The Transcription Factor <scp>OsWRKY64</scp> Interacts With <scp>OsART1</scp> to Positively Regulate Al Resistance in Rice

Rice (Oryza sativa) exhibits notable aluminium (Al) tolerance, as the C₂H₂-transcription factor OsART1 plays a crucial role in regulating Al tolerance in rice by modulating the expression of specific genes. However, the posttranscriptional regulation of OsART1 remains poorly understood. In this study, we identified and characterised OsWRKY64, which interacts with OsART1. OsWRKY64 was localised in the nucleus. Tissue expression analysis indicated that OsWRKY64 was primarily expressed in the roots, and its transcription and protein accumulation were significantly induced by Al. The oswrky64 mutants and the OsWRKY64 overexpression lines exhibited decreased or increased expression of genes regulated by OsART1, which consequently led to phenotypes sensitive or resistant to Al, respectively. Further analysis indicated that OsWRKY64 interacted with OsART1 to promote its stability, thereby accelerating the transcriptional activity of OsART1 on its downstream genes. Additionally, OsART1 upregulated OsWRKY64 expression by directly binding to the OsWRKY64 promoter, thus forming a negative feedback loop between OsART1 and OsWRKY64. Overall, our results demonstrated that OsWRKY64 influences Al resistance by interacting with and stabilising OsART1 to accelerate its transcriptional activity.