Speciation and paraphyly in western Mediterranean hares (Lepus castroviejoi, L. europaeus, L. granatensis, and L. capensis) revealed by mitochondrial DNA phylogeny.

Mitochondrial DNA (mtDNA) variation among specimens of the northwestern African hare (Lepus capensis schlumbergeri) and three European hares sampled in Spain (L. castroviejoi and L. granatensis, which are endemic to the Iberian Peninsula, and L. europaeus) was analyzed using seven restriction endonucleases. Fourteen haplotypes were found among the 34 animals examined. Restriction site maps were constructed and the phylogeny of the haplotypes was inferred. mtDNA of L. capensis was the most divergent, which is consistent with its allopatric African distribution and with an African origin of European hares. We estimated that mtDNA in hares diverges at a rate of 1.5-1.8% per MY assuming that the European and African populations separated 5-6 MYBP. Maximum intraspecies nucleotides divergences were 1.3% in L. capensis, 2.7% in L. castroviejoi, and 2.3% in L. granatensis but 13.0% in L. europaeus. The latter species contained two main mtDNA lineages, one on the branch leading to L. castroviejoi and the other on that leading to L. granatensis. The separation of these two lineages from the L. castroviejoi or L. granatensis lineages appears to be much older than the first paleontological record of L. europaeus in the Iberian peninsula. This suggests that the apparent polyphyly of L. europaeus is due not to secondary introgression, but to the retention of ancestral polymorphism in L. europaeus. The results suggest that L. europaeus either has evolved as a very large population for a long time or has been fractionated. Such a pattern of persistence of very divergent lineages has also been reported in other species of highly mobile terrestrial mammals. As far as mtDNA is concerned. L. europaeus appears to be the common phylogenetic trunk which has diversified during dispersion over the European continent and from which L. castroviejoi and L. granatensis speciated separately in southwest Europe.