Activation Of Cyclic Electron Transport Research Articles

The yellow-cotyledon gene (ATYCO) is a crucial factor for thylakoid formation and photosynthesis regulation in Arabidopsis

Chloroplast development underpins plant growth, by facilitating not only photosynthesis but also other essential biochemical processes. Nonetheless, the regulatory mechanisms and functional components of chloroplast development remain largely uncharacterized due to their complexity. In our study, we identified a plastid-targeted gene, ATYCO/RP8/CDB1, as a critical factor in early chloroplast development in Arabidopsis thaliana. YCO knock-out mutant (yco) exhibited a seedling-lethal, albino phenotype, resulting from dysfunctional chloroplasts lacking thylakoid membranes. Conversely, YCO knock-down mutants produced a chlorophyll-deficient cotyledon and normal leaves when supplemented with sucrose. Transcription analysis also revealed that YCO deficiency could be partially compensated by sucrose supplementation, and that YCO played different roles in the cotyledons and the true leaves. In YCO knock-down mutants, the transcript levels of plastid-encoded RNA polymerase (PEP)-dependent genes and nuclear-encoded photosynthetic genes, as well as the accumulation of photosynthetic proteins, were significantly reduced in the cotyledons. Moreover, the chlorophyll-deficient phenotype in YCO knock-down line can be effectively suppressed by inhibition of PSI cyclic electron transport activity, implying an interaction between YCO and PSI cyclic electron transport. Taken together, our findings de underscore the vital role of YCO in early chloroplast development and photosynthesis.

NADH dehydrogenase-like complex L subunit improves salt tolerance by enhancing photosynthetic electron transport

Cyclic electron transport (CET) around photosystem I (PSI) mediated by the NADH dehydrogenase-like (NDH) complex is closely related to plant salt tolerance. However, whether overexpression of a core subunit of the NDH complex affects the photosynthetic electron transport under salt stress is currently unclear. Here, we expressed the NDH complex L subunit (Ndhl) genes ZmNdhl1 and ZmNdhl2 from C4 plant maize (Zea mays) or OsNdhl from C3 plant rice (Oryza sativa) using a constitutive promoter in rice. Transgenic rice lines expressing ZmNdhl1, ZmNdhl2, or OsNdhl displayed enhanced salt tolerance, as indicated by greater plant height, dry weight, and leaf relative water content, as well as lower malondialdehyde content compared to wild-type plants under salt stress. Fluorescence parameters such as post-illumination rise (PIR), the prompt chlorophyll a fluorescence transient (OJIP), modulated 820-nm reflection (MR), and delayed chlorophyll a fluorescence (DF) remained relatively normal in transgenic plants during salt stress. These results indicate that expression of ZmNdhl1, ZmNdhl2, or OsNdhl increases cyclic electron transport activity, slows down damage to linear electron transport, alleviates oxidative damage to the PSI reaction center and plastocyanin, and reduces damage to electron transport on the receptor side of PSI in rice leaves under salt stress. Thus, expression of Ndhl genes from maize or rice improves salt tolerance by enhancing photosynthetic electron transport in rice. Maize and rice Ndhl genes played a similar role in enhancing salinity tolerance and avoiding photosynthetic damage.

Effect of Acclimation to High Temperatures on the Mechanisms of Drought Tolerance in Species with Different Types of Photosynthesis: Sedobassia sedoides (C3–C4) and Bassia prostrata (C4-NADP)

The effect of drought on the morphophysiological, biochemical, and molecular genetic parameters of plants Sedobassia sedoides (Pall.) Freitag G. Kadereit with an intermediate C3–C4-type of photosynthesis and Bassia prostrata (L.) A.J. Scott with a C4-NADP type of photosynthesis grown at different temperatures (25 and 30°C) was studied. A decrease in the biomass, water content, and effective quantum yield (ΦPSII) of PSII, as well as an increase in the expression of the psbA gene encoding the PSII D1 protein under the action of drought, was observed in both species regardless of the growing temperature. Both species showed a decrease in the content of photosynthetic enzymes ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and phosphoenolpyruvate carboxylase (PEPC) under drought conditions at 25°С, which was accompanied by a significant increase in the expression of the rbcL and PPDK genes in S. sedoides. Acclimation of S. sedoides plants to elevated temperatures led to an increase in the activity of cyclic electron transport around PSI, to mitigation of the negative effect of drought on the light reactions of photosynthesis (reduction in NPQ) and the content of the PEPC enzyme, as well as to a shift in the ionic balance caused by a decrease in the potassium content. B. prostrata showed greater drought resistance and was characterized by greater thermolability of photosynthetic enzymes, changes in the content and ratio of which allowed this species to maintain growth in drought conditions at different temperatures.

