A sensitive and rapid RNA silencing suppressor activity assay based on alfalfa mosaic virus expression vector

Plant viruses express RNA silencing suppressor (RSS) proteins to counteract plant defence mechanisms. Here, we describe a method to assess the RSS activity based on an alfalfa mosaic virus (AMV) RNA 3 expression vector and transgenic Nicotiana tabacum plants that express the P1 and P2 subunits of the AMV replicase (P12 plants). Inoculation of P12 plants with different AMV RNA 3 constructs expressing different HC-Pro mutants that differ in their RSS capabilities, revealed a perfect correlation between necrotic lesions on inoculated leaves and RSS activity. Protoplast analysis showed that the RSS activity correlated with the accumulation of the AMV RNAs. A direct comparison between three RSS activity assays and the AMV-P12 system revealed that the coat protein of carnation mottle virus displays RSS activity with the four assays and reduced the accumulation of the siRNAs.

Bibliographic Details

Main Authors:	Martínez-Pérez, Mireya, Navarro, José A., Pallás, Vicente, Sánchez-Navarro, J. A.
Other Authors:	Dirección General de Investigación Científica y Técnica, DGICT (España)
Format:	artículo biblioteca
Published:	Elsevier 2019-10-15
Subjects:	RSS assay, Alfamovirus, P12 system,
Online Access:	http://hdl.handle.net/10261/210842 http://dx.doi.org/10.13039/501100008737 http://dx.doi.org/10.13039/501100003359 http://dx.doi.org/10.13039/501100011033
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