FIGURE 5.

Depletion of N⁶ methylation of adenosine (m⁶A) increases the rice black streaked dwarf virus (RBSDV) titres. (a) RBSDV titres in insects injected with either dsLsMETTL3+14 or dsGFP. Each dot represents one RBSDV‐infected small brown planthopper (SBPH) that had been injected with either dsLsMETTL3+14 or dsGFP and collected at 6 days. Data were collected from three independent experiments. dsGFP was used as an RNA interference (RNAi) control. RBSDV titres shown as copies/10⁻⁴ actin. Significant differences between mean copies of RBSDV in insects injected with either dsLsMETTL3+14 or dsGFP were determined by Prism 8 using Student's t test (***p < 0.001). (b) Western blots of insects injected with either dsLsMETTL3+14 or dsGFP. Total protein was extracted and treated with anti‐RBSDV P10 antibody and anti‐actin antibody, respectively. (c) Fluorescence intensity of m⁶A and RBSDV in midgut epithelial cells. Excised midguts were incubated with anti‐m⁶A (green label), anti‐RBSDV (red label) antibodies, and DyLight 633 phalloidin (blue, labelling actin), then observed with a confocal laser‐scanning microscope. Scale bars, 20 μm. (d) Fluorescence intensity of m⁶A and RBSDV. Ten midgut epithelial cells of SBPHs injected with dsLsMETTL3+14 or dsGFP were evaluated. Significant differences between mean relative intensity of m⁶A or RBSDV in epithelial cells of SBPHs injected with dsLsMETTL3+14 or dsGFP were determined using Student's t test (**p < 0.01)