Efficient virus-induced gene silencing in Arabidopsis.
Virus-induced gene silencing (VIGS) is a plant RNA-silencing technique that uses viral vectors carrying a fragment of a gene of interest to generate double-stranded RNA, which initiates the silencing of the target gene. Several viral vectors have been developed for VIGS and they have been successfully used in reverse genetics studies of a variety of processes occurring in plants. This approach has not been widely adopted for the model dicotyledonous species Arabidopsis (Arabidopsis thaliana), possibly because, until now, there has been no easy protocol for effective VIGS in this species. Here, we show that a widely used tobacco rattle virus-based VIGS vector can be used for silencing genes in Arabidopsis ecotype Columbia-0. The protocol involves agroinfiltration of VIGS vectors carrying fragments of genes of interest into seedlings at the two- to three-leaf stage and requires minimal modification of existing protocols for VIGS with tobacco rattle virus vectors in other species like Nicotiana benthamiana and tomato (Lycopersicon esculentum). The method described here gives efficient silencing in Arabidopsis ecotype Columbia-0. We show that VIGS can be used to silence genes involved in general metabolism and defense and it is also effective at knocking down expression of highly expressed transgenes. A marker system to monitor the progress and efficiency of VIGS is also described.
Burch-Smith
Tessa M
TM
Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, Connecticut 06520-8103, USA.
Schiff
Michael
M
Liu
Yule
Y
Dinesh-Kumar
S P
SP
D017360:Arabidopsis / Q000235:genetics* / Q000821:virology; D029681:Arabidopsis Proteins / Q000235:genetics; D015415:Biomarkers; D018744:DNA, Plant; D020868:Gene Silencing; D020043:Genes, Essential; D017343:Genes, Plant; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D010088:Oxidoreductases / Q000235:genetics; D010935:Plant Diseases; D010942:Plant Viruses / Q000235:genetics*; D019076:Transgenes
D029681:Arabidopsis Proteins; D015415:Biomarkers; D018744:DNA, Plant; D049452:Green Fluorescent Proteins; D010088:Oxidoreductases; C022398:phytoene dehydrogenase