Rate Of Cyclic Electron Flow Research Articles

Role of melatonin in promoting plant growth by regulating carbon assimilation and ATP accumulation

Melatonin (MT) is a phytohormone important in mediating diverse plant growth processes. In this study, we performed transcriptomic, qRT-PCR, physiological and biochemical analyses of Brassica rapa seedlings in order to understand how MT promotes plant growth. The results showed that exogenous MT increased the rate of cyclic electron flow around photosystem (PS) I, fluorescence quantum yield, and electron transport efficiency between PSII and PSI to promote the vegetative growth of B. rapa seedlings without affecting oxidative stress level, as compared to control. However, MT treatment significantly reduced photosynthetic rate (Pn), transpiration rate (Tr), and stomatal conductance (Gs) by 2.25-, 1.23- and 3.50-fold at 0.05 level, respectively. This occurred in parallel with the down-regulation of the genes for carbon fixation in photosynthetic organisms in a KEGG pathway enrichment. More accelerated plant growth despite the reduced photosynthesis rate and the enhanced electron transport rate suggested that NADPH and adenosine triphosphate (ATP) were preferentially diverted into other anabolic reactions than the Calvin cycle upon MT application. MT treatment increased ATP level and facilitated carbon assimilation into primary metabolism that led to a significant enhancement of soluble protein, sucrose, and fructose, but a significant decrease in glucose content. MT-induced carbon assimilation into primary metabolism was driven by up-regulation of the genes for glutathione metabolism, Krebs cycle, ribosome, and DNA replication in a KEGG pathway enrichment, as well as down-regulation of the genes for secondary metabolites. Our results provide an insight into MT-mediated metabolic adjustments triggered by coordinate changes in a wide range of gene expression profiles to help improve the plant functionality.

Glycinebetaine mitigates tomato chilling stress by maintaining high-cyclic electron flow rate of photosystem I and stability of photosystem II.

Glycinebetaine alleviates chilling stress by protecting photosystems I and II in BADH-transgenic and GB-treated tomato plants, which can be an effective strategy for improving crop chilling tolerance. Tomato (Solanum lycopersicum) is one of the most cultivated vegetables in the world, but is highly susceptible to chilling stress and does not naturally accumulate glycinebetaine (GB), one of the most effective stress protectants. The protective mechanisms of GB on photosystem I (PSI) and photosystem II (PSII) against chilling stress, however, remain poorly understood. Here, we address this problem through exogenous GB application and generation of transgenic tomatoes (Moneymaker) with a gene encoding betaine aldehyde dehydrogenase (BADH), which is the key enzyme in the synthesis of GB, from spinach. Our results demonstrated that GB can protect chloroplast ultramicrostructure, alleviate PSII photoinhibition and maintain PSII stability under chilling stress. More importantly, GB increased the electron transfer between QA and QB and the redox potential of QB and maintained a high rate of cyclic electron flow around PSI, contributing to reduced production of reactive oxygen species, thereby mitigating PSI photodamage under chilling stress. Our results highlight the novel roles of GB in enhancing chilling tolerance via the protection of PSI and PSII in BADH transgenic and GB-treated tomato plants under chilling stress. Thus, introducing GB-biosynthetic pathway into tomato and exogenous GB application are effective strategies for improving chilling tolerance.

A Cytosolic Bypass and G6P Shunt in Plants Lacking Peroxisomal Hydroxypyruvate Reductase.

The oxygenation of ribulose 1,5-bisphosphate by Rubisco is the first step in photorespiration and reduces the efficiency of photosynthesis in C3 plants. Our recent data indicate that mutants in photorespiration have increased rates of photosynthetic cyclic electron flow around photosystem I. We investigated mutant lines lacking peroxisomal hydroxypyruvate reductase to determine if there are connections between 2-phosphoglycolate accumulation and cyclic electron flow in Arabidopsis (Arabidopsis thaliana). We found that 2-phosphoglycolate is a competitive inhibitor of triose phosphate isomerase, an enzyme in the Calvin-Benson cycle that converts glyceraldehyde 3-phosphate to dihydroxyacetone phosphate. This block in metabolism could be overcome if glyceraldehyde 3-phosphate is exported to the cytosol, where cytosolic triose phosphate isomerase could convert it to dihydroxyacetone phosphate. We found evidence that carbon is reimported as glucose-6-phosphate, forming a cytosolic bypass around the block of stromal triose phosphate isomerase. However, this also stimulates a glucose-6-phosphate shunt, which consumes ATP, which can be compensated by higher rates of cyclic electron flow.

