Mol Biol Rep (2012) 39:43834390

DOI 10.1007/s11033-011-1226-z

Molecular characterization of intra-population variability

of Jatropha curcas L. using DNA based molecular markers
Shaik G. Mastan Pamidimarri D. V. N. Sudheer
H. Rahman A. Ghosh Mangal S. Rathore
Ch. Ravi Prakash J. Chikara

Received: 21 January 2011 / Accepted: 3 May 2011 / Published online: 14 September 2011
Springer Science+Business Media B.V. 2011

Abstract Jatropha curcas L. (Euphorbiaceae) has

acquired a great importance as a renewable source of
energy with a number of environmental benefits. Very few
attempts were made to understand the extent of genetic
diversity of J. curcas germplasm. In the present study,
efforts were made to analyze the genetic diversity among
the elite germplasms of J. curcas, selected on the basis of
their performance in field using random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP) and simple sequence repeats (SSR). The
plants were selected on the basis of height, canopy circumference, number of seeds per fruit, weight of 100 seeds,
seed yield in grams per plant and oil content. Out of 250
RAPD (with 26 primers), 822 AFLP (with 17 primers) and
19 SSR band classes, 141, 346 and 7 were found to be
polymorphic, respectively. The percentage polymorphism
among the selected germplasms using RAPD, AFLP and
SSR was found to be 56.43, 57.9, and 36.84, respectively.
The Jaccards similarity coefficient was found 0.91, 0.90
and 0.91 through RAPD, AFLP and SSR marker systems,
respectively. Principle component analysis (PCA) and
dendrogarm analysis of genetic relationship among the
germplasm using RAPD, AFLP and SSR data showed a
good correlation for individual markers. The germplasm

Electronic supplementary material The online version of this

article (doi:10.1007/s11033-011-1226-z) contains supplementary
material, which is available to authorized users.
S. G. Mastan  P. D. V. N. Sudheer  H. Rahman  A. Ghosh 
M. S. Rathore  Ch. Ravi Prakash  J. Chikara (&)
Discipline of Wasteland Research, Central Salt and Marine
Chemicals Research Institute, Council of Scientific and
Industrial Research, G.B. Badheka Marg, Bhavnagar,
Gujarat 364002, India
e-mail: jchikara@csmcri.org

JCC-11, 12, 13, 14 and 15 whose yield found to be high

were clustered together in dendrogram and PCA analysis
though JCC11 is geographically distinct from others. In
overall analysis JCC6 (in RAPD), JCC8 (in AFLP) and
JCC 6 and JCC10 (in SSR) were found genetically diverse.
Characterization of geographically distinct and genetically
diverse germplasms with varied yield characters is an
important step in marker assisted selection (MAS) and it
can be useful for breeding programs and QTL mapping.
Keywords Jatropha curcas  Diversity analysis 
Bio-diesel  Marker assisted selection

Introduction
The current era of energy crisis, due to continuous depletion of conventional sources of fuel and global warming,
rekindled the interest in promotion of non-conventional
sources of energy as an alternative. Plant borne oil is being
gaining importance as a viable option for the conventional
petro-diesel. A number of plant species have been
suggested as potential source and Jatropha curcas L.
(a member of Euphorbiaceae) is one among them. It is
native to South America and widely distributed in South
and Central America, Africa and Asia [1]. It is emerging as
an important source of bio-fuel because of its seed oil
which can be converted to biodiesel whose performance
demonstrated to be superior to petro-diesel. The short
gestation period, easy adaptation to different kinds of
marginal lands, drought endurance and avoidance by animals make the plant species more attractive. However, the
crop is characterized by variable and unpredictable yield
for reasons that have not been identified [2] which limit the
large scale cultivation and warrants need for genetic

