基于DNA条形码和二级结构鉴定菟丝子及其易混伪品
Identification of Cuscutae semen and Its Miscible Counterfeit Based on DNA Barcode and Secondary Structure
周凯伦 1王雨艨 2许晴 1赵薇薇 1马进进 1张圣阳 1王默含 1吴婷婷 1胡建建 1王孟虎 1孙一帆3
作者信息
- 1. 亳州学院,安徽 亳州 236800
- 2. 亳州市食品药品检验中心,安徽 亳州 236800
- 3. 亳州职业技术学院,安徽 亳州 236800
- 折叠
摘要
目的:利用DNA条形码ITS2碱基序列及其二级结构对菟丝子及其混伪品金灯藤进行鉴定.方法:从安徽亳州药材市场收集10份菟丝子、5份南方菟丝子、5份金灯藤样品,提取基因组DNA后,利用ITS2序列扩增引物进行PCR及测序,并从GenBank下载菟丝子、南方菟丝子、金灯藤ITS2序列.利用MEGA 11.0对序列进行比对分析,计算种内、种间遗传距离,利用邻接法(NJ)构建系统聚类树,通过数据库预测ITS2二级结构.结果:菟丝子、南方菟丝子、金灯藤构建的NJ树分别聚为一支,基于ITS2碱基序列的种间遗传距离能准确鉴别菟丝子、南方菟丝子、金灯藤;菟丝子、南方菟丝子、金灯藤的二级结构在整体上有较大区别,能够明显区分.结论:基于ITS2序列可以准确地鉴别菟丝子及其混伪品.
Abstract
Objective:To identify Cuscutae semen and its miscible counterfeit Cuscuta japonica choisy using DNA barcode ITS2 base sequence and its secondary structure.Methods:Ten samples of Cuscutae semen,five samples of Cuscuta australis R.Br.and five samples of Cuscuta japonica choisy were collected from the Bozhou herbal market in Anhui Province,China.After extracting genomic DNA,ITS2 sequence amplification primers were used for PCR and sequencing;and the ITS2 sequence of Cuscutae semen,Cuscuta australis R.Br.and Cuscuta japonica choisy were downloaded from GenBank.The sequences were compared and analysed using MEGA 11.0,and the intraspecific and interspecific genetic distances were calculated.The neighbor-joining(NJ)method was used to construct the phylogenetic clustering tree,and the ITS2 secondary structure was predicted through the database.Results:The NJ tree was formed by Cuscutae semen,Cuscuta australis R.Br.and Cuscuta japonica choisy,and each of them is clustered into one branch.The interspecific genetic distance based on ITS2 base sequence could accurately identify Cuscutae semen,Cuscuta australis R.Br.and Cuscuta japonica choisy.The secondary structure of Cuscutae semen,Cuscuta australis R.Br.and Cuscuta japonica choisy were quite different on the whole and could be distinguished obviously.Conclusion:Based on ITS2 sequences,Cuscutae semen and its miscible counterfeit can be accurately identified.
关键词
菟丝子/DNA条形码/ITS2序列/二级结构Key words
Cuscutae semen/DNA barcoding/ITS2 sequence/Secondary structure引用本文复制引用
基金项目
毫州学院学科团队立项项目(2020)(BYZXKTD202004)
出版年
2024
国家科技期刊平台
NSTL
万方数据