Photosynthesis Research (v.117, #1-3)

Photosynthesis-related quantities for education and modeling by Taras K. Antal; Ilya B. Kovalenko; Andrew B. Rubin; Esa Tyystjärvi (1-30).
A quantitative understanding of the photosynthetic machinery depends largely on quantities, such as concentrations, sizes, absorption wavelengths, redox potentials, and rate constants. The present contribution is a collection of numbers and quantities related mainly to photosynthesis in higher plants. All numbers are taken directly from a literature or database source and the corresponding reference is provided. The numerical values, presented in this paper, provide ranges of values, obtained in specific experiments for specific organisms. However, the presented numbers can be useful for understanding the principles of structure and function of photosynthetic machinery and for guidance of future research.
Keywords: Photosynthesis; Plant; Chloroplast; Education; Modeling

May photoinhibition be a consequence, rather than a cause, of limited plant productivity? by William W. Adams III; Onno Muller; Christopher M. Cohu; Barbara Demmig-Adams (31-44).
Photoinhibition in leaves in response to high and/or excess light, consisting of a decrease in photosynthesis and/or photosynthetic efficiency, is frequently equated to photodamage and often invoked as being responsible for decreased plant growth and productivity. However, a review of the literature reveals that photoinhibited leaves characterized for foliar carbohydrate levels were invariably found to possess high levels of sugars and starch. We propose that photoinhibition should be placed in the context of whole-plant source–sink regulation of photosynthesis. Photoinhibition may represent downregulation of the photosynthetic apparatus in response to excess light when (1) more sugar is produced in leaves than can be utilized by the rest of the plant and/or (2) more light energy is harvested than can be utilized by the chloroplast for the fixation of carbon dioxide into sugars.
Keywords: D1 protein; Oxygen-evolving complex; Photochemical efficiency; Photoprotection; Photosystem II; Source–sink balance

Diffusional conductances to CO2 as a target for increasing photosynthesis and photosynthetic water-use efficiency by Jaume Flexas; Ülo Niinemets; Alexander Gallé; Margaret M. Barbour; Mauro Centritto; Antonio Diaz-Espejo; Cyril Douthe; Jeroni Galmés; Miquel Ribas-Carbo; Pedro L. Rodriguez; Francesc Rosselló; Raju Soolanayakanahally; Magdalena Tomas; Ian J. Wright; Graham D. Farquhar; Hipólito Medrano (45-59).
A key objective for sustainable agriculture and forestry is to breed plants with both high carbon gain and water-use efficiency (WUE). At the level of leaf physiology, this implies increasing net photosynthesis (A N) relative to stomatal conductance (g s). Here, we review evidence for CO2 diffusional constraints on photosynthesis and WUE. Analyzing past observations for an extensive pool of crop and wild plant species that vary widely in mesophyll conductance to CO2 (g m), g s, and foliage A N, it was shown that both g s and g m limit A N, although the relative importance of each of the two conductances depends on species and conditions. Based on Fick’s law of diffusion, intrinsic WUE (the ratio A N/g s) should correlate on the ratio g m/g s, and not g m itself. Such a correlation is indeed often observed in the data. However, since besides diffusion A N also depends on photosynthetic capacity (i.e., V c,max), this relationship is not always sustained. It was shown that only in a very few cases, genotype selection has resulted in simultaneous increases of both A N and WUE. In fact, such a response has never been observed in genetically modified plants specifically engineered for either reduced g s or enhanced g m. Although increasing g m alone would result in increasing photosynthesis, and potentially increasing WUE, in practice, higher WUE seems to be only achieved when there are no parallel changes in g s. We conclude that for simultaneous improvement of A N and WUE, genetic manipulation of g m should avoid parallel changes in g s, and we suggest that the appropriate trait for selection for enhanced WUE is increased g m/g s.
Keywords: Photosynthesis; Water-use efficiency; Stomatal conductance; Mesophyll conductance; Meta-analysis

