基于DNA条形码SNP鉴别东方泽泻、泽泻及市售泽泻药材  

SNP Identification of Alisma orientale,Alisma plantago-aquatica and Alismatis Rhizoma in the Market Based on DNA Barcoding

作　　者：马晓冲[1] 姚辉[1] 辛天怡[1] 陈晓辰[1] 宋经元[1,2] 

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193 [2]重庆市药物种植研究所,重庆408435

出　　处：《中国药学杂志》2015年第17期1474-1478,共5页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金资助项目(81373922)

摘　　要：目的建立基于单核苷酸多态性(SNP)分子标记鉴定泽泻药材的方法。方法采用改良的试剂盒法提取药材基因组DNA,PCR扩增ITS2序列,通过分析比对东方泽泻和泽泻ITS2序列,确定SNP位点,并对商品泽泻药材进行鉴定分析。结果东方泽泻和泽泻ITS2序列长度为311 bp,二者均无种内变异,但种间在165 bp有稳定的A-T变异,NJ树鉴定结果亦表明二者可以准确区分;基于SNP分子条形码鉴定和NJ树鉴定市售泽泻药材,仅有7%药材样本为东方泽泻,93%的市售药材样本均非《中国药典》规定基原。结论基于SNP标记能准确鉴定中药材泽泻,为保护泽泻种质资源和指导药材生产种植提供可靠依据。OBJECTIVE To establish a method for the rapid identification of Alismatis Rhizoma based on single nucleotide polymorphism （SNP）molecular marker. METHODS Total genomic DNA was extracted using improved DNA extraction kit, and the internal tran scribed spacer 2 （ITS2） regions were amplified and annotated by using the hidden Markov model（HMM）. In addition, the ITS2 sequences of Alisma orientale and Alisma plantago-aquatica were aligned through Clustal-W, and then SNP was detected to identify Alismatis Rhizoma. RESULTS The length of A. orientale and A. plantago-aquatica ITS2 sequences was 311 bp. No intra-specific variation was found among the samples of two species, respectively. One stable SNP （A/T） was detected at 165 bp in ITS2 region which was useful for identification of the two species. The result was confirmed by NJ tree method. Furthermore, authentication of commercial Alismatis Rhizoma by SNP molecular marker and NJ tree method indicated that only two medicinal samples （7%） were A. orientale and the others （93%） were A. plantago-aquatica which was not recorded in the Chinese Pharmacopoeia. CONCLUSION SNP molecular marker can stably and accurately distinguish Alismatis Rhizoma in the market and can offer scientific basis for protec- tion of germplasm resources and cultivation of Alisma.

关 键 词：泽泻药材 SNP ITS2 市售药材 分子鉴定 

分 类 号：R282[医药卫生—中药学]