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Development and Characterization of Microsatellite Markers (SSR) in Sesamum (Sesamum indicum L.) Species
Authors:B Spandana  V Prathap Reddy  G John Prasanna  G Anuradha  S Sivaramakrishnan
Institution:1. Institute of Biotechnology, A.N.G.R.A.U, Rajendranagar, Hyderabad, 500 030, India
2. Biotechnology Division, Directorate of Rice Research (D.R.R), Rajendranagar, Hyderabad, 500 030, India
Abstract:Microsatellites, also known as simple sequence repeats (SSRs), are the class of repetitive DNA sequences present throughout the genome of many plant and animal species. Recent advances in molecular genetics had been the introduction of microsatellite markers to investigate the genetic structuring of natural plant populations. We have employed an enrichment strategy for microsatellite isolation by using multi-enzymes digestion, microsatellite oligoprobes, and streptavidin magnetic beads in Sesamum (Sesamum indicum L.). More than 200 SSR motifs were detected (SSR motifs ??2 repeat units or 6?bp); 80?% of the clones contained SSR motifs. When regarding SSRs with four or more repeat units and a minimum length of 10?bp, 132 of them showed repeats. Eighteen SSR markers were initially characterized for optimum annealing temperature using a gradient PCR technique. Among the 18 SSR markers characterized, five were found to be polymorphic and used to analyze 60 Sesamum germplasm accessions. The maximum number of alleles detected was four with a single primer and the least number of two alleles with three primers with an average PIC value of 0.77. SSRs are a valuable tool for estimating genetic diversity and analyzing the evolutionary and historical development of cultivars at the genomic level in sesame breeding programs.
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