Relationship between leaf apoplast pH and iron chlorosis of sunflower (Helianthus annuus L.)

A hypothesis has been presented and tested that bicarbonate (HCO₃) and nitrate (NO₃) are the most important anions inducing iron (Fe) chlorosis because these anions increase the pH of leaf apoplast which in turn depresses ferric‐iron Fe(III]) reduction, and hence, the uptake of Fe into the symplasm. Experiments with young sunflower (Helianthus annuus) plants showed that nutrition with NO₃ as the sole nitrogen (N) source induced chlorosis whereas ammonium nitrate (NH₄NO₃) did not. Monohydrogen carbonate (bicarbonate) also favoured the development of chlorosis. The degree of chlorosis was not related to the Fe concentration in the leaves. Both anion species, NO₃ and HCO₃, increased the pH of the leaf apoplast which was measured by means of the fluorescence dye 5‐carboxyfluorescein. A highly significant negative correlation between leaf apoplast pH and chlorophyll concentration in the leaves (r = ‐0.97) was found. Ferric‐Fe reduction in the apoplast—measured by means of ferrocene—provided evidence that a low leaf apoplast pH, obtained with ammonium (NH₄) supply, favoured the reduction of Fe(III) as compared with a higher leaf apoplast pH obtained with NO₃ supply. These results support the hypothesis tested.