tailieunhanh - báo cáo khoa học: " Development of genic-SSR markers by deep transcriptome sequencing in pigeonpea [Cajanus cajan (L.) Millspaugh]"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Development of genic-SSR markers by deep transcriptome sequencing in pigeonpea [Cajanus cajan (L.) Millspaugh] | Dutta et al. BMC Plant Biology 2011 11 17 http 1471-2229 11 17 BMC Plant Biology RESEARCH ARTICLE Open Access Development of genic-SSR markers by deep transcriptome sequencing in pigeonpea Cajanus cajan L. Millspaugh f .1 1 1 1 1 Sutapa Dutta Giriraj Kumawat Bikram P Singh Deepak K Gupta Sangeeta Singh Vivek Dogra 1 1 Z X3 2 - 4 5 Kishor Gaikwad Tilak R Sharma Ranjeet S Raje Tapas K Bandhopadhya Subhojit Datta Mahendra N Singh Fakrudin Bashasab6 Pawan Kulwal7 KB Wanjari7 Rajeev K Varshney8 Douglas R Cook9 Nagendra K Singh1 Abstract Background Pigeonpea Cajanus cajan L. Millspaugh one of the most important food legumes of semi-arid tropical and subtropical regions has limited genomic resources particularly expressed sequence based genic markers. We report a comprehensive set of validated genic simple sequence repeat SSR markers using deep transcriptome sequencing and its application in genetic diversity analysis and mapping. Results In this study 43 324 transcriptome shotgun assembly unigene contigs were assembled from 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 followed by tri- hexa- tetra- and pentanucleotide repeat motifs. Primers were synthesized and tested for 772 of these loci with repeat lengths of 18 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 .