and
Education, Dehra Dun, 147 p
Somashekar P.V., 2007, Studies on in vivo and in vitro propagation of
Pseudoxytenanthera stocksii Munro. and Guadua aungustifolia Kunth.important Bamboo species. PhD Thesis, Forest Research Institute,
Dehradun, India
Somashekar P.V., Rathore T.S and Geetha Joshi, 2007, Infrastructure
development and vegetative propagation of Dendrocalamus stocksii
Munro, (Marihal bamboo), Silvas News letter, 332(11): 1-4
Somashekar P.V., Rathore T.S., and Shashidhar K.S., 2008, Rapid and
simplified method of micropropagation of Pseudoxytenanthera stocksii.
In: S.A. Ansari, C. Narayanan and A. K. Mandal (eds.), Forest
Biotechnology in India, Satish serial publishing house, Delhi., pp
165-182
Suyama Y., Obayashi K., and Hayashi I., 2000, Clonal structure in a dwarf
bamboo (Sasa senanensis) population inferred from amplified fragment
length polymorphism (AFLP) fingerprints, Mol. Eco., 9: 901906
http://dx.doi.org/10.1046/j.1365-294x.2000.00943.x
Veena V.B., 2011, Reproductive biology and biochemical changes
associated with flowering of Dendrocalamus stocksii and Ochlandra
International Journal of Mol. Ecol. and Conserv. 2015, Vol.5, No. 3, 1-8
http://ijmec.biopublisher.ca
travancorica. Phd thesis, Kerala Forest Research Institute Peechi,
Thrissur, Kerala, India
Viswanath S., Ajay D. Rane, Chandramouli S., and Srinivasa Rao, 2014,
Quest for an appropriate bamboo species in tropical homegardens
Can Dendrocalamus stocksii (Munro.) fid the bill? Wca 2014-1134
White T.L., Adams W.T., and Neale D.B., 2007, Forest Genetics, CABI
publishers, USA
http://dx.doi.org/10.1079/9781845932855.0000
Yang H.Q., An M.Y., Gu Z.J., and Tian B., 2012, Genetic Diversity and
Differentiation of Dendrocalamus membranaceus (Poaceae:
Bambusoideae), a Declining Bamboo Species in Yunnan, China, as
Based on Inter-Simple Sequence Repeat (ISSR) Analysis, International
journal of molecular sciences, 13(4): 444657
http://dx.doi.org/10.3390/ijms13044446
Yang W., De Oliveira A.C., Godwin I., Schertz K., and Bennetzen J.L., 1996,
Comparison of DNA marker technologies in characterizing plant
genomic diversity: Variability in Chinese sorghums, Crop Science, 36:
1669-1676
http://dx.doi.org/10.2135/cropsci1996.0011183X003600060042x
Zietkiewicz E., Rafalski A., and Labuda D., 1994, Genome finger printing
by simple sequence repeat (SSR) - anchored polymerase chain reaction
amplification, Genomics, 20: 176-183
http://dx.doi.org/10.1006/geno.1994.1151
Zsuffa Sennerby-Forsse L., Weisverger H., and Hall R.B., 1993, Clonal
forestry, In: M. R. Ahuja and W. J. Libby (eds), Springer Verlag. Vol. II,
pp 91-119