文章基本信息

标题：Simultaneous absolute quantification and sequencing of fish environmental DNA in a mesocosm by quantitative sequencing technique
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作者：Tatsuhiko Hoshino ; Ryohei Nakao ; Hideyuki Doi 等
期刊名称：Scientific Reports
电子版ISSN：2045-2322
出版年度：2021
卷号：11
期号：1
页码：4372
DOI：10.1038/s41598-021-83318-6
出版社：Springer Nature
摘要：Abstract 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.
其他摘要：Abstract 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.