Currently it remains difficult to obtain robust microsatellite markers for Lepidoptera. In an attempt to overcome the problems associated with developing microsatellite markers for this insect order we combined (i) biotin-enrichment protocol, (ii) next generation pyrosequencing (through 454 GS-FLX Titanium technology) and (iii) the use of individuals collected from eight geographically distant European populations representing three subspecies of Euphydryas aurinia. Out of 96 stringently designed primer pairs, 12 polymorphic microsatellite loci amplified without obvious evidence of null alleles in eight individuals from different subspecies. Between five and seven of these loci showed full within population applicability and three revealed to be robust and transferable between populations and sub-species, providing a first step towards the development of a valuable and robust tool for studying conservation issues and evolution in E. aurinia populations. Nevertheless, as in most studies dealing with Lepidoptera microsatellites, null alleles were detected in most of the developed markers. Our results emphasize the need for further research in order to better understand the complex evolution and organization of Lepidopteran genomes. and Melthide Sinama, Vincent Dubut, Caroline Costedoat, André Gilles, Marius Junker, Thibaut Malausa, Jean-François Martin, Gabriel Nève, Nicolas Pech, Thomas Schmitt, Marie Zimmermann, Emese Meglécz.
Geographic subspecies of several ithomiine butterflies on the lower east Andean slopes display a black and orange "melanic tiger" aposematic wing pattern that occurs from Colombia to Bolivia, while geographically adjacent lowland subspecies typically bear a coloured, "tiger" pattern. However, it is not clear whether subspecies with similar wing patterns in different regions have arisen through independent events of convergent adaptation, possibly through parapatric differentiation, or result from allopatric differentiation, as proposed by the refuge hypothesis. Here, we examine geographic patterns of divergence in the widespread and common ithomiine butterfly Hyposcada anchiala. We present phylogenetic hypotheses for 5 subspecies of H. anchiala, based on 1567 bp mitochondrial DNA. All topologies indicated that a single switch in mimetic pattern best explained the wing patterning of the H. anchiala studied here. This finding suggests that the subspecies of H. anchiala studied here result from at least two stages of differentiation, and is consistent with a single colonisation into a novel altitudinal zone coincident with a wing pattern switch, followed by subsequent divergence within, rather than across altitudinal zones. The subspecies divergences indicated diversifications were consistent with the Pleistocene. Furthermore, the lowland subspecies were more recently derived than the montane taxa, in contrast to predictions of the "Andean species pump" hypothesis.