The bioenergetic processes of photosynthesis and respiration are mutually beneficial. Their interaction extends to photorespiration, which is linked to optimize photosynthesis. The interplay of these three pathways is facilitated by two major phenomena: sharing of energy/metabolite resources and maintenance of optimal levels of reactive oxygen species (ROS). The resource sharing among different compartments of plant cells is based on the production/utilization of reducing equivalents (NADPH, NADH) and ATP as well as on the metabolite exchange. The responsibility of generating the cellular requirements of ATP and NAD(P)H is mostly by the chloroplasts and mitochondria. In turn, besides the chloroplasts, the mitochondria, cytosol and peroxisomes are common sinks for reduced equivalents. Transporters located in membranes ensure the coordinated movement of metabolites across the cellular compartments. The present review emphasizes the beneficial interactions among photosynthesis, dark respiration and photorespiration, in relation to metabolism of C, N and S. Since the bioenergetic reactions tend to generate ROS, the cells modulate chloroplast and mitochondrial reactions, so as to ensure that the ROS levels do not rise to toxic levels. The patterns of minimization of ROS production and scavenging of excess ROS in intracellular compartments are highlighted. Some of the emerging developments are pointed out, such as model plants, orientation/movement of organelles and metabolomics.
Keywords: Chloroplast; Metabolites; Transporters; Metabolomics; Mitochondria; Peroxisomes; Photorespiration

Variation in Rubisco content and activity under variable climatic factors by Jeroni Galmés; Iker Aranjuelo; Hipólito Medrano; Jaume Flexas (73-90).
The main objective of the present review is to provide a compilation of published data of the effects of several climatic conditions on Rubisco, particularly its activity, state of activation, and concentration, and its influence on leaf gas exchange and photosynthesis. The environmental conditions analyzed include drought, salinity, heavy metals, growth temperature, and elevated [O3], [CO2], and ultraviolet-B irradiance. The results show conclusive evidence for a major negative effect on activity of Rubisco with increasing intensity of a range of abiotic stress factors. This decrease in the activity of Rubisco is associated with down-regulation of the activation state of the enzyme (e.g., by de-carbamylation and/or binding of inhibitory sugar phosphates) in response to drought or high temperature. On the contrary, the negative effects of low temperature, heavy metal stress (cadmium), ozone, and UV-B stress on Rubisco activity are associated with changes in the concentration of Rubisco. Notably, in response to all environmental factors, the regulation of in vivo CO2 assimilation rate was related to Rubisco in vitro parameters, either concentration and/or carboxylation, depending on the particular stress. The importance of the loss of Rubisco activity and its repercussion on plant photosynthesis are discussed in the context of climate change. It is suggested that decreased Rubisco activity will be a major effect induced by climate change, which will need to be considered in any prediction model on plant productivity in the near future.
Keywords: CO2 ; Drought; Heavy metals; Ozone; Photosynthesis; Salinity; Temperature; Water stress

Photosynthetic gene expression in higher plants by James O. Berry; Pradeep Yerramsetty; Amy M. Zielinski; Christopher M. Mure (91-120).
Within the chloroplasts of higher plants and algae, photosynthesis converts light into biological energy, fueling the assimilation of atmospheric carbon dioxide into biologically useful molecules. Two major steps, photosynthetic electron transport and the Calvin-Benson cycle, require many gene products encoded from chloroplast as well as nuclear genomes. The expression of genes in both cellular compartments is highly dynamic and influenced by a diverse range of factors. Light is the primary environmental determinant of photosynthetic gene expression. Working through photoreceptors such as phytochrome, light regulates photosynthetic genes at transcriptional and posttranscriptional levels. Other processes that affect photosynthetic gene expression include photosynthetic activity, development, and biotic and abiotic stress. Anterograde (from nucleus to chloroplast) and retrograde (from chloroplast to nucleus) signaling insures the highly coordinated expression of the many photosynthetic genes between these different compartments. Anterograde signaling incorporates nuclear-encoded transcriptional and posttranscriptional regulators, such as sigma factors and RNA-binding proteins, respectively. Retrograde signaling utilizes photosynthetic processes such as photosynthetic electron transport and redox signaling to influence the expression of photosynthetic genes in the nucleus. The basic C3 photosynthetic pathway serves as the default form used by most of the plant species on earth. High temperature and water stress associated with arid environments have led to the development of specialized C4 and CAM photosynthesis, which evolved as modifications of the basic default expression program. The goal of this article is to explain and summarize the many gene expression and regulatory processes that work together to support photosynthetic function in plants.
Keywords: Chloroplast-encoded genes; Nuclear-encoded genes; Transcriptional control; Posttranscriptional control; Retrograde; Anterograde signaling

