A rapid screening method for Rhizobium meliloti symbiotic nitrogenase activity

A method for screening Rhizobium meliloti isolates for their symbiotic nitrogenase activity with alfalfa (Medicago sativa L.) cv. Apollo is described. The nitrogenase activity of each isolate is assessed by measuring the reduction of acetylene (C2H2) to ethylene (C2H4) by 50 intact plants grown in 10 plastic pouches for 2 weeks. The method is rapid, sensitive, reproducible, and accurate enough to differentiate 29 Rhizobium isolates and 5 authentic strains into 13 subsets. Under the experimental conditions used, nodulation occurred within 5 days of inoculation and there was a significant positive relationship between the nitrogenase activity of those isolates which reduced more than 60 nmol C2H2∙plant−1∙h−1 and the dry weight of the shoots of the nodulated plants in 2 weeks of growth.