ITS／ITS2条形码序列在花椒药材鉴定中的应用  

Application Study of ITS /ITS2 Barcodes in Identification of Zanthoxmli Pericarpium

作　　者：侯典云[1,2] 宋经元[2] 杨培[2] 周红[2] 姚辉[2] 

机构地区：[1]河南科技大学农学院,河南洛阳471000 [2]中国医学科学院北京协和医学院药用植物研究所、濒危药材繁育国家工程实验室,北京100193

出　　处：《中国药学杂志》2014年第7期534-538,共5页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金资助项目(81130069);国家高技术研究发展计划(863计划)资助项目(2012AA021602);教育部长江学者和创新团队发展计划(IRT1150)

摘　　要：目的评价ITS／ITS2条形码序列在花椒药材鉴定中的准确性与稳定性。方法以多基原药材花椒为研究对象，采用改良的试剂盒法提取药材基因组DNA，PCR扩增ITS序列，利用基于隐马尔可夫模型的HMMer注释方法获得ITS2序列。通过分析花椒药材及其同属混伪品的种内、种间变异，利用中药材DNA条形码鉴定系统和NJ系统聚类树对花椒药材进行鉴定分析。结果花椒药材的ITS序列长度为618～620bp，ITS2序列长度为224bp，两种基原物种花椒和青椒的ITS／ITS2序列种内平均K2P遗传距离均远小于其与混伪品的种间K2P遗传距离；中药材DNA条形码鉴定系统和NJ树均可以准确区分花椒药材及其同属混伪品。结论ITS／ITS2序列作为DNA条形码能稳定、准确鉴别花椒药材。OBJECTIVE To evaluate the accuracy and stability of ITS/ITS2 barcodes in identification of Zanthoxmli Periearpium. METHODS Total genomic DNAs from samples were extracted by using improved DNA extraction kits. ITS regions were amplified, and the ITS2 sequences were obtained by using the hidden Markov model （ HMM ） based on annotation method fi＇om the ITS sequences. The inter- and intra-specific variation of the Zanthoxmli Pericarpium and its adulterants were analyzed. Zanthoxmli Pericarpium was i- dentified through the Species identification System for Traditional Chinese Medicine and neighbor-joining （N J）phylogenetic trees. RE- SULTS The lengths of ITS/ITS2 sequence of Zanthoxmli Pericarpimn were 618 -620 bp and 224 bp. The intra-specifie genetic dis- tances were smaller than inter-specific ones in ITS/ITS2 regions of Z. schinifolium and Z. bungeanum. Zanthoxmli Pericarpium and its adulterants could be easily differentiated according to the nearest distance, the Species Identification System for Traditional Chinese Medicine and the NJ trees methods. CONCLUSION ITS/ITS2 regions as DNA barcodes can stably and accurately distinguish Zan- thoxmli Pericarpium and its adulterants.

关 键 词：花椒药材 ITS ITS2 DNA条形码 分子鉴定 

分 类 号：R284[医药卫生—中药学]