Molecular characterization and identification of markers for toxic and non-toxic varieties of Jatropha curcas L. using RAPD, AFLP and SSR markers

Abstract

Jatropha curcas L., a multipurpose shrub has acquired significant economic importance for its seed oil which can be converted to biodiesel, is emerging as an alternative to petro-diesel. The deoiled seed cake remains after oil extraction is toxic and cannot be used as a feed despite having best nutritional contents. No quantitative and qualitative differences were observed between toxic and non-toxic varieties of J. curcas except for phorbol esters content. Development of molecular marker will enable to differentiate non-toxic from toxic variety in a mixed population and also help in improvement of the species through marker assisted breeding programs. The present investigation was undertaken to characterize the toxic and non-toxic varieties at molecular level and to develop PCR based molecular markers for distinguishing non-toxic from toxic or vice versa. The polymorphic markers were successfully identified specific to non-toxic and toxic variety using RAPD and AFLP techniques. Totally 371 RAPD, 1,442 AFLP markers were analyzed and 56 (15.09%) RAPD, 238 (16.49%) AFLP markers were found specific to either of the varieties. Genetic similarity between non-toxic and toxic verity was found to be 0.92 by RAPD and 0.90 by AFLP fingerprinting. In the present study out of 12 microsatellite markers analyzed, seven markers were found polymorphic. Among these seven, jcms21 showed homozygous allele in the toxic variety. The study demonstrated that both RAPD and AFLP techniques were equally competitive in identifying polymorphic markers and differentiating both the varieties of J. curcas. Polymorphism of SSR markers prevailed between the varieties of J. curcas. These RAPD and AFLP identified markers will help in selective cultivation of specific variety and along with SSRs these markers can be exploited for further improvement of the species through breeding and Marker Assisted Selection (MAS).

Physical Mapping of Ribosomal DNA Genes on <i>Jatropha curcas</i> Chromosomes by Multicolor FISH

“…So far, several accessions of the plant are available, but still little data concerning genetic variability of Jatropha has been reported (Martínez-Herrera et al 2006, Pamidimarri et al 2008a. rDNAs of plants are genes coding for various ribosomal RNAs, including 5S and 45S rRNAs.…”

“…So far a few molecular characterizations of J. curcas, using RAPD, AFLP, SSR markers and phylogenetic analysis based on ITS sequence of rDNA, have been reported (Pamidimarri et al 2008a, b). However the genomic distribution of rDNA loci of Jatropha species has not been studied.…”

Molecular Characterization of Jatropha curcas Resources and Identification of Population-Specific Markers

“…Several studies on genetic variability in Jatropha populations were reported using different molecular markers, e.g., amplified fragment length polymorphism, randomly amplified polymorphic DNA (RAPD), single primer amplification reaction, inter-simple sequence repeat (ISSR), and simple sequence repeat (SSR) [1,4,5,15,23,27,37]. DNA marker (RAPD, ISSR, and SSR) analyses revealed that the Jatropha seeds commercialized worldwide consisted of few genetically identical or closely related clones and showed very low genetic variation either within or between varieties [1].…”

“…Narrow genetic bases were reported among germplasms collected in India [4] and in China [37]. Wider variability was present among genotypes collected from different geographic areas [5] and between toxic and non-toxic genotypes collected in Mexico [23]. Recently, Ovando-Medina et al [22] summarized the genetic diversity research on Jatropha as published in indexed journals.…”

Genetic similarity of Jatropha curcas accessions based on AFLP markers

“…Molecular markers were used in some studies with J. curcas, such as Random Amplification of Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR), to analyze 42 Jatropha accessions from different regions of India (Basha and Sujatha 2007) and more recently, Amplified Fragment Length Polymorphism (AFLP) and Simple Sequence Repeats (SSR), to differentiate toxic from non-toxic varieties (Pamidimarri et al 2008a). Phylogenetic studies were also performed by RAPD and AFLP for seven species of the genus (Pamidimarri et al 2008b).…”