摘要
目的:应用ITS2序列对朱砂根及其混伪品进行鉴定研究,为中药临床准确用药、市场规范化管理等提供依据和保障。方法:对朱砂根及其混伪品进行DNA提取、PCR扩增ITS2序列、双向测序,对序列进行比对、计算遗传距离。结果:朱砂根ITS2序列长度为217 bp,种内有10个变异位点,有9种单倍型,平均G+C含量为59.1%,除变种红凉伞外,朱砂根与其各混伪品的种间最小遗传距离均大于朱砂根种内最大遗传距离,所以利用ITS2序列可以鉴别朱砂根及其混伪品。结论 :实验结果表明ITS2序列可以鉴别朱砂根及其混伪品,但对于其与变种的关系需进一步研究。
Objective：This research aims to identify Ardisiae Crenatae Radix and its adulterants accurately using the ITS2 sequence.It will provide the foundation for its clinical medication and accurate market standardized management.Methods：Ardisia Crenatae Radix and its adulterants were identified and distinguished by ITS2 barcode，one marker of DNA barcoding.Genomic DNA of Ardisia crenata and its adulterants were extracted and then the ITS2 regions were obtained by PCR amplification and sequenced bidirectionally.The ITS2 sequences were aligned and the genetic distances were computed using MEGA 6.0 in accordance with the Kimura 2-parameter （K2P） model.Results：The length of A.crenata sequence is 217 bp and the average G＋C content was 59.1%.There were ten variable sites of the ITS2 regions of A.crenata and 48 ITS2 sequences of A.crenata included nine haplotypes.Except for A.crenata var.bicolor，the maximum intra-specific K2P genetic distance of Ardisiae Crenatae Radix and its adulterants was less than the minimum inter-specific K2P genetic distance.Conclusion：The experimental results indicated that the ITS2 sequence could be a good marker to identify Ardisia Crenatae Radix and its adulterants.But the relationship of A.crenata and A.crenata var.bicolor need to be further studied.
作者
陈新连
周建国
马双姣
姚辉
林余霖
王瑀
CHEN Xinlian ZHOU Jianguo MA Shuangjiao YAO Hui LIN Yulin WANG Yu(The Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Ministry of Education, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences ＆ Peking Union Medical College, Beijing 100193, China)
出处
《中国现代中药》
CAS
2017年第7期939-943,964,共6页
Modern Chinese Medicine
基金
国家科技支撑计划(2011BAI07B08)
中国医学科学院医学与健康科技创新工程(2016-I2M-3-016)
关键词
朱砂根
鉴别
ITS2序列
DNA条形码
混伪品
Ardisiae Crenatae Radix
identification
ITS2 sequence
DNA barcoding
adulterants
