海洋渔业2025，Vol.47Issue(4)：524-533,10.
基于荧光定量PCR方法的中华鲟环境DNA获取优化
Optimization of environmental DNA acquisition of Acipenser sinensis based on fluorescent quantitative PCR method
摘要
Abstract
The amount and fragment size of DNA released into the environment were different due to various life histories of species,so the enrichment and extraction effects of different eDNA need to be evaluated for specific species and water areas.In this study,the indoor mariculture Chinese sturgeon(Acipenser sinensis)was selected as the target species.Using the qPCR method,four kinds of filter materials(mixed cellulose,cellulose acetate,glass fiberl and polypropylene)with diameter of 50 mm and aperture of 0.45 μm were employed,and three kinds of DNA extraction kits(DNeasy Blood&Tissue Kit,Blood Genomic DNA Extraction Reagent,Mag-Bind Soil DNA Kit)were adopted to compare the effects of eDNA acquisition.The enrichment effects of three apertures(0.22,0.45,1.2 μm)of better materials were evaluated.The results showed that the filter material,aperture size and DNA kit had significant effects on the qualitative and quantitative identification of A.sinensis.Different aperture sizes of filter membranes should be used rationally in relation to the field environment.The 0.45 μm mixed cellulose material enriched more eDNA among four filter materials,and Tengen DNeasy Blood and Tissue Kit showed a higher eDNA yield of(457.33±35.10)copies·μL-1.A series of eDNA technology operation procedures for A.sinensis is optimized,which can provide a reference for the application of environmental DNA technology in monitoring the distribution of A.sinensis in the Yangtze River Estuary.关键词
中华鲟/滤膜材质/孔径大小/eDNA提取Key words
Acipenser sinensis/filter material/aperture size/eDNA extraction分类
农业科技引用本文复制引用
王肖静,冯广朋,王友保,朱佳志,姜伟,张涛,杨刚..基于荧光定量PCR方法的中华鲟环境DNA获取优化[J].海洋渔业,2025,47(4):524-533,10.基金项目
三峡工程鱼类资源保护湖北省重点实验室开放课题项目(2022051-ZHX) （2022051-ZHX）
WWF蔚蓝星球项目(PORO001782) （PORO001782）
农业农村部政采课题(17230156) （17230156）