Impact of Salinity, Elevated Temperature, and Their Interaction with the Photosynthetic Efficiency of Halophyte Crop Chenopodium quinoa Willd

Chenopodium quinoa is a genetically diverse crop that can adapt to a wide range of environments, including temperatures and salinities. However, only a few studies have assessed the combined effects of two or more environmental factors on C. quinoa. Here, we investigated the effects of salinity (300 mM NaCl), elevated temperature (35 °C), and their interaction with growth, water–salt balance, the efficiency of photosystem II (PSII), the activity of cyclic electron transport (CET) around photosystem I (PSI), Rubisco and PEPC enzyme content, and the expression of photosynthetic genes. We found that elevated temperature did not decrease the biomass but caused a significant increase in the water and potassium content of C. quinoa leaves. The decrease in PSII efficiency under elevated temperature was accompanied by an increase in the expression of genes encoding the components of PSII (psbA) and linear electron transport (FDI), as well as the main photosynthetic protein Rubisco (rbcL). Moreover, the strongest effect was induced by the combined effect of elevated temperature and salinity, which induced high oxidative stress (a threefold increase in MDA), a threefold decrease in the biomass, a twofold decrease in PSII efficiency, and a two- to eightfold decrease in the expression of the photosynthetic genes psbA, FDI, and rbcL. PSI was more tolerant to all forms of stress; however, the combined effect of elevated temperature and salinity downregulated the expression of PGR5 and FNR1, which may diminish the role of PGR5/PGRL1-dependent CET in favor of the NDH-dependent CET of PSI. The obtained data on the functioning of photosystems and the expression of photosynthetic genes under combined stress (elevated temperature and salinity) can make a significant contribution to understanding the mechanisms of tolerance of C. quinoa to multiple stresses under climate change conditions.

Dissection of respiratory and cyclic electron transport in Synechocystis sp. PCC 6803.

Cyclic electron transport (CET) is an attractive hypothesis for regulating photosynthetic electron transport and producing the additional ATP in oxygenic phototrophs. The concept of CET has been established in the last decades, and it is proposed to function in the progenitor of oxygenic photosynthesis, cyanobacteria. The in vivo activity of CET is frequently evaluated either from the redox state of the reaction center chlorophyll in photosystem (PS) I, P700, in the absence of PSII activity or by comparing PSI and PSII activities through the P700 redox state and chlorophyll fluorescence, respectively. The evaluation of CET activity, however, is complicated especially in cyanobacteria, where CET shares the intersystem chain, including plastoquinone, cytochrome b6/f complex, plastocyanin, and cytochrome c6, with photosynthetic linear electron transport (LET) and respiratory electron transport (RET). Here we sought to distinguish the in vivo electron transport rates in RET and CET in the cyanobacterium Synechocystis sp. PCC 6803. The reduction rate of oxidized P700 (P700+) decreased to less than 10% when PSII was inhibited, indicating that PSII is the dominant electron source to PSI but P700+ is also reduced by electrons derived from other sources. The oxidative pentose phosphate (OPP) pathway functions as the dominant electron source for RET, which was found to be inhibited by glycolaldehyde (GA). In the condition where the OPP pathway and respiratory terminal oxidases were inhibited by GA and KCN, the P700+ reduction rate was less than 1% of that without any inhibitors. This study indicate that the electron transport to PSI when PSII is inhibited is dominantly derived from the OPP pathway in Synechocystis sp. PCC 6803.

A scheme for C4 evolution derived from a comparative analysis of the closely related C3, C3-C4 intermediate, C4-like, and C4 species in the genus Flaveria.

