Extended Data Fig. 7: The effects of JA/AUX/SL phytohormone, overexpressing or knock-down/-out TCP21 on TSWV accumulation in Nicotiana benthamiana, pepper and Arabidopsis plant leaves.
From: NLR surveillance of pathogen interference with hormone receptors induces immunity
a, Phenotype of TSWV-inoculated N. benthamiana plants treated with DMSO, MeJA, 2,4-D or rac-GR24. N. benthamiana plants were sprayed with DMSO, 100 µM MeJA, 100 µM 2,4-D and 100 µM rac-GR24, respectively. At 3 d post treatment, the fresh sap from TSWV infected tissues was mechanically inoculated onto phytohormone treated leaves. The phenotype of TSWV-inoculated plants was photographed at 9 d post inoculation. b, TSWV accumulation was analysed in systemic infected leaves of N. benthamiana plants treated with DMSO, MeJA, 2,4-D and rac-GR24, respectively, at 9 dpi by Western blot using TSWV N specific antibodies. Ponceau S staining was used to estimate sample loadings. c, Phenotype of TSWV-inoculated pepper plant (-Tsw) treated with DMSO, MeJA, 2,4-D or rac-GR24. C. annum fasciculatum (-Tsw) were sprayed with DMSO, 100 µM MeJA, 100 µM 2,4-D and 100 µM rac-GR24, respectively. At 3 d post treatment, the fresh sap from TSWV infected tissues was mechanically inoculated onto phytohormone treated leaves. The phenotype of TSWV-inoculated plants was photographed at 6 d post inoculation. d, TSWV accumulation was analysed in systemic infected leaves of pepper plant (-Tsw) treated with DMSO, MeJA, 2,4-D and rac-GR24, respectively, at 6 dpi by Western blot using TSWV N specific antibodies. Immunoblot analysis of actin is used to estimate the sample loadings. **, p < 0.01; ns, no significance. Data were analysed by two-sided Student’s t-test; exact P values are shown in the Source Data. e, TSWV infection phenotype on wild-type (WT) and AtTCP21 transgenic Arabidopsis plants. The photos of infected plants were taken at 10 dpi. f, TSWV infection phenotype on WT and tcp7/21 mutant Arabidopsis plants. The photos of infected plants were taken at 12 dpi. g, Silencing of TCP21 in pepper plants (-Tsw) enhances resistance to TSWV. The fusion of 200 bp cDNA fragments each of CaTCP21-1, 3, 4 and 7 (TRV-CaTCP21-1/3/4/7) was used to infect C. annum fasciculatum (-Tsw). Phenotype of TSWV-inoculated C. annum fasciculatum silenced for CaTCP21-1/3/4/7 was photographed at 7 dpi (left). TSWV accumulation in systemic infected leaves of C. annum fasciculatum silenced for CaTCP21-1/3/4/7 at 7 dpi was analysed by immunoblot analysis using TSWV N specific antibodies (right). Immunoblot analysis of actin is used to estimate sample loadings. **, p < 0.01. Data were analysed by two-sided Student’s t-test; the exact P values are shown in the Source Data. h, Phenotype of Botrytis cinerea infection lesions on detached leaves of WT (left), TCP21 transgenic (middle) and tcp7/21 mutant (right) Arabidopsis plant leaves at 48 h post inoculation. B. cinerea lesions on leaves were quantified and shown in the right. Data are presented as mean values ± s.e.m.; n = 6 biologically independent samples. Lowercase letters a-c represent statistically different groups (one-way ANOVA with Tukey’s test, p < 0.05); exact P values great than 0.0001 (adjusted) are shown in Source Data. In box plots the center line represents the median, box edges delimit lower and upper quartiles and whiskers show the highest and lowest data points. Experiments were repeated at least three times with similar results.
