Salicylic acid regulates anthesis via CmWRKY25/CmWRKY53-CmSOC1 model in Clivia miniata

Flowering time are critical traits of Clivia miniata, but their regulatory mechanisms remain elusive. Here, metabolomic, physiological, and molecular analyses were combined to dissect these processes. Metabolomic profiling of four C. miniata tissues identified 787 metabolites with distinct tissue-specific signatures; scape-specific metabolites were enriched in lipids, phenolic acids, and salicylic acid (SA)—a plant hormone linked to flowering regulation. Exogenous SA accelerated C. miniata flowering in a concentration-dependent manner, with 100 mg/L SA advancing anthesis by 34 d. Transcriptional analyses showed two WRKY transcription factors, CmWRKY25 and CmWRKY53, were significantly upregulated by SA. Phylogenetic and sequence alignment confirmed their homology to Arabidopsis WRKYs with known flowering regulatory roles. Transient overexpression of CmWRKY25 and/or CmWRKY53 promoted C. miniata flower bud differentiation. Bimolecular fluorescence complementation (BiFC), and yeast two-hybrid (Y2H) assays demonstrated that CmWRKY25 and CmWRKY53 physically interacted, formed a functional module. Promoter analyses identified SA-responsive cis-elements in all three genes. Dual-luciferase (LUC), yeast one-hybrid (Y1H) assays demonstrated that CmWRKY25 and CmWRKY53 additively activated CmSOC1 transcription. Collectively, our findings uncover a regulatory pathway wherein SA induces formation of the CmWRKY25-CmWRKY53 module to activate CmSOC1 and promote C. miniata flowering, providing new insights into SA-mediated flowering control in C. miniata.