%0 Journal Article
%T Development of genic-SSR markers by deep transcriptome sequencing in pigeonpea [Cajanus cajan (L.) Millspaugh]
%A Sutapa Dutta
%A Giriraj Kumawat
%A Bikram P Singh
%A Deepak K Gupta
%A Sangeeta Singh
%A Vivek Dogra
%A Kishor Gaikwad
%A Tilak R Sharma
%A Ranjeet S Raje
%A Tapas K Bandhopadhya
%A Subhojit Datta
%A Mahendra N Singh
%A Fakrudin Bashasab
%A Pawan Kulwal
%A KB Wanjari
%A Rajeev K Varshney
%A Douglas R Cook
%A Nagendra K Singh
%J BMC Plant Biology
%D 2011
%I BioMed Central
%R 10.1186/1471-2229-11-17
%X In this study, 43,324 transcriptome shotgun assembly unigene contigs were assembled from 1.696 million 454 GS-FLX sequence reads of separate pooled cDNA libraries prepared from leaf, root, stem and immature seed of two pigeonpea varieties, Asha and UPAS 120. A total of 3,771 genic-SSR loci, excluding homopolymeric and compound repeats, were identified; of which 2,877 PCR primer pairs were designed for marker development. Dinucleotide was the most common repeat motif with a frequency of 60.41%, followed by tri- (34.52%), hexa- (2.62%), tetra- (1.67%) and pentanucleotide (0.76%) repeat motifs. Primers were synthesized and tested for 772 of these loci with repeat lengths of ¡Ý18 bp. Of these, 550 markers were validated for consistent amplification in eight diverse pigeonpea varieties; 71 were found to be polymorphic on agarose gel electrophoresis. Genetic diversity analysis was done on 22 pigeonpea varieties and eight wild species using 20 highly polymorphic genic-SSR markers. The number of alleles at these loci ranged from 4-10 and the polymorphism information content values ranged from 0.46 to 0.72. Neighbor-joining dendrogram showed distinct separation of the different groups of pigeonpea cultivars and wild species. Deep transcriptome sequencing of the two parental lines helped in silico identification of polymorphic genic-SSR loci to facilitate the rapid development of an intra-species reference genetic map, a subset of which was validated for expected allelic segregation in the reference mapping population.We developed 550 validated genic-SSR markers in pigeonpea using deep transcriptome sequencing. From these, 20 highly polymorphic markers were used to evaluate the genetic relationship among species of the genus Cajanus. A comprehensive set of genic-SSR markers was developed as an important genomic resource for diversity analysis and genetic mapping in pigeonpea.Pigeonpea [Cajanus cajan (L.) Millspaugh] is an important food legume predominantly cultivated in the t
%U http://www.biomedcentral.com/1471-2229/11/17