Molecular ecology and phylogenetics of the water beetle genus Ochthebius revealed multiple independent shifts to marine rockpools lifestyle

Marine rockpools represent a dynamic ecosystem where inhabiting species usually suffer changeable conditions, with consequent phenomena of local populations’ size expansions and reductions. Nevertheless, a few specialized insect groups are known to live in marine rockpools, and among them, several species belong to the water beetle family Hydraenidae. Three groups of Ochthebius sensu lato (s.l.) live in fact in marine rockpools: the putative subgenus Calobius, the former subgenus Cobalius (both mostly Mediterranean–Macaronesian) and two species of the O. capicola group from South Africa. In this work, we performed a molecular phylogeny of Ochthebius s.l., running molecular clocks, aiming to address the following questions: Are these three groups related? Which is their position within Ochthebius s.l.? How many different times and in how many lineages of Ochthebius s.l. a shift from inland waters to marine rockpools environments happened? Is the current taxonomic status of these three groups supported by genetics? We found that Calobius, Cobalius and the O. capicola group represent three distinct groups, with no sister relationships, suggesting that a shift from fresh/brackish waters to marine rockpools happened independently at least three different times along the diversification of the genus. Cobalius represents an effective separate subgenus, whereas such a rank is not supported by molecular data for Calobius, which represents a monophyletic clade within the nominal subgenus Ochthebius sensu stricto (s.str.). However, as Calobius is monophyletic and characterized by strongly peculiar and distinguishing morphology, we suggest referring to this group as the ‘Calobius’ lineage. Three Mediterranean taxa within this lineage represent likely valid new species, to be described soon. In the same way, the taxonomy of Cobalius should be revised, with two previously formally recognized species found to be paraphyletic, and the possible presence of two additional cryptic species.