絶滅危惧種を多く含む水生甲虫・水生カメムシで環境 DNA 分析に必要な参照 DNA 配列を網羅的に整備

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2025-10-21 兵庫県立大学

兵庫県立大学と兵庫県立人と自然の博物館を中心とする研究チームは、絶滅危惧種を多く含む日本産の水生甲虫・水生カメムシの環境DNA分析に必要な参照DNA配列データベースを初めて網羅的に整備した。両群の約7割の属をカバーし、計198種の16S rRNA配列を決定。これにより、環境DNA解析で多数の水生昆虫の種判定が可能となり、従来の調査では困難だった希少種の分布把握や保全計画立案に大きく貢献する。収集した標本は複数の博物館に保管され、分類学的改訂にも対応できる。研究は国内5機関の協力で行われ、外来種の侵入や農地開発で生息地が失われつつある淡水生態系の保全に重要な基盤を提供するものとなった。

絶滅危惧種を多く含む水生甲虫・水生カメムシで環境 DNA 分析に必要な参照 DNA 配列を網羅的に整備
(図) 本研究で参照 DNA 配列を決定した水生昆虫の一部。いずれも平澤桂撮影。

<関連情報>

環境 DNA メタバーコーディング分析に向けた日本産水生甲虫及びカメムシのミトコンドリア DNA の 16SrRNA 領域配列と証拠標本のコレクション Mitochondrial DNA 16S region and voucher specimen collection of Japanese aquatic Coleoptera and Hemiptera for environmental DNA metabarcoding analyses

Naoyuki Nakahama, Kei Hirasawa, Masaya Kato, Kohei Watanabe, Seikan Kurata, Masakazu Hayashi
Zookeys  Published:23 Sep 2025
DOI:https://doi.org/10.3897/zookeys.1253.146226

Abstract

Aquatic coleopteran and hemipteran insects primarily inhabit lentic waters, many of which are at risk of extinction due to development, agriculture, and invasive alien species. Environmental DNA (eDNA) analysis has recently emerged as a powerful tool for conducting comprehensive distribution surveys. The cytochrome c oxidase subunit I (COI) universal primers are conventionally used for DNA barcoding but they often result in non-specific amplification and frequent amplication failures. Primers in the mitochondrial DNA (mtDNA) 16S rRNA region that alleviate these issues have been developed and are considered helpful for eDNA analysis. It is necessary to accumulate reference sequences of the mtDNA 16S rRNA region in aquatic coleopteran and hemipteran insects. However, molecular identification at the genus or species level remains challenging, as only a few of these insect groups in Japan have registered reference DNA sequences for both the mtDNA COI and 16S rRNA. Therefore, we constructed a comprehensive dataset of the mtDNA 16S rRNA region for these insects distributed in Japan. As a result of this study, we were able to obtain partial sequences of the mtDNA 16S rRNA region from 140 coleopteran taxa (35.5% of Japanese aquatic species or subspecies) and 58 hemipteran taxa (45.3% of Japanese aquatic species or subspecies). These voucher specimens were deposited in four research institutions. The DNA sequence datasets are expected to significantly contribute as an essential database for eDNA analysis and other DNA metabarcoding studies.

生物環境工学
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