Low temperature induced modulation of photosynthetic induction in non-acclimated and cold-acclimated Arabidopsis thaliana: chlorophyll a fluorescence and gas-exchange measurements.

Cold acclimation modifies the photosynthetic machinery and enables plants to survive at sub-zero temperatures, whereas in warm habitats, many species suffer even at non-freezing temperatures. We have measured chlorophyll a fluorescence (ChlF) and CO2 assimilation to investigate the effects of cold acclimation, and of low temperatures, on a cold-sensitive Arabidopsis thaliana accession C24. Upon excitation with low intensity (40µmol photons m- 2 s- 1) ~ 620nm light, slow (minute range) ChlF transients, at ~ 22°C, showed two waves in the SMT phase (S, semisteady-state; M, maximum; T, terminal steady-state), whereas CO2 assimilation showed a linear increase with time. Low-temperature treatment (down to - 1.5°C) strongly modulated the SMT phase and stimulated a peak in the CO2 assimilation induction curve. We show that the SMT phase, at ~ 22°C, was abolished when measured under high actinic irradiance, or when 3-(3, 4-dichlorophenyl)-1, 1- dimethylurea (DCMU, an inhibitor of electron flow) or methyl viologen (MV, a Photosystem I (PSI) electron acceptor) was added to the system. Our data suggest that stimulation of the SMT wave, at low temperatures, has multiple reasons, which may include changes in both photochemical and biochemical reactions leading to modulations in non-photochemical quenching (NPQ) of the excited state of Chl, "state transitions," as well as changes in the rate of cyclic electron flow through PSI. Further, we suggest that cold acclimation, in accession C24, promotes "state transition" and protects photosystems by preventing high excitation pressure during low-temperature exposure.

Phenotyping of isogenic chlorophyll-less bread and durum wheat mutant lines in relation to photoprotection and photosynthetic capacity.

In our experiments, we examined high light responses and photosynthetic capacity of chlorophyll-less isogenic mutant lines of hexaploid bread wheat (Triticum aestivum L.) and tetraploid durum wheat (Triticum durum L.) in comparison to parental lines representing the wild type (WT), in two growth phases and two environments. In young plants, we observed a typical yellow-green phenotype with low chlorophyll content, significantly lower CO2 assimilation rate, elevated chlorophyll a-to-b ratio and insufficient regulation of linear electron transport. In the mutants grown in a moderate light in the growth chamber, a typical "chlorina" phenotype almost disappeared or, at least, was significantly alleviated in later growth stages, including the values of CO2 assimilation and the majority of the measured parameters related to photoprotective responses. On the other hand, in the case of the mutant lines grown in direct sunlight and fluctuating environment, the chlorophyll-less phenotype was evident also in latter growth phases. The chlorophyll-less phenotype was more severe in the durum wheat mutant lines compared to the bread wheat. For example, the durum wheat mutant lines grown outdoors expressed lower flexibility of photoprotective responses, including lower non-photochemical quenching and low rate of cyclic electron flow compared to WT or bread wheat mutants. Based on the analyses, we have identified a set of parameters providing information on the specific photosynthetic traits typical for the chlorophyll-less phenotype. Thus, the proposed way of phenotyping may serve for efficient selection of mutant genotypes for future research or screening activities. As a general result, we observed that the decrease of the chlorophyll content due to mutation was always associated with improper regulation of linear electron transport and a limited ability to prevent over-reduction of PSI acceptor side, regardless of the genotype, environment, and growth stage. This can partly explain why the low chlorophyll mutants were not successful in the evolution of higher plants, despite the photosynthetic capacity observed is high enough and they are fully competitive with wild-type plants in non-fluctuating controlled environment.