A comparative analysis of the genus Flaveria showed a C4 evolutionary process in which the anatomical and metabolic features of C4 photosynthesis were gradually acquired through C3-C4 intermediate stages. C4 photosynthesis has been acquired in multiple lineages of angiosperms during evolution to suppress photorespiration. Crops that perform C4 photosynthesis exhibit high rates of CO2 assimilation and high grain production even under high-temperature in semiarid environments; therefore, engineering C4 photosynthesis in C3 plants is of great importance in the application field. The genus Flaveria contains a large number of C3, C3-C4 intermediate, C4-like, and C4 species, making it a good model genus to study the evolution of C4 photosynthesis, and these studies indicate the direction for C4 engineering. C4 photosynthesis was acquired gradually through the C3-C4 intermediate stage. First, a two-celled C2 cycle called C2 photosynthesis was acquired by localizing glycine decarboxylase activity in the mitochondria of bundle sheath cells. With the development of two-cell metabolism, anatomical features also changed. Next, the replacement of the two-celled C2 cycle by the two-celled C4 cycle was induced by the acquisition of cell-selective expression in addition to the upregulation of enzymes in the C4 cycle during the C3-C4 intermediate stage. This was supported by an increase in cyclic electron transport activity in response to an increase in the ATP/NADPH demand for metabolism. Suppression of the C3 cycle in mesophyll cells was induced after the functional establishment of the C4 cycle, and optimization of electron transport by suppressing the activity of photosystem II also occurred during the final phase of C4 evolution.

Increased CO2 Relevant to Future Ocean Acidification Alleviates the Sensitivity of a Red Macroalgae to Solar Ultraviolet Irradiance by Modulating the Synergy Between Photosystems II and I.

While intertidal macroalgae are exposed to drastic changes in solar photosynthetically active radiation (PAR) and ultraviolet radiation (UVR) during a diel cycle, and to ocean acidification (OA) associated with increasing CO2 levels, little is known about their photosynthetic performance under the combined influences of these drivers. In this work, we examined the photoprotective strategies controlling electron flow through photosystems II (PSII) and photosystem I (PSI) in response to solar radiation with or without UVR and an elevated CO2 concentration in the intertidal, commercially important, red macroalgae Pyropia (previously Porphyra) yezoensis. By using chlorophyll fluorescence techniques, we found that high levels of PAR alone induced photoinhibition of the inter-photosystem electron transport carriers, as evidenced by the increase of chlorophyll fluorescence in both the J- and I-steps of Kautsky curves. In the presence of UVR, photoinduced inhibition was mainly identified in the O2-evolving complex (OEC) and PSII, as evidenced by a significant increase in the variable fluorescence at the K-step (Fk) of Kautsky curves relative to the amplitude of FJ−Fo (Wk) and a decrease of the maximum quantum yield of PSII (Fv/Fm). Such inhibition appeared to ameliorate the function of downstream electron acceptors, protecting PSI from over-reduction. In turn, the stable PSI activity increased the efficiency of cyclic electron transport (CET) around PSI, dissipating excess energy and supplying ATP for CO2 assimilation. When the algal thalli were grown under increased CO2 and OA conditions, the CET activity became further enhanced, which maintained the OEC stability and thus markedly alleviating the UVR-induced photoinhibition. In conclusion, the well-established coordination between PSII and PSI endows P. yezoensis with a highly efficient photochemical performance in response to UVR, especially under the scenario of future increased CO2 levels and OA.

OsCRP1, a Ribonucleoprotein Gene, Regulates Chloroplast mRNA Stability That Confers Drought and Cold Tolerance.

Chloroplast ribonucleoproteins (cpRNPs) are nuclear-encoded and highly abundant proteins that are proposed to function in chloroplast RNA metabolism. However, the molecular mechanisms underlying the regulation of chloroplast RNAs involved in stress tolerance are poorly understood. Here, we demonstrate that CHLOROPLAST RNA-BINDING PROTEIN 1 (OsCRP1), a rice (Oryza sativa) cpRNP gene, is essential for stabilization of RNAs from the NAD(P)H dehydrogenase (NDH) complex, which in turn enhances drought and cold stress tolerance. An RNA-immunoprecipitation assay revealed that OsCRP1 is associated with a set of chloroplast RNAs. Transcript profiling indicated that the mRNA levels of genes from the NDH complex significantly increased in the OsCRP1 overexpressing compared to non-transgenic plants, whereas the pattern in OsCRP1 RNAi plants were opposite. Importantly, the OsCRP1 overexpressing plants showed a higher cyclic electron transport (CET) activity, which is essential for elevated levels of ATP for photosynthesis. Additionally, overexpression of OsCRP1 resulted in significantly enhanced drought and cold stress tolerance with higher ATP levels compared to wild type. Thus, our findings suggest that overexpression of OsCRP1 stabilizes a set of mRNAs from genes of the NDH complex involved in increasing CET activity and production of ATP, which consequently confers enhanced drought and cold tolerance.

