Mitochondrial DNA (mtDNA) variation was investigated among 18 species in the Drosophila montium species subgroup. Based on the restriction patterns of 14 restriction enzymes, the subgroup was clearly classified into 3 species complexes, the kikkawai complex (6 species), the jambulina complex (4 species) and the auraria complex (8 species), which were previously proposed by cross experiments. The relationship between the kikkawai and the jambulina complex were proved to show much closer than that between the auraria and the two complexes by the proportion of restriction fragments which were shared among complexes. The phylogeny among members within the complexes constructed using mtDNA variation was not always consistent with other available information concerning about protein divergence and reproductive isolation.