Plasticity in roles of cyclic electron flow around photosystem I at contrasting temperatures in the chilling-sensitive plant Calotropis gigantea

Cyclic electron flow (CEF) around photosystem I is thought to balance the ATP/NADPH energy budget and protect photosynthetic apparatus. However, the plasticity in roles of CEF at contrasting temperatures is not well understood. We examined photosynthetic electron flow, non-photochemical quenching (NPQ), PSI redox state, and electrochromic shift (ECS) signal at 25°C (normal temperature) and 4°C (low temperature) for leaves of a chilling-sensitive tree species Calotropis gigantea. We found that electron flow through PSII (ETRII) was largely depressed at 4°C irrespective of light condition, whereas CEF was enhanced under light intensities below 500μmol photons m−2s−1 but suppressed under light intensities above 760μmol photons m−2s−1. Meanwhile, both NPQ and PSI donor side limitation [Y(ND)] were enhanced at 4°C. Moreover, the relationships between the rate of CEF and values for NPQ and Y(ND) changed between 4°C and 25°C. At low temperature, the proton gradient across thylakoid membranes (ΔpH) was enhanced and the thylakoid proton conductivity was depressed. Based on these results, we propose that, at normal growth temperature such as 25°C, CEF contributes mainly to the balancing of ATP/NADPH energy budget required by primary metabolism. In contrast, at low temperature such as 4°C, the main function of the CEF-dependent proton motive force is lumenal acidification, favoring photoprotection for photosynthetic apparatus. The plasticity in roles of CEF at contrasting temperatures may be regulated by the activity of thylakoid ATP synthase.

Defects in the Expression of Chloroplast Proteins Leads to H2O2 Accumulation and Activation of Cyclic Electron Flow around Photosystem I.

We describe a new member of the class of mutants in Arabidopsis exhibiting high rates of cyclic electron flow around photosystem I (CEF), a light-driven process that produces ATP but not NADPH. High cyclic electron flow 2 (hcef2) shows strongly increased CEF activity through the NADPH dehydrogenase complex (NDH), accompanied by increases in thylakoid proton motive force (pmf), activation of the photoprotective qE response, and the accumulation of H2O2. Surprisingly, hcef2 was mapped to a non-sense mutation in the TADA1 (tRNA adenosine deaminase arginine) locus, coding for a plastid targeted tRNA editing enzyme required for efficient codon recognition. Comparison of protein content from representative thylakoid complexes, the cytochrome bf complex, and the ATP synthase, suggests that inefficient translation of hcef2 leads to compromised complex assembly or stability leading to alterations in stoichiometries of major thylakoid complexes as well as their constituent subunits. Altered subunit stoichiometries for photosystem I, ratios and properties of cytochrome bf hemes, and the decay kinetics of the flash-induced thylakoid electric field suggest that these defect lead to accumulation of H2O2 in hcef2, which we have previously shown leads to activation of NDH-related CEF. We observed similar increases in CEF, as well as increases in H2O2 accumulation, in other translation defective mutants. This suggests that loss of coordination in plastid protein levels lead to imbalances in photosynthetic energy balance that leads to an increase in CEF. These results taken together with a large body of previous observations, support a general model in which processes that lead to imbalances in chloroplast energetics result in the production of H2O2, which in turn activates CEF. This activation could be from either H2O2 acting as a redox signal, or by a secondary effect from H2O2 inducing a deficit in ATP.

Impacts of genetically engineered alterations in carbon sink pathways on photosynthetic performance

