A P450-mediated trade-off: Terpenoid metabolism comes at the cost of pest resistance in Pleurotus ostreatus.

Abstract

Cytochrome P450 monooxygenases (P450s) core catalytic regulators of secondary metabolism and biotic stress defense across plants and fungi. The fungivore mite Tyrophagus putrescentiae is a major pest of cultivated mushrooms, but the molecular basis of fungal defense against this fungivore remains poorly understood. Here, we functionally identified and systematically characterized PoCYP5152C8, a CYP504 subfamily P450 gene from Pleurotus ostreatus that is transcriptionally responsive to mite infestation stress. Overexpression of PoCYP5152C8 in P. ostreatus increased mite survival, egg hatching, and population growth, while RNAi-mediated silencing enhanced fungal resistance. Integrated transcriptomic and metabolomic profiling revealed that PoCYP5152C8 specifically modulates the ubiquinone and other terpenoid-quinone biosynthesis pathway. In vitro enzymatic assays demonstrate that PoCYP5152C8 requires the reductase PoCPA15 to convert terpene substrates (FPP, GPP, and GGPP) into a suite of volatile terpenoids, including β-caryophyllene oxide, α-terpineol, and Z-citral. Behavioral assays identified several of these compounds as potent attractants for T. putrescentiae, with α-terpineol showing strong repellent activity at high concentrations. Our findings suggest that PoCYP5152C8 as a key metabolic node that redirects terpenoid flux toward attractant compounds, thereby reducing fungal defense. This study uncovers the enzymatic and molecular mechanism of a fungal neofunctionalized P450-mediated fungus-fungivore interactions, providing novel genetic targets for sustainable pest biocontrol in edible mushroom cultivation.