摘要
番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)是引起番茄黄化曲叶病毒病的病原,在生产中造成严重危害,开发快速准确的检测方法对该病害的防控具有重要意义。依据番茄黄化曲叶病毒DNA-A基因序列,设计用于重组酶聚合酶等温扩增(RPA)检测的特异性引物,建立了TYLCV的重组酶聚合酶等温扩增检测方法,并分析该方法的特异性和灵敏度。结果表明,建立的TYLCV-RPA方法可从TYLCV阳性的番茄样品中扩增出343 bp的特异性条带,反应条件为37℃恒温下40 min,扩增时间短,对设备要求低,且与番茄其他病毒无交叉反应,特异性好,灵敏度可达到PCR方法的10倍,适用于TYLCV的快速检测。
Tomato yellow leaf curl virus(TYLCV)is an etiology of Tomato yellow leaf curl disease(TYLCD),which will cause serious damages to tomato production.It is of important significance to exploit rapid and accurate detection method for controlling this disease.According to DNA-A gene sequence of TYLCV,this study designed special RPA primers and established a method named TYLCV-RPA,and analyzed the specificity and sensitivity of this method.The results indicated that TYLCV-RPA method could amplify specific band of 343 bp from sample tomatoes with positive TYLCV.The reaction condition was 37℃ for 40min.This method needed short time period for amplification with low requirement for equipment.It had no cross reaction with the other viruses and had good specificity.Its sensitivity was 10 times of the PCR method.It is suitable for rapid detection of TYLCV.
作者
周莹
杨丽梅
刘梅
黄金宝
ZHOU Ying;YANG Li-mei;LIU Mei;HUANG Jin-bao(Institute of Plant and Environment Protection,Beijing Academy of Agriculture and Forestry Sciences,Beijing 100097,China;Agricultural Technology Extension Station of Yufa Town,Daxing District,Beijing 102602,China)
出处
《中国蔬菜》
北大核心
2019年第1期36-40,共5页
China Vegetables
基金
北京市农林科学院青年科研基金项目(QNJJ201705)
北京市优秀人才培养资助项目(2016000020060G131)
北京市农林科学院创新能力建设项目(KJCX20180705)
北京市农林科学院科技创新团队项目(JNKYT201605)
关键词
番茄黄化曲叶病毒
重组酶聚合酶扩增
等温扩增
检测方法
Tomato yellow leaf curl virus
Recombinase polymerase amplification
Isothermal amplification
Detection method