Moderate drought stress stabilizes the primary quinone acceptor QA and the secondary quinone acceptor QB in photosystem II.

Drought induces stomata closure and lowers the CO2 concentration in the mesophyll, limiting CO2 assimilation and favoring 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 (TL), 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 TL AG-band and a downshift of the maximum temperatures of both, the B-band and the AG-band, were observed when leaves were illuminated under conditions that maintained the proton gradient. When leaves were frozen prior to the TL 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.

The water-water cycle facilitates photosynthetic regulation under fluctuating light in the epiphytic orchid Dendrobium officinale

Fluctuating light can cause selective photoinhibition of photosystem I (PSI) in angiosperms. However, the strategies for photosynthetic regulation under fluctuating light in wild epiphytic plants are little known. We here compared PSI and PSII performances under fluctuating light in the epiphytic orchid Dendrobium officinale and the terrestrial orchid Bletilla striata. Upon dark-to-light transition, D. officinale showed a rapid re-oxidation of P700, but this rapid re-oxidation of P700 was not observed in B. striata. Within the first 40 s after transition from dark to actinic light, B. striata showed much higher activation of cyclic electron transport (CET) than D. officinale. However, this strong activation of CET could not prevent the over-reduction of PSI. Therefore, the rapid re-oxidation of P700 in D. officinale was attributed to pseudo-cyclic electron transport (water-water cycle). Within the first 60 s after transition from low to high light, PSI was highly oxidized in D. officinale but was highly reduced in B. striata. Concomitantly, B. striata showed much higher stimulation of CET. These results indicate that water-water cycle significantly regulates PSI redox state under fluctuating light in the epiphytic orchid D. officinale. By comparison, the terrestrial orchid B. striata employs CET to cope with fluctuating light, similar to the phenotype of Arabidopsis thaliana. Taking together, these two orchids used different strategies for photosynthetic regulation to cope with fluctuating light.

Comparison of changes in water status and photosynthetic parameters in wild type and abscisic acid-deficient sitiens mutant of tomato (Solanum lycopersicum cv. Rheinlands Ruhm) exposed to sublethal and lethal salt stress

Abscisic acid (ABA) regulates many salt stress-related processes of plants such as water balance, osmotic stress tolerance and photosynthesis. In this study we investigated the responses of wild type (WT) and the ABA-deficient sitiens mutant of tomato (Solanum lycopersicum cv. Rheinlands Ruhm) to sublethal and lethal salt stress elicited by 100 mM and 250 mM NaCl, respectively. Sitiens mutants displayed much higher decrease in water potential, stomatal conductance and net CO2 assimilation rate under high salinity, especially at lethal salt stress, than the WT. However, ABA deficiency in sitiens caused more severe osmotic stress and more moderate ionic stress, higher K+/Na+ ratio, in leaf tissues of plants exposed to salt stress. The higher salt concentration caused irreversible damage to Photosystem II (PSII) reaction centres, severe reduction in the linear photosynthetic electron transport rate and in the effective quantum yields of PSII and PSI in sitiens plants. The cyclic electron transport (CET) around PSI, which is an effective defence mechanism against the damage caused by photoinhibition in PSI, decreased in sitiens mutants, while WT plants were able to increase CET under salt stress. This suggests that the activation of CET needs active ABA synthesis and/or signalling. In spite of ABA deficiency, proline accumulation could alleviate the stress injury at sublethal salt stress in the mutants but its accumulation was not sufficient at lethal salt stress.

