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In this work, we investigated the redox transients of a number of water-soluble spin labels upon their interactions with Photosystem II (PS II) core complexes isolated from spinach leaves. We have found that the reactivity of nitroxide radicals, determined by the rate of their reduction upon illumination of PS II, depends on the chemical structure of radicals and the capability of their coming close to low-potential redox centers of photoactive PS II complexes. An enhanced capability of nitroxide radicals to accept electrons from PS II correlates with their chemical structure. Nitroxide radicals NTI (2,2,5,5-tetramethyl-4-nitromethylene-3-imidazolidine-N-oxyl) and Tacet (4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl-acetate), containing polar groups, appear to be most efficient acceptors of electrons donated by PS II compared to neutral (TEMPOL, 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl) or positively charged (Tamine, 4-amino-2,2,6,6-tetramethylpiperidine-l-oxyl) spin labels. We assume that enhanced reactivities of polar nitroxide radicals, NTI and Tacet, are determined (1) by their relatively high redox potentials, providing the possibility to accept electrons from PS II, and (2) by their affinities to the closest binding sites on the surface of PS II in the vicinity of the primary plastoquinone acceptor PQA (12–14 Å) or/and in the intraprotein cavity for the secondary plastoquinone PQB (~ 22 Å).

In this study, we have compared the photosynthetic characteristics of two contrasting species of Tradescantia plants, T. fluminensis (shade-tolerant species), and T. sillamontana (light-resistant species), grown under the low light (LL, 50–125 µmol photons m−2 s−1) or high light (HL, 875–1000 µmol photons m−2 s−1) conditions during their entire growth period. For monitoring the functional state of photosynthetic apparatus (PSA), we measured chlorophyll (Chl) a emission fluorescence spectra and kinetics of light-induced changes in the heights of fluorescence peaks at 685 and 740 nm (F 685 and F 740). We also compared the light-induced oxidation of P700 and assayed the composition of carotenoids in Tradescantia leaves grown under the LL and HL conditions. The analyses of slow induction of Chl a fluorescence (SIF) uncovered different traits in the LL- and HL-grown plants of ecologically contrasting Tradescantia species, which may have potential ecophysiological significance with respect to their tolerance to HL stress. The fluorometry and EPR studies of induction events in chloroplasts in situ demonstrated that acclimation of both Tradescantia species to HL conditions promoted faster responses of their PSA as compared to LL-grown plants. Acclimation of both species to HL also caused marked changes in the leaf anatomy and carotenoid composition (an increase in Violaxanthin + Antheraxantin + Zeaxanthin and Lutein pools), suggesting enhanced photoprotective capacity of the carotenoids in the plants grown in nature under high irradiance. Collectively, the results of the present work suggest that the mechanisms of long-term PSA photoprotection in Tradescantia are based predominantly on the light-induced remodeling of pigment-protein complexes in chloroplasts.

In this work, we have compared photosynthetic performance and expression of the PsbS and Lhcb1 proteins in two contrast ecotypes of Tradescantia species, T. fluminensis (shade-tolerant) and T. sillamontana (light-resistant), grown at two intensities of light: 50-125 μmol photons m-2 s-1 (low light, LL) and 875-1000 μmol photons m-2 s-1 (high light, HL). Using the EPR method for measuring the P700 content, we have found that LL-grown plants of both species have higher (by a factor of ≈1.7-1.8) contents of PSI per fresh weight unit as compared to HL-grown plants. Acclimation of plants to LL or HL irradiation also influences the Chl(a + b) level and expression of the PsbS and Lhcb1 proteins. Immunoblotting analysis showed that acclimation to HL stimulates (by a factor of ≈1.7-1.8) the level of PsbS related to the total number of P700 centers. In light-resistant species T. sillamontana, the ratio PsbS/P700 is about 2-times higher than in shade-tolerant species T. fluminensis grown under the same conditions. This should enhance the capacity of their leaves for protection against the light stress. In agreement with these observations, the capacity of leaves for NPQ induction was enhanced during plant acclimation to HL. Kinetic studies of P700 photooxidation and light-induced changes in the yield of Chl a fluorescence also revealed that the short-term regulation of electron transport processes in chloroplasts, which manifested themselves in the kinetics of [Formula: see text] induction and the rate of Chl a fluorescence quenching, occurred more rapidly in HL-grown plants than in LL-grown plants. Thus, both factors, enhanced expression of PsbS and more rapid response of the photosynthetic electron transport chain to dark-to-light transitions should increase the capacity of HL-grown plants for their resistance to rapid fluctuations of solar light.

