Carotenoid pigments were extracted from Corynebacteriuma poinsettiae (wild type) ATCC 9682, and from 108 mutants obtained by exposure of a streptomycin resistant strain of C. poinsettiae to ultra-violet light irradiation and N-methyl- N'-nitro-N-nitrosoguanidine. The pigments were characterized by their absorption maxima, Rf-values, and partition ratios in petroleum ether and methanol. Thin layer chromatography was used to compare pigments of the wild type with those of the mutants. Possible biosynthetic pathways in carotenoid synthesis of the wild type were postulated on the basis of the observed genetic blocks. Mutants were found which suggested the existence of a linear pathway in carotenoid synthesis from the aliphatic C4 0 molecule to the bi-cyclic C50-diol. Other mutants suggested possible alternative pathways in the biosynthesis of these pigments or the presence of intermediates not detectable by thin layer chromatography.