Photosynthetic Electron Transfer Chain Research Articles

Transcriptomic and proteomic analyses of distinct Arabidopsis organs reveal high PSI-NDH complex accumulation in stems.

In addition to leaves, the main site of photosynthetic reactions, active photosynthesis also takes place in stems, siliques and tree trunks. Although non-foliar photosynthesis has a marked effect on plant growth and yield, only limited information on the expression patterns of photosynthesis-related genes and the structure of photosynthetic machinery in different plant organs has been available. Here, we report the results of transcriptomic analysis of various organs of Arabidopsis thaliana and compare the gene expression profiles of young and mature leaves with a special focus on photosynthetic genes. Further, we analyzed the composition and organization of the photosynthetic electron transfer machinery in leaves, stems and green siliques at the protein level using BN-PAGE. RNA-Seq analysis revealed unique gene expression profiles in different plant organs and showed major differences in the expression of photosynthesis-related genes in young as compared to mature rosettes. Gel-based proteomic analysis of the thylakoid protein complex organization further showed that all studied plant organs contain the necessary components of the photosynthetic electron transfer chain. Intriguingly, stems accumulate high amounts of PSI-NDH complex, which has previously been implicated in cyclic electron transfer.

Modulation of the wheat transcriptome by TaZFP13D under well-watered and drought conditions

Maintaining global food security in the context of climate changes will be an important challenge in the next century. Improving abiotic stress tolerance of major crops such as wheat can contribute to this goal. This can be achieved by the identification of the genes involved and their use to develop tools for breeding programs aiming to generate better adapted cultivars. Recently, we identified the wheat TaZFP13D gene encoding Zinc Finger Protein 13D as a new gene improving water-stress tolerance. The current work analyzes the TaZFP13D-dependent transcriptome modifications that occur in well-watered and dehydration conditions to better understand its function during normal growth and during drought. Plants that overexpress TaZFP13D have a higher biomass under well-watered conditions, indicating a positive effect of the protein on growth. Survival rate and stress recovery after a severe drought stress are improved compared to wild-type plants. The latter is likely due the higher activity of key antioxidant enzymes and concomitant reduction of drought-induced oxidative damage. Conversely, down-regulation of TaZFP13D decreases drought tolerance and protection against drought-induced oxidative damage. RNA-Seq transcriptome analysis identified many genes regulated by TaZFP13D that are known to improve drought tolerance. The analysis also revealed several genes involved in the photosynthetic electron transfer chain known to improve photosynthetic efficiency and chloroplast protection against drought-induced ROS damage. This study highlights the important role of TaZFP13D in wheat drought tolerance, contributes to unravel the complex regulation governed by TaZFPs, and suggests that it could be a promising marker to select wheat cultivars with higher drought tolerance.

2023 F. M. Becket Fellowship Fellowship – Summary Report: Illuminating Photo-enhanced Bioelectrocatalysis in Purple Bacteria

Purple bacteria are a special subclass of photosynthetic bacteria known for their metabolic versatility, resistance to salinity, and bright red-violet pigmentation responsible for photosynthesis. Previous work by the Minteer group has demonstrated purple bacteria to be a viable electrochemical solution for sustainable decontamination of saline wastewater, in addition to biosensing and bio-electrosynthesis applications. Notably, the bacteria’s mechanism of transferring electrons to electrodes is directly related to their photosynthetic electron transfer chain, and current density is significantly enhanced in the presence of light. Often, the light sources used for photo-bioelectrochemistry experimental studies are high-intensity (∼100 mW per cm2) and not wavelength-specific. This leads to uncertainty of the mechanism of photo-enhanced bioelectrocatalysis and may also lead to photo-inhibition at higher light intensities. A novel method was developed to study the effect of isolated light wavelengths on photo-enhanced current.

UbiN, a novel Rhodobacter capsulatus decarboxylative hydroxylase involved in aerobic ubiquinone biosynthesis.

