Cucumber mosaic virus coat protein modulates the accumulation of 2b protein and antiviral silencing that causes symptom recovery in planta
Fig 5
CMV CP attenuates 2b-mediated suppression of local GFP silencing.
(A) GFP fluorescence in regions of N. benthamiana leaves after agroinfiltration of sGFP reporter gene (OD600 = 0.4), in combination with different amounts of the pGD empty vector (V, OD600 = 0.4), the pGD-2b (OD600 = 0.2) and the pGD-CP vectors (OD600 = 0.4), as indicated. Photographs were taken under UV light at 5 dpi. (B) Protein gel blot analysis of samples extracted from infiltrated region shown in panel A. (C) Accumulation of GFP mRNA and siRNAs in the region shown in panel A. Methylene blue-stained rRNA and U6 RNA were used as loading controls for high and low molecular weight RNAs, respectively. The values under GFP mRNA (RA1) and siRNAs (RA2) represent the relative accumulation (RA) of GFP mRNA and GFP-derived siRNAs, respectively. The RA values of sGFP with 2b and V were set as 1. RA2/RA1 ratios under U6 detection represent the relative production of GFP-derived siRNAs versus GFP mRNA. (D) GFP fluorescence (left panel) in local patches agroinfiltrated with the sGFP vector (OD600 = 0.4), empty pGD vector, and the 2b vectors (OD600 = 0.2), either alone or in combination with CPWT or CPRA vectors (OD600 = 0–0.8), as indicated in middle panel. A protein gel blot analysis of samples extracted from the infiltrated regions is shown in the right panel. Anti-GFP, -CP, and -2b polyclonal antibodies were used to assess the GFP, CP, and 2b, accumulations, respectively. Mock-infected plants were used as the negative control. Coomassie brilliant blue (CBB) staining was used as the protein loading control.