Enumeration of Free-Living Aerobic N2-Fixing H2-Oxidizing Bacteria by Using a Heterotrophic Semisolid Medium and Most-Probable-Number Technique

摘要

A heterotrophic semisolid medium was used with two sensitive assay methods, C2H2 reduction and O2-dependent tritium uptake, to determine nitrogenase and hydrogenase activities, respectively. Organisms known to be positive for both activities showed hydrogenase activity in both the presence and absence of 1% C2H2, and thus, it was possible to test a single culture for both activities. Hydrogen uptake activity was detected for the first time in N2-fixing strains of Pseudomonas stutzeri. The method was then applied to the most-probable-number method of counting N2-fixing and H2-oxidizing bacteria in some natural systems. The numbers of H2-oxidizing diazotrophs were considerably higher in soil surrounding nodules of white beans than they were in the other systems tested. This observation is consistent with reports that the rhizosphere may be an important ecological niche for H2 transformation.