Fig. 3: OsTSN1 negatively regulates rice antiviral defense responses.

a Cell viability of rice protoplasts under 1,6-HD treatments. Data are the means ± SD from five independent biological replicates, determined using one-way ANOVA with Tukey’s test. ns, no significant statistical difference. b RT-qPCR showing the expression levels of SRBSDV P6 and P10 genes in SRBSDV-infected rice protoplasts treated with 0%, 0.5% and 1% 1,6-HD. Data are the means ± SD from three independent biological replicates, determined using one-way ANOVA with Tukey’s test. ns, no significant statistical difference, *P < 0.05, ***P < 0.001, ****P < 0.0001. c Subcellular localization of OsTSN1 in uninfected, SRBSDV-infected rice protoplasts and SRBSDV-infected rice protoplasts with 1% 1,6-HD treatment. White arrows indicate droplets. Scale bar, 5 μm. d Gene sequencing results showing a 5 bp deletion or one T insertion in ostsn1#2 and ostsn1#3 knockout mutant rice lines, respectively. e Western blot showing the expression levels of OsTSN1 in WT, two ostsn1 knockout mutant lines, and two OsTSN1-OE transgenic rice lines. The expression level of OsActin was used as normalizer. f Heights of uninfected and SRBSDV-infected WT, ostsn1 knockout mutant and OsTSN1-OE transgenic rice plants. Data are the means ± SD (n = 15) from three independent biological experiments, determined using one-way ANOVA with Tukey’s test. ns, no significant statistical difference, ****P < 0.0001. g–l Phenotypes and virus accumulations of SRBSDV-infected WT, ostsn1 knockout mutant and OsTSN1-OE transgenic rice plants. Phenotypes of uninfected or SRBSDV-infected WT, ostsn1 knockout mutant rice (g) and OsTSN1-OE transgenic plants (j) at 30 dpi. Scale bar, 10 cm. RT-qPCR showing the expression level of SRBSDV P10 gene in SRBSDV-infected WT, ostsn1 knockout mutant (h) and OsTSN1-OE transgenic plants (k) at 30 dpi. Data are the means ± SD from three independent biological replicates per treatment, determined using one-way ANOVA with Tukey’s test. ****P < 0.0001. Western blot assay showing the accumulation of SRBSDV P10 in SRBSDV-infected WT, ostsn1 knockout mutant (i) and OsTSN1-OE transgenic plants (l) at 30 dpi. The Rubisco bands are used to show equal loading. Experiments in (c, e, i, and l) were repeated three times with similar results.