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Plastid terminal oxidase (PTOX) accepts electrons from plastoquinol to reduce molecular oxygen to water. We introduced the gene encoding Chlamydomonas reinhardtii (Cr)PTOX2 into the Arabidopsis (Arabidopsis thaliana) wild-type (WT) and proton gradient regulation5 (pgr5) mutant defective in cyclic electron transport around photosystem I (PSI). The accumulation of CrPTOX2 only mildly affected photosynthetic electron transport in the WT background during steady-state photosynthesis but partly complemented the induction of nonphotochemical quenching (NPQ) in the pgr5 background. During the induction of photosynthesis by actinic light (AL) of 130 µmol photons m−2 s−1, the high level of PSII yield (Y(II)) was induced immediately after the onset of AL in WT plants accumulating CrPTOX2. NPQ was more rapidly induced in the transgenic plants than in WT plants. P700 was also oxidized immediately after the onset of AL. Although CrPTOX2 does not directly induce a proton concentration gradient (ΔpH) across the thylakoid membrane, the coupled reaction of PSII generated ΔpH to induce NPQ and the downregulation of the cytochrome b6f complex. Rapid induction of Y(II) and NPQ was also observed in the pgr5 plants accumulating CrPTOX2. In contrast to the WT background, P700 was not oxidized in the pgr5 background. Although the thylakoid lumen was acidified by CrPTOX2, PGR5 was essential for oxidizing P700. In addition to acidification of the thylakoid lumen to downregulate the cytochrome b6f complex (donor-side regulation), PGR5 may be required for draining electrons from PSI by transferring them to the plastoquinone pool. We propose a reevaluation of the contribution of this acceptor-side regulation by PGR5 in the photoprotection of PSI.

Honeysuckle (Lonicera japonica Thunb.) is a traditional medicinal plant in China which is often threatened by high temperature at midday during summer. Heat-induced effects on the photosynthetic apparatus in honeysuckle are associated with a depression of the photosystem II (PSII) photochemical efficiency. However, very limited information is available on regulation of photosynthetic electron flow in PSI photoprotection in heat-stressed honeysuckle. Simultaneous analyses of chlorophyll fluorescence and the change in absorbance of P700 showed that energy transformation and electron transfer activity in PSII decreased under heat stress, but the fraction of photo-oxidizable PSI (Pm) remained stable. With treatments at 38 and 42 °C, the photochemical electron transport in PSII was suppressed, whereas the cyclic electron flow (CEF) around PSI was induced. In addition, the levels of high energy state quenching (qE) and P700 oxidation increased significantly with increasing temperature. However, a decline of qE in antimycin A (AA)- or 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU)-treated leaves after heat treatment was observed, while P700 oxidation decreased only in the presence of AA. The results indicate that heat-induced inhibition of PSII and induction of CEF cooperatively protect PSI from ROS damages through moderate down-regulation of photosynthetic electron flow from PSII to PSI.

To examine the effect of mitochondrial function on photosynthesis, wild-type and transgenic Nicotiana tabacum with varying amounts of alternative oxidase (AOX) were treated with different respiratory inhibitors. Initially, each inhibitor increased the reduction state of the chloroplast electron transport chain, most severely in AOX knockdowns and least severely in AOX overexpressors. This indicated that the mitochondrion was a necessary sink for photo-generated reductant, contributing to the 'P700 oxidation capacity' of photosystem I. Initially, the Complex III inhibitor myxothiazol and the mitochondrial ATP synthase inhibitor oligomycin caused an increase in photosystem II regulated non-photochemical quenching not evident with the Complex III inhibitor antimycin A (AA). This indicated that the increased quenching depended upon AA-sensitive cyclic electron transport (CET). Following 12 h with oligomycin, the reduction state of the chloroplast electron transport chain recovered in all plant lines. Recovery was associated with large increases in the protein amount of chloroplast ATP synthase and mitochondrial uncoupling protein. This increased the capacity for photophosphorylation in the absence of oxidative phosphorylation and enabled the mitochondrion to act again as a sink for photo-generated reductant. Comparing the AA and myxothiazol treatments at 12 h showed that CET optimized photosystem I quantum yield, depending upon the P700 oxidation capacity. When this capacity was too high, CET drew electrons away from other sinks, moderating the P700+ amount. When P700 oxidation capacity was too low, CET acted as an electron overflow, moderating the amount of reduced P700. This study reveals flexible chloroplast-mitochondrion interactions able to overcome lesions in energy metabolism.

During photosynthesis, energy is transiently stored as an electrochemical proton gradient across the thylakoid membrane. The resulting proton motive force (pmf) is composed of a membrane potential (ΔΨ) and a proton concentration gradient (ΔpH) and powers the synthesis of ATP. Light energy availability for photosynthesis can change very rapidly and frequently in nature. Thylakoid ion transport proteins buffer the effects that light fluctuations have on photosynthesis by adjusting pmf and its composition. Ion channel activities dissipate ΔΨ, thereby reducing charge recombinations within photosystem II. The dissipation of ΔΨ allows for increased accumulation of protons in the thylakoid lumen, generating the signal that activates feedback downregulation of photosynthesis. Proton export from the lumen via the thylakoid K+ exchange antiporter 3 (KEA3), instead, decreases the ΔpH fraction of the pmf and thereby reduces the regulatory feedback signal. Here, we reveal that the Arabidopsis (Arabidopsis thaliana) KEA3 protein homo-dimerizes via its C-terminal domain. This C-terminus has a regulatory function, which responds to light intensity transients. Plants carrying a C-terminus-less KEA3 variant show reduced feed-back downregulation of photosynthesis and suffer from increased photosystem damage under long-term high light stress. However, during photosynthetic induction in high light, KEA3 deregulation leads to an increase in carbon fixation rates. Together, the data reveal a trade-off between long-term photoprotection and a short-term boost in carbon fixation rates, which is under the control of the KEA3 C-terminus., The regulatory C-terminus of the thylakoid Kexchange antiporter 3 (KEA3) is required for mitigating high light stress and protein dimerization.

Made available in DSpace on 2021-06-25T10:31:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-05-21 Despite the wide range of possible applications of nanopesticides, the mechanisms involved in their enhanced action remain largely unknown. Understanding the interaction between nanopesticides and plants is crucial for evaluating their potential safety application. Using an experimental and theoretical approach, this study aimed to investigate the target effect of paraquat-loaded chitosan/tripolyphosphate nanoparticles (ca. 200 nm) on photosystem I (PSI). Electrochemical analysis showed distinct electroactivity behaviour between the encapsulated and non-encapsulated herbicide. The amount of lipid peroxidation, photooxidizable P700 reaction centre content, and NADPH/NADP+ ratio levels were significantly decreased in spinach leaf tissue exposed to the nanoherbicide compared to those with the non-encapsulated herbicide. The data also revealed that the nanoformulation might promote oxidative stress based on changes observed in antioxidant enzymes. Also, molecular docking results showed a preferential disposition of the herbicide paraquat and paraquat-tripolyphosphate complex (TPP:PQ) in the ligand domain close to FAD and Glu312. Due to the inhibitor's strategic position in the catalytic pocket, a model of electron-capture is proposed, where the herbicide disturbs the redox process NADP+ ⇌ NADPH by capturing electrons to reduce itself. Finally, our findings provide important insights where the redox behaviour of paraquat may play a key role in the enhanced efficiency of nanoherbicides to the target binding site. Plant Resources Study Group Natural Resources Program Center for Natural Resources Study (CERNA) Mato Grosso Do sul State University (UEMS) São Paulo State University (UNESP) Department of Physics and Chemistry Faculty of Engineering Department of Pharmacology Institute of Biomedical Sciences University of São Paulo (USP) Optics and Photonics Group Institute of Physics Federal University of Mato Grosso Do sul (UFMS) School of Life Science University of Essex Colchester São Paulo State University (UNESP) Department of Physics and Chemistry Faculty of Engineering

Drought induces stomata closure and lowers the CO2 concentration in the mesophyll, limiting CO2 assimilation and favouring photorespiration. The photosynthetic apparatus is protected under drought conditions by a number of downregulation mechanisms like photosynthetic control and activation of cyclic electron transport leading to the generation of a high proton gradient across the thylakoid membrane. Here, we studied photosynthetic electron transport by chlorophyll fluorescence, thermoluminescence and P700 absorption measurements in spinach exposed to moderate drought stress. Chlorophyll fluorescence induction and decay kinetics were slowed down. Under drought conditions an increase of the thermoluminescence AG-band and a downshift of the maximum temperatures of both, the B-band and the AG-band was observed when leaves were illuminated under conditions that maintained the proton gradient. When leaves were frozen prior to the thermoluminescence measurements, the maximum temperature of the B-band was upshifted in drought-stressed leaves. This shows a stabilization of the QB /QB •- redox couple in accordance with the slower fluorescence decay kinetics. We propose that, during drought stress, photorespiration exerts a feedback control on photosystem II via the binding of a photorespiratory metabolite at the non-heme iron at the acceptor side of photosystem II. According to our hypothesis, an exchange of bicarbonate at the non-heme iron by a photorespiratory metabolite such as glycolate would not only affect the midpoint potential of the QA /QA •- couple, as shown previously, but also that of the QB /QB •- couple. This article is protected by copyright. All rights reserved.

