Morpho-Genetic Characterization and Bioactivity Evaluation of Cordyceps militaris cultivated on Various Substrates

Cordyceps militaris, a well-known medicinal mushroom, possesses diverse pharmacological properties attributed to its rich repertoire of bioactive compounds such as cordycepin, polysaccharides and phenolics. This study aimed to investigate the morphological and genetic characteristics of C. militaris and to evaluate the antimicrobial and antioxidant potential of its fruiting bodies cultivated on five different low-cost substrates: brown rice, foxtail millet, pearl millet, wheat and corn. Solid-state fermentation was employed for cultivation and morphological traits such as fruiting body length, weight and coloration were recorded. Brown rice substrate supported the highest yield in terms of fruiting body size and biomass, while foxtail millet showed the least productivity. Genetic identification using ITS sequencing confirmed the strain's alignment with C. militaris and its close phylogenetic relationship with other Cordyceps species. Ethanolic extracts exhibited the strongest antimicrobial activity, particularly those derived from foxtail millet and brown rice substrates, with inhibition zones up to 25.76 mm against S. aureus. Methanolic extracts showed moderate activity, while aqueous extracts were the least effective. The antioxidant activity was assessed using the DPPH free radical scavenging assay. Ethanolic extracts from fruiting bodies grown on foxtail millet exhibited the highest antioxidant activity with 91% radical scavenging and an IC₅₀ of 0.324 mg/mL, outperforming other substrate-derived extracts. This indicates that substrate composition significantly influences the production of bioactive compounds. Overall, this research underscores the importance of substrate selection in optimizing both the yield and medicinal value of Cordyceps militaris. The findings support the potential of C. militaris as a natural source of antimicrobial and antioxidant agents, paving the way for its application in nutraceutical and pharmaceutical industries. Further exploration of genetic mechanisms governing bioactive compound synthesis may enhance production efficiency and therapeutic efficacy.