We use extensive geographical sampling and surveys of nuclear microsatellite and mitochondrial DNA loci to investigate the phylogeographic structure of the only recognized self-fertilizing vertebrates, the mangrove killifishes, currently thought to comprise two cryptic species, Kryptolebias marmoratus and Kryptolebias hermaphroditus. All genetic markers revealed three concordant main clades. The Northern clade includes populations from Florida, northern Cuba, Bahamas, Belize and Honduras and corresponds to K. marmoratus. The Southern clade encompasses populations from Brazil and corresponds to K. hermaphroditus. This species was considered endemic to southeastern Brazil, but molecular data corroborate its occurrence in northeastern Brazil. The Central clade, not previously resolved with genetic data, includes populations from Panama and Antilles. Despite the geographic proximity of the Northern and Central clades, the latter is genetically closer to the Southern clade. The discovery of the Central clade raises some taxonomic issues -it can either be considered a distinct species or united with the Southern clade into a single species with two subspecies. Another possible taxonomic solution is a single selfing species, K. marmoratus, with three subspecies. We show that the Central and Southern clades are highly selfing (97-100%), whereas selfing rates of the Northern clade populations vary geographically (39-99%). Genetic patterns indicate that populations in SE Brazil are recent, contrary to expectations based on the known distributions of related species.
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