Determination of oxygen emission and uptake in leaves by pulsed, time resolved photoacoustics

Pulsed, time resolved photoacoustics has sufficient sensitivity to determine oxygen emission and uptake by single turnover flashes to leaves. The advantage over previous methodologies is that when combined with single turnover flashes the kinetics of the thermal and the gas signals can be resolved to 0.1 millisecond and separated. The S-state oscillations of oxygen formation are readily observed. The gas signal from common spongy leaves such as spinach (Spinacia sp.), Japanese andromeda (Pieris japonica), mock orange (Philadelphus coronarius) and viburnum (Viburnum tomentosum), after correction for instrumental rise time, show a lag of only 1 millisecond and a rise time of 5 milliseconds in the formation of oxygen. Thus a recent proposal that the formation of oxygen requires over 100 milliseconds cannot be true for choroplasts in vivo. The rapid emission is correlated with structure of the leaf. At low light flash energies a rapid gas uptake is observed. The uptake has slightly slower kinetics than oxygen evolution, and its magnitude increases with damage to the leaf. The pulse methodology shows that the uptake begins with the very first flash after dark adaption, and allows the detection of a positive signal (oxygen) on the third flash. These observations, the long wavelength of excitation (695 nanometers) and the magnitude of the signal support the contention that the gas uptake is oxygen reduction by electrons from photosystem I. These results show that important physiological aspects of a leaf can be studied by pulsed, time resolved photoacoustics.