Novel genomic and EST-derived SSR markers in Japanese chestnuts

A total of 366 novel simple sequence repeat (SSR) markers were developed in Japanese chestnut (Castanea crenata), comprising 220 genomic SSRs derived from enriched genomic libraries and 146 expressed sequence tag (EST)–SSRs obtained from large-scale EST sequencing analysis. Thirty accessions, comprising Japanese, Chinese (C. mollissima), European (Castanea sativa), and American chestnuts (Castanea dentata), were used for evaluation of SSR polymorphism and transferability across species. The EST–SSRs showed less polymorphism than the genomic SSRs and were more transferable. The mean observed heterozygosity (HO) and the mean expected heterozygosity (HE) of genomic SSRs in the Japanese chestnuts were 0.63 and 0.68, respectively; those of EST-SSRs were each 0.47. Although about 80% of the genomic SSRs were amplified in all 4 species, more than 95% of the EST–SSRs were transferable across all 4 species. The many novel SSRs developed in this study will be applicable for the construction of genetic linkage maps, QTL analysis of phenotypic traits, high-throughput genotyping of marker-assisted selection, and association genetics.

► We developed 220 genomic SSRs and 146 EST–SSRs in Japanese chestnut.
► The mean values of observed heterozygosity (HO) of genomic SSRs and EST-SSRs were 0.63 and 0.47, respectively.
► The EST–SSRs showed less polymorphism than the genomic SSRs.
► More than 95% of the EST–SSRs were transferable across 4 species.