基于COI条形码序列的蛤蚧及其混伪品的DNA分子鉴定

Identification of Gekko geeko Linnaeus and Adulterants Using the COI Barcode

目的:利用COI序列对蛤蚧药材及其常见混伪品进行DNA条形码鉴定,为蛤蚧药材鉴定提供新的方法。方法:以COI作为条形码序列,对蛤蚧实验样品进行DNA提取,PCR扩增和双向测序,用MEGA6.0对所有11个物种的103份样品进行序列比对和邻接(NJ)树构建。结果:所有实验样品均可以获得COI序列,蛤蚧COI序列种内平均K2P距离为0.005,种内最大K2P距离为0.013,基于COI序列构建的NJ树中蛤蚧单独聚在一支,与其混伪品可以相互区分。结论:运用COI条形码序列能够准确鉴定蛤蚧及其混伪品,为保障蛤蚧药材临床用药安全和市场监管提供新的方法。

DNA barcoding technology is widely opplied for species identification and discovery by using short and standard fragments of DNA sequences. In this study, the cytochrome c oxidase subunit 1 （COI） sequence as a DNA barcode was used to identify the Gekkogecko Linnaeus medicinal materials and adulterants in order to provide a new identification method of G. gecko Linnaeus. Genomic DNA was extracted from the experimental samples. The COI regions of nrDNA were amplified using polymerase chain reaction and sequenced bidirectionally. The alignment and NJ tree construction were performed in MEGA6.0. COI sequences can be obtained from all samples. The average intra-specific K2P distance of G. gecko Linnaeus was 0.005 and the maximum intra-specific distance was 0.013. All the G. gecko Linnaeus medicinal samples clustered into a clade in the NJ tree and can be distinguished from the adulterants. In a conclusion, COI can be used to correctly identify G. gecko Linnaeus, and it will be a potential DNA barcode for identification of other animal medicinal materials.