An independent origin of an annual life cycle in a North American killifish species

An annual life cycle is characterized by short-lived adults that exploit seasonally productive but temporary habitats, paired with a hardy embryo stage that survives unfavourable environmental conditions, such as winter or the dry season. This life cycle is rare amongst vertebrates but has evolved independently several times in African and South American killifishes adapted to life in seasonally ephemeral aquatic pools. A single species of North American annual killifish, with a limited geographical distribution in Mexico, also exhibits this seasonal life cycle. This enigmatic species, Millerichthys robustus, is geographically separated from the nearest South American annual killifish species by >2000 km. Millerichthys robustus exhibits morphological features that have led to several competing phylogenetic hypotheses. To date, its phylogenetic relationships are unclear because no molecular data have been available. Here, we sequence the mitochondrial genome of Millerichthys robustus and generate a molecular phylogeny of killifishes that includes this species. Our results indicate that, rather than being most closely related to South American annual killifishes, this species is sister to two non-annual killifish species from Cuba (Rivulus cylindraceus and Rivulus berovidesi). Ancestral state reconstruction strongly supports an independent origin of an annual life cycle and embryonic diapause in Millerichthys robustus.