Species identification of the mud snails Batillaria multiformis and B. attramentaria by PCR-RFLP method : the advantages of two restriction enzymes

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  • Nakai Shizuko
    Department of Marine Science and Resources, College of Bioresource Sciences Nihon University
  • Itoi Shiro
    Department of Marine Science and Resources, College of Bioresource Sciences Nihon University
  • Kanaya Gen
    National Institute for Environmental Studies
  • Itoh Hajime
    National Institute for Environmental Studies
  • Miura Osamu
    Faculty of Agriculture and Marine Science, Kochi University

書誌事項

タイトル別名
  • Species identification of the mud snails <i>Batillaria multiformis</i> and <i>B. attramentaria</i> by PCR-RFLP method—the advantages of two restriction enzymes—

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<p>The habitats of Batillaria multiformis (Near Threatened species) and Batillaria attramentaria are often overlapping on a mudflat, and young individuals of two Batillaria snails were difficult to identify by shell morphology. This is one of the causes that the study, such as population dynamics of two Batillaria species, did not advance. Recently, species identification by DNA analysis for B. multiformis and B. attramentaria was developed using a PCR-RFLP method. However, using only one restriction enzyme and universal primer may lead to misidentification. Therefore, this study examined a more precise method using two restriction enzymes for the PCR-RFLP technique. The mitochondrial DNA gene encoding cytochrome c oxidase subunit I (COI) was amplified, and two restriction enzymes, PstI and HinfI, were selected for this method. The number and position of bands after the agarose gel electrophoresis enable us to identify the two Batillaria species. After the effectiveness of this method was confirmed by adult B. multiformis and B. attramentaria that had each typical shell shape, young Batillaria snail samples that were difficult for morphological identification were identified by this PCR-RFLP method. We proved that the proposed PCR-RFLP identification method is simple and reliable in identifying the morphologically similar B. multiformis and B. attramentaria.</p>

収録刊行物

  • Plankton and Benthos Research

    Plankton and Benthos Research 18 (1), 47-51, 2023-02-28

    日本プランクトン学会、日本ベントス学会

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