Ubiquinone (UQ) is a lipophilic electron carrier that functions in the respiratory and photosynthetic electron transfer chains of proteobacteria and eukaryotes. Bacterial UQ biosynthesis is well studied in the gammaproteobacterium Escherichia coli, in which most bacterial UQ-biosynthetic enzymes have been identified. However, these enzymes are not always conserved among UQ-containing bacteria. In particular, the alphaproteobacterial UQ biosynthesis pathways contain many uncharacterized steps with unknown features. In this work, we identified in the alphaproteobacterium Rhodobacter capsulatus a new decarboxylative hydroxylase and named it UbiN. Remarkably, the UbiN sequence is more similar to a salicylate hydroxylase than the conventional flavin-containing UQ-biosynthetic monooxygenases. Under aerobic conditions, R. capsulatus ΔubiN mutant cells accumulate 3-decaprenylphenol, which is a UQ-biosynthetic intermediate. In addition, 3-decaprenyl-4-hydroxybenzoic acid, which is the substrate of UQ-biosynthetic decarboxylase UbiD, also accumulates in ΔubiN cells under aerobic conditions. Considering that the R. capsulatus ΔubiD-X double mutant strain (UbiX produces a prenylated FMN required for UbiD) grows as a wild-type strain under aerobic conditions, these results indicate that UbiN catalyzes the aerobic decarboxylative hydroxylation of 3-decaprenyl-4-hydroxybenzoic acid. This is the first example of the involvement of decarboxylative hydroxylation in ubiquinone biosynthesis. This finding suggests that the C1 hydroxylation reaction is, at least in R. capsulatus, the first step among the three hydroxylation steps involved in UQ biosynthesis. Although the C5 hydroxylation reaction is often considered to be the first hydroxylation step in bacterial UQ biosynthesis, it appears that the R. capsulatus pathway is more similar to that found in mammalians.

Effect of Different Monochromatic LEDs on the Environmental Adaptability of Spathiphyllum floribundum and Chrysanthemum morifolium.

Light-emitting diodes (LEDs) can be programmed to provide specialized light sources and spectra for plant growth. UV-A (397.6 nm), blue (460.6 nm), green (520.7 nm), and red (661.9 nm) LED light sources were used to study the effects of different monochromatic lights on the growth, antioxidant system, and photosynthetic characteristics of Spathiphyllum floribundum 'Tian Jiao' (a shade-loving species) and Chrysanthemum morifolium 'Huang Xiu Qiu' (a sun-loving species). This research revealed that green and blue light could enhance the morphological indicators, Chl a/b, photosynthetic electron transfer chain performance, and photosystem activity of S. floribundum, blue and red light could enhance the solution protein, Chl a, and photosynthetic electron transfer chain performance of C. morifolium, red and UV-A light viewed the highest SOD and CAT activities of S. floribundum (275.56 U·min·g-1; 148.33 U·min·g-1) and C. morifolium (587.03 U·min·g-1; 98.33 U·min·g-1), respectively. Blue and green light were more suitable for the growth and development of the shade-loving plant S. floribundum, while red and blue light were more suitable for the sun-loving plant C. morifolium. UV-A light could be used for their stress research. The research revealed the different adaptation mechanism of different plants to light environmental conditions.

Metabolic changes associated with dark-induced leaf senescence in Arabidopsis nadk2 mutants

ABSTRACT Arabidopsis NADK2 (NAD kinase 2) is a chloroplast-localized enzyme involved in NADP+ synthesis, which acts as the final electron acceptor in the photosynthetic electron transfer chain. The NADK2-deficient mutant (nadk2) was used to analyze the effect of NAD(P)(H) unbalance in the dark-induced leaf senescence. During senescence, WT plants and nadk2 mutants showed a similar reduction in chlorophyll content. NAD(P)(H) quantification showed that the amount of total NAD(P)(H) decreased on the day 7 in WT but on the day 3 in nadk2. The phosphorylation ratio (i.e. NADP(H)/NAD(H)) decreased on day 1 in WT. In contrast, the nadk2 showed lower phosphorylation ratio at 0 day and no change throughout the aging process. Metabolome analysis showed that the metabolic profiles of both WT plants and nadk2 mutants subjected to dark-induced senescence adopted similar patterns as the senescence progressed. However, the changes in individual metabolites in the nadk2 mutants were different from those of the WT during dark-induced senescence.

