Abstract Aim Our aim was to elucidate the postglacial migration of Viola rupestris F.W. Schmidt, with its two subspecies, rupestris and relicta Jalas, in Europe, using molecular methods. Location Norway, Sweden, Finland, England, the Netherlands, France and Switzerland. Methods Isoenzymes were analysed from 49 populations of Viola rupestris s.lat. Based on former experience of variation in the taxon, the following were included: aspartate aminotransferase, leucine/aminopeptidase, and phosphoglucomutase, yielding a total of nine interpretable and six variable (putative) loci. Results Six multilocus phenotypes were revealed (A–F). Phenotype A (only one individual) is restricted to northern Norway, B is shared by all populations in northern Norway and an isolated population on the west coast of southern Norway (Møre). Phenotype B comes close to C, co-occurring with D in England, the latter also occurring in southern France. Phenotypes E and F occur in the rest of Europe. Main conclusions Viola rupestris ssp. relicta Jalas, previously known only from northern Scandinavia (northern unicentric), was discovered on the west coast of southern Norway (Møre), making the taxon ‘bicentric’. Populations are connected through a putative migration route from the Grands Causses in southern France (Massif Central) via England and western Norway to northern Scandinavia. The Grands Causses might represent a glacial refugium.