Abstract
Next-generation Roche 454 pyrosequencing was used to rapidly identify polymorphic microsatellites from enriched DNA libraries for the pink stem borer, Sesamia inferens (Walker). A total of 1,459 simple sequence repeats (SSRs) were isolated from the microsatellite-enriched library using 454 sequencing. Thirty-nine microsatellite markers were selected to synthesize for further optimization, and 12 loci exhibited reliable amplification of a single product of expected size. The forward primer of 12 primer pairs was end labeled with a fluorescent dye. All of the 12 microsatellite loci were polymorphic, with 5–13 alleles per locus and observed heterozygosities ranging from 0.097 to 0.957. Here, we also tested these 12 SSRs for cross-species amplification in Chilo suppressalis (Walker), Tryporyza incertulas (Walker) and Cnaphalocrocis medinalis (Guenee). These polymorphic markers will be a valuable tool for analyses of population connectivity and genetic structure in this rice pest.
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