Light-induced pH and K+ changes in the apoplast of intact leaves

The K+-sensitive fluorescent dye benzofuran isophthalate (PBFI) and the pH-sensitive fluorescein isothiocyanate dextran (FITC-Dextran) were used to investigate the influence of light/dark transitions on apoplastic pH and K+ concentration in intact leaves of Vicia faba L. with fluorescence ratio imaging microscopy. Illumination by red light led to an acidification in the leaf apoplast due to light-induced H+ extrusion. Similar apoplastic pH responses were found on adaxial and abaxial sides of leaves after light/dark transition. Stomatal opening resulted only in a slight pH decrease (0.2 units) in the leaf apoplast. Gradients of apoplastic pH exist in the leaf apoplast, being about 0.5–1.0 units lower in the center of the xylem veins as compared with surrounding cells. The apoplastic K+ concentration in intact leaves declined during the light period. A steeper light-induced decrease in apoplastic K+, possibly caused by higher apoplastic K+, was found on the abaxial side of leaves concentration. Simultaneous measurements of apoplastic pH and K+ demonstrated that a light-induced decline in apoplastic K+ concentration indicative of net K+ uptake into leaf cells occurs independent of apoplastic pH changes. It is suggested that the driving force that is generated by H+ extrusion into the leaf apoplast due to H+-ATPase activity is sufficient for passive K+ influx into the leaf cells.