Figure 5.

Analysis of the role in immunity to Pst of the Agp1 SP and putative GPI anchor. A, Top: amino acid sequence of the Agp1 protein. SP sequence is highlighted in green and the GPI-anchored sequence is highlighted in blue. Schematics show the substituted amino acids or deletions of the Agp1 protein, each fused to an HA epitope tag. B, Leaves of 5-week-old N. benthamiana plants were syringe-infiltrated with Agrobacterium (1D1249) strains (OD = 0.5) carrying a binary expression vector expressing each gene. Two days later, the same agroinfiltrated spots were syringe-infiltrated with 5 × 10⁴ cfu · mL⁻¹ DC3000ΔhopQ1-1. Pst DC3000 populations were measured 2 days after the second infiltration. Cfu, colony-forming unit. Bars show means ± sd. Different letters indicate significant differences based on a one-way ANOVA followed by Student’s t test (P < 0.05). Three plants were tested with each gene in each experiment. The experiments were performed twice with similar results. C, Proteins were extracted from N. benthamiana leaves expressing each Agp1:HA variant 2 days after agroinfiltration. Proteins were detected by immunoblotting with an α-HA antibody. The upper red arrow indicates the Agp1:HA fusion protein and the lower black arrow indicates the expected mass of the Agp1:HA protein.