Mycelial physiological maturation of Lentinula edodes associated with a potential interaction network of reactive oxygen species, amino acids and carbon flux

• Potential physiological maturation mechanism of L. edodes mycelium is firstly outlined. • A method for determining the optimal time of mycelial maturity is established. • Some amino acid metabolism is sensitive to physiological state of mycelium. • Glutamate and pyruvate metabolism may be two core pathways for mycelial maturity. • Carbon flux from cellulose degradation is crucial for mycelial maturity. Mycelial physiological maturation during the browning stage is poorly understood process and is crucial for vegetative and reproductive transitions in Lentinula edodes cultivation. To elucidate the underlying molecular events, dynamic changes in browning and fruiting characteristics, along with comprehensive transcriptomic and metabolomic profiles, were investigated across eight culturing times of KS11. D95, characterized by a 100 % fruiting rate, high yield, and well-formed fruiting bodies, was identified as the optimal time for KS11 mycelial maturation. Multi-omics analyses revealed temporally coordinated dynamics among reactive oxygen species (ROS) metabolism, amino acid homeostasis, and carbon flux. Based on these correlations, we propose an integrative working model wherein ROS dynamics may act as a signaling hub, amino acid metabolism could serve as a signal-metabolic interface, and carbon supply is likely channeled into both energy production and dominant storage forms like mannitol. The model provides a testable framework for understanding the transition from mycelial growth to fruiting body development. Furthermore, it implies that targeting key nodes within this network, such as modulating ROS signaling through oxygen supply or influencing carbon/nitrogen flux via substrate formulation, could provide practical levers to optimize maturation timing and overall production efficiency.