Significance Statement Developmental acclimation to light intensity in plants involves changes in the organisation and regulation of photosynthetic electron transfer chain components in the chloroplast thylakoid membrane. Here, using mass spectrometry, we quantified changes in the thylakoid proteome and correlated these with key photosynthetic parameters to gain insight into the process., Summary Photosynthetic acclimation, the ability to adjust the composition of the thylakoid membrane to optimise the efficiency of electron transfer to the prevailing light conditions, is crucial to plant fitness in the field. While much is known about photosynthetic acclimation in Arabidopsis, to date there has been no study that combines both quantitative label‐free proteomics and photosynthetic analysis by gas exchange, chlorophyll fluorescence and P700 absorption spectroscopy. Using these methods we investigated how the levels of 402 thylakoid proteins, including many regulatory proteins not previously quantified, varied upon long‐term (weeks) acclimation of Arabidopsis to low (LL), moderate (ML) and high (HL) growth light intensity and correlated these with key photosynthetic parameters. We show that changes in the relative abundance of cytb 6 f, ATP synthase, FNR2, TIC62 and PGR6 positively correlate with changes in estimated PSII electron transfer rate and CO2 assimilation. Improved photosynthetic capacity in HL grown plants is paralleled by increased cyclic electron transport, which positively correlated with NDH, PGRL1, FNR1, FNR2 and TIC62, although not PGR5 abundance. The photoprotective acclimation strategy was also contrasting, with LL plants favouring slowly reversible non‐photochemical quenching (qI), which positively correlated with LCNP, while HL plants favoured rapidly reversible quenching (qE), which positively correlated with PSBS. The long‐term adjustment of thylakoid membrane grana diameter positively correlated with LHCII levels, while grana stacking negatively correlated with CURT1 and RIQ protein abundance. The data provide insights into how Arabidopsis tunes photosynthetic electron transfer and its regulation during developmental acclimation to light intensity.

Effects of low concentrations of cadmium ions on growth, photosynthesis, and cell ultrastructure of microalga Ankistrodesmus sp. B-11 were investigated. The addition of cadmium to growth medium at concentrations of 0.005–0.02 mg/L led to a significant decrease in the number of Ankistrodesmus sp. B-11 cells relatively to that in the untreated culture. The addition of cadmium at concentrations >0.05 mg/L completely stopped cell growth. Cadmium ions induced ultrastructural changes in the arrangement of thylakoids within the stroma, the detachment of thylakoid membranes with the formation of void interthylakoid spaces, and a significant increase in vacuolization of microalgal cells. Simultaneous measurements of fluorescence induction curves and redox transformations of photosystem I components on a microsecond time scale by means of a M‑PEA-2 fluorometer revealed that cadmium ions inhibit electron transport in photosystem II (PSII). The quantum yield of electron transport in PSII (φEo) and the performance index (PIABS) were found to decrease; the photoreduction of P700 pigment was decelerated, while energy dissipation (DI0/RC) and ΔpH-dependent nonphotochemical quenching (qE) increased significantly under the action of cadmium. The performance index (PIABS) was the most sensitive parameter; it can be used for the detection of early toxic effects of cadmium ions on algae.

Leaf-type ferredoxin-NADP(H) oxidoreductase, encoded by AtLFNR1 gene in Arabidopsis, extensively exists in chloroplast stroma and thylakoids and is responsible for the reduction of NADP+ to NADPH in PSI. To investigate the relationship between the characteristics of chlorophyll a fluorescence transients OJIP and the function of AtLFNR1 protein, the fluorescence rise kinetics OJIP curves of AtLFNR1 mutant leaves were determined. It is observed that AtLFNR1 mutant has a lower level of variable fluorescence intensity relative to wild type, especially the J-step and the IP-phase. The JIP-test analysis suggests that the loss of AtLFNR1 function decreases clearly the oxidation rate of plastocyanin as well as PSI reaction center, damages slightly PSII antenna size and pigment assemblies, but stimulates distinctly the photosynthetic activity of PSII. Such multiple effects contribute to the change of shape characteristics of the OJIP transient curve in AtLFNR1 mutant.

The analysis of electron and proton transport in chloroplasts of higher plants has been carried out on the basis of a mathematical model that takes into account the pH-dependent regulation of electron transport and the operation of the ATP synthase. Numerical experiments aimed at simulation of these processes under pseudocyclic electron transport (water–water cycle) have shown good agreement with experimental data on the kinetics of electron transfer to photosystem 1 (PS1) in class B chloroplasts in metabolic states corresponding to high (state 3) and low (state 4) ATP synthase activity. The simulation of electron transport processes that took into account the Calvin–Benson cycle (CBC), cyclic electron transport around PS1, pH-dependent heat dissipation of energy in photosystem 2 (PS2), and nonphotochemical quenching (NPQ) made it possible to estimate the contribution of these factors to the kinetics of induction phenomena in chloroplasts in situ. It has been shown that the multiphase kinetics of the photooxidation of P700 (a primary electron donor in PS1) reflects the redistribution of electron flows between cyclic and non-cyclic electron transfer pathways, caused by the activation of CBC due to the alkalization of the stroma, as well as the change of the limiting stage in the electron transport chain, induced by a decrease in the intrathylakoid pH (pHin). The electron flux between PS2 and PS1 decelerates with pHin decrease, which may be caused by the reduced rate of plastoquinol oxidation and attenuated activity of PS2 due to NPQ.

Upon a sudden transition from high to low light, the rate of CO2 assimilation (AN) in some plants first decreases to a low level before gradually becoming stable. However, the underlying mechanisms remain controversial. The activity of chloroplast ATP synthase (gH+) is usually depressed under high light when compared with low light. Therefore, we hypothesize that upon a sudden transfer from high to low light, the relatively low gH+ restricts ATP synthesis and thus causes a reduction in AN. To test this hypothesis, we measured gas exchange, chlorophyll fluorescence, P700 redox state, and electrochromic shift signals in Bletilla striata (Orchidaceae). After the transition from saturating to lower irradiance, AN and ETRII decreased first to a low level and then gradually increased to a stable value. Within the first seconds after transfer from high to low light, gH+ was maintained at low levels. During further exposure to low light, gH+ gradually increased to a stable value. Interestingly, a tight positive relationship was found between gH+ and ETRII. These results suggested that upon a sudden transition from high to low light, AN was restricted by gH+ at the step of ATP synthesis. Taken together, we propose that the decline in AN upon sudden transfer from high to low light is linked to the slow kinetics of chloroplast ATP synthase.

Cyanobacteria living in the harsh environment of the desert have to protect themselves against high light intensity and prevent photodamage. These cyanobacteria are in a desiccated state during the largest part of the day when both temperature and light intensity are high. In the desiccated state, their photosynthetic activity is stopped, whereas upon rehydration the ability to perform photosynthesis is regained. Earlier reports indicate that light-induced excitations in Leptolyngbya ohadii are heavily quenched in the desiccated state, because of a loss of structural order of the light-harvesting phycobilisome structures (Bar Eyal et al. in Proc Natl Acad Sci 114:9481, 2017) and via the stably oxidized primary electron donor in photosystem I, namely P700+ (Bar Eyal et al. in Biochim Biophys Acta Bioenergy 1847:1267–1273, 2015). In this study, we use picosecond fluorescence experiments to demonstrate that a third protection mechanism exists, in which the core of photosystem II is quenched independently. Electronic supplementary material The online version of this article (10.1007/s11120-019-00675-0) contains supplementary material, which is available to authorized users.

