Figure 1. HrpL-FLAG is functional.
(A) HrpL-FLAG is recognized by anti-FLAG antibody in a Western Blot. (B) hrpL-FLAG and WT DC3000 strains evoke the hypersensitive response while it is abolished in the ΔhrpL strain. Bacteria were infiltrated via a blunt syringe into three independent leaves at 3×108 CFU/ml. Photos were taken after 3 days. Symptoms are identical among three replicates. (C) Expression of a plasmid-based (pBS181) β-glucuronidase (GUS) reporter driven by a hrp promoter (hrpJ) in hrpL-FLAG and WT DC3000 backgrounds. Three different media were used: HMM (hrp-minimal medium for highest induction of hrp promoters), MG (Mannitol-Glutamate medium for intermediate induction of hrp promoters but better bacterial growth), and KB (King’s B rich medium for repression of HrpL regulon expression). All plates contain appropriate antibiotics to maintain plasmids and X-Gluc, a substrate of GUS. (D-E) Relative fold change of transcript levels for hrpL and hopQ1-1 (PSPTO_0877) after medium shift from KB to MG (supplemented with ferric iron at 50 µM final concentration) over 9 hours.