Background: The Candida glabratadoes not develop into a pathogenic hiphal form; however, it has become the second most common pathogen of fungal infections in humans, partly because of its adhesion ability and virulence. Objectives: The present study aimed to determine whether Flo8, a transcription factor that plays an important role in the virulence and drug resistance in Candida albicans, has a similar role in C. glabrata. Methods: We constructed FLO8null strains of a C. glabratastandard strain and eight clinical strains from different sources, and a FLO8complemented strain. Real-time quantitative PCR, biofilm formation assays, hydrophobicity tests, adhesion tests, Caenorhabditis eleganssurvival assay, and drug-susceptibility were then performed. Results: Compared with the wild-type strains, the biofilm formation, hydrophobicity, adhesion, and virulence of the FLO8-deficient strains decreased, accompanied by decreased expression of EPA1, EPA6, and EPA7. On the other hand, it showed no changes in antifungal drug resistance, although the expression levels of CDR1, CDR2, and SNQ2increased after FLO8deletion. Conclusions: These results indicated that Flo8 is involved in the adhesion and virulence of C. glabrata, with FLO8deletion leading to decreased expression of EPA1, EPA6, and EPA7and decreased biofilm formation, hydrophobicity, adhesion, and virulence.