基于DNA条形码技术的新疆民族药材天山堇菜基原研究
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摘要
目的:明确市售药材天山堇菜的植物基原,为药材鉴定提供方法依据。方法:使用DNA条形码ITS2序列,建立了民族药材天山堇菜及同属126份基原植物的DNA条形码ITS2序列标准数据库,分析市售18份"天山堇菜"药材的基原。结果:成功建立了堇菜属126份植物的ITS2序列数据库,通过此数据库对18份市售药材的鉴定结果表明,8份为天山堇菜(44.4%),3份为香堇菜(16.7%),6份为堇菜属植物(33.3%),1份为非堇菜属植物(5.6%)。结论:市售民族药材天山堇菜来源于多种植物,其中不足一半与标准中基原一致,多数为混伪品,其是否均能作为天山堇菜使用,还有必要结合用药历史考证。
Objective: We surveyed the species composition of commercial V. kunawarensis products using DNA barcoding technology, which would provide method for the identification of medicinal materials. Methods: DNA barcode ITS2 sequence was used to establish a standard database which included 126 Viola genus plants, the database were used to identify 18 samples of V. kunawarensis that were purchased from drug stores and hospitals. Results: The standard database of 126 Viola genus plants were establish successfully, the identification result showed that eight sequences(44.4%) were authentic as V. kunawarensis, whereas the other samples were all adulterants. Among the adulterants, three sequences(16.7%) were authenticated as V. odorata, six sequences(33.3%) were authenticated Viola genus plants, and one sequence were authenticated as a plants that was not belong to Viola genus. Conclusion: This result indicates that V.kunawarensis were available in the market have complex origins, only less than fifty-percent were quality products, and most of them were adulterants, we need to textual criticism its medication history to ensure whether the adulterants can be used as Viola kunawarensis.
Objective: We surveyed the species composition of commercial V. kunawarensis products using DNA barcoding technology, which would provide method for the identification of medicinal materials. Methods: DNA barcode ITS2 sequence was used to establish a standard database which included 126 Viola genus plants, the database were used to identify 18 samples of V. kunawarensis that were purchased from drug stores and hospitals. Results: The standard database of 126 Viola genus plants were establish successfully, the identification result showed that eight sequences(44.4%) were authentic as V. kunawarensis, whereas the other samples were all adulterants. Among the adulterants, three sequences(16.7%) were authenticated as V. odorata, six sequences(33.3%) were authenticated Viola genus plants, and one sequence were authenticated as a plants that was not belong to Viola genus. Conclusion: This result indicates that V.kunawarensis were available in the market have complex origins, only less than fifty-percent were quality products, and most of them were adulterants, we need to textual criticism its medication history to ensure whether the adulterants can be used as Viola kunawarensis.
引文
1中华人民共和国卫生部药典委员会.中华人民共和国药品标准(维药分册).乌鲁木齐:新疆科技卫生出版社, 1999:8.
2 新疆民族自治区食品药品监督管理局.新疆民族自治区中药民族药饮片炮制规范.乌鲁木齐:新疆人民卫生出版社, 2010:244.
3 刘勇民.民族药志,上册.乌鲁木齐:新疆科技卫生出版社, 1999:35.
4 孙雪芹,刘玫,孙天航,等.东北堇菜属植物叶形态结构的研究及其分类学价值的探讨.草业学报, 2014, 23(2):223-234.
5 梁冠欣,邢福武.堇菜属组间的系统发育与亲缘关系:基于trnLtrnF、psbA-trnH、rpL16、ITS序列细胞学以及形态学证据.云南植物研究, 2010, 32(6):477-488.
6 陈又生.中国堇菜属的分类修订.北京:中国科学院研究生院(植物研究所)博士研究生学位论文, 2006.
7 苏来曼·哈力克,孙磊,沈晓丽,等.民族药质量标准现状分析及发展建议.中国药事, 2015, 12:1256-1262.
8 樊丛照,徐建国,李晓瑾,等.基于DNA条形码技术的民族药材桑葚基原鉴定研究.中国中药杂志, 2017, 42(16):3219-3224.
9 杨珺,樊丛照,李晓瑾.基于ITS2序列的民族药材神香草鉴定研究.中国现代中药, 2017, 19(5):625-629.
10 杨珺,樊丛照,李晓瑾,等.民族药材红豆杉植物基原研究.生物资源, 2017, 39(1):30-35.