Impact of cytotoxic plant naphthoquinones, juglone, plumbagin, lawsone and 2-methoxy-1,4-naphthoquinone, on Chlamydomonas reinhardtii reveals the biochemical mechanism of juglone toxicity by rapid depletion of plastoquinol

Hydrophilic, untethered 1,4-naphthoquinones (1,4-NQs) are plant secondary metabolites that are often excreted into the environment and play a role in various plant–microbial, plant–fungal, plant–insect and plant–plant interactions. The biological activity of 1,4-NQs is mainly related to their redox properties, i.e. the ability to undergo redox cycling in cells. These compounds may also undergo electrophilic addition to thiol-containing compounds. The aim of this study was to compare the impact of juglone, plumbagin, lawsone and 2-methoxy-1,4-naphthoquinone (2-met-NQ) on the antioxidant response of the green microalga Chlamydomonas reinhardtii. The algae were incubated with the examined compounds under low light for 6 h and the content of photosynthetic pigments, prenyllipid antioxidants, ascorbate, soluble thiols, proline, and superoxide dismutase activity was assessed. To examine the interaction between photosynthetic activity and naphthoquinone toxicity, we carried out the second experiment, in which C. reinhardtii was incubated with 1,4-NQs for 1 h under high light or in darkness. The pro-oxidant action of the examined 1,4-NQs depended on their reduction potentials, which decrease in order: juglone > plumbagin > 2-met-NQ > lawsone. Lawsone did not display pro-oxidant properties. Exposure to high light strongly enhanced the pro-oxidant effect of juglone, plumbagin, and 2-met-NQ, which is thought to result from the interception of the electrons from photosynthetic electron transfer chain. Only juglone was able to cause a fast depletion of plastoquinol, which may be an important mode of action of this allelochemical, responsible for its high toxicity to plants.

Cryo-EM structure of the four-subunit Rhodobacter sphaeroides cytochrome bc1 complex in styrene maleic acid nanodiscs

Cytochrome bc1 complexes are ubiquinol:cytochrome c oxidoreductases, and as such, they are centrally important components of respiratory and photosynthetic electron transfer chains in many species of bacteria and in mitochondria. The minimal complex has three catalytic components, which are cytochrome b, cytochrome c1, and the Rieske iron-sulfur subunit, but the function of mitochondrial cytochrome bc1 complexes is modified by up to eight supernumerary subunits. The cytochrome bc1 complex from the purple phototrophic bacterium Rhodobactersphaeroides has a single supernumerary subunit called subunit IV, which is absent from current structures of the complex. In this work we use the styrene-maleic acid copolymer to purify the R. sphaeroides cytochrome bc1 complex in native lipid nanodiscs, which retains the labile subunit IV, annular lipids, and natively bound quinones. The catalytic activity of the four-subunit cytochrome bc1 complex is threefold higher than that of the complex lacking subunit IV. To understand the role of subunit IV, we determined the structure of the four-subunit complex at 2.9 Å using single particle cryogenic electron microscopy. The structure shows the position of the transmembrane domain of subunit IV, which lies across the transmembrane helices of the Rieske and cytochrome c1 subunits. We observe a quinone at the Qo quinone-binding site and show that occupancy of this site is linked to conformational changes in the Rieske head domain during catalysis. Twelve lipids were structurally resolved, making contacts with the Rieske and cytochrome b subunits, with some spanning both of the two monomers that make up the dimeric complex.

PH-dependent redox properties of epigallocatechin gallate (EGCG) and its effect on respiration, photosynthesis and cell death in pea

The redox properties of the green tea component epigallocatechin gallate (EGCG) in vitro were studied and its effect on plant (pea) cells was tested. EGCG exhibited both pro- and antioxidant properties. In solutions, EGCG was oxidized by oxygen at physiological (slightly alkaline) pH values. Lowering the pH slowed down this process. Oxidation of EGCG was accompanied by the formation of O2-• and H2O2. In addition, EGCG functioned as an electron donor for peroxidase and, in combination with it, utilized H2O2. Exposure of EGCG to pea cells (leaf cuts or epidermis) suppressed respiration, reduced the transmembrane electric potential difference (Δψ) in mitochondria, and inhibited electron transfer in the photosynthetic electron transfer chain. Among the sites of the photosynthetic redox chain, Photosystem II was the least sensitive to the action of EGCG. EGCG reduced the rate of formation of reactive oxygen species in the epidermis, which was induced by NADH treatment and was registered using 2′,7′-dichlorofluorescine diacetate. In vivo, EGCG at concentrations from 10 µM to 1 mM suppressed KCN-induced death of guard cells in the epidermis, which was determined from the destruction of cell nuclei. EGCG at a concentration of 10 mM disrupted the barrier function of the guard cell plasma membrane, increasing its permeability to propidium iodide.

