Ineffectiveness of viomycin-resistant mutants of Rhizobium meliloti

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Abstract
Under clearly defined conditions one-step acquisition of viomycin resistance by a normally effective strain of Rhizobium meliloti resulted in one-step acquisition of ineffectiveness in nitrogen fixation, which probably occurred with a one-gene change in the R. meliloti genome. Two-step mutants retained their ability to produce root nodules but such nodules also were ineffective. Increased sensitivity of the viomycin-resistant mutants to glycine and D-alanine was not noted. Bacteroids were not seen in nodules formed by the viomycin-resistant mutants on their homologous host plant. Nitrogenase activity was not detected, by acetylene reduction, in detached ineffective nodules, whereas effective nodules formed 10.6 μmoles of ethylene per hour per gram of nodules. Growth of the effective parent strain in a low concentration of viomycin resulted in elongation and swelling of the cells so that they appeared as artificially produced bacteroids. Viomycin-resistant mutants did not undergo this transformation. Antigens could be readily extracted by hot- and cold-saline extraction of wet packed cells of both resistant and sensitive cultures but antigenic differences, which may have indicated cell wall differences, were not noted.

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