To investigate the genetic differentiation between the Bonin (Ogasawara) Islands' freshwater goby Rhinogobius sp. Bonin Island (BI) form (Ogasawara-yoshinobori) and the Japan-Ryukyu Archipelago relatives, the mitochondrial DNA (mtDNA) phylogeny of Japanese Rhinogobius species was inferred from partial nucleotide sequences of the mitochondrial NADH dehydrogenase 5 subunit (ND5) gene (945 bp). The resultant tree showed that the Bonin Islands group separated first from the other Japanese lineage, and a test calculation indicated the divergence date to be approximately 3 million years BP. Although it is necessary to use a more reliable estimate to confirm the divergence date, Rhinogobius sp. BI has retained its mtDNA lineage in the islands for millions of years.Key words Rhinogobius 路 mtDNA 路 Genetic divergence 路 Geographical isolation entiation between Rhinogobius sp. BI and the JapanRyukyu Archipelago relatives by using mitochondrial DNA (mtDNA) analysis, information on phylogenetic relationships, and genetic divergences based on the mtDNA sequences providing useful evidence.
Materials and MethodsA total of 16 individuals of Rhinogobius sp. BI were collected from three localities on Chichijima Island [Kiyose River (n = 1), Minamifukurosawa (n = 3), and Yatsuse River (n = 6)] and two on Hahajima Island [Kitako (n = 3) and Okiko (n = 3)]. The specimens were deposited in the National Science Museum, Tokyo (catalogue number ). Nine other species of Rhinogobius were also used in the study: Rhinogobius giurinus [Okinawa Island (Okinawa)], Rhinogobius duospilus (bought from an aquarium retailer), Rhinogobius flumineus [Shigenobu River (Ehime), Yasu River (Shiga), and Fuefuki River (Yamanashi)], Rhinogobius sp. crossband (CB) form [Shigenobu River (Ehime) and Okinawa Island (Okinawa)], Rhinogobius sp. cobalt (CO) form [Nezugaseki River (Yamagata)], Rhinogobius sp. dark (DA) form [Kushimoto (Wakayama) and Iriomote Island (Okinawa)], Rhinogobius sp. depressed large-dark (DL) form [Iriomote Island (Okinawa)], Rhinogobius sp. large-dark (LD) form [Abe River (Shizuoka)], and Rhinogobius sp. orange (OR) form [Akigawa River (Tokyo) and Lake Biwa (Shiga)]. The abbreviations of each species except R. giurinus, R.