Nuclear and mitochondrial phylogeography of the European fire-bellied toads Bombina bombina and Bombina variegata supports their independent histories.

Exact location and number of glacial refugia still remain unclear for many European cold-blooded terrestrial vertebrates. We performed a fine-scaled multilocus phylogeographic analysis of two Bombina species combining mitochondrial variation of 950 toads from 385 sites and nuclear genes (Rag-1, Ncx-1) from a subset of samples to reconstruct their colonization and contemporary variation patterns. We identified the lowlands northwest of the Black Sea and the Carpathians to be important refugial areas for B. bombina and B. variegata, respectively. This result emphasizes the importance of Central European refugia for ectothermic terrestrial species, far north of the Mediterranean areas regarded as exclusive glacial refugia for the animals. Additional refugia for B. variegata have been located in the southern Apennines and Balkans. In contrast, no evidence for the importance of other east European plains as refugial regions has been found. The distribution of mtDNA and Ncx-1 variation suggests the presence of local refugia near the Black Sea for B. bombina; however, coalescent simulations did not allow to distinguish whether one or two refugia were present in the region. Strong genetic drift apparently accompanied postglacial expansions reducing diversity in the colonization areas. Extended sampling, coupled with the multilocus isolation with migration analysis, revealed a limited and geographically restricted gene flow from the Balkan to Carpathian populations of B. variegata. However, despite proximity of inferred B. bombina and B. variegata refugia, gene exchange between them was not detected.

Divergence in the face of gene flow: the case of two newts (amphibia: salamandridae).

Understanding the process of divergence requires the quantitative characterization of patterns of gene flow between diverging taxa. New and powerful coalescent-based methods give insight into these processes in unprecedented details by enabling the reconstruction of the temporal distribution of past gene flow. Here, we use sequence variation at eight nuclear markers and mitochondrial DNA (mtDNA) in multiple populations to study diversity, divergence, and gene flow between two subspecies of a salamander, the smooth newt (Lissotriton vulgaris kosswigi and Lissotriton vulgaris vulgaris) in Turkey. The ranges of both subspecies encompass mainly the areas of this important glacial refugial area. Populations in refugia where species have been present for a long time and differentiated in situ should better preserve the record of past gene flow than young populations in postglacial expansion areas. Sequence diversity in both subspecies was substantial (nuclear pi(sil) = 0.69% and 1.31%). We detected long-term demographic stability in these refugial populations with large effective population sizes (N(e)) of the order of 1.5-3 x 10(5) individuals. Gene trees and the isolation with migration (IM) analysis complemented by tests of nested IM models showed that despite deep, pre-Pleistocene divergence of the studied newts, asymmetric introgression from vulgaris to kosswigi has occurred, with signatures of recent gene flow in mtDNA and an anonymous nuclear marker, and evidence for more ancient introgression in nuclear introns. The distribution of migration times raises the intriguing possibility that even the initial divergence may have occurred in the face of gene flow.