Puffinus shearwaters (Family Procellariidae) are a diverse genus of pelagic seabirds or petrels with a worldwide distribution. Five subgroups of species have been recognized on the basis of osteological and external morphological characters. However, phylogenetic relationships among extant taxa are poorly understood, and evolutionary scenarios to explain the current overlapping distribution of the species subgroups are speculative. Phylogenetic analyses of partial mitochondrial cytochrome b gene sequences were used to examine the relationships among 19 species and subspecies of Puffinus shearwaters. In general, the molecular data support previous morphologically based phylogenetic hypotheses. However, the subgroup Neonectris appears to be polyphyletic with Puffinus nativitatis more closely related to species in the subgroup Puffinus. In addition the molecular data revealed a phylogenetic split between the Puffinus subgroup, with a worldwide distribution, and the remaining four subgroups which have an essentially southern hemisphere distribution. This suggests an evolutionary history in which the Puffinus ancestor was split between two geographic regions, from which dispersal and vicariant events resulted in the evolution and distribution of extant taxa.