P700 Oxidation Research Articles

Regulation of photosystems II and I depending on N partitioning to Rubisco in rice leaves: a study using Rubisco-antisense transgenic plants.

We have previously suggested that in rice (Oryza sativa L.) leaves of different ages and N nutrition statuses, photosystems II and I (PSII and PSI, respectively) are regulated depending on N partitioning to Rubisco, which can determine the magnitude of unutilized light energy. The robustness of this mechanism was tested using Rubisco-antisense transgenic rice plants, in which reduced N partitioning to Rubisco markedly increases unutilized light energy. In wild-type plants, N partitioning to Rubisco tended to be smaller in the leaves at lower positions owing to leaf senescence. In the transgenic plants, N partitioning to Rubisco was generally smaller than in the wild-type plants and was relatively constant among leaf positions. The quantum efficiency of PSII [Y(II)] and quantum yield of non-photochemical quenching [Y(NPQ)] correlated positively and negatively, respectively, with N partitioning to Rubisco irrespective of leaf position or genotype. The oxidation levels of the reaction center chlorophyll of PSI (P700) [Y(ND)] negatively correlated with N partitioning to Rubisco. However, in mature and early senescent leaves of the transgenic plants, Y(ND) was markedly lower than expected from N partitioning to Rubisco. These results suggest that in the transgenic plants, the regulation depending on N partitioning to Rubisco is robust for PSII but fails for PSI in mature and early senescing leaves. In these leaves, the magnitudes of P700 oxidation were found to be less than expected from the Y(II) and Y(NPQ) values. The mechanistic reasons and physiological implications of these phenomena are discussed.

Differential FeS cluster photodamage plays a critical role in regulating excess electron flow through photosystem I.

The photosynthetic electron flux from photosystem I (PSI) is mainly directed to NADP+ and CO2 fixation, but a fraction is always shared between alternative and cyclic electron transport. Although the electron transfer from P700 to ferredoxin, via phylloquinone and the FeSX, FeSB and FeSA clusters, is well characterized, the regulatory role of these redox intermediates in the delivery of electrons from PSI to NADP+, alternative and cyclic electron transport under environmental stress remains elusive. Here we provide evidence for sequential damage to PSI FeS clusters under high light and subsequent slow recovery under low light in Arabidopsis thaliana. Wild-type plants showed 10-35% photodamage to their FeSA/B clusters with increasing high-light duration, without much effect on P700 oxidation capacity, FeSX function or CO2 fixation rate, and without additional oxygen consumption (O2 photoreduction). Parallel FeSA/B cluster damage in the pgr5 mutant was more pronounced at 50-85%, probably due to weak photosynthetic control and low non-photochemical quenching. Such severe electron pressure on PSI was also shown to damage the FeSX clusters, with a concomitant decrease in P700 oxidation capacity and a decrease in thylakoid-bound ferredoxin in the pgr5 mutant. The results from wild-type and pgr5 plants reveal controlled damage of PSI FeS clusters under high light. In wild-type plants, this favours electron transport to linear over alternative pathways by intact PSI centres, thereby preventing reactive oxygen species production and probably promoting harmless charge recombination between P700+ and FeSX- as long as the majority of FeSA/B clusters remain functional.

Manganese deficiency alters photosynthetic electron transport in Marchantia polymorpha

Manganese (Mn) is considered as an essential element for plant growth. Mn starvation has been shown to affect photosystem II, the site of the Mn4CaO5 cluster responsible for water oxidation. Less is known on the effect of Mn starvation on photosystem I. Here we studied the effects of Mn deficiency in vivo on redox changes of P700 and plastocyanin (Pc) in the liverwort Marchantia polymorpha using the KLAS-NIR spectrophotometer. Far-red illumination is used to excite preferentially photosystem I, thus facilitating cyclic electron transport. Under Mn starvation, we observed slower oxidation of P700 and a decrease in the Pc signal relative to P700. The lower Pc content under Mn deficiency was confirmed by western blots. Re-reduction kinetics of P700+ and Pc+ were faster in Mn deficient thalli than in the control. The above findings show that the kinetics studied under Mn deficiency not only depend on the number of available reductants but also on how quickly electrons are transferred from stromal donors via the intersystem chain to Pc+ and P700+. We suggest that under Mn deficiency a structural reorganization of the thylakoid membrane takes place favoring the formation of supercomplexes between ferredoxin, cytochrome b6f complex, Pc and photosystem I, and thus an enhanced cyclic electron transport.

