The paramount influence of nitrate in increasing apoplastic pH of young sunflower leaves to induce Fe deficiency chlorosis, and the re-greening effect brought about by acidic foliar sprays

Abstract

The main objective of the present work was to clarify the causal relationship between leaf apoplastic pH increase and Fe chlorosis under alkaline growth conditions. It has been shown that nitrate supply in contrast to ammonium supply induced a pH increase in the apoplast of young green leaves of Helianthus annuus which was followed within 12 hours by leaf yellowing. Hence nitrate nutrition is the primary cause of a high leaf apoplastic pH which induces Fe deficiency chlorosis and not the impaired provision of ATP for plasmalemma H + pumps in yellow leaves. Supply of bicarbonate in physiological concentrations had virtually no influence on leaf apoplastic pH. Spraying leaves with diluted acids (citric acid, sulphuric acid) resulted in a decrease of apoplastic pH followed by leaf re-greening. Interestingly, the Fe concentrations remained the same in the yellow control leaves and in the sprayed green leaves. From this it follows that Fe efficiency in leaves is mainly related to the Fe distribution between apoplast and symplast. It was demonstrated that Fe chlorosis induced by nitrate nutrition begins from the base of the youngest leaves, presumably from growing interveinal microsites showing high nitrate uptake rates. Leaf yellowing spread gradually from the leaf base to the tip and after seven days of nitrate supply the leaf was almost completely yellow (98%). Leaf yellowing was measured by means of a video imaging technique. Leaf apoplastic pH recordings were conducted after loading the fluorescent dye FITC-Dextran (4000 D) into the leaf apoplast of intact plants thus simulating in vivo conditions. It was also shown using the new loading technique that the fluorescent dye did not penetrate the leaf symplast.