Cyanobacterial photosystem I (PSI) functions as a light-driven cyt c(6)-ferredoxin/oxidoreductase located in the thylakoid membrane. In this work, the energy and charge transfer processes in PSI complexes isolated from Thermosynechococcus elongatus via conventional n-dodecyl-β-D-maltoside solubilization (DM-PSI) and a, to our knowledge, new detergent-free method using styrene-maleic acid copolymers (SMA-PSI) have been investigated by pump-to-probe femtosecond laser spectroscopy. In DM-PSI preparations excited at 740 nm, the excitation remained localized on the long-wavelength chlorophyll forms within 0.1–20 ps and revealed little or no charge separation and oxidation of the special pair, P700. The formation of ion-radical pair P700(+)A1(−) occurred with a characteristic time of 36 ps, being kinetically controlled by energy transfer from the long-wavelength chlorophyll to P(700). Quite surprisingly, the detergent-free SMA-PSI complexes upon excitation by these long-wave pulses undergo an ultrafast (

Fluctuating light (FL) and drought stress usually occur concomitantly. However, whether drought stress affects photosynthetic performance under FL remains unknown. Here, we measured gas exchange, chlorophyll fluorescence, and P700 redox state under FL in drought-stressed tomato (Solanum lycopersicum) seedlings. Drought stress significantly affected stomatal opening and mesophyll conductance after transition from low to high light and thus delayed photosynthetic induction under FL. Therefore, drought stress exacerbated the loss of carbon gain under FL. Furthermore, restriction of CO2 fixation under drought stress aggravated the over-reduction of photosystem I (PSI) upon transition from low to high light. The resulting stronger FL-induced PSI photoinhibition significantly supressed linear electron flow and PSI photoprotection. These results indicated that drought stress not only affected gas exchange under FL but also accelerated FL-induced photoinhibition of PSI. Furthermore, drought stress enhanced relative cyclic electron flow in FL, which partially compensated for restricted CO2 fixation and thus favored PSI photoprotection under FL. Therefore, drought stress has large effects on photosynthetic dark and light reactions under FL.

Photosystem I (PSI) of photosynthetic organisms is a multi-subunit pigment-protein complex and functions in light harvesting and photochemical charge-separation reactions, followed by reduction of NADP to NADPH required for CO2 fixation. PSI from different photosynthetic organisms has a variety of chlorophylls (Chls), some of which are at lower-energy levels than its reaction center P700, a special pair of Chls, and are called low-energy Chls. However, the site of low-energy Chls is still under debate. Here, we solved a 2.04-Å resolution structure of a PSI trimer by cryo-electron microscopy from a primitive cyanobacterium Gloeobacter violaceus PCC 7421, which has no low-energy Chls. The structure showed absence of some subunits commonly found in other cyanobacteria, confirming the primitive nature of this cyanobacterium. Comparison with the known structures of PSI from other cyanobacteria and eukaryotic organisms reveals that one dimeric and one trimeric Chls are lacking in the Gloeobacter PSI. The dimeric and trimeric Chls are named Low1 and Low2, respectively. Low2 does not exist in some cyanobacterial and eukaryotic PSIs, whereas Low1 is absent only in Gloeobacter. Since Gloeobacter is susceptible to light, this indicates that Low1 serves as a main photoprotection site in most oxyphototrophs, whereas Low2 is involved in either energy transfer or energy quenching in some of the oxyphototrophs. Thus, these findings provide insights into not only the functional significance of low-energy Chls in PSI, but also the evolutionary changes of low-energy Chls responsible for the photoprotection machinery from photosynthetic prokaryotes to eukaryotes.

Photosynthetic electron transfer comprises a series of light-induced redox reactions catalysed by multiprotein machinery in the thylakoid. These protein complexes possess cofactors susceptible to redox modifications by reactive small molecules. The gaseous radical nitric oxide (NO), a key signalling molecule in green algae and plants, has earlier been shown to bind to Photosystem (PS) II and obstruct electron transfer in plants. The effects of NO on cyanobacterial bioenergetics however, have long remained obscure. In this study, we exposed the model cyanobacterium Synechocystis sp. PCC 6803 to NO under anoxic conditions and followed changes in whole-cell fluorescence and oxidoreduction of P700 in vivo. Our results demonstrate that NO blocks photosynthetic electron transfer in cells by repressing PSII, PSI, and likely the NDH dehydrogenase-like complex 1 (NDH-1). We propose that iron‑sulfur clusters of NDH-1 complex may be affected by NO to such an extent that ferredoxin-derived electron injection to the plastoquinone pool, and thus cyclic electron transfer, may be inhibited. These findings reveal the profound effects of NO on Synechocystis cells and demonstrate the importance of controlled NO homeostasis in cyanobacteria.

The transport of electrons along photosynthetic and respiratory chains involves a series of enzymatic reactions that are coupled through redox mediators, including proteins and small molecules. The use of native and synthetic redox probes is key to understanding charge transport mechanisms and to the design of bioelectronic sensors and solar energy conversion devices. However, redox probes have limited tunability to exchange charge at the desired electrochemical potentials (energy levels) and at different protein sites. Herein, we take advantage of electrochemical scanning tunneling microscopy (ECSTM) to control the Fermi level and nanometric position of the ECSTM probe in order to study electron transport in individual photosystem I (PSI) complexes. Current–distance measurements at different potentiostatic conditions indicate that PSI supports long-distance transport that is electrochemically gated near the redox potential of P700, with current extending farther under hole injection conditions.

The kinetics of charge recombination in Photosystem I P700-FA/FB complexes and P700-FX cores lacking the terminal iron‑sulfur clusters were studied over a temperatures range of 310 K to 4.2 K. Analysis of the charge recombination kinetics in this temperature range allowed the assignment of backward electron transfer from the different electron acceptors to P700+. The kinetic and thermodynamic parameters of these recombination reactions were determined. The kinetics of all electron transfer reactions were activation-less below 170 K, the glass transition temperature of the water-glycerol solution. Above this temperature, recombination from [FA/FB]− in P700-FA/FB complexes was found to proceed along two pathways with different activation energies (Ea). The charge recombination via A1A has an Ea of ~290 meV and is dominant at temperatures above ~280 K, whereas the direct recombination from FX− has an Ea of 22 meV and is prevalent in the 200 K to 270 K temperature range. Charge recombination from the FX cluster becomes highly heterogeneous at temperatures below 200 K. The conformational mobility of Photosystem I was studied by molecular dynamics simulations. The FX cluster was found to ‘swing’ by ~30° along the axis between the two sulfur atoms proximal to FA/FB. The partial rotation of FX is accompanied by significant changes of electric potential within the iron‑sulfur cluster, which may induce preferential electron localization at different atoms of the FX cluster. These effects may account for the partial arrest of forward electron transfer and for the heterogeneity of charge recombination observed at the glass transition temperature.

Photosynthetic induction, a gradual increase in photosynthetic rate on a transition from darkness or low light to high light, has ecological significance, impact on biomass accumulation in fluctuating light and relevance to photoprotection in strong light. However, the experimental quantification of the component electron fluxes in and around both photosystems during induction has been rare. Combining optimized chlorophyll fluorescence, the redox kinetics of P700 [primary electron donor in Photosystem I (PSI)] and membrane inlet mass spectrometry in the absence/presence of inhibitors/mediator, we partially estimated the components of electron fluxes in spinach leaf disks on transition from darkness to 1,000 �mol photons�m−2�s−1 for up to 10 min, obtaining the following findings: (i) the partitioning of energy between both photosystems did not change noticeably; (ii) in Photosystem II (PSII), the combined cyclic electron flow (CEF2) and charge recombination (CR2) to the ground state decreased gradually toward 0 in steady state; (iii) oxygen reduction by electrons from PSII, partly bypassing PSI, was small but measurable; (iv) cyclic electron flow around PSI (CEF1) peaked before becoming somewhat steady; (v) peak magnitudes of some of the electron fluxes, all probably photoprotective, were in the descending order: CEF1 > CEF2 + CR2 > chloroplast O2 uptake; and (vi) the chloroplast NADH dehydrogenase-like complex appeared to aid the antimycin A-sensitive CEF1. The results are important for fine-tuning in silico simulation of in vivo photosynthetic electron transport processes; such simulation is, in turn, necessary to probe partial processes in a complex network of interactions in response to environmental changes.

