The yeast-to-hyphal morphotype transition and subsequent biofilm formation are important virulence factors of Candida albicans and are closely associated with ergosterol biosynthesis. Flo8 is an important transcription factor that determines filamentous growth and biofilm formation in C. albicans. However, the relationship between Flo8 and regulation of the ergosterol biosynthesis pathway remains elusive. Here, we analyzed the sterol composition of a -deficient C. albicans strain by gas chromatography-mass spectrometry and observed the accumulation of the sterol intermediate zymosterol, the substrate of Erg6 (C-24 sterol methyltransferase). Accordingly, the transcription level of was reduced in the -deficient strain. Yeast one-hybrid experiments revealed that Flo8 physically interacted with the promoter. Ectopic overexpression of in the -deficient strain partially restored biofilm formation and virulence in a Galleria mellonella infection model. These findings suggest that Erg6 is a downstream effector of the transcription factor Flo8 that mediates the cross talk between sterol synthesis and virulence factors in C. albicans. Biofilm formation by C. albicans hinders its eradication by immune cells and antifungal drugs. Flo8 is an important morphogenetic transcription factor that regulates the biofilm formation and virulence of C. albicans. However, little is known about how Flo8 regulates biofilm formation and fungal pathogenicity. Here, we determined that Flo8 directly binds to the promoter of to positively regulate its transcriptional expression. Consistently, loss of results in the accumulation of the substrate of Erg6. Moreover, ectopic overexpression of at least partially restores the biofilm formation and virulence of the -deficient strain both and . This work provides a new perspective on the metabolic link between transcription factors and morphotypes in C. albicans.