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Using thermoluminescence, PAM-fluorometry, and electron paramagnetic resonance (EPR) for assaying electron transport processes in chloroplasts in situ, we have compared photosynthetic characteristics in Tradescantia fluminensis leaves grown under low light (LL, 50–125 µmol photons m−2 s−1) or high light (HL, 875–1000 µmol photons m−2 s−1) condition. We found differences in the thermoluminescence (TL) spectra of LL- and HL-acclimated leaves. The LL and HL leaves show different proportions of the Q (~ 0 °C) and B (~ 25–30 °C) bands in their TL spectra; the ratios of the “light sums” of the Q and B bands being SQ/SB ≈ 1/1 (LL) and SQ/SB ≈ 1/3 (HL). This suggests the existence of different redox states of electron carriers on the acceptor side of PSII in LL and HL leaves, which may be affected, in particular, by different capacities of their photo-reducible PQ pools. Enhanced content of PQ in chloroplasts of LL leaves may be the reason for an efficient performance of photosynthesis at low irradiance. Kinetic studies of slow induction of Chl a fluorescence and measurements of P700 photooxidation by EPR demonstrate that HL leaves have faster (about 2 times) response to switching on actinic light as compared to LL leaves grown at moderate irradiation. HL leaves also show higher non-photochemical quenching (NPQ) of Chl a fluorescence. These properties of HL leaves (faster response to light and generation of enhanced NPQ) reflect the flexibility of their photosynthetic apparatus, providing sustainability and rapid response to fluctuations of environmental light intensity and solar stress resistance. Analysis of time-courses of the EPR signals of $${\text{P}}_{700}^{ + }$$ induced by far-red (λmax = 707 nm), exciting predominantly PSI, and white light, exciting both PSI and PSII, suggests that there is a contribution of cyclic electron flow around PSI to electron flow through PSI in HL leaves. The data obtained are discussed in terms of photosynthetic apparatus sustainability of HL and LL leaves under variable irradiation conditions.

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, we investigated electron transport around the photosynthetic pigment-protein complex of Photosystem I (PS I) mediated by external high-potential electron carrier 2,3-dichloro-1,4-naphtoquinone (Cl2 NQ) and ascorbate. It has been demonstrated that the oxidized species of Cl2 NQ and ascorbate serve as intermediates capable of accepting electrons from the iron-sulfur cluster FX of PS I. Reduced species of Cl2 NQ and ascorbate are oxidized by photooxidized PS I primary donor P700+ and/or by molecular oxygen. We have found the synergistic effect of Cl2 NQ and ascorbate on the rate of P700+ reduction. Accelerated electron flow to P700+, observed in the presence of both Cl2 NQ and ascorbate, is explained by an increase in the reduced species of Cl2 NQ due to electron transfer from ascorbate.

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.

This work is devoted to critical analysis and reexamination of the problem of lateral heterogeneity of the trans-thylakoid pH difference (ΔpH = pHout − pHin) in thylakoid membranes of chloroplasts. Correct measurements of ΔpH may be complicated by nonuniform partitioning of the protons pumped into the lumen of granal (stacked) and stroma-exposed thylakoids. We have compared results of ΔpH estimations in isolated bean chloroplasts by two different methods. One of the methods is based on the use of pH-sensitive spin-probes. Another approach relies on the analysis of pH-dependent post-illumination reduction of P700+ - oxidized reaction center of photosystem I (PSI). Both methods lead to virtually the same values of ΔpH, as measured in the state of photosynthetic control when the ATP synthase complexes are inactive (ΔpH ~ 2.3–2.5 at pHout ~ 8). Under the photophosphorylation conditions (metabolic state 3), ΔpH decreases due to the proton drain from the lumen to stroma via active ATP synthases (ΔpH ~ 1–2 at pHout ~ 8). In this state, ΔpH values derived from kinetic data are smaller than ΔpH measured with the pH-probing amines. Such a discrepancy can be explained by the coexistence of thylakoids with different pHin established in granal and stromal thylakoids. The kinetic method is equivalent to the use of a “local pH-meter”, which is sensitive to less significant decrease in pHin inside the stroma-exposed thylakoids. Otherwise, pH-indicating probes give information on pHin values averaged out at both granal and stromal thylakoids. Experimental results have been analyzed within the framework of our mathematical model developed for simulation of electron and proton transport processes in laterally heterogeneous thylakoids. The model provides a reasonable description of experimental data, supporting the notion that the long-range diffusion of protons within the lumen and obstructed diffusion of mobile electron carriers (PQH2 and Pc) influence the lateral profiles of pH along the thylakoid membranes. The model predicts significant alkalization of the inter-thylakoid gap and the establishment of nonuniform lateral profiles of ΔpH under the photophosphorylation conditions. These results are discussed in the context of the problem of energy coupling in laterally heterogeneous lamellar system of chloroplasts.

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.

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.

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.

In this work, we investigated electron transport processes in the cyanobacterium Synechocystis sp. PCC 6803, with a special emphasis focused on oxygen-dependent interrelations between photosynthetic and respiratory electron transport chains. Redox transients of the photosystem I primary donor P700 and oxygen exchange processes were measured by the EPR method under the same experimental conditions. To discriminate between the factors controlling electron flow through photosynthetic and respiratory electron transport chains, we compared the P700 redox transients and oxygen exchange processes in wild type cells and mutants with impaired photosystem II and terminal oxidases (CtaI, CydAB, CtaDEII). It was shown that the rates of electron flow through both photosynthetic and respiratory electron transport chains strongly depended on the transmembrane proton gradient and oxygen concentration in cell suspension. Electron transport through photosystem I was controlled by two main mechanisms: (i) oxygen-dependent acceleration of electron transfer from photosystem I to NADP+, and (ii) slowing down of electron flow between photosystem II and photosystem I governed by the intrathylakoid pH. Inhibitor analysis of P700 redox transients led us to the conclusion that electron fluxes from dehydrogenases and from cyclic electron transport pathway comprise 20–30% of the total electron flux from the intersystem electron transport chain to P700+.

The kinetics of the light-induced redox changes of the photosystem 1 (PS 1) primary donor P700 in whole cells of the cyanobacteria Synechocystis sp. PCC 6803 were studied by the electron paramagnetic resonance method. It was shown that the linear photosynthetic electron transport in cyanobacteria was controlled by two main mechanisms: (i) oxygen-dependent acceleration of electron transfer from PS 1 to NADP+ due to activation of the Calvin cycle reactions and (ii) retardation of electron flow between two photosystems governed by a transmembrane proton gradient. In addition to the linear photosynthetic electron transport, cyanobacteria were capable of maintaining alternative pathways involving cyclic electron transfer around PS 1 and respiratory chains.