Effects of sampling strategies and DNA extraction methods on eDNA metabarcoding: A case study of estuarine fish diversity monitoring

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Publicat a:Zoological Research vol. 43, no. 2 (Mar 2022), p. 192
Autor principal: Ruan, Hui-Ting
Altres autors: Wang, Rui Li, Li, Hong-Ting, Liu, Li, Kuang, Tian-Xu, Li, Min, Zou, Ke-Shu
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Kunming Institute of Zoology, The Chinese Academy of Sciences
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024 7 |a 10.24272/j.issn.2095-8137.2021.331  |2 doi 
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100 1 |a Ruan, Hui-Ting  |u University Joint Laboratory of Guangdong Province, Hong Kong and Macao Region on Marine Bioresource Conservation and Exploitation, College of Marine Science, South China Agricultural University, Guangzhou, Guangdong 510642, China 
245 1 |a Effects of sampling strategies and DNA extraction methods on eDNA metabarcoding: A case study of estuarine fish diversity monitoring 
260 |b Kunming Institute of Zoology, The Chinese Academy of Sciences  |c Mar 2022 
513 |a Case Study Journal Article 
520 3 |a Environmental DNA (eDNA) integrated with metabarcoding is a promising and powerful tool for species composition and biodiversity assessment in aquatic ecosystems and is increasingly applied to evaluate fish diversity. To date, however, no standardized eDNA-based protocol has been established to monitor fish diversity. In this study, we investigated and compared two filtration methods and three DNA extraction methods using three filtration water volumes to determine a suitable approach for eDNA-based fish diversity monitoring in the Pearl River Estuary (PRE), a highly anthropogenically disturbed estuarine ecosystem. Compared to filtration-based precipitation, direct filtration was a more suitable method for eDNA metabarcoding in the PRE. The combined use of DNeasy Blood and Tissue Kit (BT) and traditional phenol/chloroform (PC) extraction produced higher DNA yields, amplicon sequence variants (ASVs), and Shannon diversity indices, and generated more homogeneous and consistent community composition among replicates. Compared to the other combined protocols, the PC and BT methods obtained better species detection, higher fish diversity, and greater consistency for the filtration water volumes of 1 000 and 2 000 mL, respectively. All eDNA metabarcoding protocols were more sensitive than bottom trawling in the PRE fish surveys and combining two techniques yielded greater taxonomic diversity. Furthermore, combining traditional methods with eDNA analysis enhanced accuracy. These results indicate that methodological decisions related to eDNA metabarcoding should be made with caution for fish community monitoring in estuarine ecosystems. 
610 4 |a China Agricultural University 
651 4 |a China 
651 4 |a South China Sea 
651 4 |a Pearl River Estuary 
653 |a Community composition 
653 |a Chloroform 
653 |a Environmental monitoring 
653 |a Fish 
653 |a Estuaries 
653 |a Species composition 
653 |a Phenols 
653 |a Brackishwater environment 
653 |a Species diversity 
653 |a Biodiversity 
653 |a Estuarine dynamics 
653 |a Water purification 
653 |a Bottom trawling 
653 |a Aquatic ecosystems 
653 |a Ecological monitoring 
653 |a Decision analysis 
653 |a Diversity indices 
653 |a Methods 
653 |a Composition 
653 |a Filtration 
653 |a Ecosystems 
653 |a Agricultural production 
653 |a Estuarine fisheries 
653 |a Anthropogenic factors 
653 |a Deoxyribonucleic acid--DNA 
653 |a Surveys 
653 |a Nucleotide sequence 
653 |a Trawling 
653 |a Estuarine ecosystems 
653 |a River ecology 
653 |a Taxonomy 
653 |a Fishing 
653 |a Brackishwater fishes 
653 |a Environmental DNA 
653 |a Commercial fishing 
653 |a Fisheries 
653 |a Laboratories 
653 |a Environmental 
700 1 |a Wang, Rui Li  |u University Joint Laboratory of Guangdong Province, Hong Kong and Macao Region on Marine Bioresource Conservation and Exploitation, College of Marine Science, South China Agricultural University, Guangzhou, Guangdong 510642, China 
700 1 |a Li, Hong-Ting  |u Key Laboratory for Sustainable Utilization of Open-sea Fishery, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong 510300, China 
700 1 |a Liu, Li  |u University Joint Laboratory of Guangdong Province, Hong Kong and Macao Region on Marine Bioresource Conservation and Exploitation, College of Marine Science, South China Agricultural University, Guangzhou, Guangdong 510642, Chinac 
700 1 |a Kuang, Tian-Xu  |u University Joint Laboratory of Guangdong Province, Hong Kong and Macao Region on Marine Bioresource Conservation and Exploitation, College of Marine Science, South China Agricultural University, Guangzhou, Guangdong 510642, China 
700 1 |a Li, Min 
700 1 |a Zou, Ke-Shu 
773 0 |t Zoological Research  |g vol. 43, no. 2 (Mar 2022), p. 192 
786 0 |d ProQuest  |t Science Database 
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