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Tobacco rattle virus-based virus-induced gene silencing in Nicotiana benthamiana

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Title Tobacco rattle virus-based virus-induced gene silencing in Nicotiana benthamiana
 
Creator Senthil-Kumar, Muthappa
Mysore, Kirankumar S.
 
Description Tobacco rattle virus (TRV)-based virus-induced gene silencing (VIGS) is widely used in various plant species to downregulate
the expression of a target plant gene. TRV is a bipartite, positive-strand RNA virus with the TRV1 and TRV2 genomes. To induce
post-transcriptional gene silencing (PTGS), the TRV2 genome is genetically modified to carry a fragment of the target gene and
delivered into the plant (along with the TRV1 genome) by agroinoculation. TRV1- and TRV2-carrying Agrobacterium strains are then
co-inoculated into 3-week-old plant leaves by one of three methods: a needleless syringe, the agrodrench method or by pricking
with a toothpick. Target gene silencing occurs in the newly developed noninoculated leaves within 2–3 weeks of TRV inoculation.
The TRV-VIGS protocol described here takes only 4 weeks to implement, and it is faster and easier to perform than other gene
silencing techniques that are currently available. Although we use Nicotiana benthamiana as an example, the protocol is adaptable
to other plant species.
This project was funded by The Samuel Roberts Noble Foundation. We thank J. Gallaway for excellent plant care. TRV constructs were obtained from S.P. Dinesh-Kumar, University of California Davis. We thank B. Stearns and S. McNeill for making and editing the video. We thank K. Brown for artwork in the figures and J. Kelley for editing the manuscript.
 
Date 2015-12-23T11:00:53Z
2015-12-23T11:00:53Z
2014
 
Type Article
 
Identifier Nature Protocols, 9(7): 1549-1562
http://172.16.0.77:8080/jspui/handle/123456789/454
http://www.nature.com/nprot/journal/v9/n7/full/nprot.2014.092.html
10.1038/nprot.2014.092
 
Language en_US
 
Publisher Nature Publishing Group