Enhanced Reduction of Ferredoxin in PGR5-Deficient Mutant of Arabidopsis thaliana Stimulated Ferredoxin-Dependent Cyclic Electron Flow around Photosystem I.

The molecular entity responsible for catalyzing ferredoxin (Fd)-dependent cyclic electron flow around photosystem I (Fd-CEF) remains unidentified. To reveal the in vivo molecular mechanism of Fd-CEF, evaluating ferredoxin reduction-oxidation kinetics proves to be a reliable indicator of Fd-CEF activity. Recent research has demonstrated that the expression of Fd-CEF activity is contingent upon the oxidation of plastoquinone. Moreover, chloroplast NAD(P)H dehydrogenase does not catalyze Fd-CEF in Arabidopsis thaliana. In this study, we analyzed the impact of reduced Fd on Fd-CEF activity by comparing wild-type and pgr5-deficient mutants (pgr5hope1). PGR5 has been proposed as the mediator of Fd-CEF, and pgr5hope1 exhibited a comparable CO2 assimilation rate and the same reduction-oxidation level of PQ as the wild type. However, P700 oxidation was suppressed with highly reduced Fd in pgr5hope1, unlike in the wild type. As anticipated, the Fd-CEF activity was enhanced in pgr5hope1 compared to the wild type, and its activity further increased with the oxidation of PQ due to the elevated CO2 assimilation rate. This in vivo research clearly demonstrates that the expression of Fd-CEF activity requires not only reduced Fd but also oxidized PQ. Importantly, PGR5 was found to not catalyze Fd-CEF, challenging previous assumptions about its role in this process.

Effects of drought stress on the oxidation of the reaction center chlorophyll of photosystem I and grain yield in paddy-field grown rice plants

ABSTRACT Drought is a serious abiotic stress that leads to decreased yield. In pot experiments, the reaction center chlorophyll of photosystem I (PSI) (P700) has been reported to be oxidized in response to drought stress. In the present study, we examined whether drought stress that decreases grain yield was detected by P700 oxidation in paddy-field-grown rice (Oryza sativa L.) plants. The P700 reduction and SPAD values were also evaluated. Drought stress was imposed by the prolonged mid-season drainage in 2019 and 2020. P700 oxidation was measured at saturated CO2 levels before heading. P700 oxidation was not stimulated in the 2019-drought plots. In contrast, P700 oxidation was stimulated in the 2020-drought plot. However, this phenomenon in 2020 was primarily accounted for by decreases in total leaf-N levels, as previously observed in the rice plants grown under low N availability. Rough rice yields and ratios of filled spikelets tended to decrease in the drought plots. Therefore, drought stress responsible for a decrease in grain yields could not be directly detected by P700 oxidation in paddy-field-grown rice plants, whereas the P700 oxidation responded to changes in total leaf-N levels due to the drought stress treatment. The same conclusion was drawn for the P700 reduction and SPAD value.

Cyclophilin 37 maintains electron transport via the cytochrome b6/f complex under high light in Arabidopsis.

Plants have evolved multiple mechanisms to cope with diverse types of light stress, particularly the regulation of the electron transport chain (ETC). Under high light (HL) conditions, the balance of electron flux in the ETC is disturbed, which leads to the overaccumulation of reactive oxygen species (ROS) and results in photodamage and photoinhibition. The cytochrome (Cyt) b6/f complex, which coordinates electron transfer between photosystems I and II (PSI and PSII), plays an essential role in regulating the ETC and initiating photoprotection. However, how the Cyt b6/f complex is maintained under HL conditions remains unclear. Here, we report that the activity of the Cyt b6/f complex is sustained by thylakoid-localized cyclophilin 37 (CYP37) in Arabidopsis (Arabidopsis thaliana). Compared with wild-type plants, cyp37 mutants displayed an imbalance in electron transport from Cyt b6/f to PSI under HL stress, which led to increased ROS accumulation, decreased anthocyanin biosynthesis, and increased chlorophyll degradation. Surprisingly, CYP37's role in regulating ETC balance was independent of photosynthesis control, which was indicated by a higher Y (ND), an indicator of P700 oxidation in PSI. Furthermore, the interaction between CYP37 and photosynthetic electron transfer A (PetA), a subunit of the Cyt b6/f complex, suggests that the central function of CYP37 is to maintain Cyt b6/f complex activity rather than to serve as an assembly factor. Our study provides insights into how plants balance electron flow between PSII and PSI via Cyt b6/f complex under HL.

