High genetic diversity at the extreme range edge: nucleotide variation at nuclear loci in Scots pine (Pinus sylvestris L.) in Scotland

Nucleotide polymorphism at 12 nuclear loci was studied in Scots pine populations across an environmental gradient in Scotland, to evaluate the impacts of demographic history and selection on genetic diversity. At eight loci, diversity patterns were compared between Scottish and continental European populations. At these loci, a similar level of diversity (θsil=~0.01) was found in Scottish vs mainland European populations, contrary to expectations for recent colonization, however, less rapid decay of linkage disequilibrium was observed in the former (ρ=0.0086±0.0009, ρ=0.0245±0.0022, respectively). Scottish populations also showed a deficit of rare nucleotide variants (multi-locus Tajima's D=0.316 vs D=−0.379) and differed significantly from mainland populations in allelic frequency and/or haplotype structure at several loci. Within Scotland, western populations showed slightly reduced nucleotide diversity (πtot=0.0068) compared with those from the south and east (0.0079 and 0.0083, respectively) and about three times higher recombination to diversity ratio (ρ/θ=0.71 vs 0.15 and 0.18, respectively). By comparison with results from coalescent simulations, the observed allelic frequency spectrum in the western populations was compatible with a relatively recent bottleneck (0.00175 × 4Ne generations) that reduced the population to about 2% of the present size. However, heterogeneity in the allelic frequency distribution among geographical regions in Scotland suggests that subsequent admixture of populations with different demographic histories may also have played a role

Plant Responses to Extreme Climatic Events: A Field Test of Resilience Capacity at the Southern Range Edge

The expected and already observed increment in frequency of extreme climatic events may result in severe vegetation shifts. However, stabilizing mechanisms promoting community resilience can buffer the lasting impact of extreme events. The present work analyzes the resilience of a Mediterranean mountain ecosystem after an extreme drought in 2005, examining shoot-growth and needle-length resistance and resilience of dominant tree and shrub species (Pinus sylvestris vs Juniperus communis, and P. nigra vs J. oxycedrus) in two contrasting altitudinal ranges. Recorded high vegetative-resilience values indicate great tolerance to extreme droughts for the dominant species of pine-juniper woodlands. Observed tolerance could act as a stabilizing mechanism in rear range edges, such as the Mediterranean basin, where extreme events are predicted to be more detrimental and recurrent. However, resistance and resilience components vary across species, sites, and ontogenetic states: adult Pinus showed higher growth resistance than did adult Juniperus; saplings displayed higher recovery rates than did conspecific adults; and P. nigra saplings displayed higher resilience than did P. sylvestris saplings where the two species coexist. P. nigra and J. oxycedrus saplings at high and low elevations, respectively, were the most resilient at all the locations studied. Under recurrent extreme droughts, these species-specific differences in resistance and resilience could promote changes in vegetation structure and composition, even in areas with high tolerance to dry conditions.This study was supported by Ministerio de Ciencia e Innovación (Spanish Government) Projects CGL2008-04794 and CGL2011-29910 to R.Z., and by grant FPU-MEC (AP2005-1561) to A. H

Species-specific chloroplast DNA polymorphism in the trnV-rbcL region in Pinus sylvestris and P. mugo

Four cpDNA regions were analyzed with the use of PCR-RFLP technique and nucleotide sequences of two mtDNA regions were characterized in order to find P. sylvestris and P. mugo species specific markers useful for studies of the species hybridization. The difference in the restriction fragment patterns of trnV-rbcL region after digestion with MvaI endonuclease was detected. The analyses of the species representatives from various geographic regions revealed that the observed polymorphism is species specific. No differences have been disclosed in the analyzed trnS-trnT, trnK1-trnK2, trnC-trnD cpDNA regions. The P. sylvestris and P.mugo mtDNA sequences of orf25 and coxI regions proved to be identical

Genetic evaluation of seeds of highly endangered Pinus uliginosa Neumann from Wegliniec Reserve for ex-situ conservation program

Peat-bog pine Pinus uliginosa Neumann has become extinct or rare in many parts of Europe. We have investigated the levels of genetic variation and inbreeding in seeds collected from a highly endangered reserve of this species in Poland, using allozymes as genetic markers. Generally, a high level of genetic variation was observed. The mean expected heterozygosity was 0.376, while average (Na) and effective (Ne) numbers of alleles per locus were 2.45 and 1.67, respectively. Nevertheless, we have detected relatively low levels of outcrossing, and potential biparental inbreeding. The population-wide multilocus outcrossing rate was estimated to be 0.706 (±0.091), while the minimum variance mean of single-locus estimates was distinctly lower (ts=0.611). The estimates of outcrossing calculated for individual trees ranged widely from 0.051 to 1.017, indicating the complexity of outcrossing patterns. The investigated population of P. uliginasa from Węgliniec is small and surrounded by extensive forest stands of P. sylvestris. Our three-year records of phenological observations demonstrated that flowering periods for P. uliginosa and P. sylvestris overlap, allowing for cross-pollination. The possibility of P. uliginosa pollination by P. sylvestris creates a potential danger of genetic erosion of the P. uliginosa gene pool. Nonetheless, based on a species specific cpDNA marker we have found that among 533 seedlings of P. uliginosa there were only six seedlings carrying cpDNA marker specific for P. sylvestris, indicating that such hybridization seems to be rare

Shikimate dehydrogenase (E.C. 1.1.1. 25 ShDH) alleles as potential markers for flowering phenology in Pinus sylvestris

The aims of this study were 1) to determine the variability in the flowering phenology of Scots pine (Pinus sylvestris L.) clones in a seed orchard and 2) to compare the genetic structure and genetic markers (13 isozyme loci and 5 chloroplast and 3 nuclear DNA microsatellite loci) among groups of clones that are differentiated by flowering phenology. Using the timing of male inflorescence development, 57 plus trees represented by their clones in a seed orchard were classified into three phenological groups: early-, intermediate-, and late-flowering. The microsatellites showed no significant differences in the genetic structure of the analyzed phenological groups. However, the frequency of allele 2 at the shikimate dehydrogenase A locus (ShDH A 2) differed significantly between the groups of early- and late-flowering trees and between the groups of intermediate- and late-flowering trees. In addition, a significant difference in the frequencies of the genotype ShDH A 11 was observed between the intermediate- and late-flowering groups. Nei’s genetic distance indicated that the late-flowering group was the most genetically distant among the phenological groups. These results suggest that the ShDH A locus might be considered as isoenzymatic marker that differentiates these flowering groups of Scots pine clones. At several isozyme and DNA loci, the presence of private alleles in each group of pines was observed. However, these alleles cannot serve as markers of Scots pine flowering time because of their low frequencies