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. 1985 Aug;50(2):537–539. doi: 10.1128/aem.50.2.537-539.1985

Determination of the Hydrogenase Status of Individual Legume Nodules by a Methylene Blue Reduction Assay

Grant R Lambert 1,*, F Joe Hanus 1, Sterling A Russell 1, Harold J Evans 1
PMCID: PMC238657  PMID: 16346869

Abstract

We adapted a method for the rapid screening of colonies of free-living Rhizobium japonicum for hydrogenase activity to determine the hydrogenase status of individual soybean nodules. Crude bacteroid suspensions from nodules containing strains known to be hydrogen uptake positive (Hup+) caused a localized decolorization of filter paper disks, whereas suspensions from nodules arising from inoculation with hydrogen uptake-negative (Hup) mutants or strains did not decolorize the disks. The reliability of the method was demonstrated by its successful application to 29 slow-growing rhizobia. The Hup phenotype on methylene blue filters agreed with that determined amperometrically with either methylene blue or oxygen as the electron acceptor.

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Selected References

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