Higher Reduced State of Fe/S-Signals, with the Suppressed Oxidation of P700, Causes PSI Inactivation in Arabidopsis thaliana.

Environmental stress increases the risk of electron accumulation in photosystem I (PSI) of chloroplasts, which can cause oxygen (O2) reduction to superoxide radicals and decreased photosynthetic ability. We used three Arabidopsis thaliana lines: wild-type (WT) and the mutants pgr5hope1 and paa1-7/pox1. These lines have different reduced states of iron/sulfur (Fe/S) signals, including Fx, FA/FB, and ferredoxin, the electron carriers at the acceptor side of PSI. In the dark, short-pulse light was repetitively illuminated to the intact leaves of the plants to provide electrons to the acceptor side of PSI. WT and pgr5hope1 plants showed full reductions of Fe/S during short-pulse light and PSI inactivation. In contrast, paa1-7/pox1 showed less reduction of Fe/S and its PSI was not inactivated. Under continuous actinic-light illumination, pgr5hope1 showed no P700 oxidation with higher Fe/S reduction due to the loss of photosynthesis control and PSI inactivation. These results indicate that the accumulation of electrons at the acceptor side of PSI may trigger the production of superoxide radicals. P700 oxidation, responsible for the robustness of photosynthetic organisms, participates in reactive oxygen species suppression by oxidizing the acceptor side of PSI.

Distinct contribution of two cyclic electron transport pathways to P700 oxidation.

Cyclic electron transport (CET) around Photosystem I (PSI) acidifies the thylakoid lumen and downregulates electron transport at the cytochrome b6f complex. This photosynthetic control is essential for oxidizing special pair chlorophylls (P700) of PSI for PSI photoprotection. In addition, CET depending on the PROTON GRADIENT REGULATION 5 (PGR5) protein oxidizes P700 by moving a pool of electrons from the acceptor side of PSI to the plastoquinone pool. This model of the acceptor-side regulation was proposed on the basis of the phenotype of the Arabidopsis (Arabidopsis thaliana) pgr5-1 mutant expressing Chlamydomonas (Chlamydomonas reinhardtii) plastid terminal oxidase (CrPTOX2). In this study, we extended the research including the Arabidopsis chlororespiratory reduction 2-2 (crr2-2) mutant defective in another CET pathway depending on the chloroplast NADH dehydrogenase-like (NDH) complex. Although the introduction of CrPTOX2 did not complement the defect in the acceptor-side regulation by PGR5, the function of the NDH complex was complemented except for its reverse reaction during the induction of photosynthesis. We evaluated the impact of CrPTOX2 under fluctuating light intensity in the wild-type, pgr5-1 and crr2-2 backgrounds. In the high-light period, both PGR5- and NDH-dependent CET were involved in the induction of photosynthetic control, whereas PGR5-dependent CET preferentially contributed to the acceptor-side regulation. On the contrary, the NDH complex probably contributed to the acceptor-side regulation in the low-light period but not in the high-light period. We evaluated the sensitivity of PSI to fluctuating light and clarified that acceptor-side regulation was necessary for PSI photoprotection by oxidizing P700 under high light.

A tomato chloroplast-targeted DnaJ protein, SlDnaJ20 maintains the stability of photosystem I/II under chilling stress

ABSTRACT DnaJ proteins are key molecular chaperones that act as a part of the stress response to stabilize plant proteins, thereby maintaining protein homeostasis under stressful conditions. Herein we used transgenic plants to explore the role of the tomato (Solanum lycopersicum) SlDnaJ20 chloroplast DnaJ protein in to the resistance of these proteins to cold. When chilled, transgenic plants exhibited superior cold resistance, with reduced growth inhibition and cellular damage and increased fresh mass and chlorophyll content relative to control. These transgenic plants further exhibited increased Fv/Fm, P700 oxidation, φRo, and δRo relative to control plants under chilling conditions. Under these same cold conditions, these transgenic plants also exhibited higher levels of core proteins in the photosystem I (PSI) and II (PSII) complexes (PsaA and PsaB; D1 and D2) relative to control wild-type plants. Together these results suggested that the overexpression of SlDnaJ20 is sufficient to maintain PSI and PSII complex stability and to alleviate associated photoinhibition of these complexes, thereby increasing transgenic plant resistance to cold stress.

