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1.北京中医药大学 中药学院,北京 100102
2.北京中医药大学 北京中医药研究院,北京 100029
3.北京中医药大学 东直门医院,北京 100700
[第一作者] 罗凌龙,在读硕士,从事药用植物与分子生药学研究,E-mail:luolinglong1995@163.com
*王学勇,博士,教授,从事药用植物与分子生药学研究,E-mail: wxyph.d@163.com
收稿日期:2019-03-26,
网络出版日期:2019-10-12,
纸质出版日期:2020-01-20
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罗凌龙, 张弛, 邓博, 等. 基于DNA条形码技术的俄罗斯特色药用植物资源调查[J]. 中国实验方剂学杂志, 2020,26(2):175-180.
Ling-long LUO, Chi ZHANG, Bo DENG, et al. Russian Characteristic Medicinal Plant Resources Based on DNA Barcode Technology[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(2): 175-180.
罗凌龙, 张弛, 邓博, 等. 基于DNA条形码技术的俄罗斯特色药用植物资源调查[J]. 中国实验方剂学杂志, 2020,26(2):175-180. DOI: 10.13422/j.cnki.syfjx.20200211.
Ling-long LUO, Chi ZHANG, Bo DENG, et al. Russian Characteristic Medicinal Plant Resources Based on DNA Barcode Technology[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(2): 175-180. DOI: 10.13422/j.cnki.syfjx.20200211.
目的:
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调查俄罗斯高加索、阿尔泰野外地区药用植物的分布现状,明确该地区药用植物的种类、功效信息,深入挖掘一带一路沿线国家地区的药用植物新资源、新功效。
方法:
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在野外搜寻并采集药用植物制成蜡叶标本运回国,提取腊叶标本的植物总DNA,使用内转录间隔区(ITS)序列通用引物进行聚合酶链式反应(PCR)扩增,PCR产物送双向测序,测序结果经软件进行拼接,根据美国国立生物技术信息中心(NCBI)数据库BLAST所获相似度最高物种的同属ITS序列,经DNAman软件对待鉴定物种的ITS序列和同属ITS序列进行相似度分析,利用MEGA 7软件作进化树,采用Kimura-2参数遗传距离,采用邻接法构建邻接法(NJ)系统树,发育树各分支的置信度用自举检验法检验,共进行2 000次循环,根据聚类结果进行鉴定。在此基础上对俄罗斯高加索、阿勒泰野外地区的重点药用植物进行归纳与分析。
结果:
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经过NCBI数据库BLAST比对,各标本ITS序列与NCBI数据库上登录序列聚为一支,与外类群明显分开,结合植物标本性状特征,确定了待鉴定标本的种属分类。该次野外收集的标本中共鉴定出51种植物,覆盖17科44属,有明确功效记载的植物共有29种。查阅俄罗斯联邦国家药品目录和《中国药典》,分析国内常用药用植物,总结出20种中俄常见药材其在当地具有不同药用功效。该研究对扩大中药用药范围和临床经验具有重要的借鉴意义,也为当地加强物种保护和开发利用野生药用植物资源提供了依据。
Objective:
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To investigate the distribution status of medicinal plants in the wild areas of Russian Caucasus and Altai
and clarify the types and efficacy information of medicinal plants in the area
so as to dig deep into new resources and new functions of medicinal plants in the countries along the Belt and Road.
Method:
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Medicinal plants in the wild were searched and collected to make waxy specimens
and sent back to the country to extract the total DNA of the leaves of the leaves. Internal Transcribed Spacer(ITS)sequence universal primers were used for Polymerase Chain Reaction (PCR)amplification. The PCR products were sent for the two-way sequencing
and the sequencing results are spliced by software according to National Center for Biotechnology Information(NCBI). The same ITS sequence of the highest similarity species obtained by database BLAST was analyzed by DNAman software to identify the ITS sequence of the species and the ITS sequence of the same species. The MEGA 7 software was used as the phylogenetic tree
and the Kimura-2 parameter genetic distance was used to construct the neighbor joining(NJ) phylogenetic tree by the neighbor-joining method. The confidence of each branch of the development tree was tested by the bootstrap test method. A total of 2 000 cycles were performed
and the results were identified based on the clustering results. On this basis
the key medicinal plants in the Russian Caucasus and Altay wild areas were summarized and analyzed.
Result:
2
After BLAST alignment in NCBI database
the ITS sequences of each specimen were clustered with the login sequences on the NCBI database
which were separated from the outer group. The species classification of the specimens to be identified was determined by combining the characteristics of the specimens. A total of 51 plants were identified from the specimens collected in the field
covering 44 genera of 17 families
and 29 plants had clear efficacy records. The National Drug List of the Russian Federation and the Chinese Pharmacopoeia were retrieved to summarize commonly used medicinal plants in China and conclude that 20 kinds of Chinese and Russian common medicinal materials have different medicinal effects in local areas. This study has a reference significance for expanding the scope and clinical experience of traditional Chinese medicines
and provides a basis for strengthened local species conservation
development and utilization of wild medicinal plant resources.
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The State Register of Medicinal Drugs . Official publication in 2 volumes [M]. Moscow : Medicine , 2004 .
国家药典委员会 . 中华人民共和国药典.一部 [M]. 北京 : 中国医药科技出版社 , 2015 .
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