(P700+ – P700) Fourier transform visible and infrared difference spectra (DS) have been obtained using photosystem I (PSI) complexes isolated from cells of Fischerella thermalis PCC 7521 grown under white light (WL) or far-red light (FRL). PSI from cells grown under FRL (FRL-PSI) contain ~8 chlorophyll f (Chl f) molecules (Shen et al., Photosynth. Res. Jan. 2019). Both the visible and infrared DS indicate that neither the PA or PB pigments of P700 are Chl f molecules, but do support the conclusion that at least one of the A–1 cofactors is a Chl f molecule. The FTIR DS indicate that the hydrogen bond to the 131-keto C O group of the PA pigment of P700 is weakened in FRL-PSI, as might be expected given that the proteins that bind the P700 pigments are substantially different in FRL-PSI (Gan et al., Science 345, 1312–1317, 2014). The FTIR DS obtained using FRL-PSI display a band at 1664 cm−1 that is assigned (based on density functional theory calculations) to the 21-formyl C O group of Chl f, that upshifts 5 cm−1 upon P700+ formation. This is much less than expected for a cation-induced upshift, indicating that the Chl f molecule is not one of the pigments of P700. In WL-PSI the A–1 cofactor is a Chl a molecule with 131-keto and 133-methylester C O mode vibrations at 1696 and 1750 cm−1, respectively. In FRL-PSI the A–1 cofactor is a Chl f molecule with 131-keto and 133-methylester C O mode vibrations at 1702 and 1754 cm−1, respectively.

Mechanisms of pharmaceuticals action on biochemical and physiological processes in plants that determine plant growth and development are still mostly unknown. This study deals with the effects of non-steroidal anti-inflammatory drug diclofenac (DCF) on photosynthesis as an essential anabolic process. Changes in primary and secondary photosynthetic processes were assessed in chloroplasts isolated from Lemna minor exposed to 1, 10, 100, and 1000 μM DCF. Decreases in the potential and effective quantum yields of photosystem II (FV/FM by 21%, ΦII by 44% compared to control), changes in non-photochemical fluorescence quenching (NPQ), and a substantial drop in Hill reaction activity (by 73%), especially under 1000 μM DCF, were found. Limitation of electron transport through photosystem II was confirmed by increased fluorescence signals in steps J and I (by 50% and 23%, respectively, under 1000 μM DCF) in OJIP fluorescence transient. Photosystem I exhibited changes only in the redox state of P700 reaction centres (decrease in Pm by 10%, increase in reduced P700 by 5% under 1000 μM DCF). Similarly, RuBisCO activity was only lowered by 30% under 1000 μM DCF. In contrast, a significant increase in reactive oxygen and nitrogen species (by 116% and 157%, respectively) was observed under 10 μM DCF, and lipid peroxidation increased even at 1 μM DCF (by nearly seven times compared to the control). Results demonstrate the ability of environmentally relevant DCF concentrations to induce oxidative stress in isolated duckweed chloroplasts; however, photosynthetic processes were affected considerably only by the highest DCF treatments.

The regulation of photosystem I (PSI) redox state under fluctuating light was investigated for four species using P700 measurement and electrochromic shift analysis. Species included the angiosperms Camellia japonica, Bletilla striata and Arabidopsis thaliana and the fern Cyrtomium fortunei. For the first seconds after transition from low to high light, all species showed relatively low levels of the proton gradient (ΔpH) across the thylakoid membranes. At this moment, PSI was highly oxidized in C. japonica and C. fortunei but was over-reduced in B. striata and A. thaliana. In B. striata and A. thaliana, the redox state of PSI was largely dependent on ΔpH. In contrast, the rapid oxidation of P700 in C. japonica was relatively independent of ΔpH, but was mainly dependent on the outflow of electrons to O2 via the water-water cycle. In the fern C. fortunei, PSI redox state was rapidly regulated by the fast photo-reduction of O2 rather than the ΔpH. These results indicate that mechanisms regulating PSI redox state under fluctuating light differ greatly between species. We propose that the water-water cycle is an important mechanism regulating the PSI redox state in angiosperms.

Despite photosystem I (PSI) is susceptible to photoinhibition under fluctuating light in wild-type angiosperms, the underlying mechanism is not well known. Because proton gradient (ΔpH) across the thylakoid membranes plays a crucial role in protecting PSI, we hypothesized that PSI photoinhibition under fluctuating light may be linked to the formation of ΔpH. To test this hypothesis, we measured chlorophyll fluorescence, PSI redox state, and the electrochromic shift signal during transition from low to high light in two angiosperms Bletilla striata (Orchidaceae) and Arabidopsis thaliana. The measurement of fast P700 redox kinetics indicated that the fast re-oxidation of P700 mediated by photo-reduction of O2 was absent in B. striata. Furthermore, the redox state of PSI is highly determined by ΔpH in both species. For the first 20 s after transition from low to high light, both species could not build up a sufficient ΔpH, which was accompanied by the over-reduction of P700. During prolonged illumination at high light, the sufficient ΔpH made PSI to be highly oxidized. These results demonstrated that PSI photoinhibition under fluctuating light in wild-type angiosperms occurred mainly at the first 20 s after an increase in light intensity, which was caused by the insufficient ΔpH.

The effect of drought on the prompt chlorophyll a fluorescence (PF) transient (OJIP), delayed chlorophyll a fluorescence (DF), modulated 820-nm reflection (MR), energy conversion efficiencies in photosystems (PS) I and II, and cyclic electron flow (CEF) activity in two maize hybrids with contrasting drought tolerance was investigated. Our aim was to identify the target site of drought stress on the photosynthetic electron transport chain and investigate the relevance of the CEF pathway to the drought tolerance of maize plants. The OJIP analysis showed that drought stress, depending on its duration, decreased FP, increased FJ, and induced a pronounced K-band and a positive L-band. Moreover, OJIP parameters, including PIABS, RC/CSO, TRO/ABS, and ETO/TRO, were significantly reduced. The DF analysis showed that the values of I1 and I2 in the induction curve and L1 and L2 derived from the decay curve decreased progressively with the duration of drought stress. The MR analysis showed that drought stress inactivated both the fast decrease and slow increase phases of the MR transient, resulting in a gradual decrease in both VPSI and VPSII-PSI. The energy conversion analysis showed that drought stress decreased the PSI photochemical quantum yield Y(I) and PSII photochemical quantum yield Y(II). Compared to the tolerant hybrid, the drought-induced changes in the sensitive hybrid were stronger and appeared at an earlier treatment stage. The CEF activity analysis showed that the CEF pathway under drought stress operated for a longer time in the tolerant hybrid than that in the sensitive hybrid. The above results indicate that drought stress damaged the donor and acceptor sides of PSII, the PSII reaction center and the acceptor side of PSI and decreased the efficiency of both PSI and PSII and the capacity of electron transfer. The CEF pathway might play an important role in the tolerance of the maize photosynthetic electron transport chain to drought stress.

PSI has the potential to generate reactive oxygen species and be oxidatively inactivated by the reactive oxygen species. The photo-oxidative damage of PSI (also called PSI photoinhibition) causes the inhibition of the plant growth and is a lethal event for plants. It has been reported that PSI photoinhibition does not occur as long as the reaction-center chlorophyll (P700) remains oxidized, even in excess light conditions. This process is termed P700 oxidation and is supported by various regulatory mechanisms and likely also by the stoichiometric quantities of photosynthetic apparatus. In this study, we assessed how decreased photochemically active PSI in Arabidopsis (Arabidopsis thaliana) affected a variety of photosynthetic parameters, including P700 oxidation. Inactivation of PSI was rapidly and selectively induced by repetitive short-pulse illumination. PSI photoinhibition correlated linearly with decreases in effective quantum yield of PSII and nonphotochemical quenching; however, the photosynthetic CO(2) assimilation rate was less affected, as exemplified by ∼50% of the normal CO(2) assimilation rate maintained with an 80% loss in PSI photochemical activity. In contrast, effective quantum yield of PSI was enhanced following PSI photoinhibition, mainly owing to a decrease in the electron donor-side limitation of PSI. Based on these results, we propose that the stoichiometric quantity of PSI is optimized to induce P700 oxidation for dissipating excess light energy in PSI, thus avoiding inhibition of photosynthetic CO(2) assimilation caused by PSI photoinhibition.

