Mutants of Rhodospirillum rubrum Obtained After Long-Term Anaerobic, Dark Growth

Rhodospirillum rubrum S(1) cells were grown for more than 100 generations under strict anaerobic, dark conditions in liquid medium with sodium pyruvate. During this time, growth became nonpigmented. When cells were streaked onto the surface of solid growth medium in anaerobic bottles and placed in the dark, a few light-red colonies developed, but the majority was nonpigmented. Mutants were obtained from colonies selected on the basis of pigmentation and bacteriochlorophyll a content. The growth, ultrastructure, and light reactivity of two mutants were examined. Mutant C synthesized bacteriochlorophyll a (7.2 mumoles per mg of protein), altered membrane structures, and chromatophores during dark growth. Examination of light-induced changes of the absorption spectrum of this mutant suggested that only an electron transport pathway, which included the low potential cytochrome-like pigment C428, could be detected. Mutant C grew anaerobically in the light, whereas mutant G1 was light sensitive and produced only trace amounts of bacteriochlorophyll a (0.6 mumole per ml of protein). Poorly pigmented G1 cells contained unusual membrane structures. When dark-grown G1 colonies were placed in the light, deep-red colored papillae developed in the nonpigmented colonies. During anaerobic, dark growth with sodium pyruvate, both C and G1 synthesized poly-beta-hydroxybutyrate and produced acetate, carbon dioxide, and hydrogen gas.