We analysed the polymorphism of cross-amplified microsatellite loci in two endangered butterflies of the genus Phengaris, which inhabit warm grasslands. Specimens of P. arion and P. 'rebeli' collected in Poland showed contrasting levels of variability in the investigated loci. All six tested microsatellites were highly variable in P. arion, whereas in P. 'rebeli' one locus was monomorphic and the other four loci showed a rather low level of polymorphism. Genetic differentiation was small or moderate among P. arion populations and high among P. 'rebeli' populations. We hypothesise that these differences reflect: (1) the relatively high genetic variability of P. arion compared with other members of the genus, indicated by previous studies on allozymes; (2) the small, in the majority of cases, genetic differentiation among populations of P. arion, which suggests recent gene flow; (3) a decrease in the genetic variability and increased isolation of peripheral populations of P. 'rebeli'. In both species, the occurrence of "null" alleles was suggested for some loci, which should be taken into consideration in future studies of the population structure. Additional analysis performed on the corrected frequency of alleles indicated that "null" alleles constitute only a minor problem when estimating genetic differentiation, using FST and AMOVA, however they strongly influence estimates of heterozygosity and FIS, and inferences about the number of genetic groups and assignment of individuals, especially in the case of 'P. rebeli'.
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.