基于DNA条形码鉴别青葙子与鸡冠花子及其遗传多样性分析  

DNA barcode screening and genetic diversity analysis of the Celosiae semen and Celosia cristatae semen

作　　者：宋叶 李国卫 兰小勇 钟春琳 谭斯尹 刘闪闪 潘礼业 陈向东 孙冬梅 SONG Ye;LI Guowei;LAN Xiaoyong;ZHONG Chunlin;TAN Siyin;LIU Shanshan;PAN Liye;CHEN Xiangdong;SUN Dongmei(Guangdong Yifang Pharmaceutical Co.,Ltd.,Guangdong Provincial Key Laboratory of Traditional Chinese Medicine Formula Granule,Foshan 528244,Guangdong Province,China)

机构地区：[1]广东一方制药有限公司,广东省中药配方颗粒企业重点实验室,广东佛山528244

出　　处：《药学前沿》2025年第1期73-81,共9页China Pharmacist

基　　金：佛山市南海区重点领域科技攻关项目(南科﹝2023﹞20号-18);产业技术基础公共服务平台项目(2022-230-221)。

摘　　要：目的应用分子生物学鉴定技术,筛选出应用于鉴定青葙子与鸡冠花子的最佳DNA条形码,建立快速、准确、便捷的青葙属植物鉴定方法。方法收集青葙子、鸡冠花子样品共计21份,提取样品总DNA,对内部转录间隔区(ITS)、psbA-trnH、matK、rbcL和trnL-trnF序列进行扩增和测序,采用MEGA-X软件对数据进行分析处理,计算Kimura-2-parameter遗传距离,建立邻接聚类进化树,进行对比分析,基于TaxonDNA计算BestMatch、BestCloseMatch以评估DNA条形码的鉴别能力,利用ITS2数据库和RNAfold数据库预测条形码序列二级结构。结果条形码ITS、matK、psbA-trnH、rbcL、trnL-trnF序列均扩增成功且具有较高的测序成功率,其中psbA-trnH具有最多的变异位点,ITS次之。psbA-trnH、ITS2、trnL-trnF具有较明显的条形码间隙,以psbA-trnH最为显著(100%)。系统进化树显示,IITS、ITS2、psbA-trnH、matK、trnL-trnF序列均可将青葙子与鸡冠花子各自聚为一支,各分支点的支持率均高于60%,以psbA-trnH、trnL-trnF的各分支点的支持率最高(99%)。分子方差分析结果显示psbA-trnH序列的群体遗传分化指数最高,适用于区分青葙子和鸡冠花子物种间差异。除matK外,青葙子与鸡冠花子psbA-trnH、ITS2、trnL-trnF序列的二级结构均有差异。结论以psbA-trnH为主、ITS和trnL-trnF为辅,可实现青葙子和鸡冠花子的准确鉴定。Objective To screen the optimal DNA barcode for identifying Celosiae semen and Celosia cristatae semen using the molecular biology identification techniques,and to establish a rapid,accurate,and convenient approach for identifying species in the Celosia L..Methods A total of 21 samples of Celosiae semen and Celosia cristatae semen were collected,and total DNA was extracted from the samples.The sequences of ITS,psbA-trnH,matK,rbcLand trnL-trnF were amplified and sequenced,MEGA-X software was used to analyze and process the data,the Kimura-2-parameter genetic distance was calculated,an neighbor-joining method cluster evolutionary tree was established,and the comparative analysis was conducted.The BestMatch and BestCloseMatch was calculate based on TaxonDNA to evaluate the discriminative ability of DNA barcodes.The ITS2 database and RNAfold database were used to predict the secondary structure of the barcode sequence.Results The barcode ITS,matK,psbA-trnH,rbcL,and trnL-trnF sequences were successfully amplified and had high sequencing success rates.Among them,the psbA-trnH had the most mutation sites,followed by ITS.The psbA-trnH,ITS2 and trnL-trnF had significant barcoding gap,and psbA-trnH was the most significant(100%).According to the phylogenetic tree,the IITS,ITS2,psbA-trnH,matK and trnL-trnF sequences could cluster Celosiae semen and Celosia cristatae semen into a separate branch,the support rates of each branch point were above 60%,with psbA-trnH and trnL-trnF having the highest support rate(99%).Analysis of molecular variance results showed that the psbA-trnH had the highest population genetic differentiation index and was the most suitable for distinguishing differences between Celosiae semen and Celosia cristatae semen.Except for matK,there were differences in the secondary structure of psbA-trnH,ITS2 and trnL-trnF sequences between Celosiae semen and Celosia cristatae semen.Conclusion With psbA-trnH as the main component ITS and trnL-trnF as the auxiliary ones,accurate identification of Celosiae semen an

关 键 词：青葙子 鸡冠花子 DNA条形码 物种鉴定 遗传距离 系统发育树分析 二级结构 

分 类 号：R282.5[医药卫生—中药学]