基于环境DNA技术的红鳍笛鲷生物量评估方法研究  

Study on biomass evaluation method of Lutjanus erythropterus based on eDNA technology

作　　者：辛益 邢晓东 郭禹 秦传新[2,3,4,5] 于刚 马振华[2,3,4,5] 王兴强 XIN Yi;XING Xiaodong;GUO Yu;QIN Chuanxin;YU Gang;MA Zhenhua;WANG Xingqiang(School of Marine Science and Fisheries,Jiangsu Ocean University,Lianyungang Jiangsu 222005,China;Tropical Aquatic Research and Development Center,South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Lingshui Hainan 572426,China;Key Laboratory of Efficient Utilization and Processing of Marine Fishery Resources of Hainan Province,Sanya Tropical Fisheries Research Institute,Sanya Hainan 572018,China;Key Laboratory of Marine Ranch of the Ministry of Agriculture and Rural Affairs,South China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510300,China;National Agricultural Experimental Station for Fishery Resources and Environment Dapeng,Shenzhen Guangdong 518120,China;Sinohydro Engineering Bureau 4 Company Limited,Xining 810000,China)

机构地区：[1]江苏海洋大学海洋科学与水产学院,江苏连云港222005 [2]中国水产科学研究院南海水产研究所热带水产研究开发中心,海南陵水572426 [3]三亚热带水产研究院,海南省深远海渔业资源高效利用与加工重点实验室,海南三亚572018 [4]中国水产科学研究院南海水产研究所,农业农村部海洋牧场重点实验室,广州510300 [5]国家渔业资源环境大鹏实验站,广东深圳518120 [6]中国水利水电第四工程局有限公司,西宁810000

出　　处：《海洋渔业》2024年第3期275-285,共11页Marine Fisheries

基　　金：海南省自然科学基金创新团队(321CXTD446);中国水产科学研究院南海水产研究所基本科研业务费(2020SY01);防城港市重点研发计划(防科AB21014015)。

摘　　要：环境DNA可以定量评估水生生物丰度,但其准确性因引物与构建模式差异而有所不同。以岩礁性鱼类红鳍笛鲷(Lutjanus erythropterus)为对象,采用室内实验和实时荧光定量PCR(qPCR)的方法对红鳍笛鲷引物和探针进行设计与开发,建立DNA浓度与生物量间的关系,解析环境DNA(eDNA)持久性和衰减的过程,初步探究了eDNA技术对红鳍笛鲷生物量评估的可行性。结果显示,当引物退火温度为60℃时,引物扩增效果最好;红鳍笛鲷eDNA浓度与生物量间呈显著正相关(R^(2)=0.93);移出红鳍笛鲷后的20 d内,eDNA衰减浓度与时间呈显著负相关(R^(2)=0.98)。对红鳍笛鲷增殖放流前后的水环境检测结果表明,放流后水体内红鳍笛鲷eDNA浓度显著增加,表明本研究所设计的引物在检测红鳍笛鲷生物量方面的可行性,可为应用eDNA技术开展红鳍笛鲷生物量的准确评估提供理论基础。Reliable abundance information is the foundation for managing aquatic resources.But it is full of challenges to capture the distribution of rare species through traditional methods.Environmental DNA(eDNA)can produce quantitative estimates of fish abundances,but the precision varies greatly depending on the species and system.Therefore,it is necessary to evaluate its performance and investigate how fish biomass affects eDNA dynamics on a case-by-case basis.Lutjanus erythropterus is an important marine economic fish in the South China Sea,and its wild resources have been declining greatly in recent years.Environmental DNA technology is a new technology for sensitive,low-cost and non-invasive species monitoring.It has great potential for applications in the detection of endangered and invasive species.In order to establish a real-time quantitative PCR(qPCR)method to detect Lutjanus erythropterus,eDNA primers and TaqMan probes were designed based on Cyt b gene sequence in mtDNA in this study.The qPCR method for detecting Lutjanus erythropterus was established by PCR amplifying Cyt b gene sequence and constructing standard plasmid with serial dilution of standard plasmid as template.Subsequently,the sensitivity,specificity and application of the method were evaluated.The results showed a strong linear relationship(R^(2)=0.9994)between the cycle threshold value(Ct)of the qPCR assay and the logarithm of standard plasmid concentration.The amplification specificity analysis showed that the method could specifically detect Lutjanus erythropterus.Then,a positive correlation(R^(2)=0.56-0.87)was detected between eDNA concentration and biomass of Lutjanus erythropterus in water samples in aquariums with different biomass of Lutjanus erythropterus.In addition,after the removal of Lutjanus erythropterus,the copies of eDNA were negatively correlated with time,and its retention time in water environment was about 20.The results of water samples before and after the enhancement and release of Lutjanus erythropterus showed that the

关 键 词：红鳍笛鲷 环境DNA 特异性引物 生物量评估 

分 类 号：S931[农业科学—渔业资源]