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Fish eDNA detection and its technical optimization: A case study of Acanthopagrus latus

文献类型： 外文期刊

作者： Sun, Meijing 1 ; Guo, Yu 2 ; Zhao, Naiqian 4 ; Zhang, Shuo 1 ; Pei, Kun 5 ; Qin, Chuanxin 2 ;

作者机构： 1.Shanghai Ocean Univ, Coll Marine Sci, Shanghai 201306, Peoples R China

2.Chinese Acad Fishery Sci, South China Sea Fisheries Res Inst, Guangzhou 510300, Peoples R China

3.Natl Agr Expt Stn Fishery Resources & Environm Da, Shenzhen 518120, Peoples R China

4.Fangchenggang Fishery Radio, Fangchenggang 538001, Peoples R China

5.Fangchenggang Fisheries Technol Extens Stn, Fangchenggang 538001, Peoples R China

关键词： Environmental DNA; Detection technology; Water filter; Preservation method; Water bathing time

期刊名称：MARINE ENVIRONMENTAL RESEARCH （ 影响因子：3.737； 五年影响因子：4.338 ）

ISSN： 0141-1136

年卷期： 2022 年 176 卷

页码：

收录情况： SCI

摘要： The development of a standardized eDNA detection process is the primary step in improving the accuracy and efficiency of eDNA detection. In this study, primers and probes with high specificity were selected to identify the COI gene of Acanthopagrus latus. Through experiments on the influence of different water quantities, methods of water sample preservation and water bathing times on the result of eDNA detection, the accuracy of this method for extracted water samples was improved. Specifically, a water bathing time of 6 h provided an optimal eDNA concentration from the water sample. After 6 h, the concentration began to decrease, so 6 h was determined to be the best water bathing time for A. latus. Five water extraction volumes (250 mL, 500 mL, 1 L, 2 L, and 3 L) were tested, and there was a positive correlation between water extraction volume and the DNA concentration in the water sample. Different water sample preservation methods were also compared, and it was found that at <= 7 d, the concentration obtained with the cryopreservation method for different water extraction volumes was higher than that obtained with the ethanol preservation method. In this study, we established and optimized a technical procedure for eDNA-based detection of A. latus in aquatic environments. We hope to apply this method in field investigations and provide a reference for the study of eDNA in other fishes.

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