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目的以三峡库区沿江口岸捕获的鼠类为研究对象,应用DNA条形码技术进行鼠类鉴定,分析各种鼠类的分子鉴定特征。方法在三峡库区沿江口岸采集鼠类标本,并保存整只鼠体,制作标本用于形态学鉴定,采集肝脏或肌肉组织,提取DNA,采用通用引物PCR法扩增细胞色素C氧化酶亚基Ⅰ(COⅠ)基因,并测序。将测序结果与Gen Bank中其他鼠类的DNA条码进行Blast比对,并构建分子进化树。结果共捕获4种61只鼠类,其中42份样本通过PCR扩增获得特异性COⅠ基因条带,所构建的分子进化树结果与形态学鉴定结果一致,但不同口岸鼠类在分子进化水平存在一定差异。结论分子鉴定技术能够应用于对三峡库区口岸鼠类进行有效的物种鉴定。
OBJECTIVES: To identify rodents based on the DNA barcoding technique and to identify the molecular characteristics of various rodents. Methods Rodent samples were collected from the Yangtze River estuary in the Three Gorges Reservoir area and the whole rat body was preserved. Specimens were prepared for morphological identification, and liver and muscle tissues were collected for DNA extraction. The universal primer PCR method was used to amplify the cytochrome C oxidase subunit Ⅰ (COⅠ) gene and sequenced. The sequencing results were compared with the DNA barcodes of other murine GenBanks to construct a phylogenetic tree. Results A total of four 61 rat species were captured. Among them, 42 samples were amplified by PCR to obtain specific COⅠ gene bands. The results of molecular phylogenetic tree were consistent with the results of morphological identification. However, Some difference. Conclusion The molecular identification technique can be used to validate the species of rodents at the port of the Three Gorges Reservoir.