Triplet states in the reaction center of Photosystem II.

In oxygenic photosynthesis sunlight is harvested and funneled as excitation energy into the reaction center (RC) of Photosystem II (PSII), the site of primary charge separation that initiates the photosynthetic electron transfer chain. The chlorophyll ChlD1 pigment of the RC is the primary electron donor, forming a charge-separated radical pair with the vicinal pheophytin PheoD1 (ChlD1+PheoD1-). To avert charge recombination, the electron is further transferred to plastoquinone QA, whereas the hole relaxes to a central pair of chlorophylls (PD1PD2), subsequently driving water oxidation. Spin-triplet states can form within the RC when forward electron transfer is inhibited or back reactions are favored. This can lead to formation of singlet dioxygen, with potential deleterious effects. Here we investigate the nature and properties of triplet states within the PSII RC using a multiscale quantum-mechanics/molecular-mechanics (QM/MM) approach. The low-energy spectrum of excited singlet and triplet states, of both local and charge-transfer nature, is compared using range-separated time-dependent density functional theory (TD-DFT). We further compute electron paramagnetic resonance properties (zero-field splitting parameters and hyperfine coupling constants) of relaxed triplet states and compare them with available experimental data. Moreover, the electrostatic modulation of excited state energetics and redox properties of RC pigments by the semiquinone QA- is described. The results provide a detailed electronic-level understanding of triplet states within the PSII RC and form a refined basis for discussing primary and secondary electron transfer, charge recombination pathways, and possible photoprotection mechanisms in PSII.

How to tune the absorption spectrum of chlorophylls to enable better use of the available solar spectrum

Photon capture by chlorophylls and other chromophores in light-harvesting complexes and photosystems is the driving force behind the light reactions of photosynthesis. Excitation of photosystem II allows it to receive electrons from the water-oxidizing oxygen-evolution complex and to transfer them to an electron-transport chain that generates a transmembrane electrochemical gradient and ultimately reduces plastocyanin, which donates its electron to photosystem I. Subsequently, excitation of photosystem I leads to electron transfer to a ferredoxin which can either reduce plastocyanin again (in so-called “cyclical electron-flow”) and release energy for the maintenance of the electrochemical gradient, or reduce NADP+ to NADPH. Although photons in the far-red (700–750 nm) portion of the solar spectrum carry enough energy to enable the functioning of the photosynthetic electron-transfer chain, most extant photosystems cannot usually take advantage of them due to only absorbing light with shorter wavelengths. In this work, we used computational methods to characterize the spectral and redox properties of 49 chlorophyll derivatives, with the aim of finding suitable candidates for incorporation into synthetic organisms with increased ability to use far-red photons. The data offer a simple and elegant explanation for the evolutionary selection of chlorophylls a, b, c, and d among all easily-synthesized singly-substituted chlorophylls, and identified one novel candidate (2,12-diformyl chlorophyll a) with an absorption peak shifted 79 nm into the far-red (relative to chlorophyll a) with redox characteristics fully suitable to its possible incorporation into photosystem I (though not photosystem II). chlorophyll d is shown by our data to be the most suitable candidate for incorporation into far-red utilizing photosystem II, and several candidates were found with red-shifted Soret bands that allow the capture of larger amounts of blue and green light by light harvesting complexes.

Compensatory Transcriptional Response of Fischerella thermalis to Thermal Damage of the Photosynthetic Electron Transfer Chain.