123

4384

improvement of the species. For the genetic improvement

of any species preliminary information about its genetic
back ground and characterized germplasm is very essential.
Molecular diversity analysis, germplasm characterization
through DNA fingerprinting techniques like random
amplified polymorphic DNA (RAPD), amplified fragment
length polymorphism (AFLP) and simple sequence repeats
(SSRs) have been well established and studied to some
extent in Jatropha to understand the extent of diversity that
exist; however, studies were limited only to natural population and/or germplasms of narrow geographical area [3, 4].
Assessment of genetic diversity among elite germplasms
has important implications for breeding programs and
conservation of plant genetic resources. The characterized
germplasms and identified polymorphic markers are good
source of plant genetic resources and can be further
exploited for genetic improvement of the species through
marker assisted breeding and QTL analysis. Till date there
is very limited information about molecular characterization of J. curcas germplasms, selected on the basis of
performance in field which is an important parameter for
genetic improvement of the plant species for yield attributing characters through marker assisted breeding. Therefore, in present study attempts were made to assess genetic
diversity using molecular markers viz. RAPD, AFLP and
SSRs among different selected germplasms collected from
different geographical areas and whose performance is
evaluated in the field experiments.

Mol Biol Rep (2012) 39:43834390

phenol was taken and extracted further with chloroform:isoamyl alcohol (24:1) twice, and precipitated with
80% of ethanol. The genomic DNA was air dried and
dissolved in 100 ll of Milli Q water. The genomic DNA
was quantified spectrophotometrically (Analytical spectrophotometer, U.K.) and diluted to the final concentration
of 1015 ng/ll.
RAPD analysis
Amplification of RAPD fragments was performed according to Williams et al. [6], using decamer arbitrary primers
(Table 2 supplementary) (Operon technologies Inc, USA;
IDT, USA). The reaction was carried out in 25 ll volume of
reaction mixture containing final concentration of 10 mM
TrisHCl (pH 9.0), 50 mM KCl, 0.1 Triton X-100, 0.2 mM
each dNTPs, 3.0 mM MgCl2, 0.4 lM primer, 25 ng template, 1 U Taq DNA polymerase (Biogene, USA). Amplification was performed in programmed thermal cycler
(Master cycle epgradient S, eppendorf, Germany) with
program of initial denaturation at 94C for 3 min, 42 cycles
of denaturation at 94C for 30 s, primer annealing at 32C
for 1 min, extension at 72C for 2.5 min and final extension
at 72C for 4 min. Amplification products were electrophoresed in 1.5% agarose in 19 TBE buffer. The gels were
stained with ethidium bromide and documented using gel
documentation system (Syngene, UK).
AFLP analysis

Materials and methods

Plant materials and genomic DNA extraction
The plant material for present study was collected from 15
selected germplams of J. curcas growing in CSMCRI
experimental field station (Chorvadla, Gujarat at an altitude
21400 N, 071470 E). The plants were selected on the basis
of their performance in the field trials (Table 1 supplementary) viz. plant height, canopy circumference, seeds per
fruit, 100 seed weight, seed yield in grams per plant and oil
yield in percentage of the year 20082009. Genomic DNA
was extracted using CTAB protocol with slight modification [5]. 0.1 g of leaf tissue was grinded in liquid nitrogen
and taken into a 2.0 mL microcentrifuge tube. To the
grinded tissues, 0.5 mL of extraction buffer (2% CTAB,
100 mM TrisHCl, 3.5 M NaCl, 20 mM EDTA, 0.2 M bMercaptoethanol, 2% PVP, pH 8.0.) was added and incubated at 65C for 90 min. The samples were extracted with
equal volume of chloroform:isoamyl alcohol (24:1) and
supernatant was transferred into a new tube. These samples
were treated with RNase and extracted with Tris saturated
phenol. The supernatant after extraction with Tris saturated

