Characterization of 16 novel microsatellite loci for Ephippiger diurnus (Orthoptera: Tettigoniidae) using pyrosequencing technology and cross-species amplification
2016
Esquer Garrigos, Yareli | Greenfield, Michael D. | Party, Virginie | Streiff, Rejane | Centre de Biologie pour la Gestion des Populations (UMR CBGP) ; Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut National de la Recherche Agronomique (INRA)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Université de Montpellier (UM)-Institut de Recherche pour le Développement (IRD [Occitanie])-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro) | Institut de recherche sur la biologie de l'insecte (IRBI) ; Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS) | Diversité, Génomes & Interactions Microorganismes - Insectes [Montpellier] (DGIMI) ; Institut National de la Recherche Agronomique (INRA)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université de Montpellier (UM)
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 species
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