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Journal of Virology, March 2004, p. 3149-3154, Vol. 78, No. 6
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.6.3149-3154.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Cell-to-Cell, but Not Long-Distance, Spread of RNA Silencing That Is Induced in Individual Epidermal Cells

Eugene V. Ryabov,1* Rene van Wezel,1 John Walsh,2 and Yiguo Hong1*

Horticulture Research International, East Malling, West Malling, Kent ME19 6BJ,1 Horticulture Research International, Wellesbourne, Warwick CV35 9EF, United Kingdom2

Received 26 August 2003/ Accepted 17 November 2003

A Turnip crinkle virus (TCV)-based system was devised to discriminate cell-to-cell and systemic long-distance spread of RNA silencing in plants. Modified TCV-GFP{Delta}CP, constructed by replacing the coat protein (CP) gene with the green fluorescent protein (GFP) gene, replicated in single epidermal cells but failed to move from cell to cell in Nicotiana benthamiana. Mechanical inoculation of TCV-GFP{Delta}CP induced effective RNA silencing in single epidermal cells which spread from cell to cell to form silenced foci on inoculated leaves, but no long-distance systemic spread of RNA silencing occurred. Agroinfiltration of TCV-GFP{Delta}CP was, however, able to induce both local and systemic RNA silencing. TCV coinfection arrested TCV-GFP{Delta}CP-mediated local induction of RNA silencing. Possible mechanisms involved in cell-to-cell and long-distance spread of RNA silencing are discussed.

* Correspondence author. Mailing address: Horticulture Research International, Wellesbourne, Warwick CV35 9EF, United Kingdom. Phone: 44 178 947 0382. Fax: 44 178 947 0552. E-mail: eugene.ryabov{at}hri.ac.uk (E. V. Ryabov) and yiguo.hong{at}hri.ac.uk (Y. Hong).

Journal of Virology, March 2004, p. 3149-3154, Vol. 78, No. 6
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.6.3149-3154.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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