DNA条形码技术在河北省坝上地区夜行鼠鉴定中的应用

Application of DNA barcoding technique in identification of nocturnal rodent species in Bashang area of Hebei province, China

目的以河北省坝上地区捕获的夜行鼠为对象,应用DNA条形码技术进行鼠类鉴定。方法在河北省坝上地区采集鼠肝脏标本,并保存整只鼠,制作标本,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 36份样本均能通过PCR扩增出特异性COⅠ基因条带,所构建的分子进化树结果与形态学鉴定结果有所不同,经头骨鉴定,进化树结果正确无误,更正形态学鉴定结果。其中2只夜行鼠未能在Gen Bank中找到同源性较高的鼠类基因,且头骨受到严重损坏,暂无法确定鼠种。结论 DNA条形码技术能够对鼠类进行有效的物种鉴定。

Objective To identify nocturnal rodent species in the Bashang area of Hebei province, China, using DNA barcoding.MethodsLiver tissue samples of nocturnal rodents were collected from the study area. The whole mice wer ealso preserved and dissected. Genomic DNA was extracted for PCR amplification of the mitochondrial cytochrome c oxidase subunitⅠ（COⅠ） gene with consensus primers, followed by sequencing. Blast comparisons were performed between the obtained sequencing data and the DNA bar codes of other rodent species in the Gen Bank. A molecular evolutionary tree was built.ResultsAll the 36 samples produced specific COⅠ gene bands by PCR amplification. The result of molecular evolutionary tree was slightly different from that of morphological identification. Skull identification revealed that the molecular evolutionary tree was correct, and the result of morphological identification was corrected. High-homology genes were not found in the Gen Bank for two nocturnal rodents. Since their skulls were seriously damaged, we temporally could not determine the species of these two individuals.Conclusion DNA barcoding can be used to effectively identify rodent species.