中国中药杂志

2015, v.40(10) 1914-1918

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基于trnL-trnF序列的人参和西洋参DNA条形码鉴定研究
DNA barcoding identification of Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix based on trnL-trnF sequences

宋晓娜;顾选;刘春生;李妍芃;张雪;张媛;刘勇;马长华;
SONG Xiao-na;GU Xuan;LIU Chun-sheng;LI Yan-peng;ZHANG Xue;ZHANG Yuan;LIU Yong;MA Chang-hua;School of Chinese Materia Medica,Beijing University of Chinese Medicine;

摘要(Abstract):

探讨trnL-trnF序列的相似度、特异位点、遗传距离和系统发育树这4个分子鉴定指标对人参和西洋参药材的鉴定能力,优化人参和西洋参分子鉴定指标。从人参和西洋参药材中提取DNA,以trnL-trnF通用引物进行扩增测序,利用NCBI数据库中的BLAST功能计算相似度,利用DNAman软件比较特异位点,利用Mega软件计算遗传距离,构建系统发育树。结果表明,人参和西洋参的种内相似度均为100%,种间相似度为99.6%;人参和西洋参分别在G153A,T463A,C732G,T818C有特异位点;种内遗传距离(0)小于种间遗传距离(0.004);系统发育树中人参和西洋参均分为2个单系分支。因此,利用trnL-trnF序列,相似度、特异位点、遗传距离和系统发育树指标均可以区分人参和西洋参。综合分析,trnL-trnF序列的G153A,T463A,C732G,T818C特异位点指标是人参和西洋参的最佳鉴定指标。
To optimize indices of molecular identification for authentication of Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix,four indices,including sequence similarity,specific positions,genetic distance and phylogenetic tree,were compared based on trn L-trn F sequences. Total DNA was extracted from Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix,and trn L-trn F sequences were amplified and sequenced. Sequence similarity was calculated by BLAST analysis. Specific positions were compared by DNAman software. Genetic distance and phylogenetic tree were analyzed by Mega software. The results showed that the inter-specific and intra-specific similarity of P. ginseng and P. quinquefolius respectively was 100% and 99. 6%. There were four specific positions at G153 A,T463A,C732 G and T818 C. The inter-specific genetic distance( 0) of trn L-trn F sequences was lower than intra-specific genetic distance( 0. 004). P. ginseng can be distinguished from P. quinquefolius based on the phylogenetic tree. It is concluded that Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix can be authenticated by identification indices of sequence similarity,specific positions,genetic distance and phylogenetic tree. Index of specific positions based on trn L-trn F sequences is the most efficient index to authenticate Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix.

关键词(KeyWords): 鉴定指标;人参;西洋参;trnL-trnF
identification index;Ginseng Radix et Rhizoma;Panacis Quinquefolii Radix;trnL-trnF

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基金项目(Foundation): 国家科技支撑计划项目(2012BAI29B02);; 北京市自然科学基金项目(7142101)

作者(Author): 宋晓娜;顾选;刘春生;李妍芃;张雪;张媛;刘勇;马长华;
SONG Xiao-na;GU Xuan;LIU Chun-sheng;LI Yan-peng;ZHANG Xue;ZHANG Yuan;LIU Yong;MA Chang-hua;School of Chinese Materia Medica,Beijing University of Chinese Medicine;

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