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大黄与易混伪品土大黄、虎杖原植物的ITS2序列鉴定
引用本文:李美妮,韩蕊莲,韩建萍,姚辉,罗焜,宋经元. 大黄与易混伪品土大黄、虎杖原植物的ITS2序列鉴定[J]. 环球中医药, 2012, 5(3): 185-189
作者姓名:李美妮  韩蕊莲  韩建萍  姚辉  罗焜  宋经元
作者单位:1. 712100,陕西杨凌,西北农林科技大学生命科学学院;中国医学科学院,北京协和医学院药用植物研究所中草药物质基础与资源利用教育部重点实验室
2. 西北农林科技大学生命科学学院,陕西杨凌,712100
3. 中国医学科学院,北京协和医学院药用植物研究所中草药物质基础与资源利用教育部重点实验室
基金项目:国家自然科学基金(81001608)
摘    要:目的分析大黄与其易混伪品原植物的rDNA上的ITS2序列信息,探索鉴定大黄及其混伪品的新方法。方法提取DNA模板后对其ITS2区进行PCR扩增、测序;拼接序列;计算种内种间K2P距离,最后基于K2P距离建立NJ树。结果大黄与其混伪品的最小种间K2P距离(38.6%)大于最大种内K2P距离(8.0%),NJ树中大黄三个基原的不同来源样品聚为一支。结论因此ITS2序列可以作为鉴定大黄与易混伪品土大黄、虎杖的候选条形码序列,DNA条形码技术在中药鉴定中具有极大的前景。

关 键 词:大黄  混伪品  鉴定  ITS2序列

Identification of original plants of Radix et Rhizoma Rhei from its adulterants Rhizoma et Radix Polygoni Cuspidati and Radix Rumicis Obtusifolii by ITS2 sequences
Affiliation:LI Mei-ni, HAN Rui-lian, HAN flan-ping,et al. (College of Life Science, Northwest Science and Technology University of Agriculture and Forestry, Shaanxi 712100, China)
Abstract:Objective The ITS2 sequence of rDNA was utilized as a novel technique to discriminate three original plants of Radix et Rhizoma Rhei from its adulterants. Methods DNA was extracted from each sample as template, of which the ITS2 region was amplified in a PCR and then sequenced. Sequences were assembled. Inter-and intra-specific K2P distances were calculated, based on which NJ tree was performed. Results The K2P distance of the minimum inter-specific 38.6% was higher than that of the maximum intra-specific 8.0%. The different samples of three original plants of Radix et Rhizoma Rhei were clustered into one elade. Conclusion The ITS2 fragment could be used as a candidate bareode for identifying original plants of Radix et Rhizoma Rhei from its adulterants Rhizoma et Radix Polygoni Cuspidati and Radix Rumicis Obtusifolii, and DNA barcoding has a wide prospect in authenticating Traditional Chinese Medicine.
Keywords:Radix et Rhizoma Rhei  Adulterants  Identification  ITS2 sequence
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