Genetic engineering of photosynthetic organisms typically redirects native metabolism towards desirable products, which thereby represent new metabolic sinks. There is limited information on how these modifications impact the evolved mechanisms of photosynthetic energy metabolism and cellular growth. Two engineered strains of Synechocystis sp. PCC 6803 with altered carbon sink capacity were assayed for their photosynthetic and CO2 concentrating mechanism properties in conditions of high and low inorganic carbon (Ci) availability. In the ΔglgC mutant, glycogen cannot be synthesized and a carbon sink pathway has been effectively removed. The JU547 strain has been engineered by integration of the Pseudomonas syringae ethylene forming enzyme and provides a new sink. When cultured under high carbon conditions, ΔglgC displayed diminished photochemical efficiency, a more reduced NADPH pool, delayed initiation of the Calvin-Benson-Bassham cycle, and impairment of linear and cyclic electron flows. It also exhibited a large decrease in photochemical quenching indicative of the accumulation of QA−, normally associated with a reduced PQ pool, but appears instead to be the result of an undefined dissipative mechanism to spill excess energy. In the case of carbon sink integration, JU547 displayed slightly more oxidized PQ and NADPH pools and increased rates of cyclic electron flow and an enhanced demand for inorganic carbon as suggested by increase in the expression of the bicarbonate transporter, SbtA. Overall, the results highlight the importance of the native regulatory network of autotrophic metabolism in governing photosynthetic performance and provide cogent examples of both predicable and difficult to predict phenotypic consequences upon installation of new pathways in autotrophs.

Effect of mono- and dichromatic light quality on growth rates and photosynthetic performance of Synechococcus sp. PCC 7002.

Synechococcus sp. PCC 7002 was grown to steady state in optically thin turbidostat cultures under conditions for which light quantity and quality was systematically varied by modulating the output of narrow-band LEDs. Cells were provided photons absorbed primarily by chlorophyll (680 nm) or phycocyanin (630 nm) as the organism was subjected to four distinct mono- and dichromatic regimes. During cultivation with dichromatic light, growth rates were generally proportional to the total incident irradiance at values <275 μmol photons m−2 · s−1 and were not affected by the ratio of 630:680 nm wavelengths. Notably, under monochromatic light conditions, cultures exhibited similar growth rates only when they were irradiated with 630 nm light; cultures irradiated with only 680 nm light grew at rates that were 60–70% of those under other light quality regimes at equivalent irradiances. The functionality of photosystem II and associated processes such as maximum rate of photosynthetic electron transport, rate of cyclic electron flow, and rate of dark respiration generally increased as a function of growth rate. Nonetheless, some of the photophysiological parameters measured here displayed distinct patterns with respect to growth rate of cultures adapted to a single wavelength including phycobiliprotein content, which increased under severely light-limited growth conditions. Additionally, the ratio of photosystem II to photosystem I increased ~40% over the range of growth rates, although cells grown with 680 nm light only had the highest ratios. These results suggest the presence of effective mechanisms which allow acclimation of Synechococcus sp. PCC 7002 acclimation to different irradiance conditions.

Redox and ATP control of photosynthetic cyclic electron flow in Chlamydomonas reinhardtii: (II) Involvement of the PGR5–PGRL1 pathway under anaerobic conditions

In oxygenic photosynthesis, cyclic electron flow around photosystem I denotes the recycling of electrons from stromal electron carriers (reduced nicotinamide adenine dinucleotide phosphate, NADPH, ferredoxin) towards the plastoquinone pool. Whether or not cyclic electron flow operates similarly in Chlamydomonas and plants has been a matter of debate. Here we would like to emphasize that despite the regulatory or metabolic differences that may exist between green algae and plants, the general mechanism of cyclic electron flow seems conserved across species. The most accurate way to describe cyclic electron flow remains to be a redox equilibration model, while the supramolecular reorganization of the thylakoid membrane (state transitions) has little impact on the maximal rate of cyclic electron flow. The maximum capacity of the cyclic pathways is shown to be around 60 electrons transferred per photosystem per second, which is in Chlamydomonas cells treated with 3(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) and placed under anoxic conditions. Part I of this work (aerobic conditions) was published in a previous issue of BBA-Bioenergetics (vol. 1797, pp. 44–51) (Alric et al., 2010).

