The phylogenetic relationship between two Drosophila species of the takahashii subgroup, D. takahashii and D. pseudotakahashii, was determined through the study of male genitalia, metaphase and salivary chromosomes, mating behaviour and interspecific hybridization. From all lines of evidence it is concluded that the species are very closely related, and now constitute a pair of allopatric sibling species.