The intraspecific variability of the Aspergillus viridinutans species was examined using various techniques including morphological examinations, carbon-source-utilization tests, restriction-enzyme analysis of the mitochondrial and nuclear DNA, and sequence analysis of part of the beta-tubulin gene. Although the genetic distances between the isolates were higher than between, e.g., A. fumigatus and Neosartorya fischeri, most A. viridinutans isolates, together with N. aureola and N. udagawae strains, were found to belong to a single cluster based on sequence data. Strain FRR 1266, which was earlier classified as a highly divergent A. fumigatus isolate, was found to belong to the A. viridinutans species. The ochratoxin A producing A. viridinutans strain IMI 306135 was most closely related to an asexual isolate. These two latter strains were more closely related to A. fumigatus and N. fischeri than to any A. viridinutans strains, and possibly represent a new species in the Aspergillus section Fumigati. The dendrogram based on carbon source utilization data and results of restriction analysis of the mitochondrial and nuclear DNA of the strains supported most of the evolutionary relationships observed on the basis of sequence data. The results indicate that the presence or absence of nodding conidial heads is not an unequivocal morphological character for the identification of species within the Aspergillus section Fumigati.