OsACL‐A2 negatively regulates cell death and disease resistance in rice

ATP‐citrate lyases (ACL) play critical roles in tumour cell propagation, foetal development and growth, and histone acetylation in human and animals. Here, we report a novel function of ACL in cell death‐mediated pathogen defence responses in rice. Using ethyl methanesulphonate (EMS) mutagenesis and map‐based cloning, we identified an Oryza sativa ACL‐A2 mutant allele, termed spotted leaf 30‐1 (spl30‐1), in which an A‐to‐T transversion converts an Asn at position 343 to a Tyr (N343Y), causing a recessive mutation that led to a lesion mimic phenotype. Compared to wild‐type plants, spl30‐1 significantly reduces ACL enzymatic activity, accumulates high reactive oxygen species and increases degradation rate of nuclear deoxyribonucleic acids. CRISPR/Cas9‐mediated insertion/deletion mutation analysis and complementation assay confirmed that the phenotype of spl30‐1 resulted from the defective function of OsACL‐A2 protein. We further biochemically identified that the N343Y mutation caused a significant degradation of SPL30(N343Y) in a ubiquitin‐26S proteasome system (UPS)‐dependent manner without alteration in transcripts of OsACL‐A2 in spl30‐1. Transcriptome analysis identified a number of up‐regulated genes associated with pathogen defence responses in recessive mutants of OsACL‐A2, implying its role in innate immunity. Suppressor mutant screen suggested that OsSL, which encodes a P450 monooxygenase protein, acted as a downstream key regulator in spl30‐1‐mediated pathogen defence responses. Taken together, our study discovered a novel role of OsACL‐A2 in negatively regulating innate immune responses in rice.