Characterisation of sixteen additional polymorphic microsatellite loci for the spreading but locally rare European butterfly, Brenthis ino (Lepidoptera: Nymphalidae)

Abstract

Whilst the overall geographic range of the lesser marbled fritillary, Brenthis ino (Rottemburg, 1775), is currently expanding, this species is patchily distributed at a local spatial scale due to its use of flower rich semi-natural meadows and the aggregated distribution of its host plant. Therefore, understanding the dispersal patterns of this butterfly and the effect of increasing fragmentation of the landscapes in central and Western Europe on its population dynamics is key to determining whether this patchy distribution can lead to metapopulation structuring and dynamics. One way to determine the degree to which local populations are isolated from one another is to use high resolution molecular genetic markers and thence quantify gene flow and genetic drift. Eleven microsatellite loci have previously been developed for this species, but six showed evidence of null alleles, effectively violating key assumptions of the models used to infer gene flow. We therefore developed a set of new primer pairs to amplify a suite of 16 highly polymorphic microsatellite loci (number of alleles ranged from 2-30) of which nine were found to conform to the Hardy-Weinberg's expectations, whilst at the same time not showing any clear signature for the presence of null alleles. We further describe how these primers were optimized for landscape and metapopulation genetics studies in Belgian Ardenne.