Barren stalks and kernel abortion are the major obstacles that hinder maize production. After many years of inbreeding, our group produced a pair of barren stalk/non-barren stalk near-isogenic lines SN98A/SN98B. Under weak light stress, the barren stalk rate is up to 98 % in SN98A but zero in SN98B. Therefore, we consider that SN98A is a weak light-sensitive inbred line whereas SN98B is insensitive. In the present study, the near-isogenic lines SN98A/SN98B were used as test materials to conduct cytological and photosynthetic physiological analyses of the physiological mechanism associated with the differences in maize barren stalk induced by weak light stress. The results showed that weak light stress increased the accumulation of reactive oxygen species (ROS), decreased the function of chloroplasts, destroyed the normal rosette structure, inhibited photosynthetic electron transport, and enhanced lipid peroxidation. The actual photochemical quantum efficiency for PSI (Y(I)) and PSII (Y(II)), relative electron transfer rate for PSI (ETR(I)) and PSII (ETR(II)), and the P700 activities decreased significantly in the leaves of SN98A and SN98B under weak light stress, where the decreases were greater in SN98A than SN98B. After 10 days of shading treatment, the O2·- production rate, H2O2 contents, the yield of regulated energy dissipation (Y(NPQ)), the donor side restriction for PSI (Y(ND)) and the quantum efficiency of cyclic electron flow photochemistry were always higher in SN98A than SN98B, and the antioxidant enzyme activities were always lower in SN98A than those in SN98B. These results show that SN98B has a stronger ability to remove ROS at its source, and maintain the integrity of the structure and function of the photosynthetic system. This self-protection mechanism is an important physiological reason for its adaptation to weak light.

Under natural field conditions, mosses experience fluctuating light intensities combined with temperature stress. Alternative electron flow mediated by flavodiiron proteins (FLVs) and cyclic electron flow (CEF) around photosystem I (PSI) allow mosses to growth under fluctuating light conditions. However, little is known about the roles of FLVs and CEF in the regulation of photosynthesis under temperature stress combined with fluctuating light. Here, we measured chlorophyll fluorescence and P700 redox state under fluctuating light conditions at 4 °C, 20 °C, and 35 °C in three mosses with different light requirements. Upon a sudden increase in light intensity, electron flow from photosystem II initially increased and then gradually decreased at 20 °C and 35 °C, indicating that the operation of FLV-dependent flow lasted much longer than previously thought. Furthermore, the absolute rates of FLV-dependent flow and CEF were enhanced under fluctuating light at 35 °C, pointing to their important roles in photoprotection when exposed to fluctuating light at moderate high temperature. Furthermore, the downregulation of FLV activity at 4 °C was partially compensated for by enhanced CEF activity. These results suggested the subtle coordination between FLV activity and CEF under fluctuating light and temperature stress. Racomitrium japonicum and Hypnum plumaeforme, which usually grow under relatively high light levels, exhibited higher FLV activity and CEF than the shade-grown moss Plagiomnium ellipticum. Based on our results, we conclude that photosynthetic acclimation to fluctuating light and temperature stress in different mosses is largely linked to the adjustment of FLV activity and CEF.

Photosystem I (PSI) is the primary target of photoinhibition under fluctuating light (FL). Photosynthetic organisms employ alternative electron flows to protect PSI under FL. However, the understanding of the coordination of alternative electron flows under FL at temperature stresses is limited. To address this question, we measured the chlorophyll fluorescence, P700 redox state, and electrochromic shift signal in leaves of Dendrobium officinale exposed to FL at 42 °C, 25 °C, and 4 °C. Upon a sudden increase in illumination at 42 °C and 25 °C, the water–water cycle (WWC) consumed a significant fraction of the extra reducing power, and thus avoided an over-reduction of PSI. However, WWC was inactivated at 4 °C, leading to an over-reduction of PSI within the first seconds after light increased. Therefore, the role of WWC under FL is largely dependent on temperature conditions. After an abrupt increase in light intensity, cyclic electron flow (CEF) around PSI was stimulated at any temperature. Therefore, CEF and WWC showed different temperature responses under FL. Furthermore, the enhancement of CEF and WWC at 42 °C quickly generated a sufficient trans-thylakoid proton gradient (ΔpH). The inactivation of WWC at 4 °C was partially compensated for by an increased CEF activity. These findings indicate that CEF and WWC coordinate to protect PSI under FL at temperature stresses.

Upon exposure to environmental stress, the primary electron donor in photosystem I (PSI), P700, is oxidized to suppress the production of reactive oxygen species that could oxidatively inactivate the function of PSI. The illumination of rice leaves with actinic light induces intrinsic fluctuations in the opening and closing of stomata, causing the net CO2 assimilation rate to fluctuate. We examined the effects of these intrinsic fluctuations on electron transport reactions. Under atmospheric O2 conditions (21 kPa), the effective quantum yield of photosystem II (PSII) (Y(II)) remained relatively high while the net CO2 assimilation rate fluctuated, which indicates the function of alternative electron flow. By contrast, under low O2 conditions (2 kPa), Y(II) fluctuated. These results suggest that photorespiration primarily drove the alternative electron flow. Photorespiration maintained the oxidation level of ferredoxin (Fd) throughout the fluctuation of the net CO2 assimilation rate. Moreover, the relative activity of photorespiration was correlated with both the oxidation level of P700 and the magnitude of the proton gradient across the thylakoid membrane in 21 kPa O2 conditions. These results show that photorespiration oxidized P700 by stimulating the proton gradient formation when CO2 assimilation was suppressed by stomatal closure.

The inhibitory effects of cadmium (CdSO4 ) on the primary photosynthetic processes were studied in vivo in Pisum sativum by using Multi-function Plant Efficiency Analyser (M-PEA-2). Photosynthetic parameters related to photosystem (PS) II, PS I and intersystem electron carriers were calculated from the light-induced kinetics of prompt chlorophyll a fluorescence (OJIP transient), delayed chlorophyll a fluorescence (DF), and 820 nm modulated reflection (MR). Low-dose exposure to cadmium (20 μm CdSO4 for 48 h) reduced probability of electron transfer from plastoquinones to the terminal electron acceptors of PSI (δRo ) accompanied by a decrease in the rate of P700+ and PC reduction (Vred ) and the magnitude of the I2 step on the DF kinetics. Electron transport through PSI remained unaltered. The obtained results allowed us to propose existence of the potential site of inhibition of photosynthetic electron flow by cadmium between PSII and PSI. We propose to use parameters δRo , Vred , and I2 /I1 as sensitive indicators of an early contamination by heavy metals.

Natural light intensities can rise several orders of magnitude over subsecond time spans, posing a major challenge for photosynthesis. Fluctuating light tolerance in the green alga Chlamydomonas reinhardtii requires alternative electron pathways, but the role of nonphotochemical quenching (NPQ) is not known. Here, fluctuating light (10 min actinic light followed by 10 min darkness) led to significant increase in NPQ/qE-related proteins, LHCSR1 and LHCSR3, relative to constant light of the same subsaturating or saturating intensity. Elevated levels of LHCSR1/3 increased the ability of cells to safely dissipate excess light energy to heat (i.e., qE-type NPQ) during dark to light transition, as measured with chlorophyll fluorescence. The low qE phenotype of the npq4 mutant, which is unable to produce LHCSR3, was abolished under fluctuating light, showing that LHCSR1 alone enables very high levels of qE. Photosystem (PS) levels were also affected by light treatments, constant light led to lower PsbA levels and Fv/Fm values, while fluctuating light led to lower PsaA and maximum P700+ levels, indicating that constant and fluctuating light induced PSII and PSI photoinhibition, respectively. Under fluctuating light, npq4 suffered more PSI photoinhibition and significantly slower growth rates than parental wild type, whereas npq1 and npq2 mutants affected in xanthophyll carotenoid compositions had identical growth under fluctuating and constant light. Overall, LHCSR3 rather than total qE capacity or zeaxanthin is shown to be important in C. reinhardtii in tolerating fluctuating light, potentially via preventing PSI photoinhibition.

