基于叶绿体DNA条形码的披麻草药材及其混伪品的分子鉴定  

Molecular Distinguishment of Original Species of Pimacao from the Adulterants:Based on Chloroplast DNA Barcoding

作　　者：时瑞梓 田星 赵禹 李荣姣 李国栋 SHI Rui-zi;TIAN Xing;ZHAO Yu;LI Rong-jiao;LI Guo-dong(Yunnan Medical Health College,Kunming 650500,China;Yunnan Key Laboratory of Dai and Yi Medicines,Yunnan University of Chinese Medicine,Kunming 650500,China)

机构地区：[1]云南医药健康职业学院,云南昆明650500 [2]云南中医药大学云南省傣医药与彝医药重点实验室,云南昆明650500

出　　处：《中国现代中药》2023年第1期31-37,共7页Modern Chinese Medicine

基　　金：云南省教育厅科学研究基金项目(2021J1366)。

摘　　要：目的:分析trnV-atpE、psaC-ndhE序列变异特点,并通过该序列鉴别披麻草不同基原植物与其混伪品。方法:分别提取披麻草3种基原植物及其混伪品的总DNA,以trnV-atpE、psaC-ndhE为DNA条形码候选序列进行聚合酶链式反应(PCR)扩增和测序。采用DnaSP、MEGA软件统计其片段长度、鸟嘌呤(G)+胞嘧啶(C)占比、变异位点个数、简约信息位点、插入或缺失位点,并计算样品K2-P(Kimura 2-parameter)遗传距离和构建邻接法(NJ)系统发育树。结果:trnV-atpE、psaC-ndhE序列长度分别为373~381、491~534 bp,简约信息位点分别为18、15个。trnV-atpE和psaC-ndhE序列中披麻草3种基原植物最大遗传距离均小于与混伪品之间的最小遗传距离。基于2个序列构建的NJ系统发育树显示,披麻草与其混伪品能明显区分。结论:综合分析序列特点和种内、种间变异性,trnV-atpE和psaC-ndhE序列均可将披麻草3种基原植物与其混伪品区分开。Objective:To analyze the variation characteristics of trnV-atpE and psaC-ndhE sequence fragments and distinguish the original species(Veratrum taliense,Veratrum stenophyllum,Veratrum mengtzeanum)of Pimacao(a medicinal derived from the roots or rhizome/roots of the three species)from the adulterants with the sequences.Methods:The genomic DNA was extracted from the original species and the adulterants,respectively,and polymerase chain reaction amplification and sequencing were performed with trnV-atpE and psaC-ndhE as candidate sections for DNA barcoding.DnaSP and MEGA were used to measure the fragment length,guanine(G)+cytosine(C)content,number of variant sites,parsimony informative sites,and InDel sites.The K2-P(Kimura 2-parameter)genetic distance of the samples was calculated,and the phylogenetic tree was constructed with the Neighbor-Joining(NJ)method.Results:The trnV-atpE and psaC-ndhE were 373-381 bp and 491-534 bp,separately,with 18 and 15 parsimony informative sites,respectively.Based on trnV-atpE and psaC-ndhE sequences,the maximum genetic distance among original species was smaller than the minimum genetic distance between the original species and the adulterants.The phylogenetic tree clearly distinguished the original species from the adulterants.Conclusion:The original species of Pimacao and the adulterants can be clearly distinguished by trnV-atpE and psaC-ndhE DNA sequences based on comprehensive analysis of sequence characteristics and intraspecific and interspecific variation.

关 键 词：披麻草 叶绿体DNA trnV-atpE序列 psaC-ndhE序列 遗传距离 系统发育树 分子鉴定 

分 类 号：R282[医药卫生—中药学]