The relationship between thylakoid stacking and salt induced chlorophyll fluorescence changes.

Salt induced chlorophyll fluorescence increase and thylakoid stacking have been measured under various conditions. 1. Aging of pea chloroplasts led to a loss of salt induced chlorophyll fluorescence increase and thylakoid stacking which is suggested to be due to a decrease in membrane fluidity as measured by 1,6-diphenylhextriene fluorescence polarization. 2. The aging treatment was accompanied by a decreased in surface charge density as indicated by chloroplast electrophoretic mobility measurements. 3. Lowering of the temperature to about 0 degrees C retarded the time courses of salt induced stacking and chlorophyll fluorescence increase. 4. Like aging, addition of linolenic acid led to an inhibition of the salt induced fluorescence and stacking phenomena but in this case there was a concomitant increase in electrophoretic mobility without any detectable change in the polarization of 1,6-diphenylhextriene fluorescence. 5. Maximum stacking occurred in both aged and fresh chloroplasts in a low salt medium at about pH 4.3 and the time course for the pH induced process was rapid and relatively temperature insensitive when compared with salt induced stacking. 6. The chlorophyll a/chlorophyll b ratio was lower for salt induced 'grana' than for pH induced 'grana'. 7. The results are discussed in terms of the hypothesis that changes in the lateral interaction of membrane pigment-protein complexes underlie the salt induced chlorophyll fluorescence increase and thylakoid stacking. It is argued that electrostatic screening by cations leads to the formation of domains of low-charge, fluorescent pigment-protein complexes, seggregated from domains of high-charge, quenching complexes, resulting in a increase in chlorophyll fluorescence yield and stacking at low-charge regions on adjacent membranes. In contrast to this, it is argued that the pH induced stacking occurs because of electrostatic neutralization, a mechanism which would not be expected to induce domain formation and associated chlorophyll fluorescence changes.