Metabolomics analysis reveals the formation mechanism of Cordyceps militaris fruiting bodies in liquid culture

Cordyceps militaris is a functional fungus renowned for its significant nutritional and therapeutic value. However, the mechanisms governing its growth and fruiting body formation in liquid culture remain poorly understood. In this study, we employed UHPLC-MS/MS-based untargeted metabolomics to profile the metabolome of C. militaris across key developmental stages. A total of 2288 metabolites were identified in C. militaris. Among these, amino acids and derivatives accounted for the highest proportion, with 894 types identified. Organic acids are the second most abundant compounds, with 244 types identified. Metabolite profiles varied significantly across growth stages. Amino acids, organic acids, and lipids may serve as potential markers for distinguishing developmental stages. Notably, significant alterations were observed in six key metabolic pathways during mycelial proliferation, primordium differentiation, and fruiting body maturation. Our findings elucidate the metabolic drivers of C. militaris growth in liquid culture, providing a mechanistic basis for strain improvement and the industrial-scale production of bioactive metabolites.