Photorespiration and carbon concentrating mechanisms: two adaptations to high O2, low CO2 conditions by James V. Moroney; Nadine Jungnick; Robert J. DiMario; David J. Longstreth (121-131).
This review presents an overview of the two ways that cyanobacteria, algae, and plants have adapted to high O2 and low CO2 concentrations in the environment. First, the process of photorespiration enables photosynthetic organisms to recycle phosphoglycolate formed by the oxygenase reaction catalyzed by ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). Second, there are a number of carbon concentrating mechanisms that increase the CO2 concentration around Rubisco which increases the carboxylase reaction enhancing CO2 fixation. This review also presents possibilities for the beneficial modification of these processes with the goal of improving future crop yields.
Keywords: Calvin–Benson–Bassham cycle; Carbon concentrating mechanism; Carbonic anhydrase; Cyanobacteria; Photorespiration; Rubisco

CO2-concentrating mechanism in cyanobacterial photosynthesis: organization, physiological role, and evolutionary origin by Elena V. Kupriyanova; Maria A. Sinetova; Sung Mi Cho; Youn-Il Park; Dmitry A. Los; Natalia A. Pronina (133-146).
The cellular and molecular organization of the CO2-concentrating mechanism (CCM) of cyanobacteria is reviewed. The primary processes of uptake, translocation, and accumulation of inorganic carbon (Ci) near the active site of carbon assimilation by the enzyme ribulose-1,5-bisphosphate carboxylase in the C3 cycle in cyanobacteria are described as one of the specialized forms of CO2 concentration which occurs in some photoautotrophic cells. The existence of this form of CO2 concentration expands our understanding of photosynthetic Ci assimilation. The means of supplying Ci to the C3 cycle in cyanobacteria is not by simple diffusion into the cell, but it is the result of coordinated functions of high-affinity systems for the uptake of CO2 and bicarbonate, as well as intracellular CO2/HCO3 interconversions by carbonic anhydrases. These biochemical events are under genetic control, and they serve to maintain cellular homeostasis and adaptation to CO2 limitation. Here we describe the organization of the CCM in cyanobacteria with a special focus on the CCM of relict halo- and alkaliphilic cyanobacteria of soda lakes. We also assess the role of the CCM at the levels of the organism, the biosphere, and evolution.
Keywords: Carbonic anhydrase; CO2-concentrating mechanisms; Cyanobacteria; Evolutionary origin of the CCM; Inorganic carbon transport; Relict cyanobacteria

The biochemistry and leaf anatomy of plants using C4 photosynthesis promote the concentration of atmospheric CO2 in leaf tissue that leads to improvements in growth and yield of C4 plants over C3 species in hot, dry, high light, and/or saline environments. C4 plants like maize and sugarcane are significant food, fodder, and bioenergy crops. The C4 photosynthetic pathway is an excellent example of convergent evolution, having evolved in multiple independent lineages of land plants from ancestors employing C3 photosynthesis. In addition to C3 and C4 species, some plant lineages contain closely related C3–C4 intermediate species that demonstrate leaf anatomical, biochemical, and physiological characteristics between those of C3 plants and species using C4 photosynthesis. These groups of plants have been extremely useful in dissecting the modifications to leaf anatomy and molecular biology, which led to the evolution of C4 photosynthesis. It is now clear that great variation exists in C4 leaf anatomy, and diverse molecular mechanisms underlie C4 biochemistry and physiology. However, all these different paths have led to the same destination—the expression of a C4 CO2 concentrating mechanism. Further identification of C4 leaf anatomical traits and molecular biological components, and understanding how they are controlled and assembled will not only allow for additional insights into evolutionary convergence, but also contribute to sustainable food and bioenergy production strategies.
Keywords: C3–C4 intermediate; C4 leaf anatomy; C4 photosynthetic enzyme; C2 photosynthesis; C4 photosynthesis; Evolution of C4 photosynthesis

