Luminol-based ROS Detection in Plants

Luminol-based ROS production in leaf tissue was performed as described [38] (link) using indicated PAMP concentrations or using Pto DC3000 or Pto hrcC⁻[42] (link). For inhibitor treatments, leaf tissue was pre-treated by floating leaf discs in water-containing 30 µM Wortmannin (Wm; Sigma-Aldrich; St. Louis, MO) for one hour prior to subsequent flg22-elicitation [13] (link). For ROS assays in six-to-seven-day old plants, cotyledons were cut in half, and the two halves were placed into a 96-well microplate for elicitation. All ROS experiments shown in a same panel were performed in the same 96-well plate at the same time to allow for direct comparison.