The OsWRKY7-OsVQ3 module regulates seedling-stage cold tolerance in rice

Cold stress poses a significant threat to rice productivity and quality, threatening food security globally. While WRKY transcription factors represent one of the largest plant-specific gene families, their functional mechanisms in cold response remain poorly understood. Here, we identify OsWRKY7 as a negative regulator of cold tolerance in rice and elucidate the molecular mechanism of the OsWRKY7-OsVQ3 module-mediated cold stress regulation. OsWRKY7-knockout mutants exhibited enhanced cold tolerance in rice seedlings, whereas OsWRKY7 overexpression lines showed increased cold sensitivity. Transcriptome profiling revealed that OsWRKY7 disruption induces extensive transcriptional reprogramming under cold stress, suppressing cold-sensitive genes while activating reactive oxygen species (ROS)-scavenging pathways. OsPP2C27 and OsXLG4, two cold-sensitive genes, were identified as direct downstream targets of OsWRKY7. Biochemical analyses demonstrated that OsVQ3 physically interacts with OsWRKY7, antagonizing its transcriptional activation and DNA binding ability. Consistently, OsVQ3-knockout mutants displayed impaired cold tolerance at both the seedling and shooting stages. Natural variation in the OsWRKY7 promoter defines distinct haplotypes associated with latitude-specific adaptation in rice. The japonica-specific Hap3, enriched in high-latitude regions, confers enhanced cold tolerance through reduced OsWRKY7 transcriptional responsiveness to cold stress. Our findings uncover a key regulatory module governing cold response and provide potential targets for breeding cold-resistant rice varieties.