Isolation and characterization of microsatellite loci in Rhododendron ferrugineum (Ericaceae) using pyrosequencing technology

Abstract

Microsatellite primers were developed for Rhododendron ferrugineum (Ericaceae) to evaluate genetic diversity, population genetic structure, and mating system of this self-compatible species. The new-generation 454 FLX Titanium pyrosequencing technology was used to isolate 102 novel microsatellite loci. Two multiplex PCR sets were optimized to genotype nine polymorphic microsatellite loci. The level of genetic diversity was assessed in two populations from the Pyrenees (France). The mean number of alleles per locus ranged from 1.5 to 7. The mean observed and expected heterozygosities ranged from 0 to 0.76 and from 0.03 to 0.66, respectively. Cross-species amplification was successful for 13 other Rhododendron species and two additional genera of Ericaceae, with an average of seven pairs of primers amplifying per species. These markers will facilitate further studies on the evolutionary history of the large Ericaceae family and Rhododendron in particular.