Early changes of the pH of the apoplast are different in leaves, stem and roots of Vicia faba L. under declining water availability.

Changes in pH of the apoplast have recently been discussed as an important factor in adjusting transpiration and water relations under conditions of drought via modulatory effect on abscisic acid (ABA) concentration. Using Vicia faba L., we investigated whether changes in the root, shoot and leaf apoplastic pH correlated with (1) a drought-induced reduction in transpiration and with (2) changes in ABA concentration. Transpiration, leaf water potential and ABA in leaves were measured and correlated with root and shoot xylem pH, determined by a pH microelectrode, and pH of leaf apoplast quantified by microscopy-based in vivo ratiometric analysis. Results revealed that a reduction in transpiration rate in the early phase of soil drying could not be linked with changes in the apoplastic pH via effects on the stomata-regulating hormone ABA. Moreover, drought-induced increase in pH of xylem or leaf apoplast was not the remote effect of an acropetal transport of alkaline sap from root, because root xylem acidified during progressive soil drying, whereas the shoot apoplast alkalized. We reason that other, yet unknown signalling mechanism was responsible for reduction of transpiration rate in the early phase of soil drying.

High apoplastic solute concentrations in leaves alter water relations of the halophytic shrub, Sarcobatus vermiculatus.

Predawn plant water potential (Psi(w)) is used to estimate soil moisture available to plants because plants are expected to equilibrate with the root-zone Psi(w). Although this equilibrium assumption provides the basis for interpreting many physiological and ecological parameters, much work suggests predawn plant Psi(w) is often more negative than root-zone soil Psi(w). For many halophytes even when soils are well-watered and night-time shoot and root water loss eliminated, predawn disequilibrium (PDD) between leaf and soil Psi(w) can exceed 0.5 MPa. A model halophyte, Sarcobatus vermiculatus, was used to test the predictions that low predawn solute potential (Psi(s)) in the leaf apoplast is a major mechanism driving PDD and that low Psi(s) is due to high Na+ and K+ concentrations in the leaf apoplast. Measurements of leaf cell turgor (Psi(p)) and solute potential (Psi(s)) of plants grown under a range of soil salinities demonstrated that predawn symplast Psi(w) was 1.7 to 2.1 MPa more negative than predawn xylem Psi(w), indicating a significant negative apoplastic Psi(s). Measurements on isolated apoplastic fluid indicated that Na+ concentrations in the leaf apoplast ranged from 80 to 230 mM, depending on salinity, while apoplastic K+ remained around 50 mM. The water relations measurements suggest that without a low apoplastic Psi(s), predawn Psi(p) may reach pressures that could cause cell damage. It is proposed that low predawn apoplastic Psi(s) may be an efficient way to regulate Psi(p) in plants that accumulate high concentrations of osmotica or when plants are subject to fluctuating patterns of soil water availability.

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.