Multiomics Analysis across the Life Cycle Identifies Zn ₂ Cys ₆ _61 as a Target for Enhancing Triterpenoid Production in <i>Ganoderma lucidum</i>

Ganoderic acids (GAs) are high-value lanostane-type triterpenoids derived from <i>Ganoderma lucidum</i> (<i>G. lucidum</i>) with broad applications in functional foods and nutraceuticals, yet their low natural abundance limits industrial production. In this study, an integrated life-cycle multiomics analysis combining metabolomics, transcriptomics, and proteomics was conducted across six developmental stages in four <i>G. lucidum</i> strains to elucidate regulatory mechanisms governing GA biosynthesis. Weighted gene coexpression network analysis identified candidate cytochrome P450 enzymes and transcription factors associated with GA accumulation. A Zn₂Cys₆-type transcription factor, Zn₂Cys₆_61, was identified as a central regulator and functionally validated through overexpression and RNA interference. Genetic manipulation of Zn₂Cys₆_61 expression significantly altered GA levels, with overexpression markedly enhancing GA accumulation. Further analysis demonstrated that Zn₂Cys₆_61 directly binds to and activates the promoter of squalene synthase, a key enzyme in triterpenoid backbone biosynthesis. Together, these findings identify Zn₂Cys₆_61 as an effective engineering target and provide a transcription factor-based strategy for improving GA production in medicinal mushrooms.