首页|多基原药材大黄叶绿体matK基因序列分析及鉴定研究

多基原药材大黄叶绿体matK基因序列分析及鉴定研究

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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.
Sequence analysis and identification of a chloroplast matK gene in Rhei Rhizoma from different botanical origins
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 matK gene sequences of different Rheum species and authenticate inspected species,the matK 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 matK gene sequences were analyzed by ContingExpress,DNAman and MEGA5.0.The length of matK 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.

RheummatK gene sequencemolecular authenticationgenotype

张晓芹、刘春生、闫兴丽、程小丽、刘娟、王秋玲、刘凯、魏胜利

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北京中医药大学中药学院,北京100102

重庆市九龙坡区第一人民医院,重庆400050

中国医学科学院、北京协和医学院药用植物研究所,北京100193

大黄 matK基因序列 分子鉴定 基因型

国家自然科学基金国家自然科学基金教育部留学回国人员启动基金资助项目科技基础性工作专项重点项目北京中医药大学自主选题(藏医药类)课题资助项目

309738803117030703291/5372007FY1106002013-zyy-02

2013

药学学报
中国药学会 中国医学科学院药物研究所

药学学报

CSTPCDCSCD北大核心
影响因子:1.274
ISSN:0513-4870
年,卷(期):2013.48(11)
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