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大型河流中鱼类组成的eDNA监测效率:以长江武汉江段为例
引用本文:杨海乐,吴金明,张辉,杜浩,李君轶,王成友,沈丽,刘志刚,危起伟.大型河流中鱼类组成的eDNA监测效率:以长江武汉江段为例[J].中国水产科学,2021,28(6):796-807.
作者姓名:杨海乐  吴金明  张辉  杜浩  李君轶  王成友  沈丽  刘志刚  危起伟
作者单位:中国水产科学研究院长江水产研究所, 农业农村部淡水生物多样性保护重点实验室, 湖北 武汉 430223
基金项目:农业农村部财政专项(CJDC-2017-14); 中国水产科学研究院基本科研业务费项目(2020JBF01); 中国水产科学研究院基本科研业务费创新团队项目(2020TD08).
摘    要:为了探讨大型河流中的 eDNA 监测效率, 以长江为大型河流的代表, 以鱼类为水生生物的代表, 分析传统捕捞监测和 eDNA 监测结果的差异, 研究 eDNA 监测技术对长江武汉江段鱼类组成的监测能力、监测效率、平行样设置等问题。结果显示: (1) 用 1 对 eDNA 宏条形码引物(mlCOIintF/jgHCO2198R)共监测到 89 种鱼类, 其中 30 种可与历史捕捞调查记录互相确认, 另外 59 种需要更完善的条形码数据库来解决序列比对注释问题; (2) 9 月在武汉监测断面, 可用单引物(mlCOIintF/jgHCO2198R) eDNA 监测到的物种最优估计约 99 种, 单样品 eDNA 监测的鱼类物种检出能力约为 26 种, 检出效率约为 25.8%; (3) 在 80%的检出度目标下, 需要约 10 个平行样, 在 95%的检出度目标下, 需要约 17 个平行样。本研究在长江武汉江段鱼类 eDNA 监测效率和平行样设置的相关量化结果可为长江其他断面及其他大型河流的 eDNA 监测提供量化参考。同时, 本研究表明, 更完善的 DNA 宏条形码数据库和合适的平行样设置是未来 eDNA 监测作为常规监测手段进行运用的前提。

关 键 词:长江    鱼类    种类组成    eDNA  宏条形码    eDNA  监测    监测效率

Environmental DNA metabarcoding utilization efficiency in monitoring large river fish species composition: a case study in the Wuhan transect of the Yangtze River
Yang Haile,Wu Jinming,Zhang Hui,Du Hao,Li Jundie,Wang Chengyou,Shen Li,Liu Zhigang,Wei Qiwei.Environmental DNA metabarcoding utilization efficiency in monitoring large river fish species composition: a case study in the Wuhan transect of the Yangtze River[J].Journal of Fishery Sciences of China,2021,28(6):796-807.
Authors:Yang Haile  Wu Jinming  Zhang Hui  Du Hao  Li Jundie  Wang Chengyou  Shen Li  Liu Zhigang  Wei Qiwei
Institution:Key Laboratory of Freshwater Biodiversity Conservation, Ministry of Agriculture and Rural Affairs, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223 , China
Abstract:Large rivers are the key regions of aquatic biodiversity. Recently, most large rivers are under significant anthropogenic pressure, and the aquatic biodiversity is being heavily and negatively affected. Consequently, effective biodiversity conservation and management would be required, which depends on accurate, timely, and reliable state assessments, as well as changes in the aquatic communities. However, a major limitation of past assessment methods was their high cost, methodological diversity across taxonomic groups, and the inability of spatiotemporal upscaling of the methods. Therefore, novel technologies would be crucially required in the field. The use of environmental DNA (eDNA) metabarcoding in biodiversity monitoring and surveys is an example of such a novel technology advancement, which would be a game-changer for large river bioassessment and biodiversity monitoring. Until recently, certain studies are available on the origin, state, transport, and fate of the eDNA. However, less is known about the efficiency of the eDNA metabarcoding monitoring aquatic biodiversity in large rivers, which determined the processes of using eDNA in biodiversity monitoring. Therefore, this study focused on the efficiency of the eDNA metabarcoding monitoring in large rivers, taking a transect of Yangtze River in Wuhan as a case study. In this case study, we surveyed the fish species composition using both traditional fishing (15 days with a single boat) and eDNA survey (13 parallel samples with a single metabarcoding) in September 2020. The results of the eDNA survey showed the detection of 89 fish species (13 eDNA samples detected using the primers mlCOIintF and jgHCO2198R), in which 30 fish species were recorded in the historical traditional fishing survey and 59 species were not. In other words, more reference sequences of the fishes in the Yangtze River would be necessary for the DNA metabarcoding database. In this case study, 68.6% of the fish species recorded in the traditional fishing (15 days) were detected by using the eDNA survey (13 eDNA samples). Nearly 26 fish species could be detected in a single eDNA sample. Based on the species accumulation curves, we estimated that the upper limit of the fish species that could be detected by eDNA metabarcoding (using the primers mlCOIintF and jgHCO2198R) was nearly 99. The efficiency of the eDNA metabarcoding monitoring by a single eDNA sample was nearly 25.8 %. With the goals of detecting 80% and 95% of the fish species in this transect, 10 and 17 parallel eDNA samples would be necessary, respectively. Considering the discharge of the Yangtze River in September and the Fishing Ban of the Yangtze River since 2021, no more parallel eDNA samples would be required in this transect in the future. In other transects of the Yangtze River, even other large rivers, the need for parallel eDNA samples could be roughly estimated referring to the number of the parallel samples in the Wuhan transect of the Yangtze River based on the discharge, fish diversity, and abundance. Our results suggested that a complete DNA metabarcoding database and suitable parallel sample setting would be pivotal for using eDNA as a routine biodiversity monitoring method in the future.
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