Phylogenetic relationships of mosquitoes in the subfamily Anophelinae are presented based on a cladistic analysis of 163 morphological characters from females, males, fourth-instar larvae, and pupae of 64 species. Species examined include one Chagasia Cruz, three Bironella Theobald, and 60 species representing all six subgenera of Anopheles Meigen. Uranotaenia lowii Theobald and Aedeomyia squamipennis (Lynch Arribalzaga) are used as outgroups. This analysis indicates that Anophelinae is monophyletic, that Chagasia is the earliest-diverged lineage within Anophelinae, and that the genus Anopheles, as currently defined, is paraphyletic because it excludes Bironella. Four nonhomoplastic synapomorphies support the monophyly of the clade composed of the genera Bironella and Anopheles. Three major lineages are recognized within this clade. The basal lineage (lineage 1) contains An. (Nyssorhynchus), An. (Kerteszia), and An. implexus (Theobald). This analysis supports the monophyly of the sister groups Nyssorhynchus Blanchard and Kerteszia Theobald, but finds the Albimanus, Argyritarsis, and Myzorhynchella sections of the subgenus Nyssorhynchus to be paraphyletic. The second lineage (lineage 2) contains species of the subgenus Cellia Theobald, which is monophyletic. Except for the Cellia series, however, all other series of the subgenus Cellia are paraphyletic. The third lineage (lineage 3) contains species of the subgenera Anopheles Meigen, Stethomyia Theobald, Lophopodomyia Antunes, and of the genus Bironella. Bironella and Stethomyia are monophyletic sister groups. The Lophoscelomyia and Arribalzagia series are monophyletic. The Arribalzagia and Cycloleppteron series are sister groups nested within the Myzorhynchus series. We conclude that there is no support for the generic recognition of Bironella, nor the subgeneric rank of Lophopodomyia and Stethomyia.