Functional characterization of transcription factors Gl Zn 2 Cys 6 ₈2 regulating ganoderic acids in Ganoderma lucidum

Ganoderic acids (GAs), the major lanostane-type triterpenoids in Ganoderma lucidum (G. lucidum), possess diverse pharmacological activities but are produced at low levels, limiting their applications. Transcriptional regulation of GA biosynthesis remains poorly understood. In this study, we functionally characterized the GlZn2Cys6-type transcription factor GlZn2Cys6₈2 and elucidated its role in GA biosynthesis. Overexpression of GlZn2Cys6₈2 significantly increased total triterpenoids during the mycelial stage and promoted the accumulation of multiple individual GAs at the primordia stage, whereas silencing resulted in markedly reduced GA levels. qRT-PCR analyses revealed that GlZn2Cys6₈2 positively regulates key biosynthetic genes, including HMGR, SQS, and LS, with SQS showing the strongest transcriptional induction. Subcellular localization assays demonstrated that GlZn2Cys6₈2 is a nuclear protein, and yeast one-hybrid analyses confirmed its direct binding to the SQS promoter, establishing SQS as a downstream target. Comparative transcriptome profiling further showed that GlZn2Cys6₈2 modulates genes involved in the mevalonate pathway and secondary metabolism. Collectively, our findings identify GlZn2Cys6₈2 as a positive regulator of GA biosynthesis and provide a basis for metabolic engineering to increase GA production.