中国中药杂志

2012, v.37(08) 1107-1113

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ITS2和psbA-trnH序列鉴别马鞭草科药用植物
DNA bacording of Verbenaceae medicinal plant by using ITS2 and psbA-trnH region

陈倩;刘义梅;刘震;陈士林;陈科力;
CHEN Qian 1,LIU Yimei 1,LIU Zhen 1,CHEN Shilin 2,CHEN Keli 1(1.Key Laboratory of Ministry of Education on Traditional Chinese Medicine Resource and Compound Prescription & Hubei University of Chinese Medicine,Wuhan 430065,China; 2.Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100193,China)

摘要(Abstract):

目的:评价几条热点DNA条形码候选序列对马鞭草科药用植物的鉴别能力。方法:选择psbA-trnH,rbcL,matK,ITS2和ITS序列进行评价,使用各序列的通用引物进行扩增和测序,用扩增及测序成功率,种内种间差异,barcod-ing gap,基于BLAST 1和Nearest Distance方法的鉴定成功率等指标来评价各序列的鉴定能力。结果:对马鞭草科32个种55个样本的序列分析发现,matK的扩增成功率太低,未纳入后续分析;ITS,ITS2,psbA-trnH以及rbcL的扩增及测序成功率分别为83.6%,83.6%,96.4%,98.2%,其鉴定成功率除rbcL为77.8%,75.9%外都为100%,但ITS2序列的种间,种内差异,barcoding gap较psbA-trnH,ITS有明显优势。ITS2序列进一步纳入网上163个样品的数据后其鉴定成功率依然可以达到89.5%,87.6%。结论:考虑到psbA-trnH较高的测序成功率,ITS2和psbA-trnH是适合马鞭草科植物鉴别的一个较好DNA条形码序列组合。
Objective: To evaluate the ability of several hotspot candidate sequence of DNA barcodes for identifying medicinal plant species in Verbenaceae.Method: Using universal primers,three chloroplast sequences,psbA-trnH,rbcL,matK,two nuclear ribosomal DNA ITS2 and ITS were amplified and sequenced.PCR amplification and sequencing efficiency,intra-and inter-specific variation,barcoding gap and identification efficiency(with BLAST 1 and Nearest Distance methods) were used to evaluate these loci.Result: The rate of successful amplification using matK was too low to further analyze,and the rate of successful amplification and sequencing using psbA-trnH,ITS,and ITS2 and rbcL sequence was 83.6%,83.6%,96.4%,98.2%,respectively.The rate of successful identification using psbA-trnH,ITS,and ITS2 was 100% at the species level except that rbcL was 77.8%,75.9% for 55 samples belonging to 32 species,but ITS2 did better in intra-and inter-specific variation,barcoding gap than the other loci.The rate of successful identification of ITS2 was 89.5%,87.6% even when joining the date of 165 samples from GenBank.Conclusion: It proposes that the combination of ITS2 and psbA-trnH senquence is promising for the identification of the species in Verbenaceae.

关键词(KeyWords): DNA条形码;ITS2;ITS;rbcL;psba-trnH;马鞭草科
DNA barcoding;ITS2;ITS;rbcL;psbA-trnH;Verbenaceae

Abstract:

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基金项目(Foundation): 国际科技合作项目(2007DFA30990);; 卫生行业科研专项(200802043)

作者(Author): 陈倩;刘义梅;刘震;陈士林;陈科力;
CHEN Qian 1,LIU Yimei 1,LIU Zhen 1,CHEN Shilin 2,CHEN Keli 1(1.Key Laboratory of Ministry of Education on Traditional Chinese Medicine Resource and Compound Prescription & Hubei University of Chinese Medicine,Wuhan 430065,China; 2.Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100193,China)

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