Molecular evidence for the existence of five cryptic species within the Japanese species of marphysa (Annelida: Eunicidae) known as “iwa-mushi”

Molecular phylogenetic analyses were conducted to clarify how genetically homogeneous the common Japanese species of Marphysa known as “Iwa-mushi” is. This is a well-known polychaete used as a fishing bait that was first described as Marphysa iwamushi Izuka, 1907 (type locality: Japan and Taiwan) and later synonymized to Marphysa sanguinea (Montagu, 1813) (type locality: England). The nucleotide sequences of a nuclear gene (18S rRNA) and two mitochondrial genes (16S rRNA and cytochrome c oxidase subunit I (COI)) were compared between specimens newly collected from 14 localities in Japan including commercially sold fishing baits and DDBJ/ENA/GenBank data for congeneric species. Our results show that the Japanese “Iwa-mushi” is not a single species but a species complex comprising five genetically well-separated clades that were tentatively designated as five undetermined species (Marphysa spp. A, B, C, D, and E). It is unclear whether any of these species corresponds to M. iwamushi. The COI nucleotide sequence of Marphysa sp. A was almost identical to that of M. victori Lavesque, Daffe, Bonifácio & Hutchings, 2017 (type locality: France) and M. bulla Liu, Hutchings & Kupriyanova, 2018 (type locality: China), suggesting that they are conspecific and supporting the hypothesis of Lavesque et al. (2017) that the population in France was introduced from Japan. The COI sequence of Marphysa sp. B corresponds to that of M. maxidenticulata Liu, Hutchings & Kupriyanova, 2018 (type locality: China). Marphysa sp. E was found only in Tokyo Bay and may be an alien species that was introduced by importing live fishing bait. Our results indicate that the European species M. sanguinea is not distributed in Japanese waters.