Fig. 8: Arabidopsis LHT1 is required for flg22-induced depletion of apoplastic proline and maximal pattern-triggered immunity to P. syringae. | Nature Communications

Fig. 8: Arabidopsis LHT1 is required for flg22-induced depletion of apoplastic proline and maximal pattern-triggered immunity to P. syringae.

From: Transporter-mediated depletion of extracellular proline directly contributes to plant pattern-triggered immunity against a bacterial pathogen

Fig. 8

a–c Col-0 and lht1-7 leaves were syringe-infiltrated with 100 nM flg22 or a mock treatment. a Proline levels in apoplastic wash fluid (AWF) isolated from leaves eight hours post-treatment. Graphed are means ± SE of normalized peak areas for proline, n = 3. b 13C-proline was infiltrated into treated leaves eight hours post-treatment, then AWF isolated from the treated leaves at timepoints indicated. Graphed are means ± SE of normalized peak area for 13C-proline in AWF, n = 3. c P. syringae DC3000 carrying a hrpLpromotor:gfp or empty gfp plasmid were incubated for 10 h in AWF isolated from leaves eight hours post-treatment. Graphed are means ± SE of normalized GFP fluorescence, n = 3. d 1 × 106 cfu/mL of P. syringae DC3000 or DC3000ΔputA was syringe-infiltrated into Col-0 and lht1-7 leaves pre-treated for six hours with 100 nM flg22 or a mock treatment. Leaf bacteria were enumerated on day 3 by serial dilution and plating of leaf extracts. Graphed are log-transformed means ± SE of colony-forming units (cfu) of bacteria isolated from infected tissue, n = 12 and 20 for DC3000 ΔputA -and DC3000-infected samples, respectively. e Proline levels in apoplastic wash fluid (AWF) isolated from Col-0 and lht1-7 plants eight hours after syringe-infiltration with 100 nM flg22 or a mock treatment. Graphed are means ± SE of normalized peak areas for proline, n = 5. f 1 × 106 cfu/mL of P. syringae DC3000 was syringe-infiltrated into sid2-1 and sid2-1/lht1-7 leaves pre-treated for six hours with 100 nM flg22 or a mock treatment. Leaf bacteria were enumerated on day 3 by serial dilution and plating of leaf extracts. Graphed are log-transformed means ± SE of colony-forming units (cfu) of bacteria isolated from infected tissue, n = 18. Lower case letters in all panels denote significance groupings based upon ANOVA with Tukey’s HSD, p < 0.05. Box plots in d and f show median, interquartile range, min, max, black boxes are the mean values. Data in all panels were pooled from three independent experiments.

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