The IRE1-bZIP60 branch of Unfolded Protein Response is required for Arabidopsis immune response to B. cinerea

Abstract

The Unfolded Protein Response (UPR) is a retrograde signalling pathway which is activated when endoplasmic reticulum (ER) proteostasis is disturbed. Here, we have investigated by reverse genetics the contribution of such pathway in Arabidopsis thaliana response to two necrotrophic fungi of agricultural importance, Botrytis cinere a which is responsible for the development of grey mold disease, and Alternaria brassicicola which triggers black spot disease. We found that the branch of UPR dependent on the INOSITOL-REQUIRING ENZYME 1 (IRE1) and the transcription factor (TF) bZIP60 is required to restrict foliar necrotic symptoms induced by both fungi. Accordingly, focussing on B. cinerea, we provided evidence for the production of the active bZIP60 form during infection. This activation was accompanied by an increased expression of UPR-responsive genes coding for ER-localized chaperones and co-chaperones that belong to the ER-Quality Control (ER-QC) system. Furthermore, mutants deficient for two ER-QC components were also more susceptible to infection. By contrast, investigating the involvement of CELL DIVISION CYCLE 48 (CDC48) AAA+-ATPAses that assist ER-Associated Degradation (ERAD) pathway for disposal of luminal unfolded proteins, we showed that a series of mutants and transgenics are more resistant to grey mold disease. Seeking for molecular insights into how the ER could shape Arabidopsis immune response to B. cinerea, we quantified the expression of defence gene and cell death markers in single bzip60 and double ire1 mutants. However, none of those genes were mis-regulated in mutant genetic backgrounds, indicating that IRE1-bZIP60 branch of UPR modulates the Arabidopsis response to B. cinerea by a yet-to-be-identified mechanism. Interestingly, we identified the NAC053/NTL4 TF as a potential actor of this unknown mechanism, linking the UPR and proteasome stress regulon. Author summaryNecrotrophic fungi are one of the most economically significant plant pathogens worldwide, inflicting massive pre- and post-harvest losses on a wide range of fruit and vegetable crops. They adopt a necrotrophic lifestyle, deriving their nutrients predominantly from dead plant tissues to complete their life cycle. Botrytis cinerea is the causal agent of grey mold and no plant shows complete resistance towards this pathogen. The use of genetic models such as the plant Arabidopsis thaliana has partially enabled the understanding of the immunity mechanisms involved in the plants response to B. cinerea. Our work provides new insights into the cellular mechanisms of how plants cope with this pathogen. In this context, by means of a reverse genetic approach, we explored the role of the Unfolded Protein Response (UPR), a cell signalling pathway regulating protein homeostasis within the endoplasmic reticulum (ER) and thus protecting cells from a harmful over-accumulation of aberrant or misfolded proteins.