To investigate the origins and phylogenetic relationships of domestic sheep, mitochondrial DNA (mtDNA) from 243 sheep of five European, one African, and four Asian breeds and several mouflon (Ovis musimon), urial (O. vignei bochariensis), and argali (O. ammon nigrimontana, O. a. collium) were assayed for restriction fragment length polymorphisms (RFLP). Twenty haplotypes were identified which occurred in three major phylogenetic groups: urial/argali, mouflon/domestic, and domestic sheep. From the branches that contain mouflon and domestic sheep, two major domestic sheep lineages are apparent. One lineage, termed European lineage, contains the majority of haplotypes detected among European domestic sheep. These mtDNAs resemble mouflon haplotypes. The other lineage, termed Asian lineage, consists of haplotypes found in central Asian and some European domestic sheep. The mean sequence difference between these two lineages (0.72%) is of similar magnitude as that between two argali subspecies. To accurately estimate sequence differences between the European and Asian mtDNA types, the mitochondrial control region of one animal from each lineage and of one mouflon and urial were completely sequenced. Sequence comparisons show that Asian and European domestic sheep lineages differ by 4.43%. The mouflon sequence diverges from the Asian type by 4.52%, but by only 1.36% from the European type. Our data supports the hypothesis that some modern domestic sheep and European mouflon derive from a common ancestor and provide evidence of an additional wild ancestor, other than the urial and argali groups, which has yet to be identified.