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. 2013 Mar 4;14(3):265–278. doi: 10.1111/mpp.12002

Figure 2.

figure

Silencing suppression activity of Chinese wheat mosaic virus (CWMV) cysteine‐rich protein (CRP) in a viral movement complementation assay. (A) Schematic representation of recombinant Potato virus X‐green fluorescent protein (PVX‐GFP) and P25 mutants used in trans‐complementation assay (not to scale). Broken lines represent the deleted region of the P25 gene (ΔP25). The P25 mutant (P25/T117A) which contains the substitution of tyrosine‐117 by alanine (marked with an asterisk) is defective in silencing suppression, but retains its virus movement function. 35S and Nos represent cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase terminator sequences, respectively. (B–D) Leaves of Nicotiana benthamiana plants infiltrated with mixtures of Agrobacterium cultures (1 : 1 : 1) harbouring pGR106:PVX(ΔP25)‐GFP (diluted 10 000‐fold), pBin‐P25/T117A and pBin‐p19, pBin‐HC‐Pro, pBin‐CRP or CRP mutants (see Fig. 3A). GFP fluorescence was observed using long‐wavelength UV light, and photographed (B) or observed using confocal laser scanning microscopy (C, D) at 5 days after inoculation (dai). Bars, 200 μm.