定量PCR を用いた有害ラフィド藻Chattonella marina およびHeterosigma akashiwo 定量法の検討

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定量PCR を用いた有害ラフィド藻Chattonella marina およびHeterosigma akashiwo 定量法の検討

フォーマット:
雑誌論文
Kyushu Univ. Production 九州大学成果文献
タイトル(他言語):
Development of a Quantification Method of Harmful Raphidophytes Chattonella marina and Heterosigma akashiwo by the quantitative PCR assay
責任表示:
向井, 幸樹(九州大学大学院農学研究院水産生物環境学研究室)
Mukai, Koki(Laboratory of Marine Environmental Science, Faculty of Agriculture, Kyushu University)
太田, 耕平(九州大学大学院農学研究院水産生物環境学研究室 | 九州大学大学院農学研究院海洋生物学研究室 : 准教授)

Ohta, Kohei(Laboratory of Marine Environmental Science, Faculty of Agriculture, Kyushu University | Laboratory of Marine Biology, Faculty of Agriculture, Kyushu University : Associate Professor)
島崎, 洋平(九州大学大学院農学研究院水産生物環境学研究室 : 准教授)
Shimasaki, Yohei(Laboratory of Marine Environmental Science, Faculty of Agriculture, Kyushu University : Associate Professor)
鵜木(加藤), 陽子(九州大学大学院農学研究院水産生物環境学研究室)
Unoki-Kato, Yoko(Laboratory of Marine Environmental Science, Faculty of Agriculture, Kyushu University)
大嶋, 雄治(九州大学大学院農学研究院水産生物環境学研究室 : 教授)
Oshima, Yuji(Laboratory of Marine Environmental Science, Faculty of Agriculture, Kyushu University : Professor)

全て表示
本文言語:
日本語
出版情報:
九州大学大学院農学研究院学芸雑誌. 72, (2), pp. 39-46, 2017-09-08. 九州大学大学院農学研究院
バージョン:
Publisher
概要:
定量PCR を用いて,海水中に存在する有害ラフィド藻Chattonella marinaおよびHeterosigma akashiwo遺伝子の検出法の検討を行った.プライマーおよびプロー ブ(TaqMan プローブ) は,C. marinaについてはmtDNA のcyt c 領域に,H. akashiwo についてはrRNA遺伝子のITS 領域に設定した.標的遺伝子の希釈系列を鋳型に用いた定量PCR の結果,高い増幅効率(C.marina:80.0 %; H. akashiwo:82.6 %)が確認され,さらにCt 値と遺伝子コピー数との間に高い正の相関(C. marina:R^2 = 0 . 991; H. akashiwo:R^2 = 0 . 998)が認められた.また,他種藻類(渦鞭毛藻,珪藻等)からDNeasy Plant Mini Kit(QIAGEN)を用いて抽出したDNA に対して非特異的増幅を示さなかった.さらに両種を対数増殖期から衰退期まで培養し,検鏡および定量PCR による細胞密度の定量を行った結果,両定量値間には有意(p<0.05)な正の相関(C. marina:R^2 =0.84;H. akashiwo:R^2 =0.96)が観察された.以上の結果から,DNA 抽出から定量PCR に至る本法の有用性が強く示唆された. 続きを見る
目次:
はじめに / 材料と方法 / 結果 / 考察 / 謝辞 / 要約
はじめに / 材料と方法 / 結果 / 考察 / 謝辞 / 要約

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