Plant Physiology Preview
Published on June 30, 2006; 10.1104/pp.106.084624
Received June 2, 2006
Accepted June 20, 2006
Efficient virus induced gene silencing in Arabidopsis thaliana
Tessa M. Burch-Smith , Michael Schiff , Yule Liu , and S.P Dinesh-Kumar *
Department of Molecular, Cellular, and Developmental Biology, 219 Prospect Street, Yale University, New Haven, CT 06520-8103, USA
* Corresponding author; email: savithramma.dinesh-kumar{at}yale.edu.
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 the 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 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 (TRV)-based VIGS vector can be used for silencing genes in A. thaliana ecotype Columbia-0. The protocol involves the Agro-infiltration of VIGS vectors carrying fragments of genes of interest into seedlings at the two to three leaves stage and requires minimal modification of existing protocols for VIGS with TRV vectors in other species like Nicotiana benthamiana and tomato. The method described here gives efficient silencing in A. thaliana ecotype Columbia-0. We show that VIGS can be used to silence genes involved in general metabolism and defense, and it also effective at knocking down expression of highly expressed transgenes. A marker system to monitor the progress and efficiency of VIGS is also described.
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