An open question in developmental biology is how groups of dividing cells can generate specific numbers of segments or organs. We describe the phenotypic effects of mutations in PERIANTHIA, a gene specifically required for floral organ patterning in Arabidopsis thaliana. Most wild-type Arabidopsis flowers have 4 sepals, 4 petals, 6 stamens, and 2 carpels. Flowers of perianthia mutant plants most commonly show a pentamerous pattern of 5 sepals, 5 petals 5 stamens, and 2 carpels. This pattern is characteristic of flowers in a number of plant families, but not in the family Brassicaceae, which includes Arabidopsis. Unlike previously described mutations affecting floral organ number, perianthia does not appear to affect apical or floral meristem sizes, nor is any other aspect of vegetative or floral development severely affected. Floral organs in perianthia arise in a regular, stereotypical pattern similar to that in distantly related species with pentamerous flowers. Genetic analysis shows that PERIANTHIA acts downstream of the floral meristem identity genes and independently of the floral meristem size and floral organ identity genes in establishing floral organ initiation patterns. Thus PERIANTHIA acts in a previously unidentified process required for organ patterning in Arabidopsis flowers.