The beta 1-, beta 2-, and beta 3-tubulin genes have been mapped by in situ hybridization on the polytene chromosomes of 11 selected species (15 strains) belonging to the Drosophila montium subgroup. Although the hybridization pattern among the strains of the same species does not differ, this pattern is significantly different among the species. The beta-tubulin genes in the montium subgroup seem to be organized in a cluster, or in a semicluster or are completely dispersed. The clustered arrangements is found in the North-Oriental sibling species D. auraria, D. triauraria, and D. quadraria. The semiclustered arrangement, wherein the beta 1 and beta 2 genes are located at the same locus while beta 3 is at a different one, appears in the South-Oriental species D. bicornuta, D. serrata, and D. birchii, as well as in the Afrotropical species D. diplacantha and D. seguyi. The complete separation of the genes is observed in the Indian species D. kikkawai and D. jambulina and in the Afrotropical species D. vulcana. Based on the above results, a possible mode of evolution of the beta-tubulin genes in the montium subgroup is attempted. In addition, phylogenetic relationships among the montium species are discussed.