Potato virus Y coat protein involvement in viral movement

Potato virus Y (PVY) is one of the most important plant viruses worldwide, responsible for major potato yield losses. We constructed different PVY N terminal mutants to determine the crucial regions for cell-to-cell and systemic movement. Viral cell-to-cell movement was followed with confocal microscopy, and the movement dynamics of different deletion mutants was observed by Whole plant imaging system Newton 7.0 BIO (Vilber). With detailed spatio-temporal foci analysis, we showed that viral cell-to-cell spread is dependent on the length of the CP N terminal region, as with increased number of deleted amino acid residues at CP N terminal region, we observed slower cell-to-cell viral spread. As expected, slower cell-to-cell viral spread impacted viral spread in systemic tissue. Furthermore, with introducing point mutations in the identified region, the PVY movement was prevented (S21G) or delayed (G20G). This dataset is deposit of the raw confocal microscopy, whole plant imaging system images, transmission electron microscopy micrographs and supplementary datasets S1-S12. Additional information about deposited dataset: All confocal microscopy images exported from Leica LAS X software are systematically organized into folders by mutant as observed in the experiments at different days post bombardment (dpb): ΔN14-CP and ΔN23-CP_ΔN20-CP _ΔN19-CP at 5dpb, ΔN26-CP at 5, 10, 14 dpb, ΔN40-CP at 5 and 10 dpb, ΔN50-CP at 12 and 13dpb and point mutants (D14A, E18A and G20P_S21G_P24A) at 5 dpb. For mutants observed at multiple timepoints, additional subfolders indicate the specific timepoints (dpb) at which images were taken. All pictures are maximum projections from Z-stacks after excitation with 488 nm laser and unidirectionally scanning speed 400 Hz and frame average 1 or 2. Emissions were recorded between 505 and 530 nm. Detailed properties of each image are explained in the caption including information about the objective, zoom and gain. Whole plant imaging system images exported from Vilber EvolutionCapt edge software are systematically organized into folders based on viral multiplication area on inoculated (folder viral cell-to-cell spread) and systemic leaves (folder systemic viral spread) of constructed deletion and point mutants (see above). Each folder contains subfolders for individual experiments. Within each experiment folder, additional subfolders correspond to the timepoints (dpb) at which images were taken and included in the analysis. GFP emission was followed with 480 excitation channel and emission filter F-550. The subfolders captions for inoculated leaves summarize the imaging properties, including exposure time, field of view and focus. While the properties for observing viral spread on systemic leaves were the same for all timepoints, using 50 s exposure time, 20x20 FOV and focus in the range between 1871 and 1905 in different time points. All transmission electron microscopy (TEM) micrographs of bombarded Nicotiana clevelandii leaves with deletion and point mutants are systematically organized into the folders according to the mutant observed. Micrographs for point mutants are stored in a separate folder. Sampling for ΔN50-CP was at 27 days post bombardment (dpb), for ΔN40-CP at 25 dpb, for ΔN26 at 14 dpb, for ΔN23-CP, ΔN19-CP, ΔN14-CP at 5 dpb, for WT at 25 dpb, 14 dpb, 13 dpb, while sampling for all point mutants (D14A, E18A, G20P, S21G, P24A) was at 13 dpb. All supplemental datasets S1-S12 from the article are deposited in Microsoft Excel format. Note that in dataset S11 only two datapoints are displayed for E18A because the third plant was excluded from analysis due to not appropriate focus of the image.