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JXB Advance Access originally published online on May 31, 2008
Journal of Experimental Botany 2008 59(10):2803-2813; doi:10.1093/jxb/ern141
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© 2008 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details)

RESEARCH PAPER

Influence of viral genes on the cell-to-cell spread of RNA silencing

Yu Zhou *, Eugene Ryabov, Xuemei Zhang * and Yiguo Hong{dagger}

Warwick HRI, University of Warwick, Wellesbourne, Warwick, CV35 9EF, UK

{dagger} To whom correspondence should be addressed. E-mail: yiguo.hong{at}warwick.ac.uk

The turnip crinkle virus-based vector TCV–GFP{Delta}CP had been devised previously to study cell-to-cell and long-distance spread of virus-induced RNA silencing. TCV–GFP{Delta}CP, which had been constructed by replacing the coat protein (CP) gene with a green fluorescent protein (GFP) coding sequence, was able to induce RNA silencing in single epidermal cells, from which RNA silencing spread from cell-to-cell. Using this unique local silencing assay together with mutagenesis analysis, two TCV genes, p8 and p9, which were involved in the intercellular spread of virus-induced RNA silencing, were identified. TCV–GFP{Delta}CP and its p8- or p9-mutated derivatives, TCVmp8–GFP{Delta}CP and TCVmp9–GFP{Delta}CP, replicated efficiently but were restricted to single Nicotiana benthamiana epidermal cells. TCV–GFP{Delta}CP, TCVmp8–GFP{Delta}CP, or TCVmp9–GFP{Delta}CP was able to initiate RNA silencing that targeted and degraded recombinant viral RNAs in inoculated leaves of the GFP-expressing N. benthamiana line 16c. However, cell-to-cell spread of silencing to form silencing foci was triggered only by TCV–GFP{Delta}CP. Non-replicating TCVmp88–GFP{Delta}CP and TCVmp28mp88–GFP{Delta}CP with dysfunctional replicase genes, and single-stranded gfp RNA did not induce RNA silencing. Transient expression of the TCV p9 protein could effectively complement TCVmp9–GFP{Delta}CP to facilitate intercellular spread of silencing. These data suggest that the plant cellular trafficking machinery could hijack functional viral proteins to permit cell-to-cell movement of RNA silencing.

Key words: Cell-to-cell movement, Nicotiana benthamiana, RNA silencing, TCV, viral movement proteins

* Present address: Chengdu Institute of Biological Products, Sichuan 610023, China.

Received 12 March 2008; Revised 21 April 2008 Accepted 22 April 2008


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J. Virol., April 15, 2009; 83(8): 3540 - 3548.
[Abstract] [Full Text] [PDF]


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