Identification of 37 microsatellite loci for Anthophora plumipes (Hymenoptera: Apidae) using next generation sequencing and their utility in related species
Abstract
Novel microsatellite markers for the solitary bee, Anthophora plumipes, were identified and characterised using 454 GS-FLX Titanium pyrosequencing technology. Thirty seven loci were tested using fluorescently labelled primers on a sample of 20 females from Prague.
The number of alleles ranged from 1 to 10 (with a mean of 4 alleles per locus), resulting in an observed heterozygosity ranging from 0.05 to 0.9 and an expected heterozygosity from 0.097 to 0.887. None of the loci showed a significant deviation from the Hardy-Weinberg equilibrium and only two loci showed the significant presence of null alleles.
No linkage between loci was detected. We further provide information on a single multiplex PCR consisting of 11 of the most polymorphic loci.
This multiplex approach provides an effective analytical tool for analysing genetic structure and carrying out parental analyses on Anthophora populations. Most of the 37 loci tested also showed robust amplification in five other Anthophora species (A. aestivalis, A. crinipes, A. plagiata, A. pubescens and A. quadrimaculata).
The result of this study demonstrates that next generation sequencing technology is a valuable method for isolating quality microsatellites in non-model species of solitary bees.