Plebejus argus is a model species for studying the biology, population ecology and genetics of butterflies. It is patchily distributed throughout most of its European range and considered to be sedentary. Habitats of the butterfly have to encompass two vital larval-resources, i.e. specific food plants and ants, since caterpillars are obligatorily myrmecophilous. The genetic structure of nine P. argus populations (85 individuals) was studied at an intermediate geographical scale (Eastern Poland, diameter of about 400 km) using two kinds of molecular markers i.e. COI (mtDNA) and EF-1α (nuclear gene). Both markers were highly variable with as many as 16 haplotypes and 39 alleles, respectively. Great genetic differentiation in the COI gene was detected (overall FST = 0.411, P < 0.001) but little genetic differentiation in the EF-1α gene (FST = 0.021, P < 0.001). The number of COI haplotypes (ranging from one to seven) and their distribution varied considerably among P. argus populations. The possibility that this heterogeneity was related to Wolbachia was excluded as this endoparasitic bacterium was not detected in samples from any of the populations studied. PCA and SAMOVA analyses divided the sampled populations into two or three groups, which could indicate different colonization routes. Moreover, the differences in genetic differentiation with respect to mtDNA and nuclear markers may suggest male-biased dispersal of P. argus at a larger scale. The hypothesis that females are philopatric is consistent with direct observations of the restricted colonization abilities of the butterfly, while the relatively homogeneous genetic structure revealed by previous allozyme studies in some areas might be explained by the possible higher mobility of males. and Marcin Sielezniew, Donata Ponikwicka-Tyszko, Miroslaw Ratkiewicz, Izabela Dziekanska, Agata Kostro-Ambroziak, Robert Rutkowski.