Moderate photoinhibition of PSII and oxidation of P700 contribute to chilling tolerance of tropical tree species in subtropics of China

In the subtropics, a few tropical tree species are distributed and planted for ornamental and horticultural purposes; however, the photosynthesis of these species can be impaired by chilling. This study aimed to understand how these species respond to chilling. Light-dependent and CO₂ assimilation reactions of six tropical tree species from geographically diverse areas, but grown at a lower subtropical site in China, were monitored during a chilling (≤ 10°C). Chilling induced stomatal and nonstomatal effects and moderate photoinhibition of PSII, with severe effect in <i>Ixora chinensis</i>. <i>Woodfordia fruticosa</i> was little affected by chilling, with negligible reduction of photosynthesis and PSII activity, higher cyclic electron flow (CEF), and oxidation state of P700 (P700⁺). Photoinhibition of PSII thus reduced electron flow to P700, while active CEF reduced oxidative damage of PSI and maintained photosynthesis during chilling. Studied parameters revealed that coupling between light-dependent and CO₂ assimilation reactions was enhanced under chilling.

The ability of P700 oxidation in photosystem I reflects chilling stress tolerance in cucumber.

Low temperature inhibits photosynthesis and negatively affects plant growth. Cucumber (Cucumis sativusL.) is a chilling-sensitive plant, and its greenhouse production requires considerable energy during the winter. Therefore, a useful stress marker for selecting chilling-tolerant cucumber cultivars is desirable. In this study, we evaluated chilling-stress damage in different cucumber cultivars by measuring photosynthetic parameters. The majority of cultivars showed decreases in the quantum yield of photosystem (PS) II [Fv/Fm and Y(II)] and the quantity of active PS I (Pm) after chilling stress. In contrast, Y(ND)-the ratio of the oxidized state of PSI reaction center chlorophyll P700 (P700+)-differed among cultivars and was perfectly inversely correlated with Y(NA)-the ratio of the non-photooxidizable P700. It has been known that P700+ accumulates under stress conditions and protects plants to suppress the generation of reactive oxygen species. In fact, cultivars unable to induce Y(ND) after chilling stress showed growth retardation with reductions in chlorophyll content and leaf area. Therefore, Y(ND) can be a useful marker to evaluate chilling-stress tolerance in cucumber.

Paradoxes in judging the inhibition of photosynthetic electron transfer chain using P700 oxidation and dark re-reduction analyses

• P700 redox kinetics analyses to identify functions of photosynthetic inhibitors • DNP-INT is unable to block electron flow between photosystem II and photosystem I • Leakage of electrons from Cytochrome b6f to plastocyanine cause reduction of oxidized P700 • P700 redox kinetics ensures inhibition of electron flow to PSI and alternate electron flow from cytochrome b6f and PQ-pool

Oxidation of the reaction center chlorophyll of photosystem I is induced via close cooperation of photosystems II and I with progress of drought stress in soybean seedlings

ABSTRACT Drought is a serious abiotic stress that impairs plant productivity. We previously suggested that in rice (Oryza sativa L.) plants, the reaction center chlorophyll of photosystem I (PSI) (P700) is oxidized in response to drought stress via close cooperation of photosystem II (PSII) and PSI to protect PSI, whose fitness is crucial for photosynthesis and plant growth. It has rarely been reported whether such a defense system functions in other plant species. To collect another example, this subject was examined using soybean [Glycine max (L.)]. Irrigation of soil-cultured soybean seedlings was restricted to gradually impose drought stress. A decrease in the quantum efficiency of PSII [Y(II)], induction of non-photochemical quenching (NPQ) of excess light energy at PSII, reduction of the primary quinone electron acceptor of PSII (QA), and P700 oxidation proceeded before severe drought stress led to severe leaf wilting and photoinhibition of PSII and PSI. P700 oxidation, Y(II), and reduction of QA were strongly correlated in this period. These results are in accordance with those observed in rice. In contrast, such correlations disappeared when severely drought-stressed seedlings were included, as P700 oxidation was hampered while Y(II) further decreased in such seedlings. These results suggest that close cooperation of PSII and PSI leading to P700 oxidation functions to protect PSI against drought stress in soybean, and that similar defense system functions in soybean and rice unless drought stress are severe. The drought-stress responses of PSII and PSI are suggested to be regulated by acidification of thylakoid lumen in soybean, according to the changing patterns in the PSII and PSI parameters. Based on the similarity of the photochemistry of soybean and rice in response to drought stress, the effectiveness of P700 oxidation as an index for drought stress diagnosis is discussed. P700 oxidation is affected by factors other than drought stress such as N nutrition levels. However, effects of N nutrition levels are expected to be canceled by using another parameter obtained in P700 measurement as reference, suggesting the suitability of P700 oxidation for drought stress diagnosis.