The water-water cycle (WWC) has the potential to alleviate photoinhibition of photosystem I (PSI) in fluctuating light (FL) at room temperature and moderate heat stress. However, it is unclear whether WWC can function as a safety valve for PSI in FL at chilling temperature. In this study, we measured P700 redox state and chlorophyll fluorescence in FL at 25 °C and 4 °C in the high WWC activity plant Dendrobium officinale. At 25 °C, the operation of WWC contributed to the rapid re-oxidation of P700 upon dark-to-light transition. However, such rapid re-oxidation of P700 was not observed at 4 °C. Upon a sudden increase in light intensity, WWC rapidly consumed excess electrons in PSI and thus avoided an over-reduction of PSI at 25 °C. On the contrary, PSI was highly reduced within the first seconds after transition from low to high light at 4 °C. Therefore, in opposite to 25 °C, the WWC is not a major alternative sink in FL at chilling temperature. Upon transition from low to high light, cyclic electron transport was highly stimulated at 4 °C when compared with 25 °C. These results indicate that D. officinale enhances cyclic electron transport to partially compensate for the inactivation of WWC in FL at 4 °C.

Heavy-metal contaminations in natural waters, wetlands and wastewaters pose serious threats to aquatic ecosystems - mainly via targeting microalgae. In this work, we investigated the effects of toxic amounts of chromium and cadmium ions on the structure and function of the photosynthetic machinery of Chlorella variabilis cells. To halt the propagation of cells, we used high concentrations of Cd and Cr, 50-50 mg L-1, in the forms of CdCl2 x 2.5 H2O and K2Cr2O7, respectively. Both treatments led to similar, about 50% gradual diminishment of the chlorophyll contents of the cells in 48 hrs, which was, however, accompanied by a small (~10%) but statistically significant enrichment (Cd) and loss (Cr) of s-carotene. Both Cd and Cr inhibited the activity of photosystem II (PSII) – but with more severe inhibitions with Cr. On the contrary, the PsbA (D1) protein of PSII and the PsbO protein of the oxygen-evolving complex were retained more in Cr-treated cells than in the presence of Cd. These data and the higher susceptibility of P700 redox transients in Cr-treated cells suggest that, unlike with Cd, PSII is not the main target in the photochemical apparatus. These differences at the level of photochemistry also brought about dissimilarities at higher levels of the structural complexity of the photosynthetic apparati. Circular dichroism spectroscopy measurements revealed moderate perturbations in the macro-organization of the protein complexes – with more pronounced decline in Cd-treated cells than in the cells with Cr. Also, as reflected by transmission electron microscopy and small-angle neutron scattering, the thylakoid membranes suffered shrinking and were largely fragmented in Cd-treated cells, whereas no changes could be discerned with Cr. The preservation of integrity of membranes in Cr-treated cells was most probably aided by high proportion of the de-epoxidized xanthophylls, which were absent with Cd. It can thus be concluded that beside strong similarities of the toxic effects of Cr and Cd, the response of the photosynthetic machinery of C. variabilis to these two heavy-metal ions substantially differ from each other – strongly suggesting different inhibitory and protective mechanisms following the primary toxic events.

Photorespiration coupled with CO2 assimilation is thought to act as a defense system against photoinhibition caused by osmotic stress. In the present study, we examined whether such a mechanism is operative for the protection of photosystem I (PSI) in rice (Oryza sativa L.) including transgenic plants with decreased and increased Rubisco content (RBCS-antisense and RBCS-sense plants, respectively). All plants were hydroponically grown and moderate osmotic stress was imposed using hydroponic culture solutions containing poly(ethylene glycol) (PEG) at 16% or 20% (w/v) for 2 d. In wild-type plants, the rates of CO2 assimilation (A) were significantly decreased by the PEG treatment, whereas the photorespiration activity estimated from the rates of electron transport in photosystem II (PSII) and A were not affected. The maximal quantum efficiency of PSII (F v/F m) and the maximal activity of PSI (P m) were also not affected. In RBCS-antisense plants, A and the estimated photorespiration activity were considerably lower than those in wild-type plants in the presence or absence of the PEG treatment. P m and both F v/F m and P m decreased in the 16% PEG-treated and 20% PEG-treated RBCS-antisense plants, respectively. Thus, the decrease in Rubisco content led to the photoinhibition of PSI and PSII, indicating the importance of photorespiration coupled with CO2 assimilation for the protection of PSI from moderate PEG-induced osmotic stress. It was also shown that PSI was more sensitive to osmotic stress than PSII. In the PEG-treated wild-type and RBCS-antisense plants, osmotic-stress responses of the photosynthetic electron transport reactions upstream of PSI led to the oxidation of P700, which is thought to prevent PSI from over-reduction. Although such a defense system operated, it was not sufficient for the protection of PSI in RBCS-antisense plants. In addition, there were no large differences in the parameters measured between wild-type and RBCS-sense plants, as overproduction of Rubisco did not increase photorespiration activity.

Plants in their natural environment are often exposed to fluctuating light because of self-shading and cloud movements. As changing frequency is a key characteristic of fluctuating light, we speculated that rapid light fluctuation may induce rapid photosynthetic responses, which may protect leaves against photoinhibition. To test this hypothesis, maize seedlings were grown under fluctuating light with various frequencies (1, 10, and 100 cycles of fluctuations/10 h), and changes in growth, chlorophyll content, gas exchange, chlorophyll a fluorescence, and P700 were analyzed carefully. Our data show that though the growth and light-saturated photosynthetic rate were depressed by rapidly fluctuating light, photosynthesis induction was clearly speeded up. Furthermore, more rapid fluctuation of light strikingly reduced the chlorophyll content, while thermal dissipation was triggered and enhanced. The chlorophyll a fluorescence induction kinetics and P700 absorption results showed that the activities of both photosystem II and photosystem I decreased as the frequency of the fluctuating light increased. In all treatments, the light intensities of the fluctuating light were kept constant. Therefore, rapid light fluctuation frequency itself induced the acceleration of photosynthetic induction and the enhancement of photoprotection in maize seedlings, which play important roles in protecting photosynthetic apparatus against fluctuating high light to a certain extent.

Photosynthesis fixes CO2 and converts it to sugar, using chemical-energy compounds of both NADPH and ATP, which are produced in the photosynthetic electron transport system. The photosynthetic electron transport system absorbs photon energy to drive electron flow from Photosystem II (PSII) to Photosystem I (PSI). That is, both PSII and PSI are full of electrons. O2 is easily reduced to a superoxide radical (O2−) at the reducing side, i.e., the acceptor side, of PSI, which is the main production site of reactive oxygen species (ROS) in photosynthetic organisms. ROS-dependent inactivation of PSI in vivo has been reported, where the electrons are accumulated at the acceptor side of PSI by artificial treatments: exposure to low temperature and repetitive short-pulse (rSP) illumination treatment, and the accumulated electrons flow to O2, producing ROS. Recently, my group found that the redox state of the reaction center of chlorophyll P700 in PSI regulates the production of ROS: P700 oxidation suppresses the production of O2− and prevents PSI inactivation. This is why P700 in PSI is oxidized upon the exposure of photosynthesis organisms to higher light intensity and/or low CO2 conditions, where photosynthesis efficiency decreases. In this study, I introduce a new molecular mechanism for the oxidation of P700 in PSI and suppression of ROS production from the robust relationship between the light and dark reactions of photosynthesis. The accumulated protons in the lumenal space of the thylakoid membrane and the accumulated electrons in the plastoquinone (PQ) pool drive the rate-determining step of the P700 photo-oxidation reduction cycle in PSI from the photo-excited P700 oxidation to the reduction of the oxidized P700, thereby enhancing P700 oxidation.

Photosynthetic electron flux from water via photosystem II (PSII) and PSI to oxygen (water-water cycle) may act as an alternative electron sink under fluctuating light in angiosperms. We measured the P700 redox kinetics and electrochromic shift signal under fluctuating light in 11 Camellia species and tobacco leaves. Upon dark-to-light transition, these Camellia species showed rapid re-oxidation of P700. However, this rapid re-oxidation of P700 was not observed when measured under anaerobic conditions, as was in experiment with tobacco performed under aerobic conditions. Therefore, photo-reduction of O2 mediated by water-water cycle was functional in these Camellia species but not in tobacco. Within the first 10 s after transition from low to high light, PSI was highly oxidized in these Camellia species but was over-reduced in tobacco leaves. Furthermore, such rapid oxidation of PSI in these Camellia species was independent of the formation of trans-thylakoid proton gradient (ΔpH). These results indicated that in addition to ΔpH-dependent photosynthetic control, the water-water cycle can protect PSI against photoinhibition under fluctuating light in these Camellia species. We here propose that the water-water cycle is an overlooked strategy for photosynthetic regulation under fluctuating light in angiosperms.