123

AFLP fingerprinting was performed using AFLP analysis

system-II kit (Invitrogen Life Science Ltd., USA) according to Vos et al. [7]. The genomic DNA (300 ng) was
digested with EcoRI and MseI at 37C for 2 h and digested
aliquot was ligated to EcoRI and MseI specific adopters at
20C for 90 min. The ligated DNA was diluted for 1:10
and preamplified using EcoRI and MseI with one selective
nucleotide at the 30 end primer each. The preamplified
product was diluted 1:10 with sterile TrisEDTA (TE)
buffer. The diluted products were amplified using different
combinations of EcoRI and MseI primer each with three
selective nucleotides at the 50 and 30 , respectively (Table 3
supplementary). Selective amplifications were performed
using 65C as the initial annealing temperature for the first
cycle and for subsequent 11 cycles the annealing temperature was successively reduced by 0.7C. Twenty-three
cycles were run at 56C annealing temperature. To the
PCR product equal amount of formamide dye was added
and subjected to electrophoretic separation on 6% denaturing polyacrylamide gel in 19 TBE buffer in a
sequencing gel system (LKB, Sweden). The Gels were
stained with silver nitrate using silver staining kit (Sigma,
USA) and photographed for further recording.

Mol Biol Rep (2012) 39:43834390

SSR analysis
Microsatellite amplifications were carried out in a volume
of 20 ll containing 0.25 U Taq DNA polymerase (Sigma,
USA), 19 PCR buffer (10 mM TrisHCl, 50 mM KCl, 0.1
Triton X-100, pH 9.0), 0.2 mM dNTPs, 2 lM of each
primer set, 2.8 mM MgCl2 and 50 ng template DNA.
Amplification cycle is consisted of an initial denaturation at
94C for 3 min, 35 cycles of denaturation at 94C for 15 s,
specific annealing temperature for individual primer
(Table 4 supplementary) for 20 s, extension at 72C for
30 s and final extension at 72C for 4 min. Amplification
products were electrophoresed in 8% polyacrylamide gel.
The gels were stained with ethidium bromide and documented using gel documentation system (Syngene, UK).
Data analysis
Experiment was repeated at least three times with each
primer and those primers gave reproducible fingerprints
were considered for data analysis. Acquired RAPD and
AFLP fingerprints were individually scored and statistically
analyzed by assuming the fragment size as biallelic (present = 1, absent = 0) locus. A binary matrix was created.
Only those loci amplified strongly in each instance with
reproducibility were scored and included in the analyses.
Genetic similarity (GS) was calculated using Jaccards
coefficient of similarity [8] with the help of NTSYS-pc
package (version 2.2) [9]. In case of acquired SSR amplification, fingerprints were individually scored and statistically analyzed and obtained the genetic similarity based on
Nei and Li [10] definition as follows Sij = 2a/
(2a ? b ? c), where Sij is the similarity between two
individuals, i and j; a is number of bands present both in
i and j; b is number of bands present in i and absent in j; and
c is the number of bands absent in i and present in j. In all
cases (RAPD, AFLP and SSR analysis), the percentage of
polymorphism (PP) was calculated by using formula
PP = total number of polymorphic bands/total number of
bands multiplied with 100. Dendrograms were constructed
according to UPGMA (unweighted pair-group method with
arithmetic mean) method using binary data generated by
RAPD and AFLP followed by bootstrapping analysis across
the loci [11] with the help of NTSYS-pc software package.

