Figure 9.

edr4 Mutants Show Defects in FM4-64 Uptake; Focal Accumulation of EDR1 and EDR4 Requires CHC2.

(A) Delay of FM4-64 uptake in the edr4 mutants. Intracellular accumulation of FM4-64 was examined at 8 min after staining with 2 μM FM4-64 in root tip cells of 5-d-old seedlings of the wild type and edr4-1, edr4-2, and chc2-1 mutants. Bars = 10 μm.

(B) Quantification of relative FM4-64 uptake. The bars represent means and sd (n > 50 cells analyzed). Statistically significant differences among the samples are labeled with different letters (P < 0.01, one-way ANOVA). The experiments were repeated three times with similar results.

(C) Photographs and trypan blue staining of representative leaves removed from 4-week-old wild-type, chc2-1, chc2-2, and edr4-1 plants at 8 DAI with G. cichoracearum. Bars in top panels = 0.5 cm; bar in bottom panels = 100 μm.

(D) Quantitative analysis of conidiophore formation on 4-week-old wild-type, chc2-1, chc2-2, and edr4-1 plants inoculated with G. cichoracearum at 6 DAI. The bars represent means and sd. The experiment was repeated three times with similar results (n = 20). Statistically significant differences are labeled with different letters (P < 0.01, one-way ANOVA). The experiments were repeated three times with similar results.

(E) and (G) Focal accumulation of EDR1-GFP (E) or EDR4-GFP (G) at the pathogen infection site in epidermal cells of 4-week-old wild-type and chc2-1 plants after inoculation with G. cichoracearum at 24 HAI. PM, plasma membrane; PP, penetration peg. Bars = 10 μm.

(F) and (H) Quantitative analysis of powdery mildew-induced EDR1-GFP (F) or EDR4-GFP (H) focal accumulations (FA) in wild-type and chc2-1 plants. Bars represent means and sd of values obtained from three biological replicates per genotype. Statistically significant differences among the samples are labeled with different letters (P < 0.01, one-way ANOVA). The experiments were repeated three times with similar results.