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'.
Microsatellites are the most common markers used in population and conservation genetic studies. However, their isolation is laborious and expensive. In some taxa, such as Lepidoptera, it is particularly difficult to isolate microsatellite markers due to the high similarity of the flanking regions of different loci and the presence of null alleles. Here we isolated microsatellites of the endangered butterfly Boloria aquilonaris using 454 GS-FLX Titanium pyro-sequences of biotin enriched DNA libraries and tested the success of cross-amplification on the sister-species B. eunomia. Fifteen polymorphic microsatellite loci were isolated in B. aquilonaris using initially 101 stringently designed primer pairs. Unlike in many other studies of microsatellite isolation in Lepidoptera, few null alleles were detected and only at very low frequencies. Additionally, the raw data set can still be used for the isolation of other microsatellite loci. None of the selected polymorphic loci for B. aquilonaris gave clear banding patterns for B. eunomia, although about 15 other loci gave promising banding patterns for the latter species. Low intra- and inter-specific transferability of developed markers in this study also lends support to the hypothesis that the evolution of the genome of Lepidoptera is dissimilar from that of other organisms., Sofie Vandewoestijne, Camille Turlure, Michel Baguette., and Obsahuje seznam literatury