Mitochondrial DNA diversity of the alpine newt (Ichthyosaura alpestris) in the Carpathian Basin: evidence for multiple cryptic lineages associated with Pleistocene refugia

The phylogeography and molecular taxonomy of the Alpine newt, Ichthyosaura alpestris, has been intensively studied in the past. However, previous studies did not include a comprehensive sampling from the Carpathian Basin, possibly a key region in the evolution of the species. We used a 1251 bp long fragment of the mitochondrial genome to infer the species’ evolutionary history in central-eastern Europe by assigning isolated Carpathian Basin populations from 6 regions to previously defined mtDNA lineages. We also revised the morphology-based intraspecific taxonomy of the species in the light of new genetic data. Alpine newt populations from the Carpathian Basin represented two different mitochondrial lineages. The Mátra, Bükk and Zemplén Mts populations can be assigned to the Western lineage of the nominotypical subspecies. Bakony and Őrség populations showed high haplotype diversity and formed a separate clade within the Western lineage, suggesting that the Carpathian Basin might have provided cryptic refugia for Alpine newt populations in their cold-continental forest-steppe landscapes during the younger Pleistocene. Newts from Apuseni Mts were related to the Eastern lineage but formed a distinct clade within this lineage. Considering the morphological and genetic differentiation of the Bakony and Őrség populations, consistent with a long independent evolutionary history, we propose these populations be referred to as Ichthyosaura alpestris bakonyiensis (Dely, 1964). We provide a redescription of this poorly known subspecies.The project was supported by the Hungarian Scientific Research Fund (OTKA K84071) to JV and ZV and the Bolyai János Research Scholarship of the Hungarian Academy of Sciences (BO/ 00579/14/8) to JV. Project no. [KH130360] has been implemented with the support provided from the National Research, Development and Innovation Fund of Hungary, financed under the [KH_18] funding scheme to JV.Peer reviewe