11 樊丛照,坤杜孜·阿依,李晓瑾.基于ITS2序列的民族药材蜀葵及其近缘种鉴定研究.中国现代中药, 2016, 18(10):1257-1260+1284.
12 宋海龙,樊丛照,李晓瑾,等.民族药材新疆圆柏DNA条形码鉴定研究.世界科学技术-中医药现代化, 2014, 16(2):319-324.
13 国家药典委员会.中华人民共和国药典(四部).北京:中国医药科技出版社, 2015:383-384.
14 陈士林.中国药典中药材DNA条形码标准序列.北京:科学出版社,2015.
15 Tamura K, Peterson D, Peterson N, et al. MEGA5:molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol, 2011, 28(10):2731-2739.
16 Keller A, Schleicher T, Schultz J, et al. 5. 8S-28S rRNA interaction and HMM-based ITS2 annotation. Gene, 2009, 430(1):50-57.
17 Koetschan C, Hackl T, Müller T, et al. ITS2 Database IV:Interactive taxon sampling for internal transcribed spacer 2 based phylogenies. Mol Phylogenet Evol, 2012, 63(3):585-588.
18 Xin T, Yao H, Gao H, et al. Super food Lycium barbarum(Solanaceae)traceability via an internal transcribed spacer 2 barcode. Food Research International, 2013, 54(2):1699-1704.
19 Xin T Y, Li X J, Song Jingyuan, et al. Survey of commercial Rhodiola products revealed species diversity and potential safety issues. Scientific Reports, 2015, 5:8337.
2 新疆民族自治区食品药品监督管理局.新疆民族自治区中药民族药饮片炮制规范.乌鲁木齐:新疆人民卫生出版社, 2010:244.
3 刘勇民.民族药志,上册.乌鲁木齐:新疆科技卫生出版社, 1999:35.
4 孙雪芹,刘玫,孙天航,等.东北堇菜属植物叶形态结构的研究及其分类学价值的探讨.草业学报, 2014, 23(2):223-234.
5 梁冠欣,邢福武.堇菜属组间的系统发育与亲缘关系:基于trnLtrnF、psbA-trnH、rpL16、ITS序列细胞学以及形态学证据.云南植物研究, 2010, 32(6):477-488.
6 陈又生.中国堇菜属的分类修订.北京:中国科学院研究生院(植物研究所)博士研究生学位论文, 2006.
7 苏来曼·哈力克,孙磊,沈晓丽,等.民族药质量标准现状分析及发展建议.中国药事, 2015, 12:1256-1262.
8 樊丛照,徐建国,李晓瑾,等.基于DNA条形码技术的民族药材桑葚基原鉴定研究.中国中药杂志, 2017, 42(16):3219-3224.
9 杨珺,樊丛照,李晓瑾.基于ITS2序列的民族药材神香草鉴定研究.中国现代中药, 2017, 19(5):625-629.
10 杨珺,樊丛照,李晓瑾,等.民族药材红豆杉植物基原研究.生物资源, 2017, 39(1):30-35.
11 樊丛照,坤杜孜·阿依,李晓瑾.基于ITS2序列的民族药材蜀葵及其近缘种鉴定研究.中国现代中药, 2016, 18(10):1257-1260+1284.
12 宋海龙,樊丛照,李晓瑾,等.民族药材新疆圆柏DNA条形码鉴定研究.世界科学技术-中医药现代化, 2014, 16(2):319-324.
13 国家药典委员会.中华人民共和国药典(四部).北京:中国医药科技出版社, 2015:383-384.
14 陈士林.中国药典中药材DNA条形码标准序列.北京:科学出版社,2015.
15 Tamura K, Peterson D, Peterson N, et al. MEGA5:molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol, 2011, 28(10):2731-2739.
16 Keller A, Schleicher T, Schultz J, et al. 5. 8S-28S rRNA interaction and HMM-based ITS2 annotation. Gene, 2009, 430(1):50-57.
17 Koetschan C, Hackl T, Müller T, et al. ITS2 Database IV:Interactive taxon sampling for internal transcribed spacer 2 based phylogenies. Mol Phylogenet Evol, 2012, 63(3):585-588.
18 Xin T, Yao H, Gao H, et al. Super food Lycium barbarum(Solanaceae)traceability via an internal transcribed spacer 2 barcode. Food Research International, 2013, 54(2):1699-1704.
19 Xin T Y, Li X J, Song Jingyuan, et al. Survey of commercial Rhodiola products revealed species diversity and potential safety issues. Scientific Reports, 2015, 5:8337.