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. 2023 Oct 18;36(2):427–446. doi: 10.1093/plcell/koad270

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© The Author(s) 2023. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

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Figure 2. — NbADR1, NbPAD4, and NbEDS1 mediate bacterial pathogen-induced stomatal closure. A, B) Disease symptoms and bacterial populations in leaves of the Nicotiana benthamiana wild-type (WT), adr1, pad4, eds1, adr1 nrg1, nrg1, and sag101 at 4 d (A, upper panel), or at 0 and 3 d (A, lower panel) postsyringe infiltration, or at 10 d (B, upper panel), or 0 and 6 d (B, lower panel) postspray inoculation with Pseudomonas syringae pv. tomato (Pst) DC3000 ΔHopQ1 (OD₆₀₀ = 0.0001 for syringe, OD₆₀₀ = 0.4 for spray), respectively. Data are means (±Sd) of 4 biological replicates from independent plants. Letters indicate significant differences by 1-way ANOVA analysis (Tukey's post hoc test, P < 0.05). The experiments were repeated 3 times with similar results. Scale bars represent 1 cm. C, D) Stomatal apertures and images of stomata in leaves of N. benthamiana WT and the indicated mutants after 1 h of flood treatment with mock (10 mM MgCl₂) or Pst DC3000 ΔHopQ1 (OD₆₀₀ = 0.4) (C), and Pst DC3000 hrcC⁻ (OD₆₀₀ = 0.4). D) Data are means ± Sd; n = 50 stomata. Letters indicate significant differences by 1-way ANOVA analysis (Tukey's post hoc test, P < 0.05). Scale bars represent 5 μm. The experiments were repeated 3 times with similar results. E, F) Stomatal apertures E) and images of stomata F) in leaves of N. benthamiana WT and the indicated mutants with TRV-GUS or TRV-NbADR1, after 1 h of flood treatment with mock (10 mM MgCl₂) or Pst DC3000 ΔHopQ1 (OD₆₀₀ = 0.4). Data are means ± Sd; n = 50 stomata. Letters indicate significant differences by 1-way ANOVA analysis (Tukey's post hoc test, P < 0.05). Scale bars represent 5 μm. The experiments were repeated 3 times with similar results. G, H) Bacterial pathogen entry assay in leaves of N. benthamiana WT and the indicated mutants. The leaves were photographed and the luciferase (LUC) intensities were quantified by a CCD imaging system after 1 h of P. syringae pv. maculicola (Psm) ES4326-lux flood treatment (OD₆₀₀ = 0.5). Data are means (± Sd) of 3 biological replicates from independent plants. Letters indicate significant differences by 1-way ANOVA analysis (Tukey's post hoc test, P < 0.05). Scale bars represent 1 cm. The experiments were repeated 3 times with similar results. CFU, colony-forming unit; cps, counts per second.