Candida albicans has become one of the main pathogens of fungal infection, which causes great threaten to human health. Hence, it is urgent to develop new anti- C. albicans drugs. A compound named as trichoderma acid was isolated from Trichoderma spirale , which showed stronger inhibitory effect towards C. albicans even than positive control nystatin. The transcriptome sequencing and iTRAQ-based sequencing of trichoderma acid-treated C. albicans were performed to investigate the molecular mechanism of anti- C. albicans. And the most significant differential genes were verified by qRT-PCR and Western blot analysis. The results indicated that the disruption of mitochondrial membrane potential, cell membrane, endoplasmic reticulum and the ribosomal in mitochondrial and cell wall were detected in trichoderma acidtreated C. albicans, resulting in the accumulation of ROS. The expression level of supereoxidase SOD was down-regulated, thus enhancing the concentration of ROS. High concentration of ROS led to the DNA damage and destruction of cell skeleton. And the expression levels of Rho-related GTP-binding protein RND3, the asparagine synthetase ASNS, peroxidases, glutathione S-transferase and heat shock protein HSP70 were significantly up-regulated in response to the apoptotic process and toxin stimulus, which was demonstrated by Western blot analysis. The transciptomic, proteomic and cellular analysis would provide anti- C. albicans mechanism of trichoderma acid and the defensive response mechanism of C. albicans.