Role of Photorespiration and Cyclic Electron Transport in C4 Photosynthesis Evolution in the C3–C4 Intermediate Species Sedobassia sedoides

Plants from two Sedobassia sedoides (Pall.) Aschers populations (Makan and Valitovo) (Chenopodiaceae) with C2 photosynthesis (precursor of C4 photosynthesis in phylogenesis) and photorespiratory CO2-concentrating mechanism were studied. Genetic polymorphism and isotope discrimination (δ13С) levels of the plants were determined under natural conditions, and their morpho-physiological parameters such as fresh and dry biomass of the above ground parts of plants, functioning of photosystem I (PSI) and photosystem II (PSII), intensity of net photosynthesis (A), transpiration (E), photorespiration and water use efficiency (WUE) of plants were calculated under control and salinine conditions (0 and 200 mM NaCl). Results of the population-genetic analysis showed that the Makan population is polymorphic (plastic) and the Valitovo population is monomorphic (narrowly specialized). There were no significant differences between the populations based on δ13С values or growth parameters, PSII, A, E and WUE under control conditions. Under saline conditions, dry biomass accumulation decreased in the Makan population by 15% and by more than 2- fold in the Valitovo population. Population differences were revealed in terms of photorespiration intensity and P700 oxidation kinetics under control and saline conditions. Under control conditions, Makan plants were characterized by a higher photorespiration intensity, which decreased by 2-fold under saline conditions to the photorespiration level of Valitovo plants. Cyclic electron transport activity was minimal in the control Makan plants, and it increased by almost 2-fold under saline conditions to the level of that in Valitovo plants under control and saline conditions. Under control conditions, photosynthesis in Makan plants can be specified as the proto-Kranz type (transitional type from C3 to C2) and that in Valitovo plants can be specified as the C2 type (C4 photosynthesis with photorespiratory CO2-concentrating mechanism), based on their photorespiration level and cyclic electron transport activity. Under saline conditions, Makan plants exhibited features of C2 photosynthesis. Intraspecific functional differences of photosynthesis were revealed in different populations of intermediate C3–C4 plant species S. sedoides which reflect the initial stages of formation of a photorespiratory CO2-concentrating mechanism during C4 photosynthesis evolution, accompanied by decrease in salt tolerance.

Promotion of cyclic electron transport around photosystem I during the evolution of NADP–malic enzyme‐type C4 photosynthesis in the genus Flaveria

C4 plants display higher cyclic electron transport activity than C3 plants. This activity is suggested to be important for the production of ATPs required for C4 metabolism. To understand the process by which photosystem I (PSI) cyclic electron transport was promoted during C4 evolution, we conducted comparative analyses of the functionality of PSI cyclic electron transport among members of the genus Flaveria, which contains several C3, C3-C4 intermediate, C4-like and C4 species. The abundance of NDH-H, a subunit of NADH dehydrogenase-like complex, increased markedly in bundle sheath cells with the activity of the C4 cycle. By contrast, PROTON GRADIENT REGULATION5 (PGR5) and PGR5-LIKE1 increased in both mesophyll and bundle sheath cells in C4-like Flaveria palmeri and C4 species. Grana stacks were drastically reduced in bundle sheath chloroplasts of C4-like F. palmeri and C4 species; these species showed a marked increase in PSI cyclic electron transport activity. These results suggest that both the expression of proteins involved in PSI cyclic electron transport and changes in thylakoid structure contribute to the high activity of cyclic electron flow in NADP-malic enzyme-type C4 photosynthesis. We propose that these changes were important for the establishment of C4 photosynthesis from C3-C4 intermediate photosynthesis in Flaveria.

Physiological impact of mitochondrial alternative oxidase on photosynthesis and growth in Arabidopsis thaliana

The mitochondrial alternative oxidase (AOX) has been suggested to have a beneficial role in illuminated leaves, but its function has not yet been fully elucidated. In this study, we investigated the effects of a knockout of the AOX1a gene on photosynthesis and growth under several light conditions in Arabidopsis thaliana. The AOX-deficient aox1a mutant showed a lowered operating efficiency of photosystem II and an enhanced activity of cyclic electron transport around photosystem I (CET-PSI) at high irradiance. To further address the physiological association of AOX with CET-PSI, we crossed aox1a with the pgr5 mutant, which is impaired in CET-PSI activity. In the pgr5 mutant background, AOX deficiency did not affect the apparent photosynthetic efficiency, indicating that the direct contribution of AOX to photosynthesis is not so large compared with CET-PSI. Nevertheless, the growth of the aox1a pgr5 double mutant was significantly impaired depending on the light intensity under growth conditions. The possibility of a synergistic function of AOX with CET-PSI in supporting plant growth is discussed.

