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Originally published as JGV in Press, 10.1099/vir.0.011213-0 on July 1, 2009 J Gen Virol 90 (2009), 2536-2541; DOI 10.1099/vir.0.011213-0

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Short Communication

Rapid screening of RNA silencing suppressors by using a recombinant virus derived from beet necrotic yellow vein virus

H. Guilley, D. Bortolamiol, G. Jonard, S. Bouzoubaa and V. Ziegler-Graff

Institut de Biologie Moléculaire des Plantes du CNRS, Université de Strasbourg, 12 rue du Général Zimmer, 67084 Strasbourg Cedex, France

Correspondence
V. Ziegler-Graff
veronique.ziegler-graff{at}ibmp-ulp.u-strasbg.fr

To counteract plant defence mechanisms, plant viruses have evolved to encode RNA silencing suppressor (RSS) proteins. These proteins can be identified by a range of silencing suppressor assays. Here, we describe a simple method using beet necrotic yellow vein virus (BNYVV) that allows a rapid screening of RSS activity. The viral inoculum consisted of BNYVV RNA1, which encodes proteins involved in viral replication, and two BNYVV-derived replicons: rep3–P30, which expresses the movement protein P30 of tobacco mosaic virus, and rep5–X, which allows the expression of a putative RSS (X). This approach has been validated through the use of several known RSSs. Two potential candidates have been tested and we show that, in our system, the P13 protein of burdock mottle virus displays RSS activity while the P0 protein of cereal yellow dwarf virus-RPV does not.

A supplementary figure showing images of infection foci produced by RNA1 supplemented with rep5–GFP in the presence or absence of rep30 is available with the online version of this paper.






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