BACKGROUND: Legless lizards, the slow worms of the genus Anguis, are forming secondary contact zones within their Europe-wide distribution. METHODS: We examined 35 populations of A. fragilis and A. colchica to identify the level of morphological and genetic divergence in Poland. We applied a conventional study approach using metric, meristic, and categorial (coloration) features for a phenotype analysis, and two standard molecular markers, a mitochondrial (NADH-ubiquinone oxidoreductase chain 2; ND2) and a nuclear (V(D)J recombination-activating protein 1; RAG1) one. RESULTS: We found clear differences between A. fragilis and A. colchica in molecular markers and phenotype-in meristic features, e.g., ear opening, number of scales rows around the body, and higher than so far known diversity in ND2 and RAG1 haplotypes. The presence of five hybrids was detected in three populations in the Polish part of the European contact zone. In all hybrids, homozygous alleles of RAG1 were detected, which suggests a back-crossing within the genus. CONCLUSIONS: The ability to produce fertile offspring by A. fragilis x A. colchica hybrids shows inefficient mechanisms of reproductive isolation of the two legless lizards. The hybrids were indistinguishable from parental species in head proportions (principal components and discriminant analyses) but more resembling A. colchica in meristic traits.

Department of Comparative Anatomy Jagiellonian University Cracow Kraków Poland

Department of Evolutionary Biology and Conservation of Vertebrates University of Wroclaw Wroclaw Poland

Laboratory of Non Mendelian Evolution Institute of Animal Physiology and Genetics CAS Liběchov Czech Republic

Museum of Natural History University of Wroclaw Wroclaw Poland

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