The recurrent assembly of C4 photosynthesis, an evolutionary tale by Pascal-Antoine Christin; Colin P. Osborne (163-175).
Today, plants using C4 photosynthesis are widespread and important components of major tropical and subtropical biomes, but the events that led to their evolution and success started billions of years ago (bya). A CO2-fixing enzyme evolved in the early Earth atmosphere with a tendency to confuse CO2 and O2 molecules. The descendants of early photosynthetic organisms coped with this property in the geological eras that followed through successive fixes, the latest of which is the addition of complex CO2-concentrating mechanisms such as C4 photosynthesis. This trait was assembled from bricks available in C3 ancestors, which were altered to fulfill their new role in C4 photosynthesis. The existence of C4-suitable bricks probably determined the lineages of plants that could make the transition to C4 photosynthesis, highlighting the power of contingency in evolution. Based on the latest findings in C4 research, we present the evolutionary tale of C4 photosynthesis, with a focus on the general evolutionary phenomena that it so wonderfully exemplifies.
Keywords: C4 photosynthesis; Complex traits; Contingency; Convergence; Co-option; Evolution

Biochemical approaches to C4 photosynthesis evolution studies: the case of malic enzymes decarboxylases by Mariana Saigo; Marcos A. Tronconi; Mariel C. Gerrard Wheeler; Clarisa E. Alvarez; María F. Drincovich; Carlos S. Andreo (177-187).
C4 photosynthesis enables the capture of atmospheric CO2 and its concentration at the site of RuBisCO, thus counteracting the negative effects of low atmospheric levels of CO2 and high atmospheric levels of O2 (21 %) on photosynthesis. The evolution of this complex syndrome was a multistep process. It did not occur by simply recruiting pre-exiting components of the pathway from C3 ancestors which were already optimized for C4 function. Rather it involved modifications in the kinetics and regulatory properties of pre-existing isoforms of non-photosynthetic enzymes in C3 plants. Thus, biochemical studies aimed at elucidating the functional adaptations of these enzymes are central to the development of an integrative view of the C4 mechanism. In the present review, the most important biochemical approaches that we currently use to understand the evolution of the C4 isoforms of malic enzyme are summarized. It is expected that this information will help in the rational design of the best decarboxylation processes to provide CO2 for RuBisCO in engineering C3 species to perform C4 photosynthesis.
Keywords: C4 enzymes; Malic enzymes; Kinetic and structural properties; Molecular and biochemical technologies

Long-distance translocation of photosynthates: a primer by Michael Knoblauch; Winfried S. Peters (189-196).
The storage of light energy in chemical form through photosynthesis is the key process underlying life as we know it. To utilize photosynthates efficiently as structural materials or as fuel to drive endergonic processes, they have to be transported from where they are produced to where they are needed. In this primer, we provide an overview of basic biophysical concepts underlying our current understanding of the mechanisms of photosynthate long-distance transport, and briefly discuss current developments in the field.
Keywords: Cytoplasmic streaming; Münch flow; Münch hypothesis; Phloem transport; Sieve tube; Symplasmic transport

Some enzymes can be considered as a catalyst having a nanosized inorganic core in a protein matrix. In some cases, the metal oxide or sulfide clusters, which can be considered as cofactors in enzymes, may be recruited for use in other related reactions in artificial photosynthetic systems. In other words, one approach to design efficient and environmentally friendly catalysts in artificial photosynthetic systems for the purpose of utilizing sunlight to generate high energy intermediates or useful material is to select and utilize inorganic cores of enzymes. For example, one of the most important goals in developing artificial photosynthesis is hydrogen production. However, first, it is necessary to find a “super catalyst” for water oxidation, which is the most challenging half reaction of water splitting. There is an efficient system for water oxidation in cyanobacteria, algae, and plants. Published data on the Mn–Ca cluster have provided details on the mechanism and structure of the water oxidizing complex as a Mn–Ca nanosized inorganic core in photosystem II. Progress has been made in introducing Mn–Ca oxides as efficient catalysts for water oxidation in artificial photosynthetic systems. Here, in the interest of designing efficient catalysts for other important reactions in artificial photosynthesis, a few examples of our knowledge of inorganic cores of proteins, and how Nature used them for important reactions, are discussed.
Keywords: Artificial photosynthesis; Cofactor; Enzyme; Nanosized inorganic core; Photosynthesis

