The generation of the high species diversity of insects in Japan was profoundly influenced by the formation of the Japanese Archipelago. We explored the species diversification and biogeographical history of the Nemouridae Billberg, 1820 family in the Japanese Archipelago using mitochondrial DNA and nuclear DNA markers. We collected 49 species among four genera: Indonemoura Baumann, 1975; Protonemura Kempny, 1898; Amphinemura, Ris 1902 and Nemoura Latreille, 1796 in Japan, China, South Korea and North America. We estimated their divergence times—based on three molecular clock node calibrations—using Bayesian phylogeography approaches. Our results suggested that Japanese Archipelago formation events resulted in diversification events in the middle of the Cretaceous (<120 Ma), speciation in the Paleogene (<50 Ma) and intra-species diversification segregated into eastern and western Japan of the Fossa Magna region at late Neogene (20 Ma). The Indonemoura samples were genetically separated into two clades—that of Mainland China and that of Japan. The Japanese clade clustered with the Nemouridae species from North America, suggesting the possibility of a colonisation event prior to the formation of the Japanese Archipelago. We believe that our results enhanced the understanding both of the origin of the species and of local species distribution in the Japanese Archipelago.
15The generation of the high species diversity of insects in Japan was profoundly influenced by the 16 formation of the Japanese Archipelago. We explored the species diversification and biogeographical 17 history of the Nemouridae family in the Japanese Archipelago using mitochondrial DNA and nuclear 18 DNA markers. We collected 49 species among four genera: Indonemoura, Protonemura, 19Amphinemura and Nemoura in Japan, China, South Korea and North America. We estimated their 20 divergence times-based on three molecular clock node calibrations-using Bayesian phylogeography 21 approaches. Our results suggested that Japanese Archipelago formation events resulted in 22 diversification events in the middle of the Cretaceous (<120 Ma), speciation in the Paleogene (<50 Ma) 23 and intra-species diversification segregated into eastern and western Japan of the Fossa Magna region 24 at late Neogene (20 Ma). The Indonemoura samples were genetically separated into two clades-that 25 of Mainland China and that of Japan. The Japanese clade clustered with the Nemouridae species from 26North America, suggesting the possibility of a colonisation event prior to the formation of the Japanese 27Archipelago. We believe that our results enhanced the understanding both of the origin of the species 28 and of local species distribution in the Japanese Archipelago. 29 30
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