Induction kinetics of photosystem I-activated P700 oxidation in plant leaves and their dependence on pre-energization

Absorbance changes ΔA810 were measured in pea (Pisum sativum L., cv. Premium) leaves to track redox transients of chlorophyll P700 during and after irradiation with far red (FR) light under various preillumination conditions in the absence and presence of inhibitors and protonophorous uncoupler of photosynthetic electron transport. It was shown that cyclic electron transport (CET) in chloroplasts of pea leaves operates at its highest rate after preillumination of leaves with white light and is strongly suppressed after preillumination with FR light. The FR light-induced suppression was partly released during prolonged dark adaptation. Upon FR illumination of dark-adapted leaves, the induction of CET was observed, during which CET activity increased to the peak from the low level and then decreased gradually. The kinetics of P700 oxidation induced by FR light of various intensities in leaves preilluminated with white light were fit to empirical sigmoid curves containing two variables. In leaves treated with a protonophore FCCP, the amplitude of FR light-induced changes ΔA810 was strongly suppressed, indicating that the rate of CET is controlled by the pH gradient across the thylakoid membrane.

Acclimation of photosynthesis to nitrogen deficiency in Phaseolus vulgaris

Nitrogen deficiency diminishes consumption of photosynthates in anabolic metabolism. We studied adjustments of the photosynthetic machinery in nitrogen-deficient bean plants and found four phenomena. First, the number of chloroplasts per cell decreased. Chloroplasts of nitrogen starved leaves contained less pigments than those of control leaves, but the in vitro activities of light reactions did not change when measured on chlorophyll basis. Second, nitrogen deficiency induced cyclic electron transfer. The amounts of Rubisco and ferredoxin-NADP(+) reductase decreased in nitrogen starved plants. Low activities of these enzymes are expected to lead to increase in reduction of oxygen by photosystem I. However, diaminobenzidine staining did not reveal hydrogen peroxide production in nitrogen starved plants. Measurements of far-red-light-induced redox changes of the primary donor of photosystem I suggested that instead of producing oxygen radicals, nitrogen starved plants develop a high activity of cyclic electron transport that competes with oxygen for electrons. Nitrogen starvation led to decrease in photochemical quenching and increase in non-photochemical quenching, indicating that cyclic electron transport reduces the plastoquinone pool and acidifies the lumen. A third effect is redistribution of excitation energy between the photosystems in favor of photosystem I. Thus, thylakoids of nitrogen starved plants appeared to be locked in state 2, which further protects photosystem II by decreasing its absorption cross-section. As a fourth response, the proportion of non-Q(B)-reducing photosystem II reaction centers increased and the redox potential of the Q(B)/Q(B)(-) pair decreased by 25 mV in a fraction of photosystem II centers of nitrogen starved plants.

10.1007/s11183-008-2003-2

Effects of inhibitors and cofactors of cyclic and noncyclic electron transport on nonphotochemical quenching of chlorophyll fluorescence induced by action potential (AP) was investigated in Chara corallina cells. Under control conditions, energy-dependent quenching (qE) develops upon the increase in photosynthetically active radiation (PAR); it also arises and reversibly disappears after AP generation at moderate irradiances. The treatment of cells with diuron (DCMU) completely eliminated qE established at high irradiances and qE induced by AP generation. The activation of cyclic electron transport by DCMU in combination with phenazine methosulfate restored qE at high irradiances but did not restore qE imposed after AP generation. The presence in the medium of a PSI acceptor, methyl viologen at concentrations from 100 μM to 0.83 mM had no effect on fluorescence and photosynthetic activity of chloroplasts until the application of a single excitatory stimulus. Once a single AP was generated in the presence of methyl viologen, it induced irreversible qE at a wide range of irradiances, which indicates the AP-triggered redirection of a part of electron flow from the main pathway to the artificial acceptor. It is concluded that AP generation opens access for permeation of a divalent cation methyl viologen from the medium to the chloroplast stroma across two membrane barriers, the plasmalemma and the inner membrane of the chloroplast envelope.