Key organisms in the environment, such as oxygenic photosynthetic primary producers (photosynthetic eukaryotes and cyanobacteria), are responsible for fixing most of the carbon globally. However, they are affected by environmental conditions, such as temperature, which in turn affect their distribution. Globally, the cyanobacterium Fischerella thermalis is one of the main primary producers in terrestrial hot springs with thermal gradients up to 60 °C, but the mechanisms by which F. thermalis maintains its photosynthetic activity at these high temperatures are not known. In this study, we used molecular approaches and bioinformatics, in addition to photophysiological analyses, to determine the genetic activity associated with the energy metabolism of F. thermalis both in situ and in high-temperature (40 °C to 65 °C) cultures. Our results show that photosynthesis of F. thermalis decays with temperature, while increased transcriptional activity of genes encoding photosystem II reaction center proteins, such as PsbA (D1), could help overcome thermal damage at up to 60 °C. We observed that F. thermalis tends to lose copies of the standard G4 D1 isoform while maintaining the recently described D1INT isoform, suggesting a preference for photoresistant isoforms in response to the thermal gradient. The transcriptional activity and metabolic characteristics of F. thermalis, as measured by metatranscriptomics, further suggest that carbon metabolism occurs in parallel with photosynthesis, thereby assisting in energy acquisition under high temperatures at which other photosynthetic organisms cannot survive. This study reveals that, to cope with the harsh conditions of hot springs, F. thermalis has several compensatory adaptations, and provides emerging evidence for mixotrophic metabolism as being potentially relevant to the thermotolerance of this species. Ultimately, this work increases our knowledge about thermal adaptation strategies of cyanobacteria.

Utilization of light energy and photodynamic therapy based on thylakoid membranes

<p indent="0mm">The thylakoid membrane, which contains photosynthetic pigments, lipids and proteins, is the main site where the photosynthetic light reaction of green plants, eukaryotic algae and cyanobacteria occurs. Water photolysis and photosynthetic electron transport are two core processes of the light reaction. The former is responsible for converting photons into electrons, and the latter then condenses the energy from the high-potential electrons into energy carriers such as adenosine triphosphate (ATP) and reduced nicotinamide adenine dinucleotide phosphate (NADPH). Due to the electron transport and energy conversion properties of the light reaction, thylakoid membrane can be used in biophotoelectrochemical and biophotoelectrocatalytic systems. Biophotoelectrochemical systems refer to direct or indirect extraction of electrons from the photosynthetic electron transfer chain (PETC) of the thylakoid membrane and transfer the electrons into an external electrode, eventually converting light energy into electric current. Biophotoelectrocatalytic systems refer to the use of ATP and NADPH generated in the light reaction of the thylakoid membrane to drive a series of ATP- or NADPH-dependent biocatalytic reactions. These thylakoid membrane-based light energy utilization systems are featured with material renewability and environmental compatibility. In addition, they show higher energy efficiency than that of the whole photosynthetic-cell systems. Furthermore, thylakoid membranes also hold clear advantages over sub-cellular photosystems, including retention of photosynthetic complexes in their natural environment and relatively simple and fast procedures for their extraction and purification. For biophotoelectrochemical systems, scientists have optimized the electron transfer between the thylakoid membrane and electrode, for example by developing different immobilization methods, or modifying electrode surface with redox polymers and nanomaterials. As an effective platform for cofactor regeneration, the thylakoid membrane-based biophotoelectrocatalytic systems have been used to drive the enzymatic synthesis of poly(3-hydroxybutyrate) (PHB) from acetate, as well as CO₂ fixation through a synthetic crotonyl-coenzyme A (CoA)/ethylmalonyl-CoA/hydroxybutyryl-CoA (CETCH) cycle. In addition to light energy utilization, thylakoid membranes can also be used in photodynamic therapy (PDT) of tumors. Firstly, the photosensitizer porphyrin in the chlorophyll molecules can produce singlet oxygen (¹O₂) from O₂ under visible light irradiation, which in turn kills the tumor cells. Secondly, the photosynthetic oxygen evolution of thylakoid membrane can rapidly supply O₂ <italic>in situ</italic>. Moreover, thylakoid membranes contain abundant catalase, which catalyzes the decomposition of H₂O₂ into O₂, thus further increasing the supply of O₂. This paper summarizes recent progress in the research of thylakoid membranes in these three fields, focusing on the application forms and underlying mechanisms. Moreover, the difficulties and challenges in the application of thylakoid membrane are discussed, and future research directions are prospected. Specifically, developing protective structures such as hydrogels or artificial vesicles to address the issue of short half-life of isolated thylakoid membranes will be crucial. In light of their ingenious energy conversion characteristics and amenability for potential commercialization, thylakoid membrane-based biotechnological systems will have a bright future in the fields of bioenergy, biocatalysis and biomedicine.

