基于DNA条形码技术的民族药紫丹参及其近缘种鉴定研究
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摘要
目的探讨DNA条形码技术在民族药紫丹参及其近缘物种鉴定中的应用,为紫丹参药材鉴定提供参考。方法建立紫丹参及其混伪品DNA条形码参照数据库,通过比对序列、分析变异位点、计算遗传距离和构建邻接树等分析数据,并应用该数据库对市场药材进行鉴定。结果 DNA条形码技术可有效鉴别紫丹参及其混伪品。结论 ITS2序列可作为紫丹参及其混伪品的鉴定条形码,该技术在民族药物种基原鉴定、标准制定和市场监管等实际应用中具有重要的应用价值。
Objective To explore the application of DNA barcode technology in identification of ethnic medicine Salvia yunnanensis and its related species, so as to provide reference for identification of S. yunnanensis. Methods A DNA barcode database of S. yunnanensis and its adulterants was successfully constructed. Data were analyzed by aligning sequences, analyzing variation sites, calculating genetic distances, and establishing Neighbor-Joining trees. The database was used to identify 45 S.yunnanensis decoction piece samples purchased from drug stores. Results The results showed that the ITS2 barcodes can be used to identify S. yunnanensis and its adulterants. Conclusion The use of DNA barcoding for market supervision can broaden the application of this technology and provide an excellent inspection method for the supervisory institutions. This study substantially contributes to the protection of public health worldwide.
Objective To explore the application of DNA barcode technology in identification of ethnic medicine Salvia yunnanensis and its related species, so as to provide reference for identification of S. yunnanensis. Methods A DNA barcode database of S. yunnanensis and its adulterants was successfully constructed. Data were analyzed by aligning sequences, analyzing variation sites, calculating genetic distances, and establishing Neighbor-Joining trees. The database was used to identify 45 S.yunnanensis decoction piece samples purchased from drug stores. Results The results showed that the ITS2 barcodes can be used to identify S. yunnanensis and its adulterants. Conclusion The use of DNA barcoding for market supervision can broaden the application of this technology and provide an excellent inspection method for the supervisory institutions. This study substantially contributes to the protection of public health worldwide.
引文
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[3]国家中成药标准汇编内科心系分册[S].2002.
[4]李惠兰,闫彩珍,张荣平,等.滇丹参注射液对兔血小板功能的影响[J].天然产物研究与开发,2002,14(1):70-73.
[5]闫彩珍,李惠兰,张荣平,等.滇丹参、甘西鼠尾、褐毛甘西鼠尾对离体豚鼠心脏的影响[J].昆明医学院学报,2002,23(1):14-17.
[6]谢宗万.中药品种理论与应用[M].北京:人民卫生出版社,2008.
[7]李海鹰,邵林岳,林峰.丹参及其伪品牛蒡根的理化鉴别[J].中药材,1999,22(9):449-451.
[8]何广新,李忠琼.丹参和紫丹参的比较鉴别[J].中药材,1996,19(7):342-343.
[9]张晓灿,罗丹丹,陶爱恩,等.基于指纹图谱和化学计量学的丹参及紫丹参质量评价研究[J].中药材,2017,40(5):1061-1065.
[10]曹珍,谢晓亮.丹参的不同鉴别方法[J].时珍国医国药,2007,18(8):1861-1863.
[11]尚飞能,方海兰,段宝忠.基于ITS2条形码和化学指纹鉴别何首乌及其伪品[J].大理大学学报,2017,2(4):12-15.
[12]王亚丹,韩晓妮,赵玉丹,等.基于ITS2条形码鉴别市售柴胡药材及其混伪品[J].中草药,2017,48(17):3590-3596.
[13]Duan B Z,Fang H L,Li X W,et al.Survey of traditional Dai medicine reveals species confusion and potential safety concerns:a case study on Radix Clerodendri Japonicum[J].Chin J Nat Med,2017,15(6):417-426.
[14]Chen S L,Yao H,Han J P,et al.Validation of the ITS2region as a novel DNA barcode for identifying medicinal plant species[J].PLoS One,2010,5(1):e8613.
[15]张萍,黎先军,杜微波,等.基于ITS2序列的丹参及其混伪品分子鉴定研究[J].安徽农业科学,2017,45(30):122-124.
[16]陈士林,姚辉,韩建萍,等.中药材DNA条形码分子鉴定指导原则[J].中国中药杂志,2013,38(2):141-148.
[17]陈士林.中药DNA条形码分子鉴定[M].北京:人民卫生出版社,2012.
[18]中国药典[S].四部.2015.
[19]陈士林.中国药典中药材DNA条形码标准序列[M].北京:科学出版社,2015.