Figure 5
From: Unravelling the involvement of cilevirus p32 protein in the viral transport
The C-terminal region of cileviruses MP is necessary for correct co-localization with plasmodesmata. Transient expression of CiLV-C wild type (a), CiLV-CΔ228–297 (b), CiLV-CΔ225–297 (c), CiLV-C2 wild type (d), CiLV-C2Δ222–292 (e) and CiLV-C2Δ218–292 (f) MPs, carrying the GFP (green filled circle) in N. benthamiana leaves. Fluorescence signal was captured at 72 h post-infiltration. The callose deposits were stained using aniline blue (blue filled circle). The blue and green arrows indicate the callose deposits in plasmodesma and MP, respectively; while the white arrows indicate co-localization between them. The blue (aniline blue), the green (GFP), transmitted light (TL) channels and merged images are shown in the figure. Images on the left show the tubule formation of N. benthamiana protoplast transiently expressing the correspondent MP construct. The MP punctate structures co-localize at the cell periphery with the fluorochrome for the MP wt constructs (a,d), while the truncated MP constructs show no co-localization or partial co-localization (b,c,e,f). In a higher magnification image, it is shown the complete (i,iv) or partial (ii,iii,v,vi) co-localization of the MP with callose deposits in the plasmodesmata, and the chart of fluorescence intensities further confirms the co-localization. Plot shows green and blue fluorescence intensities, indicated by MP:GFP and aniline blue, in the selected region of interest (red arrows). Distance measurement starts from the base to the tip of the arrows (x axis). The mean SD of Person Correlation Coefficient (PCC) is given in the merged image. PCC was measured using the Fiji co-localization plugin for three independent images from approximately 100 individual plasmodesmata. The dotted line in the transmitted light image indicates the cell wall (CW). Cyt cytoplasm. Red and white bars correspond to 5–10 and 20 μm, respectively.