Action potential opens access for the charged cofactor to the chloroplasts of Chara corallina cells

Effects of inhibitors and cofactors of cyclic and noncyclic electron transport on nonphotochemical quenching of chlorophyll fluorescence induced by action potential (AP) was investigated in Chara corallina cells. Under control conditions, energy-dependent quenching (qE) develops upon the increase in photosynthetically active radiation (PAR); it also arises and reversibly disappears after AP generation at moderate irradiances. The treatment of cells with diuron (DCMU) completely eliminated qE established at high irradiances and qE induced by AP generation. The activation of cyclic electron transport by DCMU in combination with phenazine methosulfate restored qE at high irradiances but did not restore qE imposed after AP generation. The presence in the medium of a PSI acceptor, methyl viologen at concentrations from 100 μM to 0.83 mM had no effect on fluorescence and photosynthetic activity of chloroplasts until the application of a single excitatory stimulus. Once a single AP was generated in the presence of methyl viologen, it induced irreversible qE at a wide range of irradiances, which indicates the AP-triggered redirection of a part of electron flow from the main pathway to the artificial acceptor. It is concluded that AP generation opens access for permeation of a divalent cation methyl viologen from the medium to the chloroplast stroma across two membrane barriers, the plasmalemma and the inner membrane of the chloroplast envelope.

A Synechococcus elongatus PCC 7942 mutant with a higher tolerance toward the herbicide bentazone also confers resistance to sodium chloride stress

Following a N-methyl-N'-nitro-N-nitrosoguanidine-based mutagenesis of Synechococcus elongatus PCC 7942 wild type, we were able to select several mutants with an enhanced tolerance toward the herbicide bentazone (3-isopropyl-1H-2,1,3-benzothiadiazine-4(3H)-one 2,2-dioxide). Mutant Mu1 has in part been previously characterized. In the present paper we report on another mutant, called Mu2, which also has a higher tolerance toward bentazone. Since Mu2 showed a better growth than WT when cultivated with elevated NaCl concentrations in the growth medium and since S. elongatus WT has previously been classified to be low salt tolerant, we were especially interested in the identification of the modifications conferring this higher salt tolerance to mutant Mu2. Immunoblot analyses provided evidence that Mu2 had a constitutively higher expression of PsbO and of IsiA. In addition, in Mu2 a significantly higher concentration of IdiA was detected under salt stress as compared to WT. These three proteins most likely contribute to a better protection and/or stabilization of photosystem II. Moreover, Mu2 had a higher amount of the photosystem I reaction center proteins PsaAB under salt stress than WT. In addition, the amount of the ferredoxin:NADP+ oxidoreductase and also of the ATP synthase was constitutively higher in Mu2 than in WT. In contrast to WT the latter two proteins did not decrease under salt stress in Mu2. Therefore, it can be assumed that Mu2 could maintain a high cyclic electron transport activity around photosystem I under salt stress. It can be assumed that the combination of these modifications of the electron transport chain cause a better protection of photosystem II against oxidative damage and cause an increase of cyclic electron transport activity around photosystem I with ATP synthesis. Thus, the overall cellular energization in Mu2 relative to WT is improved. Together with putative other not yet identified modifications this seems to enable Mu2 to energize its cytoplasmic membrane-localized ion pumps more effectively than WT and, as a consequence, to keep the intracellular NaCl concentration low.

Stromal Over-reduction by High-light Stress as Measured by Decreases in P700 Oxidation by Far-red Light and its Physiological Relevance

The oxidation level of P700 induced by far-red light (DeltaA(FR)) in briefly dark-treated leaves of some sun plants decreased during the daytime and recovered at night. The dark recovery of decreased DeltaA(FR) proceeded slowly, with a half-time of about 5 h. We propose that stromal over-reduction induced by sunlight was the direct cause of the depression of DeltaA(FR). The depression of DeltaA(FR) found during the daytime was reproduced by controlled illumination with saturating light of fully dark-treated leaves. Simultaneous measurement of P700 redox and chlorophyll fluorescence showed that the depression of DeltaA(FR) was associated with dark reduction of the plastoquinone pool, which represented cyclic electron transport activity. The decrease of DeltaA(FR) in the light-stressed chloroplasts was partly reversed by treatment with 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone, an inhibitor of electron transport at the cytochrome b6/f complex, and the subsequent addition of methyl viologen, an efficient electron acceptor from photosystem I (PSI), stimulated further recovery, showing that both cyclic electron flow around PSI and the charge recombination within PSI were responsible for the light-induced depression of DeltaA(FR). The dark level of blue-green fluorescence, an indicator of NAD(P)H concentration, from intact chloroplasts was increased by high-light stress, suggesting that NADPH accumulated in stroma as a result of the high-light treatment. Possible effects on photosynthetic activity of over-reduction and its physiological relevance are discussed.