Identification of a Novel Mutation Exacerbated the PSI Photoinhibition in pgr5/pgrl1 Mutants; Caution for Overestimation of the Phenotypes in Arabidopsis pgr5-1 Mutant.

PSI photoinhibition is usually avoided through P700 oxidation. Without this protective mechanism, excess light represents a potentially lethal threat to plants. PGR5 is suggested to be a major component of cyclic electron transport around PSI and is important for P700 oxidation in angiosperms. The known Arabidopsis PGR5 deficient mutant, pgr5-1, is incapable of P700 oxidation regulation and has been used in numerous photosynthetic studies. However, here it was revealed that pgr5-1 was a double mutant with exaggerated PSI photoinhibition. pgr5-1 significantly reduced growth compared to the newly isolated PGR5 deficient mutant, pgr5hope1. The introduction of PGR5 into pgr5-1 restored P700 oxidation regulation, but remained a pale-green phenotype, indicating that pgr5-1 had additional mutations. Both pgr5-1 and pgr5hope1 tended to cause PSI photoinhibition by excess light, but pgr5-1 exhibited an enhanced reduction in PSI activity. Introducing AT2G17240, a candidate gene for the second mutation into pgr5-1 restored the pale-green phenotype and partially restored PSI activity. Furthermore, a deficient mutant of PGRL1 complexing with PGR5 significantly reduced PSI activity in the double-deficient mutant with AT2G17240. From these results, we concluded that AT2G17240, named PSI photoprotection 1 (PTP1), played a role in PSI photoprotection, especially in PGR5/PGRL1 deficient mutants.

Roles of alternative electron flows in response to excess light in Ginkgo biloba

Ginkgo biloba L., the only surviving species of Ginkgoopsida, is a famous relict gymnosperm, it may provide new insight into the evolution of photosynthetic mechanisms. Flavodiiron proteins (FDPs) are conserved in nonflowering plants, but the role of FDPs in gymnosperms has not yet been clarified. In particular, how gymnosperms integrate FDPs and cyclic electron transport (CET) to better adapt to excess light is poorly understood. To elucidate these questions, we measured the P700 signal, chlorophyll fluorescence and electrochromic shift signal under fluctuating and constant light in G. biloba. Within the first seconds after light increased, G. biloba could not build up a sufficient proton gradient (ΔpH). Concomitantly, photo-reduction of O2 mediated by FDPs contributed to the rapid oxidation of P700 and protected PSI under fluctuating light. Therefore, in G. biloba, FDPs mainly protect PSI under fluctuating light at acceptor side. Under constant high light, the oxidation of PSI and the induction of non-photochemical quenching were attributed to the increase in ΔpH formation, which was mainly caused by the increase in CET rather than linear electron transport. Therefore, under constant light, CET finely regulates the PSI redox state and non-photochemical quenching through ΔpH formation, protecting PSI and PSII against excess light. We conclude that, in G. biloba, FDPs are particularly important under fluctuating light while CET is essential under constant high light. The coordination of FDPs and CET fine-tune photosynthetic apparatus under excess light.

Physiological Roles of Flavodiiron Proteins and Photorespiration in the Liverwort Marchantia polymorpha.

Against the potential risk in oxygenic photosynthesis, that is, the generation of reactive oxygen species, photosynthetic electron transport needs to be regulated in response to environmental fluctuations. One of the most important regulations is keeping the reaction center chlorophyll (P700) of photosystem I in its oxidized form in excess light conditions. The oxidation of P700 is supported by dissipating excess electrons safely to O2, and we previously found that the molecular mechanism of the alternative electron sink is changed from flavodiiron proteins (FLV) to photorespiration in the evolutionary history from cyanobacteria to plants. However, the overall picture of the regulation of photosynthetic electron transport is still not clear in bryophytes, the evolutionary intermediates. Here, we investigated the physiological roles of FLV and photorespiration for P700 oxidation in the liverwort Marchantia polymorpha by using the mutants deficient in FLV (flv1) at different O2 partial pressures. The effective quantum yield of photosystem II significantly decreased at 2kPa O2 in flv1, indicating that photorespiration functions as the electron sink. Nevertheless, it was clear from the phenotype of flv1 that FLV was dominant for P700 oxidation in M. polymorpha. These data suggested that photorespiration has yet not replaced FLV in functioning for P700 oxidation in the basal land plant probably because of the lower contribution to lumen acidification, compared with FLV, as reflected in the results of electrochromic shift analysis.

