| 引用本文:周建国1，马双姣1，黄玉龙2，石林春1，金钺1，姚辉1.种子类药材补骨脂及其混伪品的ITS2条形码序列鉴定[J].世界中医药,2016,(05):. |
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| 种子类药材补骨脂及其混伪品的ITS2条形码序列鉴定 |
| Identification of Psoraleae Fructus and its adulterants using ITS2 sequence |
| 投稿时间：2016-04-12 |
| DOI：10.3969/j.issn.1673-7202.2016.05.008 |
| 中文关键词: 补骨脂 ITS2序列 鉴定 DNA条形码 混伪品 |
| English Keywords:Psoralea corylifolia L. ITS2 sequence Identification DNA barcoding Adulterants |
| 基金项目:重大新药创制国家科技重大专项(编号：2014ZX09304307001)；国家科技支撑计划（编号：2011BAI07B08） |
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| 中文摘要: |
| 目的：种子类药材补骨脂具有肝肾毒性，其混伪品曼陀罗子、天仙子、洋金花的种子、猪屎豆也含有毒性成分，且补骨脂与其混伪品外形相似，易导致混用误用，严重危害人体健康。本研究利用DNA条形码技术，分析补骨脂及其混伪品的ITS2序列，以期快速准确鉴定补骨脂及其混伪品，保证其用药安全及临床疗效。方法：对补骨脂及其混伪品样品进行DNA提取、PCR扩增ITS2序列，并进行双向测序，所有序列用MEGA6.0软件进行种间、种内Kimura2-parameter（K2P）遗传距离计算，并构建NJ系统聚类树。结果：补骨脂ITS2序列长度为233bp，G+C含量为69.5%，种内无变异位点。补骨脂与其混伪品种间变异位点较多，种间K2P最小距离为0.5321，远远大于其种内距离，NJ树显示可以将补骨脂及其混伪品完全分开。结论：ITS2条形码序列可准确鉴别种子类药材补骨脂及其混伪品，为保证补骨脂的临床用药安全和种质资源鉴定提供了良好的技术手段。 |
| English Summary: |
| Psoraleae Fructus has toxic effects on liver and kidney and its adulterants including Datura stramonium，Hyoscyamus niger，D.metel，Crotalaria pallid are also tested to have toxic ingredients.Psoralea corylifolia shares similar morphological characters with its adulterants，which can easily lead to misuses and greatly threaten the human health.This study aimed to identify P.corylifolia and its adulterants rapidly and accurately using DNA barcoding method based on ITS2 sequence.Methods:Genomic DNA of P.corylifolia and its adulterants were extracted and then the ITS2 sequences were obtained by direct PCR amplification and sequenced bi-directionally.The intra-and inter-specific genetic distances were analyzed using software MEGA 6.0，and the phylogenetic tree was constructed by using the neighbor-joining(NJ)method.Results:The ITS2 sequence length of P.corylifolia was 233bp，and the G+C content was 69.5%.The intra-specific variations were highly conservative.There were many variation sites of ITS2 sequences between P.corylifolia and its adulterants.The results showed that the minimum inter-specific divergence was 0.5321，which was larger than the maximum intra-specific divergence.The NJ tree indicated that P.corylifolia could be distinguished from its adulterants obviously.Conclusion:The ITS2 region can be used as a barcode for identification of P.corylifolia，which provide a new technique for the authentication of germplasm resources and clinic safety of medicinal use. |
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