The phylogenetic relationships among all members of the Drosophila virilis Sturtevant species group were inferred from mitochondrial DNA [partial 16S ribosomal RNA (rRNA), transfer RNA (tRNA)^val, and partial 12S rRNA] sequence data. These data show support for four lineages within the D. virilis species group, which are consistent with previous phylogenetic work performed on the group. However, the relationship among these four groups is not well resolved. These data also suggest that the species D. borealis Patterson is not a monophyletic assemblage, and that the disjunct distribution actually represents two genetically distinct species.