4385

of these 42 primers, 26 primers which produced clear bands

and reproducible fingerprints at each instance of repetition
were taken for the analysis (Fig. 1, Table 2 supplementary). With 26 primers, a total of 250 markers with an
average of 9.6 markers per primer were generated, out of
which 141 were polymorphic and remaining was monomorphic. The overall percentage of polymorphism among
germplasm was found 56.4. OPL7 produced the highest
number of markers [17] with 12 polymorphic loci with
70.59% of polymorphism. OPQ11 and opQ 20 generated
least number of markers [4], however opQ11 was found to
be 100% polymorphic. The number of polymorphic
markers varied 1 (opO2) to 12 (opL7) while percentage
polymorphism varied from 20% (opO2) to 100% (opQ11)
in the germplasm. Out of 26 primers used in present study,
15 produced more than 50% polymorphism in the
germplasm.
The average pair wise percentage polymorphism was
found 51.32%, ranging from 6.05 (JCC13 and JCC14) to
31.53 (JCC6 and JCC10) (Table 5 supplementary data). In
present study JCC6 was found to be the most diverged
genotype among the studied germplasm. In pair wise
comparison, average Jaccard coefficient of genetic similarity was found 0.90, ranging from 0.81 (JCC6 and
JCC10) to 0.97 (JCC13 and JCC14) (Table 6 supplementary data). Three major clusters were obtained in RAPD
dendrogram (Fig. 2). In RAPD dendrogram, least genetic
distance was observed between JCC13 and JCC14; and it
was followed by JCC1 and JCC2. JCC6 showed highest
genetic divergence from rest of the genotypes and it was
followed by JCC10. Both of them stand separate individually as well as from rest of the population in RAPD
dendrogram. PCA analysis of RAPD data resulted in
identification of four distinct groups, consisting of 1, 3, 3
and 8 genotypes. JCC6 stand separate from all the genotypes, which is in accordance to RAPD dendrogram, while
JCC10 clustered with largest group in RAPD-PCA analysis
(Fig. 3).

Results
RAPD analysis
Out of 180 RAPD primers screened initially, 42 primers
produced amplification with more than 4 markers and were
included in present study for screening of germplasm. Out

Fig. 1 RAPD finger printing of selected J. curcas germplasm using

primer opQ9; 115 JCC1JCC15 and M 1 kb marker

123

4386

Mol Biol Rep (2012) 39:43834390

Fig. 2 Dendrogram showing

genetic relationship with
bootstrapping values among the
selected germplasm of J. curcas
through RAPD

Fig. 3 PCA (principle

component analysis) showing
the genetic relationship among
the selected germplasm of
J. curcas through RAPD

AFLP analysis
Out of 21 selective primer combinations, 17 primers
producing reproducible fingerprint (Table 3 supplementary) with more than 30 markers in each instance of
repetition were used for data analysis (Fig. 4). A total of
822 AFLP markers with an average of 48.35 per primer
combination were obtained, out of these 346 were polymorphic. Numbers of amplicon ranged from 33 (P64) to
68 (P12). The overall polymorphism among germplasm
was found to be 57.9%. Highest (69.57%) polymorphism
was recorded with P11 primer combination with highest
(32) polymorphic markers while P35 reported least
(5.26%) polymorphism with least [2] number of polymorphic markers.

123

In pair wise comparison among germplasm, average

polymorphism was found to be 15.32% ranging from 3.05%
(JCC12 and JCC13) to 37.60% (JCC1 and JCC8) (Table 7
supplementary data). JCC8 was found to be the most
diverged genotype while JCC14 was least diverged according to pair wise comparisons of AFLP data. The Jaccard
coefficient of genetic similarity ranged from 0.77 (between
JCC8 and JCC1) to 0.98 (between JCC13 and JCC12) with
an average of 0.91(Table 8 supplementary data). In dendrogram constructed on the basis of AFLP data, a single
major cluster was obtained, which included all the genotypes
except JCC1, JCC6 and JCC8. JCC8 showed highest genetic
divergence in AFLP analysis and it was followed by JCC6
and JCC1, respectively (Fig. 5). Least genetic divergence
was found between JCC12 and JCC13; and it was followed