Involvement of the thylakoidal NADH-plastoquinone-oxidoreductase complex in the early responses to ozone exposure of barley (Hordeum vulgare L.) seedlings

A possible implication of the plastid NADH-plastoquinone-oxidoreductase (Ndh) complex in the response against ozone-mediated oxidative stress in barley (Hordeum vulgare L.) leaves was investigated. After a 4 h treatment, exposure of barley seedlings to moderate ozone concentrations produced leaf-age-dependent increases in lipid peroxidation, peroxidase, and Ndh complex activities in the thylakoid membranes. A significant amount and activity of the Ndh complex were detected in mature barley leaves, but not in young barley leaves. In fact, young barley leaves behaved like ndh-deficient leaves in most of the aspects studied. When plants were exposed to photo-oxidative light after ozone fumigation, the recovery of Fv/Fm was lower in young leaves than in mature leaves. Ozone treatment significantly decreased non-photochemical quenching (qN) in young leaves, but not in mature leaves. Mature leaves showed higher levels of the energy (DeltamuH+) dependent (qE) component of qN. Treatment with antimycin A, an inhibitor of cyclic electron flow, increased the decay of qN produced by ozone in young leaves, but not in mature ones. The reduction state of plastoquinone increased after ozone treatment in mature dark-adapted leaves and was strongly quenched by far red light. It is proposed that the function of the Ndh complex helps the maintenance of qN, probably through the poising of the redox steady-state level of the intersystem carriers and then by optimizing the rate of cyclic electron flow. This should constitute an age-dependent early response in barley leaves, by contributing to minimize photoinhibition in the presence of ozone and high light.

Cyclic electron flow under saturating excitation of dark-adapted Arabidopsis leaves

The rate of cyclic electron flow measured in dark-adapted leaves under aerobic conditions submitted to a saturating illumination has been performed by the analysis of the transmembrane potential changes induced by a light to dark transfer. Using a new highly sensitive spectrophotometric technique, a rate of the cyclic flow of ∼130 s −1 has been measured in the presence or absence of 3-(3,4-dichloro-phenyl)-1,1-dimethylurea (DCMU). This value is ∼1.5 times larger than that previously reported [Proc. Natl. Acad. Sci. U. S. A. 99 (2001) 10209]. We have characterized in the presence or absence of DCMU charge recombination process ( t 1/2∼60 μs) that involves P 700 + and very likely the reduced form of the iron sulfur acceptor F X. This led to conclude that, under saturating illumination, the PSI centers involved in the cyclic pathway have most of the iron sulfur acceptors F A and F B reduced. In the proposed mechanism, electrons are transferred from a ferredoxin bound to a site localized on the stromal side of the cytochrome b 6 f complex to the Q i site. Two possible models of the organization of the membrane complexes are discussed, in which the cyclic and linear electron transfer chains are isolated one from the other.

Characterization of the Light-Induced Oxygen Gas Exchange from the IC 2 Deletion Mutant of Synechocystis PCC 6803 Lacking the Photosystem II 33 kDa Extrinsic Protein

Abstract The absence of the extrinsic Mn-stabilizing 33 kDa protein in the IC 2 mutant of Synechocystis PCC 6803 disturbs the redox cycling of the water splitting system and retards the formation of its higher S-states (I. Vass, K. Cook, S. Deak, S. R. Mayes, and J. Barber, Biochim. Biophys. Acta 1102, 195-201 (1992)). We have performed analyses of the flashinduced oxygen exchange in the mutated cyanobacterium to clarify further the role of the 33 kDa protein. Under aerobic conditions, both the wild type and IC2 mutant show a relatively slow signal of oxygen rise on the first flash which is increased about twice by the addition of 10 μᴍ DCMU and significantly diminished by lowering the oxygen concentration in the medium. According to action spectra measurements, this mode of apparent oxygen release is mediated by PS I and can be attributed to a light induced inhibition of respiratory activity. In contrast to the wild type, having the usual oxygen evolution flash pattern with a periodicity of four, the IC2 mutant shows a binary oscillation pattern of flash-induced respiratory oxygen exchange at a flash frequency 10 Hz, being dampened with DCMU or by a lower flash frequency (&lt; 1 Hz). Oxygen evolution due to water splitting is clearly seen in the IC2 mutant when background far-red illumination is applied to saturate the signal due to respiratory inhibition, but a quadruple oscillatory component of flash-induced oxygen evolution appears only in the presence of artificial electron acceptors under partial aerobic conditions. The mutant possesses a higher PS I/PS II ratio compared to the wild type, as judged from both the flashinduced yields and quantum efficiencies of the steady-state rates of the oxygen exchange reactions. Estimates of antenna sizes indicate about a 20% decrease of optical cross-section at 675 nm of the PS II unit in IC2 mutants in comparison with the wild type. It is suggested that the absence of the 33 kDa protein leads to a modification of the PS II assembly and because of the slowing down of the S-state cycle, the rate of cyclic electron flow around PS II is enhanced. It seems that the absence of the 33 kDa protein in Synechocystis 6803 also disturbs energy transfer between adjacent PS II core complexes and may also alter their association with the phycobilisomes.

