Evidence on the Formation of Singlet Oxygen in the Donor Side Photoinhibition of Photosystem II: EPR Spin-Trapping Study

When photosystem II (PSII) is exposed to excess light, singlet oxygen (¹O₂) formed by the interaction of molecular oxygen with triplet chlorophyll. Triplet chlorophyll is formed by the charge recombination of triplet radical pair ³P680^•+Pheo^•−] in the acceptor-side photoinhibition of PSII. Here, we provide evidence on the formation of ¹O₂ in the donor side photoinhibition of PSII. Light-induced ¹O₂ production in Tris-treated PSII membranes was studied by electron paramagnetic resonance (EPR) spin-trapping spectroscopy, as monitored by TEMPONE EPR signal. Light-induced formation of carbon-centered radicals (R^•) was observed by POBN-R adduct EPR signal. Increased oxidation of organic molecules at high pH enhanced the formation of TEMPONE and POBN-R adduct EPR signals in Tris-treated PSII membranes. Interestingly, the scavenging of R^• by propyl gallate significantly suppressed ¹O₂. Based on our results, it is concluded that ¹O₂ formation correlates with R^• formation on the donor side of PSII due to oxidation of organic molecules (lipids and proteins) by long-lived P680^•+/TyrZ^•. It is proposed here that the Russell mechanism for the recombination of two peroxyl radicals formed by the interaction of R^• with molecular oxygen is a plausible mechanism for ¹O₂ formation in the donor side photoinhibition of PSII.