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Improved method for preparing anaerobic bacteroid suspensions of Rhizobium leguminosarum for the acetylene reduction assay

A method using ethylenediaminetetraacetate (EDTA) and toluene-treated pea bacteroid suspensions for the acetylene reduction assay is described. The high level of acetylene reduction by these bacteroids is comparable to that of intact plants. Reproducibility of the EDTA-toluene treatment is, on the average, within 5%. Preliminary experiments with soybeans indicate that the EDTA-toluene method might be applicable to other legumes as well.