Microsatellite markers derived from Calophyllum inophyllum (Clusiaceae) expressed sequence tags

 Premise of the study: Robust markers are required (inter alia) for assessing origins of Calophyllum inophyllum populations on the Bonin Islands, Japan. Therefore, informative expressed sequence tag (EST) – based microsatellite or simple sequence repeat (SSRs) markers in the species were sought.  Methods and Results: Using 135 378 ESTs derived from de novo pyrosequencing, primers for 475 EST-SSRs were developed, 48 of which were tested for PCR amplifi cation. Thirty-six of the 48 primers showed clear amplifi cation, with 23 displaying polymorphism in sampled populations. Expected heterozygosity in the samples from the Bonin Islands and Ryukyu Islands populations ranged from 0.041 to 0.697 and from 0.041 to 0.773, respectively.  Conclusions: As EST-SSRs are potentially tightly linked with functional genes, and reportedly more transferable to related species than anonymous genomic SSRs, the developed primers have utility for future studies of the origins, genetic structure, and conservation of C. inophyllum and related species.