Three new species in Tetrastemma Ehrenberg, 1828 (Nemertea, Monostilifera) from sublittoral to upper bathyal zones of the northwestern Pacific.

Monostiliferous nemerteans in the genus Tetrastemma Ehrenberg, 1828 are generally characterized as having four eyes, and they occur worldwide, from the intertidal zone to the deep-sea bottom. Recent extensive sampling of Tetrastemma has explored the high species diversity, including many undescribed forms, but phylogenic analysis has revealed non-monophyly of the genus. We herein describe three new species of the genus ( T.album sp. nov. , T.persona sp. nov. , and T.shohoense sp. nov. ) from northwestern Pacific waters based on specimens collected by dredging or by use of a remotely operated vehicle at depths of 116-455 m. Since anatomical and histological characters traditionally used in systematics of the genus are sometimes interspecifically uniform, a histology-free approach is applied for the species descriptions in this study. To confirm the generic affiliation of the new species, a molecular phylogenetic analysis based on partial sequences of cytochrome c oxidase subunit I, 16S rRNA, 18S rRNA, 28S rRNA, and histone H3 genes was performed. Our result shows that all three new species are nested in a subclade formed by species from the North Pacific and American Atlantic, inferring that geographic distribution does not reflect the cladogenesis of Tetrastemma . Furthermore, two Tetrastemma species with a cylindrical stylet basis, T.freyae Chernyshev et al., 2020 from off the coast of India and Hawaii and T.shohoense sp. nov. from Shoho Seamount, Japan, constitute a clade in the resulting tree.