The agamosporous and taxonomically critical Dryopteris affinis group was investigated as part of a cytogeographic and morphometric study of ferns in Central Europe. Material from 27 localities in the Czech Republic, Slovakia, Poland and Austria was sampled and evaluated using both morphometric multivariate and karyological analyses. Chromosome counts and flow cytometric analyses revealed the existence of two distinct triploid taxa (2n = 123) of differing genome size, which correspond to D. borreri and D. cambrensis, and of a rare pentaploid hybrid (2n = 205) D. ×critica (D. borreri × D. filix-mas). Morphometric analyses confirmed a clear separation between both triploid taxa. New quantitative characters were selected based on a discriminant analyses, and a key for the identification of the species is presented.