The effect of nitrate and nitrite on oxygen evolution and carbon-dioxide assimilation and the reduction of nitrate and nitrite by intact chloroplasts

Intact chloroplasts isolated from spinach reduced NO3 (-) and NO2 (-) in the light without the addition of either co-factors or added enzymes. The maximum rate observed, however, for the reduction of NO3 (-) was approximately 3μMoles hr(-1) mg(-1) (chlorophyll) and for NO2 (-) 6 μMoles hr(-1) mg(-1) (chlorophyll). These rates were consistent with the enzyme content of whole chloroplasts, but much lower than those found in whole leaf extracts.The addition of both NO3 (-) and NO2 (-) in low concentrations resulted in transient increases in both O2 evolution and CO2 fixation. The increases in oxygen evolution were not consistent in amount and bore no relation to the amount of substrate reduced. Similar transients were observed in a number of experiments when NaCl or NH4Cl were added.The addition of NO2 (-) at concentrations of 10(-4) M and above resulted in marked inhibition of both O2 evolution and CO2 fixation. NO2 (-) appears to inhibit by blocking the reduction of NADP. NO3 (-) at similar concentrations had no such effect.An increase in the soluble amino nitrogen content of the chloroplasts was observed when NO3 (-) or NO2 (-) was reduced. There was, however, no increase in the incorporation of (14)C from (14)CO2 into amino acids under these conditions. Even with the addition of ammonia the amount of (14)C incorporated into the amino acids was not changed from less than 5% of the total (14)C fixed. We conclude that while intact chloroplasts do have the ability to reduce both NO3 (-) and NO2 (-) at low rates, they do not synthesize appreciable amounts of amino acid directly, and this fact must be considered when formulating any pathways for nitrogen metabolism during photosynthesis.