Fig. 5: Phosphorylated P7 interacts with NbREM1.1 to reduce callose deposition and enhance virus transmission.
a LCI assay to detect interactions between P7 and mutants with NbREM1.1. Luminescence signals were recorded in N. benthamiana leaves at 48 hpi. nLUC-SGT1 and cLUC-RAR1 served as positive controls. b Pull-down analysis of the interaction between P7 and its mutants with NbREM1.1. P7/P7S59A/P7S59D-His or His was incubated with NbREM1.1-GST. Input and pull-down products were analysed by western blot with anti-His and anti-GST antibodies. The molecular weights of NbREM1.1-GST, His, P7/P7S59A/P7S59D-His are 60, 20 and 27 kDa, respectively. The experiments were repeated three times with similar results. c Callose fluorescence at PD upon aniline blue staining. (scale bar = 20 μm). PDLP5-RFP served as a PD marker. d Quantification of the callose staining intensity. The relative callose staining intensity was set as 1 for untreated WT plants. Value is the mean ± SD (one-way ANOVA with Tukey’s test, n = 3 biologically independent experiments, P-values are shown in the Source Data file). Different lowercase letters indicate significant differences (P < 0.05). e Fluorescence images of N. benthamiana leaf tissue coexpressing PVX-GFP with EV, P7-Flag, P7S59D-Flag and P7S59A-Flag. GFP fluorescence signals were recorded by an inverted fluorescence microscope at 3 dpi (scale bar = 100 μm). f Statistical analysis of PVX-GFP movement efficiency shown in (e). Value is the mean ± SD (one-way ANOVA with Tukey’s test, n = 5 biologically independent experiments, P-values are shown in the Source Data file). Different lowercase letters indicate significant differences (P < 0.01). For box plot, the horizontal lines from top to bottom represent the maximum, first quartile, median, third quartile, and minimum of the total data, respectively. Source data are provided as a Source Data file.
