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长江中游常见鱼类DNA条形码研究及其电子芯片分析
郜星晨1, 姜 伟1, 刘绍平2
1.中国长江三峡集团有限公司中华鲟研究所 三峡工程鱼类资源保护湖北省重点实验室 宜昌 443100;2.农业农村部长江中上游渔业资源环境科学观测实验站 中国水产科学研究院长江水产研究所 武汉 430223
摘要:
为建立长江中游常见鱼类的快速鉴别方法,文献调研了7目11科50属64种鱼类名录,GenBank共获取168条线粒体细胞色素c氧化酶I (COⅠ)序列,分析了序列特征、不同阶元Kimura-2- paramater (K2P)遗传距离及系统进化关系。结果显示,64种鱼类的种间遗传距离(平均值为0.084)明显大于种内(平均值为0.0079),NJ树上不同物种均能以较高支持度聚类成独立分支,以线粒体COⅠ序列作为DNA条形码可准确鉴定所研究鱼类;综合利用分子生物学软件筛选物种特异性探针,最终43种鱼类可筛选出112条物种特异性探针,物种识别率为67.2%。本研究验证了DNA条形码芯片技术在长江中游鱼类物种鉴定的可行性,可为该地区鱼类物种多样性保护提供技术支持。
关键词:  长江中游  DNA条形码  电子芯片
DOI:
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基金项目:国家重点研发计划“典型脆弱生态修复与保护研究”重点专项(2016YFC0502210)和农业农村部长江中上游渔业资源环境科学观测站共同资助
DNA Barcoding and Electronic Microarrays for Common Fish Species in the Middle Reaches of the Yangtze River
GAO Xingchen1, JIANG Wei1, LIU Shaoping2
1.Chinese Sturgeon Research Institute, China Three Gorges Corporation, Hubei Key Laboratory of Three Gorges Project for Conservation of Fishes, Yichang 443100;2.Scientific Observing and Experimental Station of Fishery Resources and Environment in the Upper and Middle Reaches of Yangtze River, Ministry of Agriculture and Rural Affairs, Yangtze River Fishery Resources Institute, Chinese Academy of Fishery Sciences, Wuhan 430223
Abstract:
To develop a method for the rapid identification and classification of fishes in the middle reaches of the Yangtze River, 168 DNA sequences of the cytochrome oxidase subunit Ⅰ (COⅠ) gene collected from GenBank. 64 common freshwater fish species in 7 orders, 11 families, and 50 genera were analyzed to test the efficacy of species identification using a DNA barcode microarray. The results showed that the intraspecific genetic distances ranged from 0 to 0.0776 (average, 0.0079). The interspecific distances ranged from 0.003 to 0.0190 (average, 0.084). The interspecific genetic distance was larger than the intraspecific distance, and all 168 sequences formed species units in a neighbor-joining phylogenetic tree, indicating that the DNA barcodes could be used to identify these 64 species. Based on the DNA microarray technology, 112 probes were selected from 43 species with high specificity, accounting for 67.2% of the total species. Thus, the DNA barcode microarray provided technical support and a new way to identify fish species in the middle reaches of Yangtze River basin.
Key words:  Middle reaches of Yangtze River  DNA barcode  Electronic microarray