Figure 4.
RESPIRATORY BURST OXIDASE HOMOLOGUE D (RBOHD) phosphorylation sites S343 and S347 are required for reactive oxygen species (ROS) production during effector-triggered immunity (ETI). (a) Immunoblot showing similar protein levels of FLAG-tagged RBOHD protein in rbohD mutants expressing 3xFLAG-RBOHD WT and the S343A/S347A variant. Coomassie stain (CBB) shows rubisco protein to demonstrate equal loading. (b) 3,3’-diaminobenzidine (DAB)-mediated H2O2 staining in rbohD and rbohD mutants expressing 3xFLAG-RBOHD WT and the S343A/S347A variant 8 h after infiltration with the bacteria. The right half of the leaves were infiltrated with 10 mM MgCl2 solution, Pseudomonas syringae pv tomato (Pto) DC3000 EV, Pto DC3000 (avrRpm1) or Pto DC3000 (avrRpt2). We repeated three times with similar results (4–5 leaves per genotype were stained each time).