Ancient genetic bottleneck and Plio-Pleistocene climatic changes imprinted the phylobiogeography of European Black Pine populations

The historical changes in European Black Pine population size across the whole natural distribution in Europe and Asia Minor were analyzed facing the Plio-Pleistocene climatic fluctuations. Thirteen chloroplast SSRs and SNPs markers have been studied under the assumptions of “neutral evolution.” Populations and meta-populations had different histories of migration routes, and they were strongly affected by complex patterns of isolation, fragmentation, speciation, expansion (1.88–4.28 Ma), purification selection (2.09–21.41 Ma) and bottleneck (1.85–21.76 Ma). A significant number of populations (min. 29–41%) were in equilibrium for very long periods. Generally, the bottleneck revealed by chloroplast DNA is weaker than the bottleneck revealed by nuclear DNA. The N _e immediately after the bottleneck reaches between 1820 and 3640 individuals. Generally, the historical effective population sizes shrink significantly for the Tertiary period from 10–15 up to 2.5 Ma in Western Europe (by 82%), followed by Asia Minor (69%) and the Balkan Peninsula (28%), likely resulting from important climatic changes. The rates and frequencies of stepwise westwards migration waves have been not sufficient to prevent isolation between the meta-populations and to suppress “sympatric speciation.” The migration was weak for the Pliocene, but was maximal for the Pleistocene, and finally silent for the present interglacial period, namely the Holocene.