ROS Burst

ROS assays were performed as previously described (Huang et al., 2013). Briefly, six leaf discs (10 mm diameter) from three 5-week-old Arabidopsis (two discs/plant) were incubated in double-distilled water in 96-well plates overnight. The following day, the water was replaced by 10 nM flg22 or elf26 in 10 mM MgSO₄ buffer or by 10 mM MgSO₄ buffer only for the mock controls containing 2 µM luminol (Sigma-Aldrich) and 10 µg/mL peroxidase (Sigma-Aldrich). The plates were analyzed every 2 min for mutants or every 2.5 min for OE lines after addition of MAMPs for a period of 30 min using a CentroLIApc LB 692 plate luminometer (Berthold Technologies).