Dissipation of the proton electrochemical gradient in chloroplasts promotes the oxidation of ATP synthase by thioredoxin-like proteins

Chloroplast FoF1-ATP synthase (CFoCF1) uses an electrochemical gradient of protons across the thylakoid membrane (ΔμH+) as an energy source in the ATP synthesis reaction. CFoCF1 activity is regulated by the redox state of a Cys pair on its central axis, that is, the γ subunit (CF1-γ). When the ΔμH+ is formed by the photosynthetic electron transfer chain under light conditions, CF1-γ is reduced by thioredoxin (Trx), and the entire CFoCF1 enzyme is activated. The redox regulation of CFoCF1 is a key mechanism underlying the control of ATP synthesis under light conditions. In contrast, the oxidative deactivation process involving CFoCF1 has not been clarified. In the present study, we analyzed the oxidation of CF1-γ by two physiological oxidants in the chloroplast, namely the proteins Trx-like 2 and atypical Cys-His-rich Trx. Using the thylakoid membrane containing the reduced form of CFoCF1, we were able to assess the CF1-γ oxidation ability of these Trx-like proteins. Our kinetic analysis indicated that these proteins oxidized CF1-γ with a higher efficiency than that achieved by a chemical oxidant and typical chloroplast Trxs. Additionally, the CF1-γ oxidation rate due to Trx-like proteins and the affinity between them were changed markedly when ΔμH+ formation across the thylakoid membrane was manipulated artificially. Collectively, these results indicate that the formation status of the ΔμH+ controls the redox regulation of CFoCF1 to prevent energetic disadvantages in plants.

Molecular mechanism of negative cooperativity of ferredoxin-NADP + reductase by ferredoxin and NADP(H): role of the ion pair of ferredoxin Arg40 of and FNR Glu154

Ferredoxin-NADP+ reductase (FNR) in plants receives electrons from ferredoxin (Fd) and converts NADP+ to NADPH at the end of the photosynthetic electron transfer chain. We previously showed that the interaction between FNR and Fd was weakened by the allosteric binding of NADP(H) on FNR, which was considered as a part of negative cooperativity. In this study, we investigated the molecular mechanism of this phenomenon using maize (Zea mays L.) FNR and Fd, as the 3D structure of this Fd:FNR complex is available. Site-specific mutants of several amino acid residues on the Fd:FNR interface were analysed for the effect on the negative cooperativity, by kinetic analysis of Fd:FNR electron transfer activity and by Fd-affinity chromatography. Mutations of Fd Arg40Gln and FNR Glu154Gln that disrupt one of the salt bridges in the Fd:FNR complex suppressed the negative cooperativity, indicating the involvement of the ion pair of Fd Arg40 and FNR Glu154 in the mechanism of the negative cooperativity. Unexpectedly, either mutation of Fd Arg40Gln or FNR Glu154Gln tends to increase the affinity between Fd and FNR, suggesting the role of this ion pair in the regulation of the Fd:FNR affinity by NADPH, rather than the stabilization of the Fd:FNR complex.

Energy status-promoted growth and development of Arabidopsis require copper deficiency response transcriptional regulator SPL7.

Copper (Cu) is a cofactor of around 300 Arabidopsis proteins, including photosynthetic and mitochondrial electron transfer chain enzymes critical for adenosine triphosphate (ATP) production and carbon fixation. Plant acclimation to Cu deficiency requires the transcription factor SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE7 (SPL7). We report that in the wild type (WT) and in the spl7-1 mutant, respiratory electron flux via Cu-dependent cytochrome c oxidase is unaffected under both normal and low-Cu cultivation conditions. Supplementing Cu-deficient medium with exogenous sugar stimulated growth of the WT, but not of spl7 mutants. Instead, these mutants accumulated carbohydrates, including the signaling sugar trehalose 6-phosphate, as well as ATP and NADH, even under normal Cu supply and without sugar supplementation. Delayed spl7-1 development was in agreement with its attenuated sugar responsiveness. Functional TARGET OF RAPAMYCIN and SNF1-RELATED KINASE1 signaling in spl7-1 argued against fundamental defects in these energy-signaling hubs. Sequencing of chromatin immunoprecipitates combined with transcriptome profiling identified direct targets of SPL7-mediated positive regulation, including Fe SUPEROXIDE DISMUTASE1 (FSD1), COPPER-DEFICIENCY-INDUCED TRANSCRIPTION FACTOR1 (CITF1), and the uncharacterized bHLH23 (CITF2), as well as an enriched upstream GTACTRC motif. In summary, transducing energy availability into growth and reproductive development requires the function of SPL7. Our results could help increase crop yields, especially on Cu-deficient soils.

