Increased Resistance to Biotrophic Pathogens in the Arabidopsis <i>Constitutive Induced Resistance 1</i> Mutant Is EDS1 and PAD4-Dependent and Modulated by Environmental Temperature

<div><p>The Arabidopsis <i>constitutive induced resistance 1</i> (<i>cir1</i>) mutant displays salicylic acid (SA)-dependent constitutive expression of defence genes and enhanced resistance to biotrophic pathogens. To further characterise the role of CIR1 in plant immunity we conducted epistasis analyses with two key components of the SA-signalling branch of the defence network, ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1) and PHYTOALEXIN DEFICIENT4 (PAD4). We demonstrate that the constitutive defence phenotypes of <i>cir1</i> require both EDS1 and PAD4, indicating that CIR1 lies upstream of the EDS1-PAD4 regulatory node in the immune signalling network. In light of this finding we examined EDS1 expression in <i>cir1</i> and observed increased protein, but not mRNA levels in this mutant, suggesting that CIR1 might act as a negative regulator of EDS1 via a post-transcriptional mechanism. Finally, as environmental temperature is known to influence the outcome of plant-pathogen interactions, we analysed <i>cir1</i> plants grown at 18, 22 or 25°C. We found that susceptibility to <i>Pseudomonas syringae</i> pv. <i>tomato</i> (<i>Pst</i>) DC3000 is modulated by temperature in <i>cir1</i>. Greatest resistance to this pathogen (relative to <i>PR-1:LUC</i> control plants) was observed at 18°C, while at 25°C no difference in susceptibility between <i>cir1</i> and control plants was apparent. The increase in resistance to <i>Pst</i> DC3000 at 18°C correlated with a stunted growth phenotype, suggesting that activation of defence responses may be enhanced at lower temperatures in the <i>cir1</i> mutant.</p>