Abstract
European brown hare, Lepus europaeus, from Central and Eastern European countries (Hungary, Poland, Serbia, Lithuania, Romania, Georgia and Italy) were sampled, and phylogenetic analyses were carried out on two datasets: 1.) 137 sequences (358 bp) of control region mtDNA; and 2.) 105 sequences of a concatenated fragment (916 bp), including the cytochrome b, tRNA-Thr, tRNA-Pro and control region mitochondrial DNA. Our sequences were aligned with additional brown hare sequences from GenBank. A total of 52 and 51 haplotypes were detected within the two datasets, respectively, and assigned to two previously described major lineages: Anatolian/Middle Eastern (AME) and European (EUR). Furthermore, the European lineage was divided into two subclades including South Eastern European (SEE) and Central European (CE). Sympatric distribution of the lineages of the brown hare in South-Eastern and Eastern Europe revealed contact zones there. BAPS analysis assigned sequences from L. europaeus to five genetic clusters, whereas CE individuals were assigned to only one cluster, and AME and SEE sequences were each assigned to two clusters. Our findings uncover numerous novel haplotypes of Anatolian/Middle Eastern brown hare outside their main range, as evidence for the combined influence of Late Pleistocene climatic fluctuations and anthropogenic activities in shaping the phylogeographic structure of the species. Our results support the hypothesis of a postglacial brown hare expansion from Anatolia and the Balkan Peninsula to Central and Eastern Europe, and suggest some slight introgression of individual haplotypes from L. timidus to L. europaeus.
Highlights
The brown hare (Lepus europaeus Pallas, 1778) is a native species to Northern, Central, Western Europe and the Western part of Asia, and it was introduced as a game into several countries (Argentina, Australia, Barbados, Brazil, Canada, Chile, Falkland Islands, New Zealand, Rèunion and the United States; [1]).The effect of translocation on hare genome was proved by previous genetic studies and they suggested that the brown hare and the Cape hare (Lepus capensis) are the same species [2]
European brown hares have been classified to two major lineages, European (EUR) and Anatolian/Middle Eastern (AME) [6, 15, 17,18] that co-exist in Republic of Northern Macedonia, North-Eastern Greece and Bulgaria [6, 10, 15]
We presented a relatively comprehensive dataset on mitochondrial DNA (mtDNA) cytochrome b, tRNA-Thr, tRNA-Pro and control region fragments of brown hares in Central-Eastern Europe, where two datasets were used in the genetic analyses; the first dataset included a 358-bp control region sequence, whereas the second dataset covered a concatenated sequence of mtDNA fragments
Summary
The brown hare (Lepus europaeus Pallas, 1778) is a native species to Northern, Central, Western Europe and the Western part of Asia, and it was introduced as a game into several countries (Argentina, Australia, Barbados, Brazil, Canada, Chile, Falkland Islands, New Zealand, Rèunion and the United States; [1]).The effect of translocation on hare genome was proved by previous genetic studies and they suggested that the brown hare and the Cape hare (Lepus capensis) are the same species [2]. Pierpaoli et al [4] showed that Italian and European hares did not share any mitochondrial haplotypes, indicating the lack of interspecific gene flow between the two species due to reproductive isolation in the course of their long separate evolutionary history. They identified two main groups of Eurasian and African hare haplotypes: Clade A (L. granatensis, L. corsicanus, L. timidus) and Clade B Mediterraneus, L. habessinicus, L. starcki, L. europaeus) These results suggest that the three species belonging to Clade A, with a common ancestor, would have colonized Europe independently of L. europaeus and would have originated by isolation during the Pleistocene glaciations in the southern or northern areas of refuge
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