Soybean is the most grown high-protein crop in the world. Despite the rapid increase of acreage and production volume, European soybean production accounts for only 34% of its consumption in Europe. This study aims to support the optimal exploitation of genetic resources by European breeding programs by investigating the genetic diversity and the genetic structure of 207 European cultivars or American introductions registered in Europe, which were genotyped by the SoySNP50K array. The expected heterozygosity (He) was 0.34 for the entire collection and ranged among countries from 0.24 for Swiss cultivars to 0.32 for American cultivars (partly reflecting differences in sample size between countries). Cluster analysis grouped all genotypes into two main clusters with eight subgroups that corresponded to the country of origin of cultivars and their maturity group. Pairwise Fstvalues between countries of origin showed the highest differentiation of Swiss cultivars from the rest of the European gene pool, while the lowest mean differentiation was found between American introductions and all other European countries. On the other hand, Fstvalues between maturity groups were much lower compared to those observed between countries. In analysis of molecular variance, the total genetic variation was partitioned either by country of origin or by maturity group, explaining 9.1% and 3.5% of the total genetic variance, respectively. On the whole, our results suggest that the European soybean gene pool still has sufficient diversity due to the different historical breeding practices in western and eastern countries and the relatively short period of breeding in Europe.