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Title

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

AuthorsMartínez-Pérez, Mireya; Navarro, José A. ; Pallás, Vicente; Sánchez-Navarro, Jesús A.
KeywordsRSS assay
Alfamovirus
P12 system
Issue Date15-Oct-2019
PublisherElsevier
CitationVirus Research 272: 197733 (2019)
AbstractPlant 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.
Publisher version (URL)http://doi.org/10.1016/j.virusres.2019.197733
URIhttp://hdl.handle.net/10261/210842
Identifiersdoi: 10.1016/j.virusres.2019.197733
issn: 0168-1702
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(IBE) Artículos
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