Extended Data Fig. 1: Flg22 represses iron uptake through FLS2.
From: Spatial IMA1 regulation restricts root iron acquisition on MAMP perception
a, Phenotype of 15-day-old Arabidopsis seedling leaves response to different iron concentrations (shown in figures) with or without low levels of flg22 (10 nM) treatment. Scale bar 1 cm. b,c, Phenotype of 15-day-old Arabidopsis seedling leaves response to sufficient iron (50 µM) or low iron (10 µM) with or without low levels of flg20 or flg22 (10 nM) treatment. b, Shoots; scale bar 1 cm. c, Total chlorophyll concentration of Col-0 shoots. Bar chart centres show means of 3 biological replicates. Error bar: s.e.m. The numbers correspond to P-values analysed by two-tailed Student t-test. d, Quantitative analysis of ferric chelate reductase activities in Col-0 roots grown for 7 days under +Fe conditions and transferred to -Fe, -Fe with flg20 and -Fe with flg22 liquid medium for 2 days. The bar chart centres show means of 5 biological replicates. Error bar: s.e.m. Different letters indicate statistically significant differences between different conditions analysed by one-way ANOVA and Tukey’s test (p < 0.05). e, Promoter activity of IRT1 in the root of pIRT1::NLS-2xYpet seedlings in response to -Fe, -Fe with flg20 and -Fe with flg22 treatment. Seedlings were grown on the +Fe medium and after 5 days transferred to the different liquid media for 24 h treatment. Green: nuclear localized Ypet; red: propidium iodide (PI) cell wall stain. For each treatment, a representative single confocal section (single image, GFP/PI), maximum intensity Z-projection (Z-max, GFP only), a single optical section of the transverse view, and the Z-projection of the transverse section are shown. Scale bar, 50 µm. f–h, Western blots showing IRT1 protein levels in Col-0 and irt1-1 roots grown in +Fe and -Fe (f) or Col-0 under +Fe, -Fe and -Fe with different concentrations of flg22 treatment (g) or Col-0 under +Fe, -Fe and -Fe with flg20 or flg22 treatment (h). Arrow indicates the IRT1 protein band. The asterisk indicates non-specific band. Tubulin protein was blotted as an internal control. i–k, Phenotype of 15-day-old Arabidopsis Col-0 and fls2 seedling leaves in response to sufficient iron (50 µM) or low iron (10 µM) with or without low levels of flg22 (10 nM) treatment. The numbers correspond to p-values that were analysed by two-tailed Student t-test. i, Shoots; scale bar 1 cm; j, total chlorophyll concentration of Col-0 shoots; k, iron concentration of Col-0 seedlings; bar chart centres show means of 3 biological replicates. Error bar: s.e.m. l, Quantitative analysis of ferric chelate reductase activities in Col-0 and fls2 roots grown for 7 days under +Fe conditions and transferred to +Fe, +Fe with flg22, -Fe and -Fe with flg22 liquid medium for 2 days. The bar chart centres show means of 5 biological replicates. Error bar: s.e.m. Different letters indicate statistically significant differences between different conditions analysed by one-way ANOVA and Tukey’s test (p < 0.05). m, Western blots showing IRT1 protein levels in Col-0 and fls2 roots in +Fe, +Fe with flg22, -Fe and -Fe with flg22 treatment. The tubulin protein was blotted as an internal control.