Moderate heat stress and fluctuating light are typical conditions in summer in tropical and subtropical regions. This type of stress can cause photodamage to photosystems I and II (PSI and PSII). However, photosynthetic responses to the combination of heat and fluctuating light in young leaves are little known. In this study, we investigated chlorophyll fluorescence and P700 redox state under fluctuating light at 25 °C and 42 °C in young leaves of tobacco. Our results indicated that fluctuating light caused selective photodamage to PSI in the young leaves at 25 °C and 42 °C. Furthermore, the moderate heat stress significantly accelerated photoinhibition of PSI under fluctuating light. Within the first 10 s after transition from low to high light, cyclic electron flow (CEF) around PSI was highly stimulated at 25 °C but was slightly activated at 42 °C. Such depression of CEF activation at moderate heat stress were unable to maintain energy balance under high light. As a result, electron flow from PSI to NADP+ was restricted, leading to the over-reduction of PSI electron carriers. These results indicated that moderate heat stress altered the CEF performance under fluctuating light and thus accelerated PSI photoinhibition in tobacco young leaves.

The main production site of reactive oxygen species (ROS) in photosynthetic organisms is photosystem (PS) I of thylakoid membranes. Unless the suppression mechanism of ROS production functions, PSI easily suffers from oxidative damages by ROS attack. Miyake group has elucidated the production and suppression mechanisms of ROS in PSI. The reaction center chlorophyll, P700, in PSI functions in P700 photo-oxidation reduction cycle. The photoexcited P700, P700*, can donate electron to O2 producing superoxide radical, ROS, with oxidized to P700+. The accumulation of the P700+ decreases the probability of the presence of P700* not to produce ROS. The present review describes the molecular mechanism to oxidize P700 and to accumulate P700+ in PSI. Tight coupling between the light and the dark reactions in photosynthesis accumulates H+ in the luminal side and electron in plastoquinone pool of thylakoid membranes on exposure to the environmental stress, which lowers the electron transport activity of Cyt b6/f-complex and suppresses the electron flux to PSI with P700+ accumulated. We discuss the molecular mechanisms to accumulate e− and H+ and its relationship with ATP synthase activity from the aspect of P700 oxidation in PSI.

International audience; The liverwort Marchantia polymorpha contains two isoforms of the plastid terminal oxidase (PTOX), an enzyme that catalyzes the reduction of oxygen to water using plastoquinol as substrate. Phylogenetic analyses showed that one isoform, here called MpPTOXa, is closely related to isoforms occurring in plants and some algae, while the other isoform, here called MpPTOXb, is closely related to the two isoforms occurring in Chlamydomonas reinhardtii. Mutants of each isoform were created in Marchantia polymorpha using CRISPR/Cas9 technology. While no obvious phenotype was found for these mutants, chlorophyll fluorescence analyses demonstrated that the plastoquinone pool was in a higher reduction state in both mutants. This was visible at the level of fluorescence measured in dark-adapted material and by post illumination fluorescence rise. These results suggest that both isoforms have a redundant function. However, when P700 oxidation and re-reduction was studied, differences between these two isoforms were observed. Furthermore, the mutant affected in MpPTOXb showed a slight alteration in the pigment composition, a higher non-photochemical quenching and a slightly lower electron transport rate through photosystem II. These differences may be explained either by differences in the enzymatic activities or by different activities attributed to preferential involvement of the two PTOX isoforms to either linear or cyclic electron flow.

This work aims to fully elucidate the effects of a trehalose glassy matrix on electron transfer reactions in cyanobacterial Photosystem I (PS I). Forward and backward electron transfer rates from A1A− and A1B− to FX, and charge recombination rates from A0−, A1B−, A1A−, FX−, and [FA/FB]− to P700+ were measured in P700–FA/FB complexes, P700–FX cores, and P700–A1 cores, both in liquid and in a trehalose glassy matrix at 11% humidity. By comparing CONTIN-resolved kinetic events over 6 orders of time in increasingly simplified versions of PS I at 480 nm, a wavelength that reports primarily A1A−/A1B− oxidation, and over 9 orders of time at 830 nm, a wavelength that reports P700+ reduction and A0− oxidation, assignments could be made for nearly all of the resolved kinetic phases. Trehalose-embedded PS I samples demonstrated partially arrested forward electron transfer. The fractions of complexes in which electron transfer did not proceed beyond A0, A1 and FX were 53%, 16% and 22%, respectively, with only 10% of electrons reaching the terminal FA/FB clusters. The ~10 μs and ~150 μs components in both liquid and trehalose-embedded PS I were assigned to recombination between A1B− and P700+ and between A1A− and P700+, respectively. The kinetics and amplitudes of these resolved kinetic phases in liquid and trehalose-embedded PS I samples could be well-fitted by a kinetic model that allowed us to calculate the asymmetrical contribution of the A1A− and A1B− quinones to the electrochromic signal at 480 nm. Possible reasons for these effects are discussed.

Time-resolved step-scan FTIR difference spectroscopy has been used to study photosystem I (PSI) with plastoquinone-9 (PQ) and two other benzoquinones (2,6-dimethyl-1,4-benzoquinone and 2,3,5,6-tetrachloro-1,4-benzoquinone) incorporated into the A1 binding site. By subtracting a (P700+A1− − P700A1) FTIR difference spectrum for PSI with the native phylloquinone (PhQ) incorporated from corresponding spectra for PSI with different benzoquinones (BQs) incorporated, FTIR double difference spectra are produced, that display bands associated with vibrational modes of the quinones, without interference from features associated with protein vibrational modes. Molecular models for BQs involved in asymmetric hydrogen bonding were constructed and used in vibrational mode frequency calculations. The calculated data were used to aid in the interpretation and assignment of bands in the experimental spectra. We show that the calculations capture the general trends found in the experimental spectra. By comparing four different FTIR double difference spectra we are able to verify unambiguously bands associated with phyllosemiquinone in PSI at 1495 and 1415 cm−1. We also resolve a previously unrecognized band of phyllosemiquinone at 1476 cm−1 that calculations suggest is due in part to a C4 − O stretching mode. For PSI with PQ incorporated, calculations and experiment taken together indicate that the C1 − O and C4 − O vibrational modes of the semiquinone give rise to bands at 1487 and 1444 cm−1, respectively. This is very distinct compared to PSI with PhQ incorporated. From the calculated and experimental spectra, we show that it is possible to distinguish between two possible orientations of PQ in the A1 protein binding site.

After transferring the dark-acclimated cyanobacteria to light, flavodiiron proteins Flv1/Flv3 serve as a main electron acceptor for PSI within the first seconds because Calvin cycle enzymes are inactive in the dark. Synechocystis PCC 6803 mutant Δflv1/Δflv3 devoid of Flv1 and Flv3 retained the PSI chlorophyll P700 in the reduced state over 10 s (Helman et al., 2003; Allahverdiyeva et al., 2013). Study of P700 oxidoreduction transients in dark-acclimated Δflv1/Δflv3 mutant under the action of successive white light pulses separated by dark intervals of various durations indicated that the delayed oxidation of P700 was determined by light activation of electron transport on the acceptor side of PSI. We show that the light-induced redox transients of chlorophyll P700 in dark-acclimated Δflv1/Δflv3 proceed within 2 min, as opposed to 1–3 s in the wild type, and comprise a series of kinetic stages. The release of rate-limiting steps was eliminated by iodoacetamide, an inhibitor of Calvin cycle enzymes. Conversely, the creation with methyl viologen of a bypass electron flow to O2 accelerated P700 oxidation and made its extent comparable to that in the wild-type cells. The lack of major sinks for linear electron flow in iodoacetamide-treated Δflv1/Δflv3 mutant, in which O2- and CO2-dependent electron flows were impaired, facilitated cyclic electron flow, which was evident from the decreased steady-state oxidation of P700 and from rapid dark reduction of P700 during and after illumination with far-red light. The results show that the photosynthetic induction in wild-type Synechocystis PCC 6803 is largely hidden due to the flavodiiron proteins whose operation circumvents the rate-limiting electron transport steps controlled by Calvin cycle reactions.

