Polymorphisms at a set of eighteen nuclear (nSSR) and chloroplast (cpSSR) microsatellite loci were investigated in sixteen populations of Scots pine (Pinus sylvestris L.) derived from the provenance trial experiment and representative of the species distribution range and climatic zones in Poland. The patterns of genetic variation were compared to the reference samples from the species distribution in Europe and Asia. A similar level of genetic variation and no evidence of population structure was found among the Polish stands. They showed genetic similarity and homogenous patterns of allelic frequency spectra compared to the Northern European populations. Those populations were genetically divergent compared to the marginal populations from Turkey, Spain and Scotland. The population structure patterns reflect the phylogeography of the species and the divergence of populations that most likely do not share recent history. As the analysed provenance trial populations from Poland are diverged in phenotypic traits but are genetically similar, they could be used to test for selection at genomic regions that influence variation in quantitative traits.