Abstract
This study aimed to develop and evaluate new hybrid strains of Pleurotus cystidiosus with enhanced functional and physiological characteristics. Hybridization between the parental strains KMCC01257 and KMCC05164 yielded four promising hybrid lines (PA-054, PA-104, PA-122, and PA-132), which were selected based on superior mycelial growth and yield performance. Morphological traits and productivity were evaluated across three developmental stages: primordium formation (C1), fruiting body development (C2), and maturation (C3). As cultivation progressed, the number of fruiting bodies decreased, whereas total yield per cultivation bag increased, indicating that late-stage management plays a critical role in maximizing productivity. Chemical analyses revealed that water extracts contained higher levels of polysaccharides, ergothioneine, and total phenolics than EtOAc extracts. Among the tested strains, PA-132 exhibited the highest phenolic content and strongest antioxidant activity, while PA-104 showed greater polysaccharide and ergothioneine accumulation than the parental strains. Antioxidant activity increased over developmental stages and was consistently higher in water extracts, whereas α-glucosidase inhibitory activity was detected primarily in EtOAc extracts with minimal variation among strains or stages. Overall, the results demonstrate that hybridization effectively enhanced the biosynthesis of bioactive metabolites and the functional properties of P. cystidiosus. The selected hybrid strains, particularly PA-132 and PA-054, represent promising candidates for the development of high-value functional mushroom cultivars and nutraceutical applications.