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

Ultrafast excited state dynamics in the monomeric and trimeric photosystem I core complex of Spirulina platensis probed by two-dimensional electronic spectroscopy.

Photosystem I (PSI), a naturally occurring supercomplex composed of a core part and a light-harvesting antenna, plays an essential role in the photosynthetic electron transfer chain. Evolutionary adaptation dictates a large variability in the type, number, arrangement, and absorption of the Chlorophylls (Chls) responsible for the early steps of light-harvesting and charge separation. For example, the specific location of long-wavelength Chls (referred to as red forms) in the cyanobacterial core has been intensively investigated, but the assignment of the chromophores involved is still controversial. The most red-shifted Chl a form has been observed in the trimer of the PSI core of the cyanobacterium Spirulina platensis, with an absorption centered at ∼740nm. Here, we apply two-dimensional electronic spectroscopy to study photoexcitation dynamics in isolated trimers and monomers of the PSI core of S. platensis. By means of global analysis, we resolve and compare direct downhill and uphill excitation energy transfer (EET) processes between the bulk Chls and the red forms, observing significant differences between the monomer (lacking the most far red Chl form at 740nm) and the trimer, with the ultrafast EET component accelerated by five times, from 500 to 100fs, in the latter. Our findings highlight the complexity of EET dynamics occurring over a broad range of time constants and their sensitivity to energy distribution and arrangement of the cofactors involved. The comparison of monomeric and trimeric forms, differing both in the antenna dimension and in the extent of red forms, enables us to extract significant information regarding PSI functionality.

True oxygen reduction capacity during photosynthetic electron transfer in thylakoids and intact leaves.

Reactive oxygen species (ROS) are generated in electron transport processes of living organisms in oxygenic environments. Chloroplasts are plant bioenergetics hubs where imbalances between photosynthetic inputs and outputs drive ROS generation upon changing environmental conditions. Plants have harnessed various site-specific thylakoid membrane ROS products into environmental sensory signals. Our current understanding of ROS production in thylakoids suggests that oxygen (O2) reduction takes place at numerous components of the photosynthetic electron transfer chain (PETC). To refine models of site-specific O2 reduction capacity of various PETC components in isolated thylakoids of Arabidopsis thaliana, we quantified the stoichiometry of oxygen production and consumption reactions associated with hydrogen peroxide (H2O2) accumulation using membrane inlet mass spectrometry and specific inhibitors. Combined with P700 spectroscopy and electron paramagnetic resonance spin trapping, we demonstrate that electron flow to photosystem I (PSI) is essential for H2O2 accumulation during the photosynthetic linear electron transport process. Further leaf disc measurements provided clues that H2O2 from PETC has a potential of increasing mitochondrial respiration and CO2 release. Based on gas exchange analyses in control, site-specific inhibitor-, methyl viologen-, and catalase-treated thylakoids, we provide compelling evidence of no contribution of plastoquinone pool or cytochrome b6f to chloroplastic H2O2 accumulation. The putative production of H2O2 in any PETC location other than PSI is rapidly quenched and therefore cannot function in H2O2 translocation to another cellular location or in signaling.

Studying Plant Stress Reactions In Vivo by PAM Chlorophyll Fluorescence Imaging.

Plant photosynthetic and mitochondrial electron transfer chains (ETCs) are delicate environmental sensors and active players in stress acclimation. The performance of photosynthetic ETC can be deduced from chlorophyll a fluorescence. This makes chlorophyll fluorescence imaging a powerful tool to study plant stress in vivo. Many stress treatments enhance production of reactive oxygen species (ROS) by photosynthetic or mitochondrial ETCs. These ROS affect cellular metabolism and signalling. Generation of ROS can be manipulated in planta by specific pharmacological treatments with methyl viologen (MV), antimycin A (AA), myxothiazol (myx), and salicylhydroxamic acid (SHAM). This chapter describes how chlorophyll fluorescence imaging together with pharmacological treatments can be employed to probe ROS-dependent plant stress reactions in vivo.