Synthesis and Accumulation of Nitrite in Soybean Nodules Supplied with Nitrate

Nodulated soybean plants (Glycine max [L.] Merr) were grown in sand culture without combined N or with a continuous supply of nitrate in nutrient solution. Moderate nitrate concentration (30 milligrams N per liter) had little effect on nodule weight/plant while high nitrate concentration (100 milligrams N per liter) depressed nodule weight/plant by 70 to 80% with harvests 30 to 60 days after planting and initiation of nitrate treatments.The effect of nitrate supply on ammonium, amino, and ureide nitrogen concentrations in nodules was small and inconsistent. In contrast, nitrate and nitrite concentrations in nodules were directly proportional to nitrate supply and inversely proportional to nodule weight/plant. Correlations between nitrate or nitrite concentration in nodules and nodule weight/plant were highly significant.Cytosol from soybean nodules was found to contain NADH-dependent nitrate reductase activity (typical activity was 0.1 micromole per milligram protein x hour). A Rhizobium japonicum mutant (derived from strain 61A76) lacking nitrate reductase was employed to show that the cytosol enzyme activity is of host origin. Growth of nodules formed by the mutant lacking nitrate reductase was inhibited by nitrate. These nodules did contain nitrite although concentrations of nitrite (about 0.3 microgram N per gram fresh weight) were low relative to nitrite concentrations (about 1.5 microgram N per gram fresh weight) in nodules formed by R. japonicum strain 61A76. The overall results support the idea that the depression of legume nodule growth by nitrate is directly related to the metabolism of nitrate in nodules.