Temporal Metabolic Reprogramming Reveals Stage-Specific Adaptations in Proso Millet Resistance Against Head Smut

BACKGROUND/OBJECTIVES: . Although partial resistance exists, the dynamic molecular mechanisms governing its defense response across developmental stages remain poorly understood. METHODS: Here, we performed untargeted metabolomics on leaf samples from Inoculated Asymptomatic (IA) and Inoculated Symptomatic (IS) plants of the partially resistant cultivar 'Chishu 13' at four key growth stages following pathogen inoculation, with group classification validated by qPCR. Using weighted metabolite co-expression network analysis (WGCNA) combined with differential metabolite screening, we identified 18 metabolites markedly enriched in the tricarboxylic acid (TCA) cycle, metabolite transport-related processes, and phenylpropanoid biosynthesis pathways. RESULTS: -coumaric acid. Our results reveal distinct temporal patterns in metabolic reprogramming that correlate with resistance outcomes in Inoculated Asymptomatic plants: early stages are characterized by differential regulation of energy metabolism, while later stages show enhanced phenylpropanoid biosynthesis. These stage-specific metabolic adaptations are strongly associated with successful defense outcomes. CONCLUSIONS: These findings elucidate stage-specific metabolic adaptations that distinguish successful defense in IA plants from susceptibility in IS plants, providing robust biomarkers and stage-targeted strategies for breeding smut-resistant millet varieties.