摘要
[目的]利用DNA条形码ITS1、ITS2碱基序列及其二级结构对巴戟天药材及其混伪品进行鉴定.[方法]利用Codon Code Aligner 17.0或SeqMan软件对测序获得的双向序列进行拼接,从GenBnak上下载巴戟天及其混伪品的ITS1、ITS2碱基序列并利用MEGA 11.0软件分别与测序获得的碱基序列进行比对分析,获得测序样品的ITS1、ITS2碱基序列.将所有ITS1、ITS2碱基序列利用MEGA 11.0软件比对分析,分别构建NJ、ML系统聚类树并计算种内、种间遗传距离及各物种种间碱基变异位点、简约信息位点,利用在线网页构建各物种ITS1、ITS2碱基序列二级结构.[结果]基于ITS1碱基序列巴戟天与大果巴戟、虎刺、鸡眼藤、假巴戟、南五味子、香巴戟、羊角藤的种间遗传距离分别为0.034、0.233、0.052、0.052、1.004、1.004、0.049,种间简约信息位点分别为6、30、9、9、90、90、10个,根据NJ系统聚类树显示虎刺与羊角藤、香巴戟与南五味子无法鉴别,ML系统聚类树显示香巴戟与南五味子无法鉴别.基于ITS2碱基序列巴戟天与大果巴戟、虎刺、鸡眼藤、假巴戟、南五味子、香巴戟、羊角藤的种间遗传距离分别为0.014、0.113、0.034、0.015、0.509、0.516、0.017,种间简约信息位点分别为7、34、15、8、91、88、9个,根据NJ系统聚类树显示除香巴戟与南五味子无法鉴别外,其他各物种均独立聚为一支,ML系统聚类树显示各物种均独立聚为一支.基于ITS2构建的二级结构能够准确对各物种进行鉴别.[结论]基于ITS2构建的ML系统发育树、二级结构均能够对巴戟天及其混伪品进行鉴别,ITS2序列的物种鉴定能力优于ITS1序列.
[OBJECTIVE]The secondary structure constructed by ITS1 and ITS2 base sequences and ITS1 and ITS2 base sequences of DNA barcodes was used to identify Morinda officinalis and its adulterants.[METHODS]Codon Code Aligner 17.0 or SeqMan software is used to splice the biaxial sequence obtained by sequencing,download the ITS1 and ITS2 base sequence of M.officinalis and its blends from GenBnak,and then compare and analyze the base sequence obtained by sequencing with MEGA 11.0 software.The ITS1 and ITS2 base sequences of sequencing samples were obtained.All ITS1 and ITS2 base sequences were comparedand analyzed by MEGA 11.0 software,NJ and ML systematic cluster trees were constructed respectively,and intraspectic and interspecific genetic distances,interspecific base variation sites and reduced information sites of each species were calculated.The secondary structure of ITS1 and ITS2 base sequences of each species was constructed by using online web pages.[RESULTS]Based on ITS1 base sequence,the genetic distance between M.officinalis,Morinda cochinchinensis,Damnacanthus indicus,Morinda parvifolia,Morinda shuanghuaensis,Kadsura longipedunculata,Schisandra propinqua,Morinda umbellata,respectively were 0.034,0.233,0.052,0.052,1.004,1.004 and 0.049.The interspecific parsimony information loci were 6,30,9,9,90,90 and 10,respectively.According to the NJ system clustering tree,it could not be identified between D.indicus and M.umbellata,and S.propinqua and K.longipedunculata,while the ML system clustering tree showed that S.propinqua and K.longipedunculata could not be identified.Based on ITS2 base sequence,the interspecific genetic distance between M.officinalis,M.cochinchinensis,D.indicus,M.parvifolia,M.shuanghuaensis,K.longipedunculata,S.M.umbellata,were 0.014,0.113,0.034,0.015,0.509,0.516,and 0.017,respectively.Interspecific parsimony information loci were 7,34,15,8,91,88 and 9,respectively.According to the NJ system clustering tree,all the other species were clustered independently into a single branch except S.propinqua a
作者
孙一帆
白华
张娣
闫岩
王孟虎
SUN Yifan;BAI Hua;ZHANG Di;YAN Yan;WANG Menghu(Bozhou Vocational and Technical College,Bozhou 236800,China;Bozhou University,Bozhou 236800,China)
出处
《商丘师范学院学报》
CAS
2024年第9期42-48,共7页
Journal of Shangqiu Normal University
基金
亳州职业技术学院项目(2023AH053216,2021bzjyxm26,KJ2020A1019,2020bzjyxm15)
亳州学院项目(bzzc2023030,KJ2019ZD80,GXXT-2023-073)。
