Ochthebiinae, with c. 650 species distributed worldwide, are the second most speciose subfamily of the aquatic beetle family Hydraenidae. They are ecologically the most diverse hydraenid subfamily, with terrestrial species as well as species in almost all types of aquatic habitats, including hypersaline waters. Ochthebiinae include the tribes Ochtheosini (four species in three genera) and Ochthebiini. We provide here the first comprehensive phylogeny of the tribe Ochthebiini, based on 186 species and four subspecies from most genera, subgenera and species groups. We obtained sequence data for a combination of mitochondrial and nuclear gene fragments including the 5 ′ and 3 ′ ends of the cytochrome c oxidase subunit 1, the 5 ′ end of 16S RNA plus the leucine tRNA transfer plus 5 ′ end of NADH dehydrogenase subunit I, and internal fragments of the large and small ribosomal units. The analyses with maximum likelihood (ML) and Bayesian probabilities consistently recovered a generally well supported phylogeny, with most currently accepted taxa and species groups as monophyletic. We provide a new classification of the tribe based on our phylogenetic results, with six genera: Meropathus Enderlein, Ochthebius Leach, Protochthebius Perkins, Prototympanogaster Perkins, Tympallopatrum Perkins and Tympanogaster Janssens. The genus Ochthebius is here divided into nine subgenera in addition to Ochthebius s.s.: (1) O. (Angiochthebius) Jäch & Ribera; (2) O. (Asiobates) Thomson; (3) O. (Aulacochthebius) Kuwert; (4) O. (Cobalius) Rey; (5) O. (Enicocerus) Stephens; (6) O. (Gymnanthelius) Perkins comb.n.; (7) O. (Gymnochthebius) Orchymont; (8) O. (Hughleechia) Perkins comb.n.; and (9) O. (Micragasma) Sahlberg. Within Ochthebius s.s., 17 species groups are proposed, five of them newly established (3, 9, 11, 13 and 16): (1sumatrensis; and (17) vandykei. We elevated to species rank two subspecies of Ochthebius: O. fallaciosus Ganglbauer stat.n. (former subspecies of O. viridis Peyron) and O. deletus Rey stat.rest. (former subspecies of O. subpictus Wollaston).
Some species of the diving beetle tribe Hygrotini (subfamily Hydroporinae) are among the few insects able to tolerate saline concentrations more than twice that of seawater. However, the phylogenetic relationships of the species of Hygrotini, and the origin and evolution of tolerance to salinity in this lineage, are unknown. In this work, we aim to reconstruct how many times salinity tolerance did evolve in Hygrotini, whether this evolution was gradual or if tolerance to hypersalinity could evolve directly from strictly freshwater (FW) species, and to estimate the probabilities of transition between habitats. We build a phylogeny with ca. 45% of the 137 species of Hygrotini, including all major lineages and almost all of the known halophile or tolerant species. We used sequence data of four mitochondrial (COI-5′, COI-3′, 16S + tRNA and NADH1) and three nuclear (28S, 18S and H3) gene fragments, plus ecological data to reconstruct the history of the salinity tolerance using Bayesian inference. Our results demonstrate multiple origins of the tolerance to salinity, although most saline and hypersaline species were concentrated in two lineages. The evolution of salinity was gradual, with no direct transitions from FW to hypersaline habitats, but with some reversals from tolerant to FW species. The oldest transition to saline tolerance, at the base of the clade with the highest number of saline species, was dated in the late Eocene-early Oligocene, a period with decreasing temperature and precipitation. This temporal coincidence suggests a link between increased aridity and the development of tolerance to saline waters, in agreement with recent research in other groups of aquatic Coleoptera.
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