Isolates of Heterobasidion annosum (Fr.) Bref. representing North American S and P and European S, P, and F intersterility groups were subjected to isozyme analysis. European S, P, and F groups had more variability than the North American S and P groups in expected hterozygosity, number of alleles per locus, and percent polymorphic loci. In contrast with the North American S and P groups, the European intersterility groups could not be distinguished from each other on the basis of individual isozyme loci, although significant differences in allele frequencies exist between European S and P groups. This suggests that evolution proceeded at different rates in the intersterility groups, or intersterility barriers appeared later in the European populations relative to the North American populations of H. annosum. Changes in climate and host species associations during the Tertiary may have been a major factor in evolution of H. annosum intersterility groups. Key words: allozymes, forest tree hosts, playnological events, evolutionary relationships, Hymenomycetes, root disease.