EDR1 associates with its homologs to synergistically regulate plant immunity in Arabidopsis.

ENHANCED DISEASE RESISTANCE 1 (EDR1), a Raf-like mitogen-activated protein kinase (MAPK) kinase kinase (MAPKKK), is a negative regulator of resistance. There are three homologs, RAF3/4/5, of EDR1 in Arabidopsis. However, the roles of RAF3/4/5 in resistance and their functional link with EDR1 in plant immunity remain unclear. Here, we showed that the raf3/4/5 triple mutant displayed wild-type-like phenotypes to the powdery mildew pathogen Golovinomyces cichoracearum UCSC1 and the bacterial pathogen Pseudomonas syringae pv. tomato (Pto) DC3000. However, the edr1 raf3/4/5 quadruple mutant exhibited enhanced resistance to G. cichoracearum UCSC1 and Pto DC3000 compared to edr1. Consistently, MPK3/6 kinase activity was more highly activated in edr1 raf3/4/5 than that in edr1. Moreover, the enhanced resistance of edr1 raf3/4/5 required SALICYLIC ACID INDUCTION DEFICIENT 2 (SID2), an isochorismate synthase required for salicylic acid (SA) synthesis. Additionally, unlike EDR1, RAF3/4/5 weakly and indirectly associated with MKK4/5, and EDR1 was directly associated with RAF3/4/5. Taken together, these data indicate that EDR1 associates with RAF3/4/5, and they may function together to synergistically suppress MAPK cascades activation, which reveal the complexity and importance of Raf-like MAPKKKs in plant immunity regulation.