BackgroundSince the discovery of deep-sea chemosynthesis-based communities, much work has been done to clarify their organismal and environmental aspects. However, major topics remain to be resolved, including when and how organisms invade and adapt to deep-sea environments; whether strategies for invasion and adaptation are shared by different taxa or unique to each taxon; how organisms extend their distribution and diversity; and how they become isolated to speciate in continuous waters. Deep-sea mussels are one of the dominant organisms in chemosynthesis-based communities, thus investigations of their origin and evolution contribute to resolving questions about life in those communities.Methodology/Principal FindingWe investigated worldwide phylogenetic relationships of deep-sea Bathymodiolus mussels and their mytilid relatives by analyzing nucleotide sequences of the mitochondrial cytochrome c oxidase subunit I (COI) and NADH dehydrogenase subunit 4 (ND4) genes. Phylogenetic analysis of the concatenated sequence data showed that mussels of the subfamily Bathymodiolinae from vents and seeps were divided into four groups, and that mussels of the subfamily Modiolinae from sunken wood and whale carcasses assumed the outgroup position and shallow-water modioline mussels were positioned more distantly to the bathymodioline mussels. We provisionally hypothesized the evolutionary history of Bathymodilolus mussels by estimating evolutionary time under a relaxed molecular clock model. Diversification of bathymodioline mussels was initiated in the early Miocene, and subsequently diversification of the groups occurred in the early to middle Miocene.Conclusions/SignificanceThe phylogenetic relationships support the “Evolutionary stepping stone hypothesis,” in which mytilid ancestors exploited sunken wood and whale carcasses in their progressive adaptation to deep-sea environments. This hypothesis is also supported by the evolutionary transition of symbiosis in that nutritional adaptation to the deep sea proceeded from extracellular to intracellular symbiotic states in whale carcasses. The estimated evolutionary time suggests that the mytilid ancestors were able to exploit whales during adaptation to the deep sea.

We sequenced the mitochondrial ND4 gene to elucidate the evolutionary processes of Bathymodiolus mussels and mytilid relatives. Mussels of the subfamily Bathymodiolinae from vents and seeps belonged to 3 groups and mytilid relatives from sunken wood and whale carcasses assumed the outgroup positions to bathymodioline mussels. Shallow water mytilid mussels were positioned more distantly relative to the vent/seep mussels, indicating an evolutionary transition from shallow to deep sea via sunken wood and whale carcasses. Bathymodiolus platifrons is distributed in the seeps and vents, which are approximately 1500 km away. There was no significant genetic differentiation between the populations. There existed high gene flow between B. septemdierum and B. brevior and low but not negligible gene flow between B. marisindicus and B. septemdierum or B. brevior, although their habitats are 5000-10 000 km away. These indicate a high adaptability to the abyssal environments and a high dispersal ability of Bathymodiolus mussels.

In order to invade and adapt to deep-sea environments, shallow-water organisms have to acquire tolerance to high hydrostatic pressure, low water temperature, toxic methane and hydrogen sulfide, and feeding strategies not relying on photosynthetic products. Our previous study showed that the "evolutionary stepping stone hypothesis", which assumes that organic falls can act as stepping-stones to connect shallow sea with deep sea, was supported in Mytilidae. However, it is not known whether other bivalves constituting chemosynthetic communities experienced the same evolutionary process or different processes from mytilid mussels. Therefore, here, we performed phylogenetic analyses by sequencing the nuclear 18S rRNA and mitochondrial COI genes of solemyid and thyasirid bivalves. In Solemyidae, the two genera Solemya and Acharax formed each clade, the latter of which was divided into three subgroups. The Solemya clade and one of the Acharax subgroups diverged in the order of shallow-sea residents, whale-bone residents, and deep-sea vent/seep residents, which supported the "evolutionary stepping stone hypothesis". Furthermore, in Thyasiridae, the two genera Thyasira and Maorithyas formed a paraphyletic group and the other genera, Adontorhina, Axinopsis, Axinulus, Leptaxinus, and Mendicula, formed a clade. The "evolutionary stepping stone hypothesis" was not seemingly supported in the other lineages of Solemyidae and Thyasiridae.