Thermoluminescence investigations on the site of action of o-phthalaldehyde in photosynthetic electron transport

Abstract

Glow curves from spinach leaf discs infiltrated with o-phthalaldehyde (OPA) show significant similarity to those obtained by DCMU treatment which is known to block the electron flow from Q_A, the stable acceptor of Photosystem II (PS II). In both the cases, the thermoluminescence (TL) peak II (Q band) was intensified significantly, whereas peaks III and IV (B band) were suppressed. Total TL yield of the glow curve remained constant even when the leaf discs were infiltrated with high concentrations of OPA (4 mM) or with DCMU (100 μM), indicating that even at these high concentrations no significant change in the number of species undergoing charge recombination in PS II occurred. However, studies with thylakoids revealed significant differences in the action of OPA and DCMU on PS II. Although OPA, at a certain concentration and time of incubation, reduced the B band intensity by about 50–70%, and completely abolished the detectable oxygen evolution, it still retained the TL flash yield pattern, and, thus, S state turnover. OPA is known to inhibit the oxidoreductase activity of in vitro Cyt b₆/f (Bhagwat et al. (1993) Arch Biochem Biophys 304: 38–44). However, in the OPA treated thylakoids the extent of inhibition of O₂ evolution was not reduced even in the presence of oxidized tetramethyl-p-phenylenediamine which accepts electrons from plastoquinol and feeds then directly to Photosystem I. This suggests that OPA inhibition is at a site prior to plastoquinone pool in the electron transport chain, in agreement with it being between Q_A and Q_B. However, an unusual feature of OPA inhibition is that even though all oxygen evolution was completely suppressed, a significant fraction of PS II centers were functional and turned over with the same periodicity of four in the absence of any added electron donor, an observation which appears to be similar to that reported by Wydrzynski (Wydrzynski et al. (1985) Biochim Biophys Acta 809: 125–136) with lauroylcholine chloride, a lipid analogue compound. The detailed chemistry of OPA inhibition remains to be studied. Since we dedicate this paper to William A. Arnold, discoverer of delayed light and TL in photosynthesis, we have also included in the Introduction, a brief history of how TL work was initiated at BARC (Bombay, India).