Figure 4

FRET-FLIM analysis of the MLO-calmodulin interaction. Barley MLO is a plant-specific calmodulin-binding protein that functions as a modulator of defence against the common powdery mildew pathogen [90]. YFP-tagged wild-type barley MLO or mutant variants thereof (W423R and L420R W423R, bearing amino acid substitutions in the calmodulin binding domain [90]) were co-expressed with CFP-tagged calmodulin in single barley leaf epidermal cells. FRET-FLIM analysis was performed as described in [32]. Donor fluorophore lifetimes are color-coded according to the scale indicated on top of the Figure. "Warmer" colors are indicative of shorter donor fluorophore lifetimes and thus interaction between MLO and calmodulin. Size bar, 20 μm.