Phospholipid biosynthetic pathways play crucial roles in the virulence of several pathogens; however, little is known about how phospholipid synthesis affects pathogenesis in fungi such as Candida albicans. A C. albicans phosphatidylserine (PS) synthase mutant, cho1 Delta/Delta, lacks PS, has decreased phosphatidylethanolamine (PE), and is avirulent in a mouse model of systemic candidiasis. The cho1 Delta/Delta mutant exhibits defects in cell wall integrity, mitochondrial function, filamentous growth, and is auxotrophic for ethanolamine. PS is a precursor for de novo PE biosynthesis. A psd1 Delta/Delta psd2 Delta/Delta double mutant, which lacks the PS decarboxylase enzymes that convert PS to PE in the de novo pathway, has diminished PE levels like those of the cho1 Delta/Delta mutant. The psd1 Delta/Delta psd2 Delta/Delta mutant exhibits phenotypes similar to those of the cho1 Delta/Delta mutant; however, it is slightly more virulent and has less of a cell wall defect. The virulence losses exhibited by the cho1 Delta/Delta and psd1 Delta/Delta psd2 Delta/Delta mutants appear to be related to their cell wall defects which are due to loss of de novo PE biosynthesis, but are exacerbated by loss of PS itself. Cho1p is conserved in fungi, but not mammals, so fungal PS synthase is a potential novel antifungal drug target.