Nitrogen and phosphorus availability limit N2 fixation in bean

Availability of nitrogen (N) and phosphorus (P) might significantly affect N2 fixation in legumes. The interaction of N and P was studied in common bean (Phaseolus vulgaris), considering their effects on nodulation and N2 fixation, nitrate reductase activity, and the composition of N compounds in xylem sap. The effect of N on the uptake of P by plants was estimated by analysing rhizospheric pH and P concentration in xylem sap and in plant shoots. Inoculated bean plants were grown in pots containing perlite/vermiculite in two experiments with different amounts of P and N. In a third experiment, bean plants were grown on two soil types or on river sand supplied with different concentrations of N. At harvest, shoot growth, number of nodules and mass, and nitrogenase activity were determined. Xylem sap was collected for the determination of ureides, amino acids, nitrate and phosphate concentration. At low nitrate concentration (1 mM), increasing amounts of P promoted both nodule formation and N2 fixation, measured as ureide content in the xylem sap. However, at high nitrate concentration (10 mM), nodulation and N2 fixation did not improve with increased P supply. Glutamine and aspartate were the main organic N compounds transported in the xylem sap of plants grown in low nitrate, whereas asparagine was the dominant N compound in xylem sap from plants grown in high nitrate. Nitrate reductase activity in roots was higher than in shoots of plants grown with low P and high N. In both soils and in the sand experiment, increased application of N decreased nodule mass and number, nitrogenase activity and xylem ureides but increased the concentration of asparagine in xylem sap. Increasing P nutrition improved symbiotic N2 fixation in bean only at low N concentrations. It did not alleviate the inhibitory effect of high nitrate concentration on N2 fixation. A decrease in plant P uptake was observed, as indicated by a lower concentration of P in the xylem sap and shoots, correlating with the amount of N supplied. Simultaneously with the specific inhibition of N2 fixation, high nitrate concentrations might decrease P availability, thus inhibiting even further the symbiotic association because of the high P requirement for nodulation and N2 fixation.