Phospholipase D1 (PLD1) mutants of Candida albicans are defective in important in vivo and in vitro virulence factors. PLD1 mutants colonize the murine alimentary tract as well as PLD1 sufficient strains. In comparison to PLD1 sufficient strains, the PLD1 mutants: (i) are unable to survive in internal organs after intravenous challenge; (ii) do not decrease the body weights of mice after oral challenge; and (iii) are not lethal for immunodeficient mice after oral challenge. In vitro, the PLD1 mutants show a drastically reduced capacity to penetrate epithelial monolayers and they fail to develop hyphae when grown on solid Spider medium. The morphogenic switch from yeast to hyphae is controlled by multiple parallel signaling pathways that couple specific stimuli to the regulation of several transcription factors. Our data suggest that PLD1 functions in at least one of these pathways regulating morphogenesis in vitro and that while the mutants are able to form hyphae in vivo, the hyphae are defective in their ability to cause oroesophageal and gastric candidiasis and to kill the C. albicans-colonized mice.