Mol Biol Rep (2012) 39:43834390

4387

[12] and five from Sun et al. [13]. Out of these, 19 primers
with reproducible results at every instance of repetition and
used for data analysis (Fig. 7). The size of loci with these
primers ranged from 92 to 459 bp (Table 4 supplementary). Seven loci out of 19 were found polymorphic. The
number of alleles at these polymorphic loci ranged from 2
(JCMNS 183 and JCDS 24 primers) to 5 (JCPS 7 and
JCMNS 292 primer). Overall 36.8% loci were found
polymorphic among the studied germplam. Average Jaccard coefficient was found 0.91, ranging from 0.78
(between JCC8 and JCC10) to 1.00 (between JCC1 and
JCC11; JCC12 and JCC14) (Table 9 supplementary data).
Three major clusters were obtained in SSR dendrogram
(Fig. 8). JCC1, 11, 5, 12, 14, 15, 13 and 7 clustered together however belong to different geographical area. JCC2
and JCC3 found to be diverse however they belong to the
same geographical area. JCC1 and JCC11; JCC12 and
JCC14 showed least genetic distance while JCC6 and
JCC10 were found highly diverse from rest of the population; however they formed same cluster. SSR PCA
analysis resulted in formation of two major groups, while
JCC2, 6, 3, 8 and 10 remained individually (Fig. 9).
Fig. 4 AFLP finger printing profle of Elite J. curcas germplasm
using primer E-ACT/M-CAG; 115 JCC1JCC15 and M 1 kb marker

by JCC2 and JCC3; JCC12/13 and JCC14. In AFLP-PCA

analysis, one major group was obtained consisting of most of
the genotypes except JCC6 and JCC8, standing individually
as accordance to the AFLP dendrogram (Fig. 6).
SSR analysis
SSR amplification was performed with primers designed
for twenty-five markers (twenty SSR from Sudheer et al.

Discussion
Molecular characterization of cultivars for the investigation
of genetic diversity and to confirm the uniformity, stability
and distinctness of different cultivars accelerated their
application in molecular breeding for the improvement of
the species [14]. Unlike the morphological and enzymatic
markers whose variations can occur due to the environmental fluctuations, the molecular marker will be stable
and reproducible [15, 16]. Thus the characterized

Fig. 5 Dendrogram showing

genetic relationship with
bootstrapping values among the
selected germplasm of J. curcas
through AFLP

123

4388

Mol Biol Rep (2012) 39:43834390

Fig. 6 PCA (principle

component analysis) showing
the genetic relationship among
the selected germplasm of
J. curcas through AFLP

germplasm and the identified markers can be a good source

of plant genetic resources and can further be exploited for
genetic improvement of the species through marker assisted breeding to obtain the better variety with high yielding
and better performing attributes. In present study all the
marker systems being employed to analyze the intra-population diversity of J. curcas germplasm were quite
informative and were able to generate unique DNA fingerprints and adequate polymorphism among the germplasm under investigation.
Beside tremendous economic benefits, there were very
few studies carried to understand the genetic diversity
using various marker systems in J. curcas. Basha and Sujatha 2007 [3] studied the extent of genetic diversity among
toxic and non-toxic varieties using RAPD and the percentage of GS is found to be 96.3. In another study Sudheer
et al. 2008 [17] reported 84.91 and 83.59% (GS) among
toxic and non-toxic J. curcas by RAPD and AFLP,
respectively and identified the specific markers of RAPD
and AFLP for both the varieties. Inter and intra-population
studies using RAPD and ISSR in 42 germplasm of
J. curcas collected from different regions in India along
with a non-toxic genotype from Mexico showed 42.00 and
37.40 PP by RAPD and ISSR, respectively [3]. Sudheer
et al. [17] studied 9.72 and 20.57 percent polymorphism in
natural germplasm of J. curcas using RAPD and AFLP,
respectively. Though there are a number of studies on
diversity analysis in J. curcas, however till date no systematic studies were made on the analysis of genetic
diversity among the selected germplasm; whose performance was evaluated in the field. Therefore, the present
study was conducted to evaluate the genetic diversity

123

Fig. 7 SSR profle of Elite J. curcas germplasm using primer

JCMNS-292; 115 JCC1JCC15 and M 1 kb marker

among the selected germplasm whose yield attributing

characters were evaluated.
In India extensive experimental field trials are being
conducted at CSMCRI (Central Salt and Marine Chemical
Research Institute) to assess the various factors like variability in morphology, seed yield, and oil content, tolerance
to biotic and abiotic stress in natural population. In the
present study taking the consideration of the yield

