From August to November 2019, 11 were set up in the Three Gorges Reservoir area and its tributaries (Taiping River, Xiangxi River, Wanzhou, Fuling, Fengdu, Fujiang, Jialing River, Jiantan River, Banan, Yunyang, Xiaojiang) At the sampling point, the selected fishing gears were ground cages (mesh 1.2cm), seal nets (diameter 1m, length 3m) and gill nets. The specifications of the gillnet are multi-mesh composite gillnets, including 12 mesh specifications: 2a=8.5cm, 4.0cm, 12.5cm, 2.0cm, 11.0cm, 1.6cm, 2.5cm, 4.8cm, 3.1cm, 1.0cm , 7.5cm and 6.0cm, each mesh gillnet is 2.5m long and 2m wide, spliced into a composite gillnet with a total length of 30m. Field fishing was carried out at each sampling point, on-site identification, and biological traits such as fish body length and weight were collected. During the sampling period of this study, 18,466 fish samples were recorded in the Three Gorges Reservoir area. The samples were preliminarily identified as 64 species, and the species were morphologically identified with reference to "Chinese Cyprinidae (Volume 1)", "Sichuan Fish Chronicles" and "Fishes of the World". All samples were preserved in 10% formaldehyde solution in intact individual form for further morphological identification, and were preserved in our laboratory (Laboratory of Animal Genetics, Jianghan University), and tissue samples were preserved in 95% ethanol. About 100 mg of fish muscle tissue or fin rays were taken, and genomic DNA was extracted by high-salt method. The COI gene sequence amplification primer sequence was: F1:5'-TCAACCAACCACAAAGACATTGGCAC3'R1:5'-TAGACTTCTGGGTGGGCCAAAGAATC3'. The PCR reaction system was 30 μL, including 10.3 μL of 2×Taq Master Mix, 1.5 μL of upstream and downstream primers (10 mM), 1.5 μL of DNA template, and 15.2 μL of double-distilled water. The PCR reaction program was as follows: 95°C high temperature pre-denaturation for 5 min; 95°C denaturation for 30s; 56°C annealing for 45s; 72°C extension for 45s, a total of 31 cycles; final extension at 72°C for 10 min; °C in the refrigerator. Finally, the results of PCR products were detected by electrophoresis. PCR products were detected by 1% agarose gel electrophoresis and then sent to a biological company for sequencing. BioEdit software was used to manually correct the forward and reverse sequencing peaks, and the DNA sequences were aligned and spliced using the splicing program SEQMAN in DNASTAR, and all sequences containing missing information and degenerate bases were eliminated. Finally, 946 COI sequences from fish were obtained in this study, and the 639bp mitochondrial COI gene sequence after homology alignment was used for barcode analysis. No deletion or insertion of bases was found in all sequences., 2019年8月-11月,在三峡库区及各支流(太平溪,香溪河,万州,涪陵,丰都,涪江,嘉陵江,箭滩河,巴南,云阳,小江)设置11个采样点,选取的渔具为地笼(网目1.2cm)、篆网(直径1m,长度3m)和刺网等。刺网的规格为多网目复合刺网,包括12种网目规格:2a=8.5cm、4.0cm、12.5cm、2.0cm、11.0cm、1.6cm、2.5cm、4.8cm、3.1cm、1.0cm、7.5cm和6.0cm,每个网目的刺网长2.5m,宽2m,拼接成总长30m的复合刺网。在各采样点进行实地捕捞,现场鉴定,采集鱼类体长及重量等生物学性状。本研究采样期间记录三峡库区鱼类样本18466尾。样品初步鉴定为64种,参考《中国鲤科鱼类志(上卷)》、《四川鱼类志》和《Fishes of the world》等对物种进行形态学鉴定。所有样本均以完整个体形式保存在10%甲醛溶液中进行进一步形态学鉴定,保存于本实验室(江汉大学动物遗传学实验室),组织样本保存在95%乙醇中。取约100mg的鱼类肌肉组织或鳍条,采用高盐法提取基因组DNA,COI基因序列扩增引物序列为:F1:5’-TCAACCAACCACAAAGACATTGGCAC3’R1:5’-TAGACTTCTGGGTGGGCCAAAGAATC3’。PCR反应体系为30 μL, 其中包括2×Taq Master Mix 10.3 μL , 上下游引物(10 mM)各1.5 μL和 DNA模板1.5μL, 和双蒸水15.2 μL。PCR反应程序为: 95℃高温预变性 5min;95℃变性30s;56℃退火 45s;72℃延伸45s,共31个循环;最后72℃延伸 10min;16℃保存20分钟,或者结束后置于4℃冰箱中。最后经电泳检测 PCR产物结果。PCR产物经1%琼脂糖凝胶电泳检测后送生物公司测序。利用BioEdit软件对正反向的测序峰图进行人工校正,利用DNASTAR中的拼接程序SEQMAN对DNA序列进行比对和拼接,剔除所有含缺失信息、兼并碱基的序列。最后本研究获取鱼类946条COI序列,同源比对后639bp的线粒体COI基因序列用于条形码分析,所有序列均未发现碱基的缺失、插入。