Exposure of cells of several strains of Staphylococcus aureus to 50 or 100 μg of N-methyl-N′-nitro-N-nitrosoguanidine for 30 to 60 minutes induced large numbers of mutants with pigment content different from that of the parent. By determination of the amount of pigment as related to the optical density of the cells, four to seven classes of pigmentation mutants could be defined. Mutants with pigment content differing from that of the parent could readily be mutated to other pigmentation states and are thus probable point mutations. In contrast, completely white mutants could not be induced by the mutagen to any degree of pigmentation and possibly represent minor deletions or cumulative single-site mutations in the chromosome. Growth of parent strains in media containing acridine dyes occasionally produced a low frequency (0.01%) of white mutants. Mutants differing in pigment content from that of the parent were unable to produce coagulase during growth, although the parent cultures elaborated considerable coagulase under identical conditions.
