An understanding of the genetic variation of the beech, especially at the edge of its natural distribution, is essential because of the change in natural distribution of the species resulting from changing climatic conditions. The main aim of the study was to determine the level of genetic diversity of European beech at the north-eastern edge of its natural range. The other aim was to check the genetic variation of beech from the two centres, the north and the south of Poland, which were identified in previous findings based on pollen analyses and phenotypic traits. The research material was the progeny of twelve beech provenances. The genetic structure of the populations was determined by ten highly variable microsatellite DNA loci. The results confirmed the high genetic diversity of beech at the north-eastern edge of its natural distribution, which infers the probability of their good adaptation to the changing climate and an extension of the range. Genetic analyses confirmed the existence of two genetic centres for beech in Poland. The populations from south-eastern Poland had a slightly higher diversity than the populations from the north-western area, which may indicate that the colonisation of Poland occurred by two routes. The results are important for creating the borders of the provenance regions and for limiting the transfer of seeds and seedlings. The choice of forest reproductive material, based on the knowledge of genetic diversity, is very important for the stability of future forests.
English yew (Taxus baccata L.) is a strictly outcrossing and dioecious species whose populations are small and isolated. It is known that sex ratios may vary in natural populations due to local environmental conditions or stochastic events. However, unbalanced sex ratios may have negative impacts on genetic diversity through enhanced genetic drift and inbreeding. The present study represents one of the first attempts to compare the genetic variation at microsatellite loci within and between populations with different gender proportions. Our results indicated that there were no significant correlations between sex ratio and the extent of genetic variation in different populations. All populations exhibited high levels of genetic diversity. Additionally, the genetic structure was characterized separately in male and female individuals. Statistical analyses of the set estimators describing the genetic structure of male and female individuals of T. baccata revealed no significant differences between the two groups. Molecular analysis verified that microsatellite nuclear loci neutrality developed for T. baccata, as there were no significant differences in the genetic variation between males and females and no evidence for any outlier loci using coalescent and hierarchical Bayesian simulations. The results demonstrate that ignoring biased sex ratios in T. baccata populations had no effect on the assessment of genetic differentiation and genetic diversity within and between populations of this species. These results are discussed with regards to the practical application of molecular markers in conservation programs.
Multiplex polymerase chain reaction (PCR) allows amplification of two or more pair of primers in parallel for amplification of multiple target sequences in a single reaction tube. In this study, we combined existing simple sequence repeat (SSR) markers (nuclear microsatellites) in the novel combination of multiplex PCR to study the population genetics of common walnut from Croatia. From twenty one tested SSR markers, eleven produced satisfactory results in one multiplex PCR. Population genetic results achieved from 15 samples of Croatian common walnut showed moderate genetic variability (average value: He 0.473; Ho 0.568). Our multiplex PCR allowed cost effective work concerning chemicals, plastic ware, device, and working time producing optimal results. The optimized multiplex PCR represented the best combination of eleven SSR primers for genotyping common walnut in a single PCR reaction.