Figure 3.

The extracellular domain of RFO1^Col−0 binds commercial and Arabidopsis cell wall-derived dmPectin.

(A) Cumulative number of vascular penetrations observed in WT (Col-0), rfo1-1, RFO1^Col−0-GFP (rfo1-1 RFO1^Col−0-GFP), and RFO1^chim-GFP (rfo1-1 RFO1^chim-GFP) roots between 3 and 7 dpt to plates containing F. oxysporum5176 pSIX1GFP microconidia. Data represent the mean ± SE of N ≥ 10 seedlings per genotype from three independent replicates. RM ANOVA with Tukey’s multi-comparison post hoc test, ∗∗p < 0.01, ∗∗∗p < 0.001. Significance shown compared with WT.

(B) Representative images of 8-day-old WT, PMEIox, rfo1-1, rfo1-1 PMEIox, rfo1-1 RFO1^Col−0-GFP PMEIox, and rfo1-1 RFO1^chim-GFP PMEIox seedlings. Scale bar, 5 mm.

(C) Relative root growth of EGCG-treated WT, rfo1-1, RFO1^Col−0-GFP, and RFO1^chim-GFP roots as a percentage of Mock. Bars represent the mean ± SE of >15 seedlings per genotype from four biological replicates. One-way ANOVA with Dunnett’s multiple-comparison post hoc test, ∗∗∗∗p < 0.0001. Significance indicated is compared with WT.

(D) Quantification of dot immunobinding assay intensities in mean gray values from membranes of immobilized commercial demethylesterified pectin (dmPectin) and methylated pectin (mPectin) probed with RFO1^ECD-SNAP or SNAP recombinant proteins as shown in Supplemental Figure 5A. Boxplots: centerlines show the medians; means are marked by +; box limits indicate the 25th and 75th percentiles; whiskers extend to the minimum and maximum. N ≥ 12 from ≥3 independent replicates, two-way ANOVA with Tukey’s multiple-comparison test, ∗∗∗∗p < 0.0001.