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DEVELOPMENT AND HORMONE ACTION

Virus-Induced Gene Silencing of Argonaute Genes in Nicotiana benthamiana Demonstrates That Extensive Systemic Silencing Requires Argonaute1-Like and Argonaute4-Like Genes¹

Department of Biology (L.J., T.K., A.E.) and Centre for Novel Agricultural Products (F.E.V.), University of York, York YO10 5YW, United Kingdom

Several distinct pathways of RNA silencing operate in plants with roles including the suppression of virus accumulation, control of endogenous gene expression, and direction of DNA and chromatin modifications. Proteins of the Dicer-Like and Argonaute (AGO) families have key roles within these silencing pathways and have distinct biochemical properties. We are interested in the relationships between different silencing pathways and have used Nicotiana benthamiana as a model system. While not being an amenable plant for traditional genetics, N. benthamiana is extensively used for RNA-silencing studies. Using virus-induced gene silencing technology we demonstrate that both NbAGO1- and NbAGO4-like genes are required for full systemic silencing but not for silencing directed by an inverted repeat transgene. Phenotypic differences between the virus-induced gene silencing plants indicate that NbAGO1 and NbAGO4 like act at different stages of the silencing pathways. Suppression of NbAGO1 expression recapitulated the hypomorphic mutant phenotype of certain Arabidopsis (Arabidopsis thaliana) ago1 alleles, however, suppression of NbAgo4 like resulted in phenotypes differing in some respects from those reported for Arabidopsis ago4. We suggest that the small interfering RNA amplification step required for full systemic silencing is dependent upon a nuclear event requiring the activity of NbAGO4 like.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Louise Jones (alj2{at}york.ac.uk).

Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.105.076109.

^* Corresponding author; e-mail alj2{at}york.ac.uk; fax 44–1904–328555.

Received December 29, 2005; returned for revision March 31, 2006; accepted April 6, 2006.

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