Detection of endangered Itasenpara bitterling using simple environmental DNA analysis

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  • 簡易的な環境DNA分析方法を用いた絶滅危惧種イタセンパラの検出
  • カンイテキ ナ カンキョウ DNA ブンセキ ホウホウ オ モチイタ ゼツメツ キグシュ イタセンパラ ノ ケンシュツ

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Abstract

<p>Environmental DNA (eDNA - genetic material released from an organism into the environment, such as water), which has recently gained attention as a new area of study, has applications in confirming the presence and estimating the biomass of target species, especially endangered fishes, as well as aiding an understanding of the general fish fauna. However, eDNA analysis requires expensive equipment and reagents, and is presently subjected to restrictions on widespread use, especially for environmental education and public awareness. For future conservation studies of the endangered Itasenpara bitterling Acheilognathus longipinnis, and subsequent application to other species, a species-specific PCR amplification and simple method of eDNA analysis was developed. Species-specific primer pairs based on nucleotide sequences were designed for the Itasenpara bitterling and closely related species. These enabled Itasenpara bitterling-specific amplification in PCR experiments on DNA samples obtained from fin tissues. Subsequently, species-specific amplification of eDNA samples obtained from a watershed containing Itasenpara bitterling habitat was confirmed using a simple PCR-based experimental method, although the amplification ratio varied, probably due to habitat conditions and bitterling growth stage.</p>

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