Attachments:

Anthophora_plumipes_primers.xls	[ ]	590 Kb
GENOSCREEN-Project3444-ANT-sequences_IE.zip	[ ]	1914 Kb

We identified and characterizes novel microsatellite markers for the solitary bee Anthophora plumipes using 454 GS-FLX Titanium pyrosequencing technology. We used commercial offer of Genoscreen Company (Lile, France). All obtained sequences together with designed primers for all detected microsatellite loci are attached. We descovered 37 microsatellite loci, from which 36 were polymorphic. All these loci were tested on 20 females from Prague. Number of alleles ranged from 1 to 10 (mean 4 alelles per locus), observed heterozygosity from 0.05 to 0.9 and expected heterozygosity from 0.097–0.887 (Tab.1). None loci showed significant deviation from the Hardy-Weinberg equilibrium, only two loci showed significant presence of null alleles. No linkage between loci was detected.

We further provide information on single multiplex PCR consisting of 11 most polymorphic loci (Tab. 2). This multiplex was tested on two distinct populations and we showed, that it provides effective analytical tool for analyzing genetic structure and performing parental analyses of Anthophora populations.

Most of 37 loci tested also showed good amplification ability on 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 of non-model solitary bee species.

All tables are from:

Černá K. & Straka J.: Identification of 37 microsatellite loci for Anthophora plumipes (Hymenoptera: Apidae) using next generation sequencing and their utility in related species. European Journal of Entomology 109: 155-160.