Figure 5. P. syringae flagellins or nonglycosylated flagellins can impair the HR elicitation by PA5ΔfliC.

(A) The macroscopic HR elicition on Nicotiana benthamiana leaves. fliC mutant (PA5ΔfliC) at 1×10⁷ cfu/ml was infiltrated simultaneously with 0.8 µM of P. syringae pv. averrhoi flagellin, nonglycosylated flagellin (NG-flagellin), A. tumefaciens flagellin (Agro-flagellin) and BSA proteins, respectively, compared to inoculation with fliC mutant (PA5ΔfliC) and complemented strain [PA5ΔfliC (pNCHU1039)]. The leaf inoculated at concentration of 3.2 µM flagellin by syringe-infiltration was served as a control. The photos were taken at 2 days post inoculation. pNCHU1039: pBBR1MCS5 carrying fliC. (B) The P. syringae pv. averrhoi flagellin protein was purified from PA5 by using ultracentrifugation as described in Materials and Methods and separated by SDS-12% PAGE. The molecular sizes of marker proteins are indicated on the left. (C) Electrolyte leakage from leaf areas inoculated with indicated inoculum. The conductivity values represent the mean and standard error of three different leaves. Significant differences (p<0.05) were indicated by asterisk (*). All experiments were repeated at least three times with similar results.