Extended Data Fig. 1: Overexpression of SNIPER1 leads to reduced expression of SARD1 and FMO1, lower SA accumulation, increased Pto DC3000 hrcC growth, but has no effect on MAPK activation and ROS production during PTI.
From: Activation of TIR signalling boosts pattern-triggered immunity
(a, b) Relative expression levels of SARD1 (a) and FMO1 (b) in WT and OX-SNIPER1 lines (L4 and L5) treated with 10 mM MgCl2 (mock) or Pto DC3000 hrcC for 12 h. Bars represent mean ± s.d. (n = 3 biologically independent samples). (c, d) Free SA (c) and SAG (d) levels in the indicated genotypes treated with 10 mM MgCl2 (mock) or Pto DC3000 hrcC (OD600 = 0.05) for 12 h. Bars represent mean ± s.d. (n = 3 biologically independent samples). (e) Growth of Pto DC3000 hrcC in the indicated genotypes. Bars represent mean ± s.d. (n = 4 plants). (f, g, i, j) flg22 and nlp20-induced MAPK activation in the indicated genotypes. Seedlings were treated with 0.1 μM flg22 (f, g) or 0.1 μM nlp20 (i, j). MAPK activation was analyzed by immunoblotting with the anti-pERK antibody. Equal loading is confirmed by Ponceau staining of Rubisco. The signals of phosphorylated MPK6 and MPK3 in samples treated with flg22 (g) or nlp20 (j) detected by western blots were normalized to the loading control. The intensity of MPK6 or MPK3 bands after elicitors treatment in WT plants was set as 1. Bars represent mean ± s.d. (n = 4 different experiments for (g), n = 3 different experiments for (j)). (h, k) ROS production in the indicated genotypes after treatment with 0.1 μM flg22 (h) or 1 μM nlp20 (k) measured as luminescence. The values show the peak of luminescence units through all the time points. Bars represent mean ± s.d. (n = 8 leaf disks from different plants). (l, m) SAG levels in the indicated genotypes treated with H2O, 1 µM nlp20 (l) or 1 µM flg22 (m). Samples were collected for SAG measurement 24 h after 1µM nlp20, or 9 h after 1 µM flg22 treatment. Bars represent mean ± s.d. (n = 3 biologically independent samples). All data were analyzed by one-way ANOVA with Tukey’s test. Exact P values are provided in Supplementary Table 5. The experiments were repeated at least three times with similar results.
