Identification and Characterization of EST‐SSR Markers in Cluster Bean (Cyamopsis spp.)
2014
Kuravadi, Nagesh Aswathnarayana | Tiwari, Pranita Bhatele | Tanwar, Umesh Kumar | Tripathi, Sangharsh Kumar | Dhugga, Kanwarpal Singh | Gill, Kulvinder S. | Randhawa, Gursharn Singh
Cluster bean, or guar, [Cyamopsis tetragonoloba (L.) Taub] is an important industrial crop. At present there are no marker resources or linkage maps available for this crop. Development of molecular markers is essential for accelerating genetic studies and breeding in guar. In the present study, mining of simple sequence repeat (SSR) sequences from 16,476 expressed sequence tags (ESTs) of guar was done using the microsatellite (MISA) identification tool. A total of 907 SSR‐containing sequences were identified. Flanking primers were designed for SSR‐containing sequences. Primers were selected on the basis of Tm, guanine‐cytosine (GC) content, and product length. A total of 362 pairs of primers were designed, out of which 224 were synthesized and used for assessment of amplification and polymorphism in five accessions of three Cyamopsis species. Out of 224 primer pairs, 187, 184, and 121 resulted in the amplification of SSR‐containing sequences producing reliable and reproducible DNA bands in C. tetragonoloba, C. serrata, and C. senegalensis, respectively. The analysis of the DNA bands revealed that 44 SSR markers were monomorphic in all the five accessions. Four SSR markers showed polymorphism in all three tested varieties of C. tetragonoloba. Our study gives the preliminary information on types, distribution, and frequencies of various SSRs present in the EST database of guar. The results provide the first set of 187 SSR markers for the guar research community.
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