Two alder species (Alnus glutinosa and A. incana) have overlapping distribution, naturally occur in Lithuania, and are considered ecologically and economically important forest tree species. The objective of our study was to estimate the likelihood of spontaneous hybridizations between native alders in natural stands of Lithuania based on leaf morphology and nuclear microsatellite markers. The sampled trees were assigned to the three taxonomic groups of A. glutinosa, A. incana, and potential hybrids based on the leaf and bark morphological traits. The genetic differentiation and potential hybridization between these three groups was tested based on 15 nSSR markers. We identified studied Alnus spp. individuals as pure species and hybrids. Two microsatellite loci were reported as discriminating well between these species. We concluded that our results showed the highest likelihood of two genetic group structures, a clear genetic differentiation between the morphology-based groups of A. glutinosa and A. incana, and rather variable likelihood values in the putative hybrid group. The results provide important implications for genetic conservation and management of Alnus spp.

Spontaneous hybrids between the native elms (genus Ulmus L.) have been observed in the forests of Europe. Gene conservation raises questions regarding the genetic background for the complex morphology and taxonomy of elms. Our objective was to dissect morphological and genetic variation in the natural swamps of Ulmus species groups in Lithuanian forests with the aid of leaf morphology and microsatellite (SSR) markers. We sampled leaves from 189 elms at 26 locations to grasp the phenotypic diversity in variable natural habitats in Lithuanian forests. We assigned the elms into six taxonomic and genetics groups based on 31 leaf morphology parameters and tested the genetic differentiation between these six groups at six nuclear SSR loci by using Bayesian and genetic distance-based clustering. The genetic and leaf morphometric analyses of putative elm hybrid swamps indicated a low genetic exchange between U. laevis Pall. and the other Ulmus groups. The genetic and morphometric data supported the differentiation of U. glabra Huds. and U. glabra (female) × U. minor Mill. (male) spontaneous hybrids. In addition, the results of the genetic analysis also confirmed the high level of genome sharing among U. minor and U. minor subsp. minor Richens., where leaf morphology failed to differentiate genetically discrete groups. For gene conservation, we would suggest considering separate gene conservation units selected based on leaf and stem morphology for U. laevis, U. glabra, U. glabra × minor, and the U. minor species complex.

We aimed to extract DNA and amplify PCR fragments at the mitochondrial DNA Nad7.1 locus and 11 nuclear microsatellite loci in nine circa 11,000-year-old individuals of Scots pine found at the bottom of the Baltic sea and test the genetic associations with the present-day gene pool of Scots pine in Lithuania. We followed a strict anticontamination protocol in the lab and, simultaneously with the aDNA specimens, tested DNA-free controls. The DNA was extracted by an ATMAB protocol from the ancient wood specimens sampled underwater from Scots pine stumps located circa 20–30 m deep and circa 12 km ashore in western Lithuania. As the references, we used 30 present-day Lithuanian populations of Scots pine with 25–50 individuals each. The aDNA yield was 11–41 ng/μL. The PCR amplification for the mtDNA Nad7.1 locus and the nDNA loci yielded reliable aDNA fragments for three and seven out of nine ancient pines, respectively. The electrophoresis profiles of all the PCR tested DNA-free controls contained the sizing standard only, indicating low likelihood for contamination. At the mtDNA Nad7.1 locus, all three ancient Scots pine individuals had the type A (300 bp) allele, indicating postglacial migration from the refugia in Balkan peninsula. The GENECLASS Bayesian assignment tests revealed relatively stringer and consistent genetic associations between the ancient Scots pine trees and the present-day southern Lithuanian populations (assignment probability 0.37–0.55) and several wetlands in Lithuania. Our study shows that salty sea water efficiently preserves ancient DNA in wood at the quality levels suitable for genetic testing of trees dated back as far as 11,000 years before present.

We studied the maternally inherited chloroplast DNA polymorphism at three microsatellite loci of 157 Quercus robur trees from 38 native populations in Lithuania. We found high diversity of eight haplotypes from the Balkan lineage A (frequency 0.75) and the “German” subbranch of the Balkan lineage A (freq. 0.12), western and eastern Italian lineages C (freq. 0.05 and 0.06, respectively), and Iberian lineage B (freq. 0.03). The haplotypes were geographically well structured (among population differentiation index PhiPT = 0.30, the p-value < 0.001) that is unexpected for such a small territory as Lithuania. We raised a hypothesis on historical overexploitation of oaks by eliminating certain haplotypes in Lithuania, following a drastic felling of oak forests over the last few centuries.