A comparison of the effects of ionic stress and an uncoupler on long-term fluorescence transients (the 'Kautsky effect') in the green alga Dunaliella tertiolecta indicated that the large quenching induced by ionic stress was caused by a pH gradient across the thylakoid membrane. This possiblity was given support by the increase in the slow phase of 3-(3',4'-dichlorophenyl)-1,1-dimethylurea-induced fluorescence relaxation in algae subjected to ionic stress. Low-temperature fluorescence emission spectra indicated that salt stress enhanced photosystem-I emission in the dark, and a comparison of simultaneous emissions at 695 and 720 nm at room temperature indicated a further increase in photosystem-I emission during the fluorescence transients. Taken together with the decrease in the fast phase of 3-(3',4'-dichlorophenyl)-1,1-dimethylurea-induced fluorescence relaxation in stressed algae, our results indicate that ionic stress stimulates cyclic electron flow, and that non-cyclic flow is inhibited. The effect of sucrose-induced osmotic stress was similar to, but less marked than, the effects of NaCl and KCl; the effect of decreasing the external salinity was small.
Guard-cell protoplasts were isolated by enzymic digestion of the epidermis peeled from the abaxial surface of leaves from Commelina communis L. The protoplasts were separated from mesophyll-cell protoplasts and other contaminants by density-gradient centrifugation, and the purity of the preparations carefully and quantitatively assessed by light microscopy. The preparations of guard-cell protoplasts were then compared with mesophyll-cell protoplasts in terms of the activity of photosystem II as assessed by a) the light-induced evolution of oxygen under both steady-state and flashing light and b) the characteristics of photosystem-II chlorophyll fluorescence. In all experiments, clear photosystem-II activity was found in guard-cell protoplasts, although some subtle distinctions between guard-cell and mesophyll-cell protoplasts were found. The contribution of any contaimination by mesophyll-cell chlorophyll to guard-cell-protoplast signals was estimated to be less than 3% in all cases. The results indicate that photosystem II is present and active in guard cells of Commelina.
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