Figure 5: NIa-Pro relocalizes to the vacuole in the presence of aphids during virus infection.
From: A viral protease relocalizes in the presence of the vector to promote vector performance
(a) Schematic representation of the recombinant TuMV-expressing GFP:NIa-Pro. Each rectangle represents individual proteins of the TuMV polyprotein. (b) Number of progeny produced by aphids on N. benthamiana infiltrated with empty agrobacterium (EV), TuMV/GFP or TuMV/GFP:NIa-Pro. (c) The image depicts a western blot from leaf extracts expressing empty vector (EV), TuMV/GFP and TuMV/GFP:NIa-Pro incubated with an antibody that binds to the coat protein (CP) of TuMV. (d) Images show confocal projection and a single section (on the right) of N. benthamiana leaves agroinfiltrated with TuMV/GFP:NIa-Pro with or without aphids for 24 h. GFP:NIa-Pro relocalizes from the nucleus (white arrow) to the vacuole (V, dashed arrow) in the presence of aphids. (e) The percentage of cells with GFP in the vacuole for N. benthamiana infected with TuMV/GFP:NIa-Pro with and without aphids. (f) The image represents a western blot from vacuole purification derived from leaves expressing EV or TuMV/GFP:NIa-Pro before or after aphid presence and incubated with a GFP antibody and an anti TuMV-CP antibody. The western blot shows that GFP is only visible in the vacuoles from leaves expressing TuMV/GFP:NIa-Pro and after aphid presence. (mean±s.e. of N=12–20 for b and N=6 for e; letters and stars represent significant differences, ANOVA and Tukey’s honest significant difference post hoc for b or Student’s t-test for e, P<0.05). Scale bars, 20 μm.
