Characterization of 16 novel microsatellite loci for Ephippiger diurnus (Orthoptera: Tettigoniidae) using pyrosequencing technology and cross-species amplification

A novel panel of 16 microsatellite markers, obtained by pyrosequencing of enriched genomic libraries, is reported for the fl ightless European bushcricket Ephippiger diurnus (Dufour) (Orthoptera: Tettigoniidae). Five multiplex and one simplex PCR protocols were optimized, and the polymorphism at the 16 loci was assessed in two natural populations from southern France. The mean allele number and (expected mean heterozygosity) were 8.94 (0.71) and 6.57 (0.70), respectively, in each population. Several loci were at Hardy-Weinberg disequilibrium (HWD), possibly due to the incidence of null alleles. The occurrence of null alleles has been previously reported for this species, and it is a common feature of microsatellite loci in Orthoptera. Cross-amplifi cation tests demonstrated the transferability of some of these loci to other ephippigerine species. The microsatellite loci reported here substantially increase the number of available loci for this species and will afford an accurate picture of E. diurnus phylogeography, the genetic structure of its populations, and an improved understanding of the evolution of male song and other sexually-selected traits in this highly variable specie