A microsatellite DNA toolkit for studying population structure in Apis mellifera

Abstract

We present a set of 18 microsatellite DNA markers that can be run in two multiplex polymerase chain reactions as standard tool for assessing molecular ecological problems in honeybees (Apis mellifera). In addition to a set of six unlinked loci testing for classical population genetic parameters, we present three sets of four tightly linked loci, each located on three different chromosomes. These linked markers are useful for determining the number of colonies in a population as well as the parentage of drones and workers. Moreover, the tool kit can test for various modes of natural selection in honeybee populations.

A survivor population of wild colonies of European honeybees in the northeastern United States: investigating its genetic structure

“…We subjected the four DNA extracts from each colony to PCR at 12 variable microsatellite loci: A24, A28, A79, A88, A107, Ap43, Ap66, Ap81, AC006, B124 (Estoup et al 1995;Garnery et al 1992;Solignac et al 2003), and HB-THE-03 and HB-THE-04 (Shaibi et al 2008), using the previously reported reaction conditions. The final reaction volume per sample was~10 μL and contained 5 μL of PCR Master Mix (Promega, Madison, WI), 1.0-2.5 μL of fluorescent dye-labeled primer, 0.9 μL of nuclease-free water, and 2 μL of DNA extract.…”

Deformed wing virus and drone mating flights in the honey bee (Apis mellifera): implications for sexual transmission of a major honey bee virus

“…Drones in the DCAs were only matched to the hives if their haplotype matched perfectly that of the reconstructed hive queens, including those represented by few hive drones. Those that could not be positively allocated to hives were allocated to unknown queens outside the study, as described before (Shaibi et al 2008;Jaffé et al 2009). …”

“…The DNA from each drone was extracted using a Chelex protocol (Walsh et al 1991). Three sets of four, tightly linked microsatellite markers were used to identify individual drone genotypes (Shaibi et al 2008). The three linkage groups are located on different chromosomes:…”

“…Microsatellite loci were amplified by multiplex polymerase chain reactions (PCRs) using fluorescence-labelled primers and the Promega PCR Master Mix (see Shaibi et al 2008 for PCR conditions). Samples containing no DNA were included in all plates as negative controls.…”

Social parasitism of queens and workers in the Cape honeybee (Apis mellifera capensis)

“…We used the three linked marker sets of the standard microsatellite tool kit for studying honeybee populations (Shaibi et al 2008). These include two loci on chromosome 3 next to the sex locus (csd) spanning 11.6 kb or 0.2 cM (HB-SEX-01, HB-SEX-03).…”