微量植物组织汁液直接RT-LAMP可视化检测马铃薯卷叶病毒方法的构建  

Establishment of the Method for the Detection of Potato Leafroll Virus by Visualization of the Product of RT-LAMP Using Micro-tissue Juice

作　　者：李姣[1] 朱晓换 古蕾 王亚男[1] LI Jiao;ZHU Xiaohuan;GU Lei;WANG Yanan(College of Life Science,Sichuan Normal University,Chengdu 610101,Sichuan)

机构地区：[1]四川师范大学生命科学学院,四川成都610101

出　　处：《四川师范大学学报（自然科学版）》2018年第6期817-821,共5页Journal of Sichuan Normal University（Natural Science）

基　　金：四川省应用基础研究重点项目(2017JY0017);四川省教育厅自然科学一般项目(15ZB0040)

摘　　要：为建立快速的马铃薯卷叶病毒检测方法以应用于脱病毒后大规模样本的检测,构建利用微量植物组织汁液直接逆转录环介导恒温扩增(RT-LAMP)可视化检测马铃薯卷叶病毒的方法,即以无菌大头针的针尖刺马铃薯植株茎尖时所粘附的汁液直接作为RNA模板,采用Bacillus stearothermophilus(Bst) DNA聚合酶进行RT-LAMP扩增,扩增产物通过肉眼观察有无白色沉淀来诊断马铃薯卷叶病毒.该方法快速、操作简单、成本低,在大规模组织培养无病毒种薯生产中具有很大的应用空间.To establish a method for rapid detection of potato leafroll virus in a large quantity of plant samples eliminated virus,we developed a visualized detection method of direct reverse transcription loopmediated isothermal amplification(RTLAMP)using microtissue juice.The template was prepared directly from the juice of plant microtissue that was adhered to the pinpoint when a pin was used to pierce the potato shoot tip.After amplified by Bacillus stearothermophilus(Bst)DNA polymerase,the product of RTLAMP could be directly detected with naked eye by seeing the appearance of the white precipitate.This method is rapid,simple and economic,and could be applied to the production of virusfree seed potato by tissue cultures in a large quantity.

关 键 词：马铃薯 马铃薯卷叶病毒 微量组织汁液直接RT-LAMP 

分 类 号：Q781[生物学—分子生物学]