Measurement of photochemical quenching of absorbed quanta in photosystem I of intact leaves using simultaneous measurements of absorbance changes at 830 nm and thermal dissipation.
The relationship between the redox state of the photosystem (PS) I primary donor, P700, and thermal energy dissipation in PSI were examined in intact leaves using simultaneous measurements of absorbance changes at 830 nm and variations of thermal emission monitored by photoacoustic (PA) spectroscopy, respectively. A strict proportionality (close to a 1:1 ratio) was found between the magnitudes of P700 oxidation and a positive variable PA signal induced by far-red light of various irradiances under conditions favoring effective electron donation from PSII to PSI. The proportionality was observed also between the ratio of reduced P700 to the total P700 content and the ratio of the variable component to the total PA signal measured with modulated light of 695 nm. Those findings clearly revealed that in intact leaves, variable thermal dissipation in PSI is determined by the fraction of P700 in the reduced state. Diuron-treated leaves exposed to 45 degrees C in which PSI received electrons not from PSII, but from soluble reductants localized in the chloroplast stroma were also used. In such leaves, the linear relationship between the ratio of reduced P700 to the total P700 content and the ratio of the variable component to the total PA signal measured with modulated light of 700 nm has been found as well, but its slope was twice smaller than in untreated leaves. This is probably related to an increased contribution of thermal emission from inactive PSII to the steady-state level of the PA signal in diuron-treated leaves exposed to high temperatures. The results demonstrated that the yield of variable thermal dissipation is strictly dependent on the redox pressure applied to the photosystem. The above illustrates the strong photochemical energy quenching occurring when the reaction centers are in open state (reduced P700).