The aim of the work is to determine the genetic diversity, genetic differentiation and genetic structure of subpopulations of the specially protected common yew in the area of South Moravian Podyjí (Czech Republic), Thayatal (Austria) and in the Svitavy region (Czech Republic) using DNA analyses (nSSR-nuclear Simple Sequence Repeats). The subjects of the analyses were yews of 1) natural occurrences in the territory of the Podyjí National Park (NP), 2) cultural origin in the Podyjí NP, 3) natural occurrences in the Thayatal NP (Austria), 4) natural occurrences outside the Podyjí NP, and 5) cultural origin in the Podyjí region outside the NP. For the comparison and better interpretation of results there also were analysed geographically more distant subpopulations, 6) natural occurrences in the Hřebečský ridge (NW Moravia), 7) cultural yew occurrences in the Svitavy region (NE Bohemia & NW Moravia), 8) natural occurrences in the Jílovské yews (N Bohemia), and 9) natural occurrences in the Moravian Karst (S Moravia). We observed higher level of genetic diversity (Ho = 0.54; He = 0.77), the FST with ENA correction ranged from 0.007 to 0.123. According to a STRUCTURE analysis, high genetic similarity was observed between samples from the Podyjí NP and Hřebečský ridge. The impact of the analysis results on the conservation management of tested populations was outlined.