一种快速检测植物病毒RNA蓄积量的竞争PCR方法  

The Method of Quantitative Competitive PCR for Detecting Accumulation of Tobacco Mosaic Virus RNA in Tobacco Leaves

作　　者：朱春玉[1] 吴元华[1] 王艳红[1] 赵秀香[1] 

机构地区：[1]辽宁大学生命科学院,辽宁沈阳110036

出　　处：《辽宁大学学报（自然科学版）》2008年第1期77-80,共4页Journal of Liaoning University：Natural Sciences Edition

基　　金：国家科技部863计划项目(2001AA247011);国家烟草专卖局科技项目(国烟科[2002]187号)

摘　　要：用Oligo6.6软件在pUC18上设计1对比目的基因多100-200bp的内参引物，摸索了目的基因和内参共同扩增的竞争PCR反应条件，测定了嘧肽霉素处理对烟草组织中烟草花叶病毒RNA相对含量的影响，同时用间接ELISA方法验证了该方法。试验结果表明：竞争PER的方法能够快速、相对准确的测定植物组织中病毒RNA蓄积量的变化情况。A new method, quantitative competitive PCR （ QC - PCR） with software of Image Master ED was described for quantitative detecting TMV - RNA in infected plant tissues. Primers pair of Internal standard contrast goal gene more than 100-200bp were designed using Oligo6.6 software on pUC18. QC-PCR reaction conditions were researched. Cytosinpeptidemycin was a newly reported antiphytoviral agricultural antibiotics. Effect on Tobacco Mosaic Virus RNA （TMV -RNA） concentration in TMV infected tobacco leaves treated with 4% Cytosinpeptidemycin was tested by this method. The results were also confirmed by Indi-ELISA technique. All the results indicted that the method of quantitative competitive PCR is applicable in quantitative determination of TMV-RNA in infected plant tissues without obvious artificial influence.

关 键 词：竞争PCR 嘧肽霉素 烟草花叶病毒 

分 类 号：O278[理学—数学] S435[农业科学—农业昆虫与害虫防治]