Aim We investigated the phylogeographical history of a clonal-sexual orchid, to test the hypothesis that current patterns of genetic diversity and differentiation retain the traces of climatic fluctuations and of the species reproductive system. Location Europe, Siberia and Russian Far East. Taxon Cypripedium calceolus L. (Orchidaceae). Methods Samples (>900, from 56 locations) were genotyped at 11 nuclear microsatellite loci and plastid sequences were obtained for a subset of them. Analysis of genetic structure and approximate Bayesian computations were performed. Species distribution modelling was used to explore the effects of past climatic fluctuations on the species range. Results Analysis of genetic diversity reveals high heterozygosity and allele diversity, with no geographical trend. Three genetic clusters are identified with extant gene pools derived from ancestral demes in glacial refugia. Siberian populations exhibit different plastid haplotypes, supporting an early divergence for the Asian gene pool. Demographic results based on genetic data are compatible with an admixture event explaining differentiation in Estonia and Romania and they are consistent with past climatic dynamics inferred through species distribution modelling. Current population differentiation does not follow isolation by distance model and is compatible with a model of isolation by colonization. Main conclusions The genetic differentiation observed today in C. calceolus preserves the signature of climatic fluctuations in the historical distribution range of the species. Our findings support the central role of clonal reproduction in the reducing loss of diversity through genetic drift. The dynamics of the clonal-sexual reproduction are responsible for the persistence of ancestral variation and stability during glacial periods and post-glacial expansion.
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