摘要
大黄为多基原药材,其混淆品种类很多,传统的药材鉴定方法无法实现对药材基原种的鉴别。本研究通过对采集于大黄主产区的正品基原以及常用混淆品药材matK基因序列的差异分析,结合已有报道的大黄matK基因序列结果,探讨从基因型角度进行大黄药材分子鉴定的可行性。采用改良试剂盒法提取DNA,以大黄属matK基因通用引物进行PCR扩增,扩增产物经回收纯化后双向测序,运用ContingExpress、DNAman、MEGA5.0等软件进行序列分析,总结大黄正品基原与混淆品的基因型对应规律。结果显示正品大黄的matK基因序列全长1 518 bp,变异位点57个。基于587、707和838位3个特异性位点可鉴别掌叶组外的混淆品与正品基原,基于基因型特异性可鉴别掌叶组内条裂大黄、六盘山鸡爪大黄和绿花唐古特大黄3种组内混淆品以及正品基原的3个种,同时也能在一定程度上实现正品基原产地的鉴别,但是对此还有待于增加药材样品进一步验证。
Rhei Rhizoma is a Chinese medicine with multiple botanical origins. There is a problem to identify it with conventional methods. To compare the characteristics of chloroplast mark gene sequences of different Rheum species and authenticate inspected species, the mark gene sequences of different species from different origins were amplified, cloned, and sequenced. Genomic DNA of Rheum plants was extracted using modified DNA extracted Kit and mark gene sequences were analyzed by ContingExpress, DNAman and MEGA5.0. The length of mark gene sequences of Rheum palmatum, R. tanguticum and R. officinale were 1 518 bp containing 57 variable loci. According to the mutation sites, R. palmatum, R. tanguticum and R. officinale were divided into different genotypes separately. Based on the established method according to the loci 587, 707, 838, we successfully identified the genuine Rheum species from its adulterants.
出处
《药学学报》
CAS
CSCD
北大核心
2013年第11期1722-1728,共7页
Acta Pharmaceutica Sinica
基金
国家自然科学基金资助项目(30973880,31170307)
教育部留学回国人员启动基金资助项目(03291/537)
科技基础性工作专项重点项目(2007FY110600)
北京中医药大学自主选题(藏医药类)课题资助项目(2013-zyy-02)