Mol Biol Rep (2012) 39:43834390

4389

Fig. 8 Dendrogram showing

genetic relationship with
bootstrapping values among the
selected germplasm of J. curcas
through SSR

Fig. 9 PCA (principle

component analysis) showing
the genetic relationship among
the selected germplasm of
J. curcas through SSR

attributing characters selected germplasm were taken for

the assessment of the molecular diversity. The mean
genetic similarity observed was found to be 0.91, 0.90 and
0.91 by RAPD AFLP and SSR, respectively which is in
accordance with previous reports [4, 18]. The dendrogram
analyses of germplasm using the three molecular markers
have given a good correlation of genetic similarity among
them. The PCA analysis of these markers showed good
correlation with dendrogram of respective marker system.
In dendrogram and PCA analysis of RAPD, AFLP and SSR
markers, JCC6 is found to be highly diverse from rest of
population however it was found to be clustered with
JCC10 in SSR dendrogram analysis.
Though JCC6, 12, 13, 14 and 15 were collected from
same geographical locations (i.e. Gujarat) and possess quite
good yield attributing characters, these germplasm clustered
together in the dendrograms and PCA analysis of all the
three marker systems except JCC6. JCC11 collected from
Orissa having highest yield (33.24% oil content) clustered

with JCC12, 13, 14, 15 (Gujarat) in RAPD, AFLP and SSR

dendrogram and also correlated with the PCA analysis of
RAPD, AFLP and SSR irrespective of geographical area.
JCC1, 2, 3, 4, 5, 6, 10, 12, 13, 14 and 15 belong to the same
geographical area (Gujarat), however they differ in yield
attributing and molecular characters. In the dendrogram
analysis JCC1, 2 and 3 clustered in the same but JCC4 found
to be in the separate cluster showing the genetic distinctness
from JCC1, 2 and 3 irrespective of geographical area. These
differences in molecular characters irrespective of geographical areas might be due to the anthropogenic activity as
reported in the previous studies [3, 4].
In the present study though the overall genetic diversity
found to be less in comparison with previous studies,
however the mean percentage of polymorphism is found to
be in accordance with Sudheer et al. [4, 17] and Basha and
Sujatha [3]. The type of genetic polymorphism, use of
different marker systems and the number of primers used
affect the correlations among different markers [19].

123

4390

Similarly the degree of genetic polymorphism detected in

the selected germplasm may be affected due to the three
marker systems and the number of primers used for study.
Variation of diversity among the germplasm using three
markers in the present study may be due to the codominant
nature of SSRs and dominant nature of RAPD and AFLP
markers. As the AFLP gives more amplified fragments than
RAPD followed by SSRs, it shows highest polymorphism
when compared with others. The finding of a slightly
higher resolution of genetic similarities by RAPDs and
AFLPs, compared to SSRs, may be due to the high polymorphism of SSRs which render them less suitable for
determining genetic relationships among cultivars [20].
The present study of diversity analysis revealed by
molecular markers and yield attributing characters among
selected germplasm can be useful in further breeding programs for generation of hybrids, in maintenance of selected
genetic stocks and for molecular ecological studies and
also further will pave way for the creation of mapping
population and linkage analysis, marker assisted selection
and QTL analysis for improvement of the species for its
yield attributing characters.
Acknowledgments The authors wish to thank Council for Scientific
and Industrial Research (CSIR), New Delhi, India for financial
support.

Mol Biol Rep (2012) 39:43834390

6.

7.

8.
9.
10.

11.
12.

13.

14.

15.

16.

