Development and application of an efficient virus-induced gene silencing system in Nicotiana tabacum using geminivirus alphasatellite  

作　　者：Chang-jun HUANG Tong ZHANG Fang-fang LI Xin-yue ZHANG Xue-ping ZHOU 

机构地区：[1]State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2011年第2期83-92,共10页浙江大学学报（英文版）B辑（生物医学与生物技术）

基　　金：supported by the National Science and Technology Major Projects of China (No. 2009ZX08009-026B);the China Postdoctoral Science Foundation (No. 20090461375);the National Basic Research Program (973) of China (No. 2006CB101903)

摘　　要：Virus-induced gene silencing (VIGS) is a recently developed technique for characterizing the function of plant genes by gene transcript suppression and is increasingly used to generate transient loss-of-function assays. Here we report that the 2mDNA1, a geminivirus satellite vector, can induce efficient gene silencing in Nicotiana tabacum with Tobacco curly shoot virus. We have successfully silenced the β-glucuronidase (GUS) gene in GUS transgenic N. tabacum plants and the sulphur desaturase (Su) gene in five different N. tabacum cultivars. These pronounced and severe silencing phenotypes are persistent and ubiquitous. Once initiated in seedlings, the silencing phenotype lasted for the entire life span of the plants and silencing could be induced in a variety of tissues and organs including leaf, shoot, stem, root, and flower, and achieved at any growth stage. This system works well between 18-32°C. We also silenced the NtEDS1 gene and demonstrated that NtEDS1 is essential for N gene mediated resistance against Tobacco mosaic virus in N. tabacum. The above results indicate that this system has great potential as a versatile VIGS system for routine functional analysis of genes in N. tabacum.Virus-induced gene silencing （VIGS） is a recently developed technique for characterizing the function of plant genes by gene transcript suppression and is increasingly used to generate transient loss-of-function assays. Here we report that the 2mDNA1, a geminivirus satellite vector, can induce efficient gene silencing in Nicotiana ta- bacum with Tobacco curly shoot virus. We have successfully silenced the #-glucuronidase （GUS） gene in GUS transgenic N. tabacum plants and the sulphur desaturase （Su） gene in five different N. tabacum cultivars. These pronounced and severe silencing phenotypes are persistent and ubiquitous. Once initiated in seedlings, the silencing phenotype lasted for the entire life span of the plants and silencing could be induced in a variety of tissues and organs including leaf, shoot, stem, root, and flower, and achieved at any growth stage. This system works well between 18-32 ~C We also silenced the NtEDS1 gene and demonstrated that NtEDS1 is essential for N gene mediated resistance against Tobacco mosaic virus in N. tabacum. The above results indicate that this system has great potential as a versatile VIGS system for routine functional analysis of genes in N. tabacum.

关 键 词：Virus induced gene silencing GEMINIVIRUS Alphasatellite Nicotiana tabacum 

分 类 号：Q93[生物学—微生物学]