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In this study,an( AG) n microsatellite-enriched library of Rhododendron simsii Planch. was constructed with FIASCO( fast Isolation by AFLP sequences containing repeats) method. Among 356 positive colonies verified by colony PCR of the( AG) n microsatellite-enriched library,233 colonies were selected for further sequencing. According to the results,202 colonies containing microsatellites were obtained,accounting for 86. 7%. Discarding sequences with miscellaneous peaks,overlapping peaks and weak signals,39 sequences were screened finally for subsequent primer exploitation,including 22 perfect type SSR loci,5 imperfect type SSR loci and 12 compound type SSR loci. The FIASCO method is efficient and feasible for constructing microsatellite-enriched libraries of Rhododendron spp.,which laid a solid foundation for isolation of microsatellite loci and investigation of genetic diversity and genetic structure of Rhododendron spp.
In this study, an (AG) n microsatellite-enriched library of Rhododendron simsii Planch. Was constructed with FIASCO (fast Isolation by AFLP sequences containing repeats) method. Among 356 positive colonies verified by colony PCR of the (AG) n microsatellite- enriched library, 233 colonies were selected for further sequencing. According to the results, 202 colonies containing microsatellites were obtained, accounting for 86.7%. Discarding sequences with miscellaneous peaks, overlapping peaks and weak signals, 39 sequences were screened finally for subsequent primer exploitation , including 22 perfect type SSR loci, 5 imperfect type SSR loci and 12 compound type SSR loci. The FIASCO method is efficient and feasible for constructing microsatellite-enriched libraries of Rhododendron spp., which laid a solid foundation for isolation of microsatellite loci and investigation of genetic diversity and genetic structure of Rhododendron spp.