Morphological markers and isozymes were used for identifying three presumed species of the Isotoma genus. Morphological traits separated three taxa of the genus. 10 isozymes determined by at least 11 loci were analysed from each taxon, and 2 loci were taxon‐specific, supporting the hypothesis that the three taxa represented three species. The genetic variation found within the taxa measured as fraction of polymorphic loci at the 99% level was 0.82, 0.55 and 0.55 with the corresponding observed heterozygosity 0.15, 0.09 and 0.12. Two populations of the same taxon from Denmark and Norway, respectively, were very closely related. Additional ecological criteria, obtained from a literature survey, also revealed pronounced differences between the three taxa. Due to these facts three distinct species are proposed, namely I. anglicana Lubbock 1862, I. riparia Nicolet 1841 and I. viridis Bourlet 1839.