| 濒危兰科药用植物DNA条形码鉴定 |
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| 引用本文： | 汤欢,向丽,李西文,孙伟,王美娜,黄云峰,叶萌.濒危兰科药用植物DNA条形码鉴定[J].中国中药杂志,2017,42(11):2058-2067. |
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| 作者姓名： | 汤欢 向丽 李西文 孙伟 王美娜 黄云峰 叶萌 |
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| 作者单位： | 四川农业大学 林学院, 四川 成都 611130;中国中医科学院 中药研究所 中药鉴定与安全性检测评估北京市重点实验室, 北京 100700,中国中医科学院 中药研究所 中药鉴定与安全性检测评估北京市重点实验室, 北京 100700,中国中医科学院 中药研究所 中药鉴定与安全性检测评估北京市重点实验室, 北京 100700,中国中医科学院 中药研究所 中药鉴定与安全性检测评估北京市重点实验室, 北京 100700,全国兰科植物种质资源保护中心 深圳市兰科植物保护研究中心, 广东 深圳 518114,广西中医药研究院, 广西 南宁 530022,四川农业大学 林学院, 四川 成都 611130 |
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| 基金项目： | 2014高校基本科研业务费专项课题（甘财教[2014]63号06课题）；甘肃教育厅科研项目（1106B-07）；科技基础性工作专项重点项目子课题（SB2007FY020）；甘肃省基础研究创新群体项目（1606RJIA323）；中央引导地方科技发展专项项目 |
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| 摘 要： | 兰科药用植物形态分类困难,此研究采用DNA条形码分子鉴定法从分子水平验证兰科药用植物的传统形态分类。以matK,psbA-trnH和ITS2序列作为DNA条形码对已进行了形态鉴定的49属135种163份兰科药用植物样品进行分子鉴定,经DNA提取,PCR扩增、双向测序及校对拼接后,将得到的序列在GenBank中进行BLAST比对,然后运用MEGA 7.0软件中的Neighbor-joining(NJ)法构建物种系统进化树。结果表明,163份样品均能成功提取DNA;matK,psbA-trnH和ITS2序列的PCR扩增效率分别为100%,100%,98.77%;共获得487条序列,其中345条序列在GenBank数据库中比对到了相应物种的序列,142条为新增序列;运用NJ法构建的兰科药用植物系统进化树中,基于matK序列所构建的物种系统进化树要优于基于psbA-trnH和ITS2序列所构建的系统进化树。matK,psbA-trnH和ITS2序列在鉴定兰科药用植物过程中互为补充,DNA条形码分子鉴定法可用于辅助兰科药用植物的分类鉴定。
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| 关 键 词： | 兰科 DNA条形码 药用植物 分子鉴定 |
| 收稿时间： | 2016/12/16 0:00:00 |
DNA barcoding identification of endangered medicinal plants of Orchidaceae |
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| TANG Huan,XIANG Li,LI Xi-wen,SUN Wei,WANG Mei-n,HUANG Yun-feng and YE Meng.DNA barcoding identification of endangered medicinal plants of Orchidaceae[J].China Journal of Chinese Materia Medica,2017,42(11):2058-2067. |
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| Authors: | TANG Huan XIANG Li LI Xi-wen SUN Wei WANG Mei-n HUANG Yun-feng and YE Meng |
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| Institution: | College of Forestry, Sichuan Agricultural University, Chengdu 611130, China;Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China,Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China,Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China,Key Laboratory of Beijing for Identification and Safety Evaluation of Chinese Medicine, Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China,The National Orchid Conservation Center of China and The Orchid Conservation & Research Center of Shenzhen, Shenzhen 518114, China,Guangxi Institute of Chinese Medicine and Pharmaceutical Science, Nanning 530022, China and College of Forestry, Sichuan Agricultural University, Chengdu 611130, China |
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| Abstract: | In this study, DNA barcoding was used to validate the traditional morphological classification of medicinal plants of Orchidaceae. The 163 samples of 135 species belong to 49 genera which have been confirmed by morphological identification were collected. Candidate sequences, including matK, psbA-trnH and ITS2 sequences, were amplified, bidirectionally sequenced, and assembled. All the sequences were blasted to GenBank database at NCBI, then analyzed using Neighbor-joining tree method by MEGA 7.0. The results showed that the DNAs of 163 samples were successfully extracted. The amplification efficiency of matK, psbA-trnH and ITS2 sequences were 100%, 100% and 98.77%, respectively. The 487 sequences were obtained, 345 sequences of which have matched corresponding sequences in the GenBank database and 142 sequences were new sequences. The topology of NJ tree which were constructed with the matK sequences was better than the trees of psbA-trnH and ITS2 sequences. In conclusion, the matK, psbA-trnH and ITS2 sequences were complementary and suitable for identification of medicinal plants of Orchidaceae. DNA barcoding can be used as an auxiliary means for identification of medicinal plants of Orchidaceae. |
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| Keywords: | Orchidaceae DNA barcoding medicinal plants molecular identification |
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