Effect of Oxygen and Malate on NO3− Inhibition of Nitrogenase in Soybean Nodules

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Abstract
Soybean (Glycine max cv Hodgson) nitrogenase activity (C2H2 reduction) in the presence or absence of nitrate was studied at various external O2 tensions. Nitrogenase activity increased with oxygen partial pressure up to 30 kilopascals, which appeared to be the optimum. A parallel increase in ATP/ADP ratios indicated a limitation of respiration rate by low O2 tensions in the nodule, and the nitrogenase activity was limited by the rate of ATP regeneration. In the presence of nitrate, the nitrogenase activity was low and less stimulated by increased pO2, although the nitrite content per gram of nodules decreased from 0.05 to 0.02 micromole when pO2 increased from 10 to 30 kilopascals. Therefore, the accumulation of nitrite inside the nodule was probably not the major cause of the inhibition. Instead, inhibition by nitrate could be due to competition for reducing power between nitrate reduction and bacteroid or mitochondrial respiration inside the nodule. This is supported by the observation of decrease in ATP/ADP ratios from 1.65, in absence of nitrate, to 0.93 in the presence of this anion at 30 kilopascals O2. Furthermore, the inhibition was suppressed by the addition, the plant nutrient solution, of 15 millimolar L-malate, a carbon substrate that is considered to be the major source of reductant for the bacteroids in the symbiosis.