Objective To identify and analyze the rodents captured in the farmland in Xinhui district, Jiangmen, Guangdong province, China, using DNA barcoding technology, and to lay a foundation for the diversity study of murine-like animals and rodent prevention and control. Methods Forty-five rodents captured in the farmland in Xinhui district, Jiangmen, Guangdong province, in December 2017, were enrolled as study subjects. Their genetic DNAs were extracted and amplified for mitochondrial cytochrome c oxidase subunit I gene fragments using consensus primers. The gene fragments were sequenced and the results were subjected to sequence homology analysis using BLAST in the National Center for Biotechnology Information. The molecular evolutionary tree was constructed by the neighbor-joining method. Genetic distance was calculated based on a two-parameter model. Results Specific gene fragments of the forty-five samples were obtained by PCR amplification. Four species of rodents (Rattus losea, R. tanezumi, R. norvegicus, and R. andamanensis) were identified through BLAST comparison. Thirty-seven samples had identification results identical to the morphological identification (R. losea), and eight samples had erroneous morphological identification results, among which five were reidentified as R. tanezumi, two were reidentified as young R. norvegicus, and one was reidentified as R. andamanensis, according to the review of the initial results. A total of 131 mutation sites were identified through alignment and analysis of the 45 sequences, and 12 different haplotypes were identified through haplotype analysis. The intraspecific genetic distance of R. losea was 0-1.2%, the interspecific genetic distance was 7.1%-11.6%, and the mean genetic distance was 10.1%±0.7%. The NJ tree showed that individuals of the same rodent species were clustered into one branch with a 100% support rate, which could distinguish among different rodent species. Conclusion The DNA barcoding technology can effectively distinguish between related species with similar morphology and correct the erroneous results of morphological identification.
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