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Geographic differentiation of Cobitis shikokuensis inferred from mtDNA RFLP analysis

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Abstract

Mitochondrial DNA divergence among populations of the Japanese spinous loach Cobitis shikokuensis, endemic to Shikoku Island, was investigated by restricted fragment length polymorphism analysis. A total of 68 restriction sites on DNA fragments from the cytochrome b to D-loop regions and from the 12S rRNA to 16S rRNA regions, amplified by PCR, were analyzed. A total of 12 haplotypes (plus 6 in outgroups) were detected in 268 specimens collected from 19 localities in seven rivers (and 41 specimens from four localities in three rivers in outgroups). Three of the seven river populations of C. shikokuensis were shown to have unique haplotypes, and four of the seven river populations were monomorphic. The nested structure of the haplotype network for populations of C. shikokuensis exhibited two large clades corresponding to (1) populations from the Shimanto River and its neighbors and (2) two genetically divergent populations in the Shigenobu and Iwamatsu Rivers. The population from the Shimanto River, the largest river inhabited by C. shikokuensis, maintains great haplotype diversity as well as the allozyme diversity previously reported. On the other hand, populations from the Hiji River, the second largest river, which exhibited the highest allozyme diversity, were monomorphic in their mtDNA. The nested clade analysis (NCA) revealed that past fragmentation between the above two clades could occur in the initial distribution process of C. shikokuensis. The large genetic divergence of two river populations from the Shigenobu and Iwamatsu Rivers was inferred to be caused by a process of long distance colonization and fragmentation. MtDNA introgression into the Hiji River population from southern river populations was suggested. Taking genetic divergence into consideration, each river population of C. shikokuensis should be conserved separately as like a distinct species, and conservation programs for the small populations showing less genetic variability should be invoked as soon as possible.

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Acknowledgments

I am grateful to Dr. H. Sakai (National Fisheries University) for valuable advice and critical reading of the manuscript. Special thanks are given to Drs. M. Nishida (University of Tokyo), K. Watanabe (Kyoto University), and A. Kawaguchi (University of Ryukyus) for teaching me the techniques of mtDNA analysis, and to Dr. Margaret Dooley Ohto for checking the English. Thanks also go to Mr. Y. Suzawa (Institute of River Biology), M. Okamoto (Ecosystem Co., Ltd.), H. Takahashi (Sumiko Techno Research Incorporated), and K. Hashimoto (Osaka Sekkei Co., Ltd.) for collecting specimens. The present study was supported in part by a consignment fund from the Fisheries Agency, Japan, to develop techniques for reproducing endangered freshwater fishes.

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  1. Ehime Prefectural Chuyo Fisheries Experimental Station, 121-3 Mori, Iyo, Ehime, 799-3125, Japan

    Takaaki Shimizu

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Correspondence to Takaaki Shimizu.

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Shimizu, T. Geographic differentiation of Cobitis shikokuensis inferred from mtDNA RFLP analysis. Ichthyol Res 55, 101–111 (2008). https://doi.org/10.1007/s10228-007-0013-2

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  • DOI: https://doi.org/10.1007/s10228-007-0013-2

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