Role of apoplast acidification by the H(+) pump : Effect on the sensitivity to pH and CO2 of iron reduction by roots of Brassica napus L.

We have studied the mechanism of the response to iron deficiency in rape (Brassica napus L.), taking into account our previous results: net H(+) extrusion maintains a pH shift between the root apoplast and the solution, and the magnitude of the pH shift decreases as the buffering power in the solution increases. The ferric stress increased the ability of roots to reduce Fe[III]EDTA. Buffering the bulk solution (without change in pH) inhibited Fe[III]EDTA reduction. At constant bulk pH, the inhibition (ratio of the Fe[III]EDTA-reduction rates measured in the presence and in the absence of buffer) increased with the rate of H(+) extrusion (modulated by the length of a pretreatment in 0.2 mM CaSO4). These results support the hypothesis that the apoplastic pH shift caused by H(+) excretion stimulated Fe[III] reduction. The shape of the curves describing the pH-dependency of Fe[III]EDTA reduction in the presence and in the absence of a buffer fitted this hypothesis. When compared to the titration curves of Fe[III]citrate and of Fe[III]EDTA, the curves describing the dependency of the reduction rate of these chelates on pH indicated that the stimulation of Fe[III] reduction by the apoplastic pH shift due to H(+) excretion could result from changes in electrostatic interactions between the chelates and the fixed chargers of the cell wall and-or plasmalemma. Blocking H(+) excretion by vanadate resulted in complete inhibiton of Fe[III] reduction, even in an acidic medium in which there was neither a pH shift nor an inhibitory effect of a buffer. This indicates that the apoplastic pH shift resulting from H(+) pumping is not the only mechanism which is involved in the coupling of Fe[III] reduction to H(+) transport. Our results shed light on the way by which the strong buffering effect of HCO 3 (-) in some soils may be involved in iron deficiency encountered by some of the plants which grow in them.