The role of nitric oxide (NO) in photosynthesis is poorly understood as indicated by a number of studies in this field with often conflicting results. As various NO donors may be the primary source of discrepancies, the aim of this study was to apply a set of NO donors and its scavengers, and examine the effect of exogenous NO on photosynthetic electron transport in vivo as determined by chlorophyll fluorescence of pea (Pisum sativum) leaves. Sodium nitroprusside-induced changes were shown to be mediated partly by cyanide, and S-nitroso-N-acetylpenicillinamine provided low yields of NO. However, the effects of S-nitrosoglutathione are inferred exclusively by NO, which made it an ideal choice for this study. Q A 2 reoxidation kinetics show that NO slows down electron transfer between Q A and Q B , and inhibits charge recombination reactions of Q A 2 with the S 2 state of the water-oxidizing complex in photosystem II. Consistent with these results, chlorophyll fluorescence induction suggests that NO also inhibits steady-state photochemical and nonphotochemical quenching processes. NO also appears to modulate reaction-center-associated nonphotochemical quenching.Plants, as well as animals, respond to ambient levels of nitric oxide (NO), and also generate NO themselves via various enzymatic and nonenzymatic pathways (Yamasaki, 2000;Neill et al., 2003;Río et al., 2004). Indeed, in the past years, a growing amount of research has provided evidence for the multiple physiological roles of this gaseous free radical in plants (for review, see Wendehenne et al., 2004;Delledonne, 2005). The turnover of NO depends on its concentration, the ambient redox status, and the concentration of target molecules. In biological systems, NO is capable of targeting thiol-and metal-containing proteins (Lamattina et al., 2003). Photosynthetic and mitochondrial electron transport chains are abundant in transition metalcontaining complexes, and NO and its derivative peroxynitrite are known to inhibit the mitochondrial electron transport chain (Millar and Day, 1996;Yamasaki et al., 2001). Yet, the effect of exogenous NO on photosynthetic activity in intact leaves has so far been poorly addressed, with often conflicting results.Previous research suggests that NO gas decreases net photosynthesis rates in oat (Avena sativa) and alfalfa (Medicago sativa) leaves (Hill and Bennett, 1970). Lum et al. (2005) have identified a number of intracellular targets of NO signalization including mitochondria, peroxisomes, and chloroplasts. They found that the NO donor sodium nitroprusside (SNP) decreases the amount of Rubisco activase and the b-subunit of the Rubisco subunit-binding protein in mung bean (Phaseolus aureus).NO is also able to influence the photosynthetic electron transport chain directly. An important action site of NO is PSII. Electron paramagnetic resonance and chlorophyll fluorescence measurements using NO gas treatment of isolated thylakoid membrane complexes have clearly demonstrated that NO can reversibly bind to several sites in PSII and...

We have studied the pH effect on the S(0) and S(2) multiline electron paramagnetic resonance (EPR) signals from the water-oxidizing complex of photosystem II. Around pH 6, the maximum signal intensities were detected. On both the acidic and alkaline sides of pH 6, the intensities of the EPR signals decreased. Two pKs were determined for the S(0) multiline signal; pK(1) = 4.2 +/- 0.2 and pK(2) = 8.0 +/- 0.1, and for the S(2) multiline signal the pKs were pK(1) = 4.5 +/- 0.1 and pK(2) = 7.6 +/- 0.1. The intensity of the S(0)-state EPR signal was partly restored when the pH was changed from acidic or alkaline pH back to pH approximately 6. In the S(2) state we observed partial recovery of the multiline signal when going from alkaline pH back to pH approximately 6, whereas no significant recovery of the S(2) multiline signal was observed when the pH was changed from acidic pH back to pH approximately 6. Several possible explanations for the intensity changes as a function of pH are discussed. Some are ruled out, such as disintegration of the Mn cluster or decay of the S states and formal Cl(-) and Ca(2+) depletion. The altered EPR signal intensities probably reflect the protonation/deprotonation of ligands to the Mn cluster or the oxo bridges between the Mn ions. Also, the possibility of decreased multiline signal intensities at alkaline pH as an effect of changed redox potential of Y(Z) is put forward.