目的 建立一种准确、高效的天山雪莲Saussurea involucrata及其易混伪品水母雪兔子及绵头雪兔子特异性DNA特征序列（DSS）标记分子鉴定方法。方法 通过CGIR植物叶绿体基因组数据库获取风毛菊属植物全部叶绿体基因组数据，利用DSS鉴定软件IdenDSS获得用于天山雪莲鉴定的特有DSS，筛选5组DSS标记并利用IdenDSS设计相应的特异性引物对，通过对样品DNA提取、PCR扩增、sanger测序以及序列比对验证，检测待测样品中是否含有目标物种天山雪莲的DSS或其反向互补序列进行基原鉴定。结果 通过比对验证5组DSS标记与天山雪莲正品来源及易混伪品水母雪兔子和绵头雪兔子序列一致性，最终确定5组DSS标记均可用于天山雪莲基原鉴定，将上述序列上传至中药分子鉴定平台（www.herbsdna.com）标准数据库中，结合数据库DSS鉴定模块，可实现对天山雪莲准确、高效的基原鉴定。结论 筛选的5组DSS标记用于天山雪莲基原鉴定特异性强、准确度高，可有效解决天山雪莲基原鉴定难题。

Objective To establish an accurate and efficient molecular identification method for specific DNA Signature Sequence Tags (DSS) of Tianshanxuelian (Saussurea involucrate) and its adulterants Shuimuxuetuzi (S. medusa) and Miantouxuetuzi (S. laniceps). Methods The Chloroplast Genome Information Resource (CGIR) were used to obtain the complete chloroplast genome sequences of Saussurea DC, the IdenDSS software was used to screen for specific DNA signature sequence tags (DSS) of the S. involucrata. Five sets of DSS markers were selected, and the corresponding specific primer pairs were designed using IdenDSS. Through DNA extraction from the sample, PCR amplification, Sanger sequencing and sequence alignment verification, the presence of DSS or its reverse complementary sequence of the target species S. involucrata in the test sample was detected for original identification. Results By comparing the selected five pairs of DSS sequences with the authentic source and its adulterants sequences of S. involucrata, all five sets of DNA Signature Sequence Tags could be used for identification of S. involucrata were finally determined. The above sequences were uploaded to the standard database of the Traditional Chinese Medicine Molecular Identification Platform (www.herbsdna.com), and combined with the DSS identification function of the database, accurate and efficient identification of S. involucrata origin can be achieved. Conclusion The five selected DNA Signature Sequence Tags had strong specificity and high accuracy, which can effectively solve the problem of original identification of S. involucrata.

R282.6

中央级公益性科研院所基本科研业务费专项资金资助（中国中医科学院中药监管科学研究项目）