The effects of long-term habitat fragmentations on genetic and population differentiation of Betula pendula Roth were investigated using chloroplast DNA (cpDNA) variations. Leaf samples were collected from four small remnant populations across the north of Iran. Three pairs of universal primers were used to amplify cpDNA, large single copy regions of trnC-trnD, trnK1-trnK2 and trnD-trnT. A total of 18 of the cpDNA haplotypes in the four populations were identified, however, no clear phylogeographic structuring of haplotypes could be detected. The total genetic diversity (HT) for all populations was high (0.932). Average intra-population genetic diversity was estimated as HS = 0.729 and average differentiation of populations GST = 0.218. Mantel tests of isolation by distance revealed a significant relationship between Wright’s inbreeding coefficient (Fst) and geographical distances for the four populations in Iran (r = 0.77, p < 0.05). The results of the hierarchical analysis of molecular variance (AMOVA) indicated that a 66% variation was partitioned within populations, whilst the variance amongst the four populations was only 34%. We suggest that significant genetic differentiation amongst populations can likely be attributed to reduced gene flow as a result of habitat fragmentation.
Pinus nigra J.F. Arnold, European black pine, is a typical component of Mediterranean and sub-Mediterranean coniferous forests with highly fragmentary distribution. Western Mediterranean populations of this species have been studied genetically to date, while eastern populations from the central Balkans, which are larger and more abundant, are still genetically understudied. We analyzed seven populations of P. nigra representing all infraspecific taxa recognized within the central Balkans (subspecies nigra with varieties nigra and gocensis Đorđević; and subspecies pallasiana (Lamb.) Holmboe with varieties pallasiana and banatica (Endl.) Georgescu et Ionescu), with three chloroplast microsatellites (cpDNA SSRs) and one mitochondrial (mtDNA) locus. Although our molecular data failed to support circumscription of studied infraspecific taxa, we found that genetic patterns at both genomes are in accordance with those found previously in westward populations of this species, that is – exceptionally high levels of genetic diversity (HT = 0.949) and low genetic differentiation (GST = 0.024) at the cpDNA level, and moderate levels of genetic diversity (HT = 0.357) and genetic differentiation (GST = 0.358) at the mtDNA level. Based on genealogical relations of mtDNA types currently present in Balkans’ and Iberian/African populations, we inferred that the ancestral gene pool of P. nigra already harbored polymorphism at position 328 prior to the divergence to two lineages currently present in westward and eastward parts of the species range distribution. Subsequent occurrence of three mutations, which distinguish these two lineages, suggests their long-term isolation.