Common root rot is a widespread cereal disease caused by a plant pathogenic fungus Bipolaris sorokiniana. The influence of fungal infection on photosynthetic light reactions in soft wheat has been studied by a simultaneous registration of fast and delayed chlorophyll fluorescence induction curves as well as the redox state of a P700 pigment. In the case of infected plants, the reduction of a quantum yield of electron transport in the photosystem II (ϕE0) and performance index on absorption basis (PIABS), as well as the increase of energy dissipation per a reaction center (DI0/RC) and ΔpH-dependent nonphotochemical fluorescence quenching (qE), has been observed. A reduction of the induction peak of delayed chlorophyll fluorescence at 10–50 ms has been revealed. Reactions of the photosystem I show a greater resistance to fungal infection as compared with photosystem II. Parameters of chlorophyll a fluorescence induction may be used for the early diagnostics of pathogen-induced changes in the physiological state of plants.

The proton motive force (pmf) across the thylakoid membranes plays a key role for photosynthesis in fluctuating light. However, the mechanisms underlying the regulation of pmf in fluctuating light are not well known. In this study, we aimed to identify the roles of chloroplastic ATP synthase and cyclic electron flow (CEF) around photosystem I (PSI) in the regulation of the pmf in fluctuating light. To do this, we measured chlorophyll fluorescence, P700 parameters, and the electrochromic shift signal in the fluctuating light alternating between 918 (high light) and 89 (low light) μmol photons m-2 s-1 every 5 min. We found that the activity of chloroplastic ATP synthase (gH+), pmf, CEF activity, non-photochemical quenching (NPQ), and the P700 redox state changed rapidly in fluctuating light. During transition from low to high light, the decreased gH+ and the stimulation of CEF both contributed to the rapid formation of pmf, activating NPQ and optimizing the redox state of P700 in PSI. During the low-light phases, gH+ rapidly increased and the pmf declined sharply, leading to the relaxation of NPQ and down-regulation of photosynthetic control. These findings indicate that in fluctuating light the gH+ and CEF are finely regulated to modulate the pmf formation, avoiding the over-accumulation of reactive intermediates and maximizing energy use efficiency.

In this work, the analysis of electron and proton transport in chloroplasts of higher plants has been carried out on the basis of a mathematical model, which takes into account the pH-dependent regulation of electron transfer and thioredoxin-dependent activation of the Calvin–Benson cycle (CBC) enzymes and the ATP synthase. The impact of reduced thioredoxin on the kinetics of electron transport, pH changes in the intrathylakoid space and ATP production has been simulated. Comparison of the computed and experimental data on the kinetics of P700 photooxidation has shown that the consideration of thioredoxin-dependent activation of the CBC and the ATP synthase provides an adequate description of the multiphase kinetics of P 700 + induction. The dynamics of electron flow through PSI and the partitioning of electron fluxes on the acceptor site of PSI has been simulated. The model predicts that at the initial stage of the induction period the alternative pathways, cyclic electron transport around PSI and electron flow to O2 (the Mehler reaction), play a significant role in photosynthetic electron transport chain, but their contribution attenuates upon the activation of the CBC reactions.

—We investigated the state of the pigments of inbred (zpp1225) and hybrid (zp341) maize (Zea mays L.) seeds exposed to ultraviolet radiation and α-irradiation using spectral methods. Exposure to different ultraviolet radiation doses from 5 to 13.39 kJ/m2 and α-irradiation from 1.5 to 3 kGy stimulated plant growth and seed germination. Exposure of seeds to a high dose of α-irradiation (15 kGy) led to inhibition of plant growth and development. Irradiation of seeds can change the state and function of the pigments present in the leaves. Irradiation of seeds induced conformational changes of carotenoid modules (increase of the length of the polyene chain and amount of molecules in 15-cis-conformation); the effect depends on the type of irradiation (the effect was larger when seeds were exposed to α-irradiation). It was shown that the photosynthetic apparatus in hybrid (zp341) maize is characterized by a relatively increased I–P phase on the induction curve of fast fluorescence and a higher degree of oxidation of the P700 reaction center, indicating a higher acceptor pool on the acceptor side of the photosystem I in the hybrid maize. Seed exposure to ultraviolet radiation and α-irradiation caused inhibition of reactions in photosystem II in leaves, observed as a decrease in the quantum yield of electron transport (φEo), an increase of non-photochemical quenching (DI0/RC and φDo) and a decrease of the energy of thylakoid membranes (higher Δψ). The highest decease caused by radiation was determined for the generalized index of photosystem II performance (PIABS). The PIABS production index can be recommended for assessing the status of plants in selection studies.

Under CO2-limited conditions such as during stomatal closure, photorespiration is suggested to act as a sink for excess light energy and protect photosystem I (PSI) by oxidizing its reaction center chlorophyll P700. In this study, this issue was directly examined with rice (Oryza sativa L.) plants via genetic manipulation of the amount of Rubisco, which can be a limiting factor for photorespiration. At low [CO2] of 5 Pa that mimicked stomatal closure condition, the activity of photorespiration in transgenic plants with decreased Rubisco content (RBCS-antisense plants) markedly decreased, whereas the activity in transgenic plants with overproduction of Rubisco (RBCS-sense plants) was similar to that in wild-type plants. Oxidation of P700 was enhanced at [CO2] of 5 Pa in wild-type and RBCS-sense plants. PSI was not damaged by excess light stress induced by repetitive saturated pulse-light (rSP) in the presence of strong steady-state light. On the other hand, P700 was strongly reduced in RBCS-antisense plants at [CO2] of 5 Pa. PSI was also damaged by rSP illumination. These results indicate that oxidation of P700 and the robustness of PSI against excess light stress are hampered by the decreased activity of photorespiration as a result of genetic manipulation of Rubisco content. It is also suggested that overproduction of Rubisco does not enhance photorespiration as well as CO2 assimilation probably due to partial deactivation of Rubisco.

Thylakoid proton motive force (pmf) plays a central role in photosynthesis. However, the regulation of pmf during photosynthetic induction is not well known. In the present study, in order to investigate the importance of cyclic electron flow (CEF) and chloroplastic ATP synthase in pmf formation during photosynthetic induction, we examined chlorophyll fluorescence, P700 signal, electrochromic shift signal for leaves of a low-light species Panax notoginseng and a high-light species Bletilla striata. During the initial induction phase at both low and high light, the activation of CEF associating with low proton conductivity of the chloroplastic ATP synthase (gH+) resulted in the rapid buildup of proton motive force (pmf) in both species. Furthermore, during photosynthetic induction at high light, the increase in gH+ in B. striata was accompanied with increases in linear electron flow (LEF) and CEF, leading to a stable pmf. Under high light, the high-light species B. striata, with much higher LEF and CEF than P. notoginseng, showed significantly lower pmf owing to the higher gH+. This result suggested that, under high light, CEF mainly contributed to ATP synthase in B. striata but favored lumenal acidification in P. notoginseng. Taking together, during photosynthetic induction, the coordination between CEF and the activity of chloroplastic ATP synthase finely regulated the pmf levels, optimizing photosynthesis and photoprotection.

Light-induced Fourier transformed infrared (FTIR) difference spectroscopy is a powerful method to study the structures and reactions of redox cofactors involved in the photosynthetic electron transport chain. So far, most of the FTIR studies of the reactions of oxygenic photosynthesis have been performed using isolated photosystem I (PSI) and photosystem II (PSII) preparations, which, however, could be modified during isolation procedures. In this study, we developed a methodology to evaluate the photosynthetic activities of thylakoids using FTIR spectroscopy. FTIR difference spectra upon successive flashes using thylakoids from spinach exhibited signals typical of the S-state cycle at the Mn4CaO5 cluster and QB reactions in PSII with period-four and -two oscillations, respectively. Similar measurement in the presence of an artificial quinone as an exogenous electron acceptor showed features specific to the S-state cycle. Simulations of the oscillation patterns provided the quantum efficiencies of the S-state cycle and electron transfer in PSII. Moreover, FTIR measurement under continuous illumination on thylakoids in the presence of DCMU showed signals due to QA reduction and P700 oxidation simultaneously. From the relative amplitudes of marker bands of QA- and P700+, the molar ratio of photoactive PSII and PSI centers in thylakoids was estimated. FTIR analyses of the photo-reactions in thylakoids, which are more intact than isolated photosystems, will be useful in investigations of the photosynthetic mechanism especially by genetic modification of photosystem proteins.