References
1. 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. Soc Automot Eng Int
26:356364
2. Ginwal HS, Phartyal SS, Rawat PS, Srivastava RL (2005) Seed
source variation in morphology, germination and seedling growth
of Jatropha crucas Linn. in Central India. Silvae Genet
54(2):7680
3. Basha SD, Sujatha M (2007) Inter and intra-population variability
of J. curcas (L.) characterized by RAPD and ISSR markers and
development of population-specific SCAR markers. Euphytica
56:375386
4. Sudheer DVN, Mastan SG, Rahman H, Reddy MP (2010)
Molecular characterization and genetic diversity analysis of
Jatropha curcas L. in India using RAPD and AFLP analysis. Mol
Biol Rep 37:22492257
5. Sudheer PDVN, Sarkar R, Meenakshi, Boricha G, Reddy MP
(2009) A simple protocol for isolation of high quality genomic

123

17.

18.

19.

20.

DNA from Jatropha curcas for genetic diversity and molecular

marker studies. Indian J Biotechnol 8:187192
Williams JG, Kubelik AR, Livak J, Rafalski J, Tingey SV (1990)
DNA polymorphism amplified by arbitrary primers are useful as
genetic markers. Nucleic Acid Res 18:65316535
Vos P, Hogers R, Bleeker M, Reijans M, de Lee V, Miranda T,
Hornes FA, Pot J, Peleman J, Kuiper M, Zabeau M (1995) AFLP:
a new technique for DNA fingerprinting. Nucleic Acid Res
23:44074414
Jaccard P (1908) Nouvelles recherche sur la distribution florale.
Bull Soc Vaud Sci Nat 44:223270
Rohlf FJ (2001) Numerical taxonomy and multivariate analysis
system. Applied Biostatistics, New York
Nei M, Li WH (1979) Mathematical model for studying genetic
variation in terms of restriction end nucleases. Proc Natl Acad Sci
USA 76:52695273
Felsenstein J (1985) Confidence limits on phylogenies: an
approach using the bootstrap. Evolution 39:783791
Sudheer PDVN, Rahman H, Mastan SG, Reddy MP (2010) Isolation of novel microsatellites using FIASCO by dual probe
enrichment from Jatropha curcas L. and study on genetic equilibrium and diversity of Indian population revealed by isolated
microsatellites. Mol Biol Rep 37:37853793
Sun QB, Lin-Feng L, Yong L, Guo-Jiang W, Xue-Jun G (2008)
SSR and AFLP markers reveal low genetic diversity in the biofuel plant Jatropha curcas in China. Crop Sci 48:18651871
E`esoniene L, Daubaras R, Gelvonauskis B (2005) Characterization of kolomikta kiwi (Actinidia kolomikta) genetic diversity by
RAPD fingerprinting. Biol Nr 3:15
Nuel G, Baril C, Robin S (2001) Varietal distinctness assisted by
molecular markers: a methodological approach. Acta Hortic
546:6571
Huang H, Wang S, Jiang Z, Zhang Z, Gong J (2003) Exploration
of Actinidia genetic resources and development of kiwifruit
industry in China. Acta Hortic 610:2943
Sudheer PDVN, Singh S, Mastan SG, Patel J, Reddy MP (2009)
Molecular characterization and identification of markers for toxic
and non-toxic varieties of Jatropha curcas L. using RAPD, AFLP
and SSR markers. Mol Biol Rep 36:13571364
Tatikoda L, Suhas PW, Seetha K, Naresh B, Thakur KS, David
AH, Prathibha D, Rajeev KV (2008) AFLP-based molecular
characterization of an elite germplasm collection of Jatropha
curcas L, a biofuel plant. Plant Sci 176(4):505513
Staub JE, Danin-Poleg Y, Fazio G, Horejsi T, Reis N, Katzir N
(2000) Comparative analysis of cultivated melon groups (Cucumis melo L.) using random amplified polymorphic DNA and
simple sequence repeat markers. Euphytica 115:225241
Belaj A, Satovic Z, Cipriani G, Baldoni L, Testolin R, Rallo L,
Trujillo I (2003) Comparative study of the discriminating
capacity of RAPD, AFLP and SSR markers and of their effectiveness in establishing genetic relationships in olive. Theor Appl
Genet 107:736744