In this work, using the EPR and PAM-fluorometry methods, we have studied induction events of photosynthetic electron transport in Hibiscus rosa-sinensis leaves. The methods used are complementary, providing efficient tools for in situ monitoring of P700 redox transients and photochemical activity of photosystem II (PSII). The induction of P700(+) in dark-adapted leaves is characterized by the multiphase kinetics with a lag-phase, which duration elongates with the dark-adaptation time. Analyzing effects of the uncoupler monensin and artificial electron carrier methylviologen (MV) on photooxidation of P700 and slow induction of chlorophyll a fluorescence (SIF), we could ascribe different phases of transient kinetics of electron transport processes in dark-adapted leaves to the following regulatory mechanisms: (i) acceleration of electron transfer on the acceptor side of PSI, (ii) pH-dependent modulation of the intersystem electron flow, and (iii) re-distribution of electron fluxes between alternative (linear, cyclic, and pseudocyclic) pathways. Monensin significantly decreases a level of P700(+) and inhibits SIF. MV, which mediates electron flow from PSI to O2 with consequent formation of H2O2, promotes a rapid photooxidation of P700 without any lag-phase peculiar to untreated leaves. MV-mediated water-water cycle (H2O→PSII→PSI→MV→O2→H2O2→H2O) is accompanied by generation of ascorbate free radicals. This suggests that the ascorbate peroxidase system of defense against reactive oxygen species is active in chloroplasts of H. rosa-sinensis leaves. In DCMU-treated chloroplasts with inhibited PSII, the contribution of cyclic electron flow is insignificant as compared to linear electron flow. For analysis of induction events, we have simulated electron transport processes within the framework of our generalized mathematical model of oxygenic photosynthesis, which takes into account pH-dependent mechanisms of electron transport control and re-distribution of electron fluxes between alternative pathways. The model adequately describes the main peculiarities of P700(+) induction and dynamics of the intersystem electron transport.

The pigment-protein complex of photosystem 1 (PS1) isolated from cyanobacterium Synechocystis sp. PCC 6803 has been adsorbed on a solid mesoporous film made from TiO2 nanoparticles. was on The TiO2 film supported on a glass substrate with a surface area of 1 cm2 adsorbs up to 0.045 nmol of PS1. PS1 molecules are distributed in the pores of the mesoporous support. Immobilization has an insignificant effect on the optical and photochemical properties of PS1. A reversible photoinduced EPR signal from the oxidized primary electron donor P700 of immobilized PS1 has been detected. It has been shown by photoelectrochemical methods that the photoexcitation of PS1 results in electron injection from PS1 to the conduction band of TiO2.

Constants of the isotropic hyperfine interaction of an unpaired electron with the hydrogen nuclei of the trimethyl-1,4-benzoquinone anion-radical were calculated using the density functional method. The EPR spectrum of the radical was simulated on the basis of these calculations, which were in good agreement with the experimental EPR spectrum of trimethyl-1,4-benzosemiquinone dissolved in dimethylsulfoxide.

