蒲黄、松花粉等花粉类药材及其混伪品的DNA条形码鉴定  

Identification of cattail pollen(Puhuang),pine pollen(Songhuafen) and its adulterants by ITS2 sequence

作　　者：马孝熙[1,2] 孙伟[2] 任伟超[2] 向丽[2] 赵博[2] 张雅琴[1] 宋明[1] 慕泽泾[3] 陈士林[2] 

机构地区：[1]武汉理工大学化学工程学院,湖北武汉430070 [2]中国中医科学院中药研究所,北京100700 [3]江西中医药大学,江西南昌330004

出　　处：《中国中药杂志》2014年第12期2189-2193,共5页China Journal of Chinese Materia Medica

基　　金：国家"重大新药创制"科技重大专项(2013ZX09301307;Z131100006513017);国家高技术研究发展计划(863)项目(2012AA021602)

摘　　要：目前市场上花粉类药材品种较多，由于在外观形态均呈粉末状，很难区分，加之市场价格较高等因素，故混用和掺伪的情况时有发生，因此花粉类药材的准确鉴定对公众的用药安全显得尤为重要。该研究采用DNA条形码技术，对蒲黄、松花粉及其混伪品的基原植物及药材共60份样本进行研究。通过primer premier 6.0设计出蒲黄ITS2特异性引物PhF-R，其扩增效率高达100%，实验结果表明蒲黄药材的ITS2序列长度为234-249 bp，种内K2P平均距离与同属种间K2P平均距离十分接近，但远小于其与混伪品的种间K2P平均距离；松花粉药材的ITS2序列长度均为247 bp，ITS2序列种内平均K2P遗传距离远小于其与混伪品种间平均K2P距离。利用ITS2序列构建的邻接（NJ）树表明蒲黄、松花粉及其混伪品可明显区分，并呈现出良好的单系性。因此，DNA条形码可对该实验材料准确鉴定，有助于花粉类药材的监管及市场流通。DNA barcoding method was conducted for the authentication of pollen materials due to difficulty of discriminating pollen materials bearing morphological similarity. In this study, a specific focus was to identify cattail pollen （Puhuang） and pine pollen （Songhuafen） samples from their adulterants which are frequently mixed-together. Regions of the internal transcribed spacer （ITS2） from 60 samples were sequenced, and new primers for cattail pollen were designed according to the sequence information. The results from the NJ trees showed that the species of pine pollen, Puhuang and their adulterants can be classified as obvious monophyly. Therefore, we propose to adapt DNA barcoding methodology to accurately distinguish cattail pollen, pine pollen and their adulterant materials. It is a great help for drug regulatory agency to supervise the quality of medicinal materials.

关 键 词：ITS2 蒲黄 松花粉 DNA条形码 

分 类 号：R282.5[医药卫生—中药学]