Heat-induced down-regulation of photosystem II protects photosystem I in honeysuckle (Lonicera japonica).

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.

Coral symbionts evolved a functional polycistronic flavodiiron gene.

Photosynthesis in cyanobacteria, green algae, and basal land plants is protected against excess reducing pressure on the photosynthetic chain by flavodiiron proteins (FLV) that dissipate photosynthetic electrons by reducing O2. In these organisms, the genes encoding FLV are always conserved in the form of a pair of two-type isozymes (FLVA and FLVB) that are believed to function in O2 photo-reduction as a heterodimer. While coral symbionts (dinoflagellates of the family Symbiodiniaceae) are the only algae to harbor FLV in photosynthetic red plastid lineage, only one gene is found in transcriptomes and its role and activity remain unknown. Here, we characterized the FLV genes in Symbiodiniaceae and found that its coding region is composed of tandemly repeated FLV sequences. By measuring the O2-dependent electron flow and P700 oxidation, we suggest that this atypical FLV is active in vivo. Based on the amino-acid sequence alignment and the phylogenetic analysis, we conclude that in coral symbionts, the gene pair for FLVA and FLVB have been fused to construct one coding region for a hybrid enzyme, which presumably occurred when or after both genes were inherited from basal green algae to the dinoflagellate. Immunodetection suggested the FLV polypeptide to be cleaved by a post-translational mechanism, adding it to the rare cases of polycistronic genes in eukaryotes. Our results demonstrate that FLV are active in coral symbionts with genomic arrangement that is unique to these species. The implication of these unique features on their symbiotic living environment is discussed.

In vivo electron donation from plastocyanin and cytochrome c6 to PSI in Synechocystis sp. PCC6803

Many cyanobacteria species can use both plastocyanin and cytochrome c6 as lumenal electron carriers to shuttle electrons from the cytochrome b6f to either photosystem I or the respiratory cytochrome c oxidase. In Synechocystis sp. PCC6803 placed in darkness, about 60% of the active PSI centres are bound to a reduced electron donor which is responsible for the fast re-reduction of P700in vivo after a single charge separation. Here, we show that both cytochrome c6 and plastocyanin can bind to PSI in the dark and participate to the fast phase of P700 reduction, but the fraction of pre-bound PSI is smaller in the case of cytochrome c6 than with plastocyanin. Because of the inter-connection of respiration and photosynthesis in cyanobacteria, the inhibition of the cytochrome c oxidase results in the over-reduction of the photosynthetic electron transfer chain in the dark that translates into a lag in the kinetics of P700 oxidation at the onset of light. We show that this is true both with plastocyanin and cytochrome c6, indicating that the partitioning of electron transport between respiration and photosynthesis is regulated in the same way independently of which of the two lumenal electron carriers is present, although the mechanisms of such regulation are yet to be understood.

Photosynthetic Linear Electron Flow Drives CO2 Assimilation in Maize Leaves.

Photosynthetic organisms commonly develop the strategy to keep the reaction center chlorophyll of photosystem I, P700, oxidized for preventing the generation of reactive oxygen species in excess light conditions. In photosynthesis of C4 plants, CO2 concentration is kept at higher levels around ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) by the cooperation of the mesophyll and bundle sheath cells, which enables them to assimilate CO2 at higher rates to survive under drought stress. However, the regulatory mechanism of photosynthetic electron transport for P700 oxidation is still poorly understood in C4 plants. Here, we assessed gas exchange, chlorophyll fluorescence, electrochromic shift, and near infrared absorbance in intact leaves of maize (a NADP-malic enzyme C4 subtype species) in comparison with mustard, a C3 plant. Instead of the alternative electron sink due to photorespiration in the C3 plant, photosynthetic linear electron flow was strongly suppressed between photosystems I and II, dependent on the difference of proton concentration across the thylakoid membrane (ΔpH) in response to the suppression of CO2 assimilation in maize. Linear relationships among CO2 assimilation rate, linear electron flow, P700 oxidation, ΔpH, and the oxidation rate of ferredoxin suggested that the increase of ΔpH for P700 oxidation was caused by the regulation of proton conductance of chloroplast ATP synthase but not by promoting cyclic electron flow. At the scale of intact leaves, the ratio of PSI to PSII was estimated almost 1:1 in both C3 and C4 plants. Overall, the photosynthetic electron transport was regulated for P700 oxidation in maize through the same strategies as in C3 plants only except for the capacity of photorespiration despite the structural and metabolic differences in photosynthesis between C3 and C4 plants.