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Mitochondrial DNA sequence divergence between “Kunimasu” Oncorhynchus kawamurae and “Himemasu” O. nerka in Lake Saiko, Yamanashi Prefecture, Japan, and their identification using multiplex haplotype-specific PCR

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Abstract

In order to develop a simplified method for discriminating between Oncorhynchus kawamurae (Kunimasu) and sympatric Oncorhynchus nerka (Himemasu), sequence variations of mitochondrial DNA control region were examined on 71 specimens of Kunimasu and 103 of Himemasu. Sequence analyses for 530 bp revealed 3 variable sites, which defined three haplotypes in Kunimasu and one in Himemasu. Based on determined nucleotide sequences, species-specific primers were designed for the multiplex haplotype-specific polymerase chain reaction, which amplified ca. 900 and 1,040 bp fragments in Kunimasu and Himemasu, respectively. This simple rapid approach can be used to distinguish between the two species without the need to sacrifice specimens.

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Acknowledgments

We thank Yasuaki Miura, Y. Watanabe, Hisashi Miura, Hiroshi Miura, Manabu Miura, K. Mitsui, T. Aoyagi, N. Kaji, M. Ohashi, S. Sasaki, M. Nozu and N. Sakai for their help in collecting specimens, and A. Tokairin and K. Kimura for their assistance in the laboratory. We also thank G. S. Hardy for his critical reading of the manuscript. This study was partly supported by the Core Stage Backup Research Fund of Kyoto University and a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (24370036).

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Authors and Affiliations

  1. Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kyoto, 606-8502, Japan

    Kouji Nakayama & Nozomu Muto

  2. The Kyoto University Museum, Kyoto University, Kyoto, 606-8501, Japan

    Tetsuji Nakabo

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  1. Kouji Nakayama
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  2. Nozomu Muto
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  3. Tetsuji Nakabo
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Correspondence to Kouji Nakayama.

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Nakayama, K., Muto, N. & Nakabo, T. Mitochondrial DNA sequence divergence between “Kunimasu” Oncorhynchus kawamurae and “Himemasu” O. nerka in Lake Saiko, Yamanashi Prefecture, Japan, and their identification using multiplex haplotype-specific PCR. Ichthyol Res 60, 277–281 (2013). https://doi.org/10.1007/s10228-013-0346-y

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  • DOI: https://doi.org/10.1007/s10228-013-0346-y

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