摘要
为鉴定食品中的三七源性成分,以内转录间隔区(internal transcribed spacer,ITS)序列作为DNA条形码来进行分析。该研究首先比较十六烷基三甲基溴化铵(cetyltrimethylammonium bromide,CTAB)法、十二烷基硫酸钠(sodium dodecyl sulfate,SDS)法和磁珠法3种常用DNA提取方法提取三七及其他植物样品DNA的效果(浓度、纯度),然后根据三七及其近缘植物ITS序列中的保守区域设计三七的特异性引物,建立一种针对三七鉴定的快速、特异、灵敏的聚合酶链式反应(polymerase chain reaction,PCR)方法。对与三七相似或近缘的植物进行特异性试验,并利用三七与常见相似植物的混合样本进行PCR扩增测试该方法的灵敏度。结果显示:3种提取方法所得DNA扩增后均可检测出清晰条带,CTAB法和磁珠法提取效果良好。利用本研究所设计的特异性引物,能够对三七进行特异性的检测,检测灵敏度为0.5%。
To identify the source components of Panax notoginseng in food,the internal transcribed spacer(ITS)sequence was analyzed as the DNA barcode. The effects(concentration and purity)of three commonly used DNA extraction methods of P. notoginseng and other plant samples were compared by cetyltrimethylammonium bromide(CTAB),sodium dodecyl sulfate(SDS),and magnetic bead extraction methods. Specific primers of P.notoginseng were designed according to the conservative region of the ITS sequence between P. notoginseng and related plants to establish a rapid,specific,and sensitive PCR method for identification. Specificity tests were performed on plants closely related to P. notoginseng. Sensitivity tests were performed by PCR amplification using mixed samples of P. notoginseng and similar plants. All three extraction methods clearly revealed bands after DNA amplification. The results obtained with CTAB and magnetic bead extraction were superior. The primers that we designed specifically detected P. notoginseng,with a detection sensitivity of 0.5%.
作者
夏晖
张芹
吴弘
季超
陈怡
原楚妍
孙星宇
郑文杰
XIA Hui;ZHANG Qin;WU Hong;JI Chao;CHEN Yi;YUAN Chu-yan;SUN Xing-yu;ZHENG Wen-jie(College of Life Sciences,Tianjin Normal University,Tianjin 300387,China;Tianjin Key Laboratory ofAnimal and Plant Resistance,Tianjin Normal University,Tianjin 300387,China;College of Plant Protection,Yunnan Agricultural University,Kunming 650201,Yunnan,China)
出处
《食品研究与开发》
CAS
北大核心
2022年第4期181-187,共7页
Food Research and Development
基金
国家大学生创新创业训练计划项目(201910065084)
云南省重大科技专项(2019ZG00901
202102AE090042)
云南绿色食品国际合作研究中心项目子课题(2019ZG00901-04)
国家现代农业产业技术体系建设专项(CARS-21)
生物技术与资源利用教育部重点实验室项目(KF2021006)。
关键词
聚合酶链式反应
ITS基因
三七
检测
鉴别
polymerase chain reaction
ITS gene
Panax notoginseng
detection
identifying
