Abstract
We developed a novel molecular genetic method using restriction fragment length polymorphisms (RFLPs) to identify the taxonomically debatable Japanese sand lances of the genus Ammodytes. Partial nucleotide sequences of the mitochondrial DNA cytochrome c oxidase subunit 1 (COI) gene of Ammodytes japonicus (n = 405), Ammodytes heian (n = 284), and Ammodytes hexapterus (n = 15) were determined and found to contain species-specific restriction sites for HindIII-HF and RsaI enzymes. The observed RFLP patterns from a combination of two enzymes were specific in each species except for specimens with rare haplotypes of A. japonicus (1.5 %) and A. heian (2.1 %), whereas detailed morphological analysis using 29 characters failed to discriminate the COI-identified specimens of these two species collected from the same locality. The developed molecular method will allow rapid and precise identification of the Japanese sand lances, especially for the sympatric A. japonicus and A. heian, thereby providing a reliable means for ecological research and stock management of these species.
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Acknowledgments
The authors thank two anonymous reviewers for their critical comments and suggestions. Dr. Tadashi Misaka of the Wakkanai Fisheries Research Institute of the Fisheries Research Department, Hokkaido Research Organization, Mr. Mugiho Ataka and Mr. Atsushi Saga of Marino-Forum 21, and Dr. Noriko Amiya of the School of Marine Biosciences, Kitasato University, kindly provided specimens for this study. Thanks are also due to the Freshwater Experiment Station of the Kanagawa Prefectural Fisheries Technology Center which kindly allowed us to use their instruments. This study was partly supported by the Japan Society for the Promotion of Science KAKENHI Grant 15K07543 to J.A.
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Tanaka, C., Aoki, R., Ida, H. et al. Molecular genetic identification of Japanese sand lances using mitochondrial DNA cytochrome c oxidase subunit 1 restriction fragment length polymorphisms. Fish Sci 82, 887–895 (2016). https://doi.org/10.1007/s12562-016-1021-9
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DOI: https://doi.org/10.1007/s12562-016-1021-9