Ancient split of major genetic lineages of European Black Pine: evidence from chloroplast DNA

Abstract : The European Black Pine (Pinus nigra Arn.) has a long and complex history. Genetic distance and frequency analyses identified three differentiated genetic groups, which corresponded to three wide geographical areas: Westerns Mediterranean, Balkan Peninsula and Asia Minor. These groups shared common ancestors (14.75 and 10.72 Ma). The most recent splits occurred after the Messinian Salinity Crisis (4.37 Ma) and the Early–Middle Pleistocene Transitions (0.93 Ma). The posterior ancestral population size (Na) is 260, 000–265,000 individuals. Each pool is further fragmented, with evidence of a phylogeographic structure (Nst > Gst) typ- ically observed in some natural populations from the Western Mediterranean region and the Balkan Peninsula. The labora- tory analysis was performed by fragment analysis—i.e. elec- trophoretic sizing of polymerase chain reaction fragments, combined with the sequencing analysis of 33 % of all individ- uals as a control. Intense sampling of chloroplast DNA poly- morphisms (3154 individuals and 13 markers: SNPs and SSRs) over the full area of the species’ natural distribution indicated moderate among-population variability (Gst(nc) ≤ 0.177) in various parts of its range. These results indicate that the natural populations have long migration his- tories that differ from one another and that they have been strongly phylogeographically affected by complex patterns of isolation, speciation and fragmentation. Long and varying climatic fluctuations in the region of the principal genetic group have been the probable cause of different forest com- munity associations with different successional patterns resulting in interglacial refugia vs. macro long-term refugia.