Patterns of genetic diversity resulting from bottlenecks in European black pine, with implications on local genetic conservation and management practices in Bulgaria

In the present study, we investigated the genetic structure and diversity of P.nigra populations in Bulgaria, using simple sequence nuclear repeats. Among-population structure was studied with distance and Bayesian frequency methods, assuming geometric distance and a “non-admixture” model. The “NJ” and “non-admixture” clusters confirm the “mountain effect” hypothesis of the black pine genetic structure in the study region. The analyses showed moderate among-population divergence (13.31 %; AMOVA) and evidence of genetic bottlenecks. The coalescent analyses suggest that P. nigra has survived for a long period (thousands of generations) under strong selection pressure and that its populations continued to be exposed to stochastic factors like climate fluctuation, forest fire and disease. The combination of recent and historic changes is responsible for the present population size and genetic diversity. Our results suggest that conservation and management practices should strive to maintain this genetic differentiation, specifically by emphasising reforestation efforts with stocks from local provenances to avoid non-local introductions

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

International audienceThe 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

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

<p>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.</p

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

<p>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.</p

Ancient genetic bottleneck and Plio-Pleisocene 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.This is a post-peer-review, pre-copyedit version of an article published in European Journal of Forest Research. The final authenticated version is available online at: [http://dx.doi.org/10.1007/s10342-017-1069-9