This study deals with effects of membrane excitation on photosynthesis and cell protection against excessive light, manifested in non-photochemical quenching (NPQ). In Chara corallina cells, NPQ and pericellular pH displayed coordinated spatial patterns along the length of the cell. The NPQ values were lower in H(+)-extruding cell regions (external pH approximately 6.5) than in high pH regions (pH approximately 9.5). Generation of an action potential by applying a pulse of electric current caused NPQ to increase within 30-60 s. This effect, manifested as a long-lived drop of maximum chlorophyll fluorescence (F(m)'), occurred at lower photosynthetic flux densities (PFD) in the alkaline as compared to acidic cell regions. The light response curve of NPQ shifted, after generation of an action potential, towards lower PFD. The release of NPQ by nigericin and the rapid reversal of action potential-triggered NPQ in darkness indicate its relation to thylakoid DeltapH. Generation of an action potential shortly after darkening converted the chloroplasts into a latent state with the F(m) identical to that of unexcited cells. This state transformed to the quenched state after turning on weak light that was insufficient for NPQ prior to membrane excitation of the cells. The ionophore, A23187, shifted NPQ plots similarly to the action potential effect, consistent with a likely role of a rise in the cytosolic Ca(2+) level in the action potential-induced quenching. The results suggest that a rapid electric signal, across the plasma membrane, might exert long-lived effects on photosynthesis and chlorophyll fluorescence through ion flux-mediated pathways.

Characean cells exposed to illumination arrange plasma-membrane H(+) fluxes and photosynthesis in coordinated spatial patterns. The limited availability of CO(2) in alkaline bands accounts for the lower effective quantum yield of photosystem II (DeltaF/F(m)') in chloroplasts of these bands compared to acidic zones. The effect of electrically triggered action potential on the spatial distribution of photosynthetic parameters (DeltaF/F(m)' and non-photochemical quenching, NPQ) and extracellular pH was studied with fluorescence imaging and pH microelectrodes. In the resting cell at a range of light intensities, the periodic profile of extracellular pH is parallel to the profile of NPQ and antiparallel to that of DeltaF/F(m)'. After triggering the action potential, the pH banding temporarily disappeared, but in contrast, the differences in effective quantum yield and NPQ patterns became more apparent. The transient changes in pH-banding, effective quantum yield and non-photochemical quenching are discussed in relation to alterations in intracellular Ca(2+) and H(+) concentrations during and after the action potential.

The influence of cell excitation and external calcium level on the dynamics of light-induced pH bands along the length of Chara corallina cells is studied in the present paper. Generation of an action potential (AP) transiently quenched these pH patterns, which was more pronounced at 0.05-0.1 mM Ca2+ than at higher concentrations of Ca2+ (0.6-2 mM) in the medium. After transient smoothing of the pH bands, some alkaline peaks reemerged at slightly shifted positions in media with low Ca2+ concentrations, while at high Ca2+ concentrations, the alkaline spots reappeared exactly at their initial positions. This Ca2+ dependency has been revealed by both digital imaging and pH microelectrodes. The stabilizing effect of external Ca2+ on the locations of recovering alkaline peaks is supposedly due to formation of a physically heterogeneous environment around the cell owing to precipitation of CaCO3 in the alkaline zones at high Ca2+ during illumination. The elevation of local pH by dissolving CaCO3 facilitates the reappearance of alkaline spots at their initial locations after temporal suppression caused by cell excitation. At low Ca2+ concentrations, when the solubility product of CaCO3 is not attained, the alkaline peaks are not stabilized by CaCO3 dissolution and may appear at random locations.

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Chlorophyll fluorescence Imaging and Microscopy PAM fluorometry were applied to study spatial dynamics of photosystem II quantum yield (ΔF/F'(m)) and non-photochemical quenching (NPQ) in resting and electrically stimulated Chara corallina cells in the absence and presence of the hydrophilic electron acceptor methyl viologen (MV) in the external medium. Electrical excitation of the plasma membrane temporarily enhanced the heterogeneity of photosynthetic patterns under physiological conditions (in the absence of MV), but irreversibly eliminated these patterns in the presence of MV. These findings suggest that the action potential (AP) of the excitable plant cell affects the spatial patterns of photosynthesis and chlorophyll fluorescence through different pathways operated in the absence and presence of MV. Based on the extent of NPQ as an indicator of MV-dependent electron flow, it is supposed that MV cannot permeate into the chloroplasts of photosynthetically active "acid cell regions" but gains an immediate access to the stroma of these chloroplasts after triggering of an AP. The AP-triggered MV-dependent non-photochemical quenching in the chloroplasts of acidic cell regions was routinely observed at 0.1 mM Ca(2+) in the medium but not at elevated (2 mM) external Ca(2+) concentration. The results are interpreted in terms of competition between two permeant divalent ion species, Ca(2+) and MV(2+), for their passage through the voltage-gated calcium channels of the plasma membrane. It is proposed that the herbicidal activity of MV in characean cells, here serving as model object, can be manipulated by triggering AP and varying Ca(2+) concentration in the environmental medium.

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