中国番茄黄化曲叶病毒利用根吸收法诱导基因沉默(VIGS)的初步研究  

Study on Chinese Tomato Yellow Leaf Curl Virus-induced Gene Silencing(VIGS) via Root Absorption

作　　者：张召军[1] 王晓彬[1] 王慧[1] 刘林[1] 张心阁 何秀霞[1] 

机构地区：[1]长春理工大学生命科学技术学院,长春130022

出　　处：《生物技术通报》2014年第1期143-146,共4页Biotechnology Bulletin

基　　金：大学生创新实验项目(2012X77);吉林省科技厅项目(20110244)

摘　　要：以含硫黄素酶Su基因的中国番茄黄化曲叶病毒卫星DNA作为载体,通过冻融法将其转化到农杆菌中,利用根部吸收法进行农杆菌侵染,对农杆菌介导的病毒诱导基因沉默体系进行了优化,探讨对病毒诱导基因沉默效率的影响。结果显示以OD600值为1.5的含质粒pBinPLUS-2mβ-Su,pBinPLUS-1.7A的农杆菌菌液1∶1混合进行根部吸收法侵染22-25 d的番茄幼苗,目的基因Su沉默导致番茄幼苗出现光漂白现象,半定量RT-PCR检测目的基因Su mRNA被显著降解,该体系的建立有利于对植物基因进行高通量功能分析。The tomato yellow leaf curl virus satellite I）NA was used as vector which containing the Su gene, and transformed the plasmid into Agrobacterium by freeze thawing method. Agrobacterium infected plants via root absorption, and virus-induced gene silencing system was optimized, twexplore the influence to virus-induced gene silencing efficiency. The results showed infected 22-25 days tomato seedling via OD 1.5 Agrobacterium-mediated root-absorption which containing plasmid pBinPLUS-2m-Su and pBinPLUS-1.7A, the silencing of Su gene resulted in photo-blcached young leaves of tomato plants. Semi-quantitative RT-PCR analysis was employed to test the effect of Su gene silencing, which showed that Su gene was degraed significantly. This gene-silencing system would benefit for the high-throughput analysis of plant gene.

关 键 词：病毒诱导基因沉默 硫黄素酶Su基因 根吸收法 

分 类 号：S436.412[农业科学—农业昆虫与害虫防治]