Biosynthesis of the photosynthetic membranes of Rhodopseudomonas sphaeroides.

The steady-state biosynthesis of the photosynthetic membrane (ICM) of Rhodopseudomonas sphaeroides has been reviewed. At moderate light intensities, 500 ft-c, preexisting ICM serves as the insertion matrix for newly synthesized membrane components. Whereas the bulk of the membrane protein, protein-pigment complexes, and pigments are inserted into preexisting ICM throughout the cell cycle, phospholipid is transferred from outside the ICM to the ICM only at the time of cell division. Because the site of cellular phospholipid synthesis is the cytoplasmic membrane, these results infer that despite the physical continuity of cytoplasmic membrane and ICM, there must exist between these membranous domains a "barrier" to the free diffusion of cellular phospholipid. The cyclical alternation in protein to phospholipid ratio of the ICM infers major structural and functional alternations, such as changes in the protein to lipid ratio of the membrane, specific density of the membrane, lipid structure within the membrane, and the rate of cyclic electron flow. When biochemical studies are correlated with detailed electron microscopic investigations we can further conclude that the number of photosynthetic units within the plane of the membrane can vary by nearly a factor of two over the course of the cell cycle. The average physical size of the photosynthetic units is constant for a given light intensity but inversely proportional to light intensity. The distribution of photosynthetic unit size classes within the membrane can be interpreted as suggesting that the "core" of the photosynthetic unit (reaction center plus fixed antenna complex) is inserted into the membrane coordinately as a structural entity. The variable antenna complex is, on the other hand, inserted independent of the "core" and randomly associates with both old and new core complexes. Finally, we conclude that there is substantial substructure to te distribution of photosynthetic units within the ICM, ie, they are highly ordered and exist in a defined spatial orientation to one another.

Cyclic electron transport in isolated intact chloroplasts. Further studies with antimycin

Antimycin has been used to study the role of cyclic electron transport in isolated intact chloroplasts maintained under aerobic conditions. At all light intensities, antimycin inhibits CO 2 fixation when assay conditions are optimal. When turnover of the Calvin cycle is inhibited, antimycin stimulates bicarbonate-dependent O 2 evolution. Energy-dependent processes such as chlorophyll a and 9-aminoacridine fluorescence quenching, and light-scattering (apparent absorption) changes are inhibited by antimycin. The results suggest that cyclic electron transport contributes to photophosphorylation under aerobic conditions and is obligatory as a source of ATP during the most active periods of CO 2 fixation in vivo. Cyclic electron transport can be stimulated either by inhibiting Photosystem II activity or increasing the turnover of Photosystem I relative to Photosystem II. These effects are interpreted in terms of the need for correct redox poising of carriers in the pathway in order to sustain maximum rates of cyclic electron flow. Binding studies indicate the presence of a high affinity antimycin binding site on chloroplast membranes. The stoichiometry and dissociation constant of the high affinity site are consistent with the idea that antimycin inhibits cyclic electron transport by binding to a b-type cytochrome in the thylakoid membrane.

Wavelength independence of light-induced cyclic electron transport in chlorella and anacystis cells

The rate of cyclic electron transport is assessed on the basis of measurements of the light-induced absorption changes at 700 nm in the presence of 3-(3′4′-dichlorophenyl)-1,1-dimethylurea (DCMU). No effects of light quality on the relative rates of cyclic electron flow are detected.