Oxygen evolution from single- and multiple-turnover light pulses: temporal kinetics of electron transport through PSII in sunflower leaves

Oxygen evolution per single-turnover flash (STF) or multiple-turnover pulse (MTP) was measured with a zirconium O₂ analyzer from sunflower leaves at 22°C. STF were generated by Xe arc lamp, MTP by red LED light of up to 18000 μmol quanta m⁻² s⁻¹. Ambient O₂ concentration was 10–30 ppm, STF and MTP were superimposed on far-red background light in order to oxidize plastoquinone (PQ) and randomize S-states. Electron (e⁻) flow was calculated as 4 times O₂ evolution. Q _A → Q _B electron transport was investigated firing double STF with a delay of 0 to 2 ms between the two. Total O₂ evolution per two flashes equaled to that from a single flash when the delay was zero and doubled when the delay exceeded 2 ms. This trend was fitted with two exponentials with time constants of 0.25 and 0.95 ms, equal amplitudes. Illumination with MTP of increasing length resulted in increasing O₂ evolution per pulse, which was differentiated with an aim to find the time course of O₂ evolution with sub-millisecond resolution. At the highest pulse intensity of 2.9 photons ms⁻¹ per PSII, 3 e⁻ initially accumulated inside PSII and the catalytic rate of PQ reduction was determined from the throughput rate of the fourth and fifth e⁻. A light response curve for the reduction of completely oxidized PQ was a rectangular hyperbola with the initial slope of 1.2 PSII quanta per e⁻ and V _m of 0.6 e⁻ ms⁻¹ per PSII. When PQ was gradually reduced during longer MTP, V _m decreased proportionally with the fraction of oxidized PQ. It is suggested that the linear kinetics with respect to PQ are apparent, caused by strong product inhibition due to about equal binding constants of PQ and PQH₂ to the Q _B site. The strong product inhibition is an appropriate mechanism for down-regulation of PSII electron transport in accordance with rate of PQH₂ oxidation by cytochrome b₆f.