Development and application of an efficient virus-induced gene silencing system in Nicotiana tabacum using geminivirus alphasatellite.
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.
State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China.
Chang-jun
CJ
Huang
Tong
T
Zhang
Fang-fang
FF
Li
Xin-yue
XY
Zhang
Xue-ping
XP
Zhou
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D017791:Geminiviridae / Q000235:genetics*; D018014:Gene Transfer Techniques; D017343:Genes, Plant; D005822:Genetic Vectors; D005966:Glucuronidase / Q000235:genetics; D010935:Plant Diseases / Q000235:genetics / Q000821:virology; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000235:genetics; D034622:RNA Interference; D012526:Satellite Viruses / Q000235:genetics*; D013696:Temperature; D014026:Nicotiana / Q000235:genetics* / Q000821:virology; D014027:Tobacco Mosaic Virus / Q000235:genetics / Q000472:pathogenicity
D017931:DNA Primers; D005966:Glucuronidase
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