Abstract
Plant viruses are obligate parasites that exploit host components for replication and spread inside the host. Transport of the viral genome is enabled by movement proteins (MPs) targeting the cell periphery to mediate passage throughout plasmodesmata (PD). Pectin methylesterase (PME) is one of the critical host factors facilitating MPs in PD gating, and a direct interaction of PME with Tobacco mosaic virus (TMV) MP is required for viral movement and in turn for virus viability. PME is a critical enzyme for host development and defence, acting via complex mechanisms involving multigenic and tissue specific isoforms and endogenous inhibitors. This composite activity of PME suggests that level and timing of protein accumulation, with respect to virus inoculation and MP expression, can be critical for the functional outcome of the PME-MP interaction and in turn for the success of a viral infection. Based on this notion, we tested different experimental conditions to evaluate the beneficial effect of the downregulation of PME gene expression on the development of TMV-induced disease and on plant protection. We used virus induced gene silencing technology (VIGS) to downregulate PME gene expression, which resulted in a 30–45 % reduction of TMV symptom severity and, correspondingly, to a 60 % reduction of TMV RNA accumulation in systemic leaves. VIGS proved to be a rapid and effective technology for PME gene silencing in functional assays and for plant defence from viral infection. Our findings indicate that N. benthamiana plants with hindered expression of PME survive a TMV infection, which kills non-silenced plants within a week.
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Acknowledgments
We thank Savithramma P. Dinesh-Kumar for the pTRV1, pTRV2 and pTRV2: PDS vectors, Giusy D’attoma and Roberto Manzari for technical assistance and Peter Palukaitis for critical reading of the manuscript. This work was supported with a grant from the Ministry of Economy and Finance to the CNR, “Integrate knowledge for Sustainability and Innovation of Made in Italy Agro-Food (CISIA, l. 191/2009), and a grant to the Public-Private Laboratory GenoPom “Integrating post-genomic platforms to enhance the tomato production chain” (GenoPOMpro, Cod. PON02_00395_3082360).
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Bubici, G., Carluccio, A.V., Cillo, F. et al. Virus-induced gene silencing of pectin methylesterase protects Nicotiana benthamiana from lethal symptoms caused by Tobacco mosaic virus . Eur J Plant Pathol 141, 339–347 (2015). https://doi.org/10.1007/s10658-014-0546-y
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DOI: https://doi.org/10.1007/s10658-014-0546-y