In this work, by using the respective advantages of W- and X-band electron paramagnetic resonance (EPR) spectroscopy techniques to investigate electron transport processes, we have studied the light-induced redox transients of the primary electron donor P700 and the secondary acceptor A1 in photosystem (PS) I complexes of intact cyanobacterial cellsSynechocystis sp. PCC 6803. We found that the kinetic behavior of the cation radical P700·+ generated by illumination with continuous light, and the EPR intensity of the radical pair P700·+A1·− generated upon laser pulse illumination strongly depend on the illumination prehistory (either the sample was frozen in the dark or during illumination). Both these processes were sensitive to the presence of electron transport inhibitors which block electron flow between the two photosystems. In line with our X-band EPR data on the kinetics of light-induced redox transients of P700, our high-field W-band EPR study of the radical-pair state P700·+A1·− shows that photosynthetic electron flow through the PS I reaction center is controlled both on the donor and on the acceptor side of PS I.

In this work, we investigated the interaction of a number of novel pH-sensitive spin probes (imidazoline- and imidazolidine-based nitroxides) with isolated chloroplasts. The light-induced changes in the electron paramagnetic resonance signals of these radicals are determined by several factors: ΔpH-driven accumulation of nitroxides inside the thylakoids, consumption (or evolution) of oxygen by chloroplasts, and irreversible loss of spin probe paramagnetism. A comparative study of these radicals characterized by different pKa allowed us to conclude that these spin probes can be used as pH indicators for monitoring the intrathylakoid pH- and light-induced changes in oxygen concentration in chloroplast suspension.

Ascorbate is one of the key participants of the antioxidant defense in plants. In this work, we have investigated the interaction of ascorbate with the chloroplast electron transport chain and isolated photosystem I (PSI), using the EPR method for monitoring the oxidized centers $$ {\text{P}}_{700}^{ + } $$ and ascorbate free radicals. Inhibitor analysis of the light-induced redox transients of P700 in spinach thylakoids has demonstrated that ascorbate efficiently donates electrons to $$ {\text{P}}_{ 7 0 0}^{ + } $$ via plastocyanin. Inhibitors (DCMU and stigmatellin), which block electron transport between photosystem II and Pc, did not disturb the ascorbate capacity for electron donation to $$ {\text{P}}_{700}^{ + } $$ . Otherwise, inactivation of Pc with CN− ions inhibited electron flow from ascorbate to $$ {\text{P}}_{700}^{ + } $$ . This proves that the main route of electron flow from ascorbate to $$ {\text{P}}_{700}^{ + } $$ runs through Pc, bypassing the plastoquinone (PQ) pool and the cytochrome b 6 f complex. In contrast to Pc-mediated pathway, direct donation of electrons from ascorbate to $$ {\text{P}}_{700}^{ + } $$ is a rather slow process. Oxidized ascorbate species act as alternative oxidants for PSI, which intercept electrons directly from the terminal electron acceptors of PSI, thereby stimulating photooxidation of P700. We investigated the interaction of ascorbate with PSI complexes isolated from the wild type cells and the MenB deletion strain of cyanobacterium Synechocystis sp. PCC 6803. In the MenB mutant, PSI contains PQ in the quinone-binding A1-site, which can be substituted by high-potential electron carrier 2,3-dichloro-1,4-naphthoquinone (Cl2NQ). In PSI from the MenB mutant with Cl2NQ in the A1-site, the outflow of electrons from PSI is impeded due to the uphill electron transfer from A1 to the iron-sulfur cluster FX and further to the terminal clusters FA/FB, which manifests itself as a decrease in a steady-state level of $$ {\text{P}}_{700}^{ + } $$ . The addition of ascorbate promoted photooxidation of P700 due to stimulation of electron outflow from PSI to oxidized ascorbate species. Thus, accepting electrons from PSI and donating them to $$ {\text{P}}_{700}^{ + } $$ , ascorbate can mediate cyclic electron transport around PSI. The physiological significance of ascorbate-mediated electron transport is discussed.

Experimentally measured and DFT calculated isotropic hyperfine splitting constants for the nitroxide radicals 4-amino-2,2,5,5-tetramethyl-3-imidazoline-N-oxyl and 4-amino-2,2,6,6-tetramethylpiperidine-N-oxyl were compared to provide the basis for a choice of an appropriate level of low-cost DFT computations of electronic characteristics of nitroxide radicals.