基于ITS2序列鉴定市售地骨皮药材及其混伪品

Identification of Commercial Lycii Cortex and Its Adulterants Using ITS2 Sequence as DNA Barcode

目的基于中药材DNA条形码分子鉴定法鉴定市售地骨皮药材及其混伪品。方法本实验对象共计137份，依照中药材DNA条形码分子鉴定法标准操作流程（DNAbarcodingSOP）获取各样本ITS2序列，构建地骨皮药材及其混伪品DNA条形码数据库，对市售地骨皮药材及饮片进行鉴定。结果地骨皮药材ITS2序列长度为212—230bp，能够明显区分地骨皮药材及其混伪品。50％市售药材及饮片能够获得可用于分析的条形码序列，应用地骨皮药材及其混伪品DNA条形码数据库鉴定结果显示所得序列均为枸杞。结论ITS2序列能够作为地骨皮药材及其混伪品鉴定的DNA条形码，本实验所建立的地骨皮药材及其混伪品DNA条形码数据库能够用于市售地骨皮药材及饮片的鉴定。

OBJECTIVE To identify the commercial medicinal materials and decoction pieces of Lycii Cortex and its adulterants using DNA barcoding technology. METHODS A total of 137 samples, including 105 voucher samples belonging to nine species, seven GenBank sequences, and 25 test samples were involved in this study. Experiments were performed in accordance with the DNA barcoding standard operating procedures （DNA barcoding SOP） to get the ITS2 sequences. A DNA barcode database of Lycii Cortex and its adulterants were successfully constructed using 112 ITS2 sequences, which were amplified from the voucher samples and down- loaded from the GenBank. This database was used to identify the commercial medicinal materials and decoction pieces of Lycii Cortex. RESULTS The lengths of the ITS2 regions of the Lycii Cortex were 212-230 bp. The ITS2 sequences could clearly distinguish Lycii Cortex and its adulterants. Fifty percent of the commercial samples gained the ideal genomic DNA for the sequence amplification. Using the established database, the above-mentioned sequences were authenticated as Lycium chinense. CONCLUSION ITS2 Sequence may be a suitable marker for the identification of Lycii Cortex and its adulterants. The DNA barcode databaseof Lyeii Cortex and its a- dulterants constructed in this study are able to successfully identify the raw materials of the commercial medicinal materials and decoc- tion pieces of Lycii Cortex that are currently available in the market.