Simultaneous absolute quantification and sequencing of fish environmental DNA in a mesocosm by quantitative sequencing technique.
Simultaneous absolute quantification and sequencing of fish environmental DNA in a mesocosm by quantitative sequencing technique.
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DOI：
10.1038/s41598-021-83318-6
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发表时间：
2021-02-23
影响因子：
4.6
通讯作者：
Minamoto T
中科院分区：
文献类型：
--
作者：
Hoshino T;Nakao R;Doi H;Minamoto T
高通量测序技术和环境DNA（eDNA）分析的结合有可能成为一个强大的工具，全面，非侵入性监测物种在环境中。为了了解eDNA丰度与自然环境中物种丰度之间的相关性，我们必须获得定量的eDNA数据，通常是通过对每个物种的单独测定。近年来发展起来的定量测序（qSeq）技术通过对首次合成DNA时添加到目标序列5′端的随机标签进行计数，可以同时对单个物种的系统发育进行鉴定和定量，本文将qSeq技术应用于eDNA分析，以检验其在生物多样性监测中的有效性。从含有5种鱼类（Hemigrammocypris neglectus、Temminckii、Oryzias latipes、Rhinogobius flumineus和Misgurnus anguillicaudatus）的水族箱中采集超过4天的水样中提取eDNA，并使用TaqMan探针通过qSeq和微流控数字PCR（dPCR）进行定量。在每个采样时间，通过qSeq定量的eDNA丰度与通过dPCR定量的eDNA丰度一致。qSeq和dPCR的相关系数分别为0.643、0.859和0.786。neglectus，O. latipes和M.这表明qSeq可以准确地定量鱼类的eDNA。
The combination of high-throughput sequencing technology and environmental DNA (eDNA) analysis has the potential to be a powerful tool for comprehensive, non-invasive monitoring of species in the environment. To understand the correlation between the abundance of eDNA and that of species in natural environments, we have to obtain quantitative eDNA data, usually via individual assays for each species. The recently developed quantitative sequencing (qSeq) technique enables simultaneous phylogenetic identification and quantification of individual species by counting random tags added to the 5′ end of the target sequence during the first DNA synthesis. Here, we applied qSeq to eDNA analysis to test its effectiveness in biodiversity monitoring. eDNA was extracted from water samples taken over 4 days from aquaria containing five fish species (Hemigrammocypris neglectus, Candidia temminckii, Oryzias latipes, Rhinogobius flumineus, and Misgurnus anguillicaudatus), and quantified by qSeq and microfluidic digital PCR (dPCR) using a TaqMan probe. The eDNA abundance quantified by qSeq was consistent with that quantified by dPCR for each fish species at each sampling time. The correlation coefficients between qSeq and dPCR were 0.643, 0.859, and 0.786 for H. neglectus, O. latipes, and M. anguillicaudatus, respectively, indicating that qSeq accurately quantifies fish eDNA.
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影响因子：
3.5
作者：
Miya M;Sato Y;Fukunaga T;Sado T;Poulsen JY;Sato K;Minamoto T;Yamamoto S;Yamanaka H;Araki H;Kondoh M;Iwasaki W
通讯作者：
Iwasaki W
影响因子：
3.7
作者：
Doi H;Uchii K;Takahara T;Matsuhashi S;Yamanaka H;Minamoto T
通讯作者：
Minamoto T
影响因子：
10.7
作者：
Sato Y;Miya M;Fukunaga T;Sado T;Iwasaki W
通讯作者：
Iwasaki W
影响因子：
3.7
作者：
Hoshino T;Inagaki F
通讯作者：
Inagaki F
影响因子：
2.9
作者：
Arezi, B;Xing, WM;Hogrefe, HH
通讯作者：
Hogrefe, HH