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Title: | A sensitive and rapid RNA silencing suppressor activity assay based on alfalfa mosaic virus expression vector |
Authors: | Martínez-Pérez, Mireya; Navarro, José A. ![]() |
Keywords: | RSS assay Alfamovirus P12 system |
Issue Date: | 15-Oct-2019 |
Publisher: | Elsevier |
Citation: | Virus Research 272: 197733 (2019) |
Abstract: | 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. |
Publisher version (URL): | http://doi.org/10.1016/j.virusres.2019.197733 |
URI: | http://hdl.handle.net/10261/210842 |
DOI: | 10.1016/j.virusres.2019.197733 |
Identifiers: | doi: 10.1016/j.virusres.2019.197733 issn: 0168-1702 |
Appears in Collections: | (IBMCP) Artículos (IBE) Artículos |
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