[Nitrate-dependent noncyclic photophosphorylation in Ankistrodesmus braunii in the absence of CO2 and O 2]

Abstract

Manometric measurements show that oxygen evolution proceeds in synchronised cells of Ankistrodesmus braunii even in an atmosphere of pure nitrogen. In this case the slow oxygen evolution is dependent on the presence of nitrate (Table 1). Light saturation is found at a low light intensity at pH 5.6, at a higher light intensity at pH 8.0 (Fig. 1). The light saturation curves are in good agreement with those of (32)P-labelling in Ankistrodesmus under the same conditions (Fig. 2).DCMU inhibition in N2 of both O2-evolution and (32)P-labelling begins only at a DCMU concentration of 5×10(-7)M or more. Complete inhibition of O2-evolution is reached only at 10(-5)M (Fig.3). In (32)P-labelling a variable percentage is still left uninhibited at 10(-5) M DCMU (Fig. 4, Table 2), which is at least partly due to cyclic photophsphorylation. Nitrate starvation for several hours causes a considerable decrease in O2-evolution and also in the sensitivity to those high concentrations of DCMU (Fig. 5), but it leads to a sensitivity to antimycin A not observed under normal conditions (Table 3). The effects of nitrate starvation thus become comparable to those of far-red light, under which noncyclic electron transport is slow or completely prevented.The inhibition by DCMU of electron transport in photosystem II is also estimated by measuring the increase in fluorescence at 684 nm in air containing additional CO2. This fluorescence is saturated only at 10(-5)M DCMU and shows that a certain percentage of photosystem II remains uninhibited at 5×10(-7)M (Fig. 6), a concentration found to be almost ineffective in inhibiting O2-evolution and (32)P-labelling in an N2-atmosphere.The results indicate that in synchronised cells of Ankistrodesmus noncyclic electron flow and noncyclic photophosphorylation can proceed in an atmosphere of pure nitrogen if nitrate is available as the electron acceptor. In this case noncyclic photophosphorylation, inspite of its low rates, still dominates over cyclic photphosphorylation. At low pH, when nitrate reduction is slow, cyclic photophosphorylation accounts for a greater part of the total phosphorylation than at high pH. Thus in the absence of CO2 and O2 cyclic photophosphorylation can be regarded as the main process of ATP formation only after nitrate starvation, in far-red light or in the presence of high concentrations of DCMU.Inhibition by DCMU, though very efficient under conditions of high photosynthetic activity, becomes rate-limiting only if the electron transport is so far reduced by DCMU that the remaining rate is of the same order as the low rate of the control or less. Therefore high concentrations of DCMU are required for the inhibition of low rates of noncyclic photophosphorylation.