Algal biofuels by Reza Razeghifard (207-219).
The world is facing energy crisis and environmental issues due to the depletion of fossil fuels and increasing CO2 concentration in the atmosphere. Growing microalgae can contribute to practical solutions for these global problems because they can harvest solar energy and capture CO2 by converting it into biofuel using photosynthesis. Microalgae are robust organisms capable of rapid growth under a variety of conditions including in open ponds or closed photobioreactors. Their reduced biomass compounds can be used as the feedstock for mass production of a variety of biofuels. As another advantage, their ability to accumulate or secrete biofuels can be controlled by changing their growth conditions or metabolic engineering. This review is aimed to highlight different forms of biofuels produced by microalgae and the approaches taken to improve their biofuel productivity. The costs for industrial-scale production of algal biofuels in open ponds or closed photobioreactors are analyzed. Different strategies for photoproduction of hydrogen by the hydrogenase enzyme of green algae are discussed. Algae are also good sources of biodiesel since some species can make large quantities of lipids as their biomass. The lipid contents for some of the best oil-producing strains of algae in optimized growth conditions are reviewed. The potential of microalgae for producing petroleum related chemicals or ready-make fuels such as bioethanol, triterpenic hydrocarbons, isobutyraldehyde, isobutanol, and isoprene from their biomass are also presented.
Keywords: Photosynthesis; Microalgae; Bioenergy; Biofuel; Biohydrogen; Biodiesel; Biomass; Photobioreactor

Stay-green plants: what do they tell us about the molecular mechanism of leaf senescence by Makoto Kusaba; Ayumi Tanaka; Ryouichi Tanaka (221-234).
A practical approach to increasing crop yields is to extend the duration of active photosynthesis. Stay-green is a term that is used to describe mutant and transgenic plants or cultivars with the trait of maintaining their leaves for a longer period of time than the wild-type or crosses from which they are derived. Analyzing stay-green genotypes contributes to our understanding of the molecular mechanism regulating leaf senescence which may allow us to extend the duration of active photosynthesis in crop plants. This article summarizes recent studies on stay-green plants and the insights they provide on the mechanism of leaf senescence. Briefly, mutations suppressing ethylene, abscisic acid, brassinosteroid, and strigolactone signal transduction or those activating cytokinin signaling often lead to stay-green phenotypes indicating a complex signaling network regulating leaf senescence. Developmentally regulated transcription factors, including NAC or WRKY family members, play key roles in the induction of leaf senescence and thus alteration in the activity of these transcription factors also result in stay-green phenotypes. Impairment in the enzymatic steps responsible for chlorophyll breakdown also leads to stay-green phenotypes. Some of these genotypes die in the middle of the process of chlorophyll breakdown due to the accumulation of toxic intermediates, while others appear to stay-green but their photosynthetic activity declines in a manner similar to wild-type plants. Alterations in certain metabolic pathways in chloroplasts (e.g., photosynthesis) can lead to a delayed onset of leaf senescence with maintenance of photosynthetic activity longer than wild-type plants, indicating that chloroplast metabolism can also affect the regulatory mechanism of leaf senescence.
Keywords: Chlorophyll; Chloroplast; Senescence; Phytohormone; Tetrapyrrole; Cell death

In cyanobacteria, the interactions among pigment–protein complexes are modified in response to changes in light conditions. In the present study, we analyzed excitation energy transfer from the phycobilisome and photosystem II to photosystem I in the cyanobacterium Arthrospira (Spirulina) platensis. The cells were grown under lights with different spectral profiles and under different light intensities, and the energy-transfer characteristics were evaluated using steady-state absorption, steady-state fluorescence, and picosecond time-resolved fluorescence spectroscopy techniques. The fluorescence rise and decay curves were analyzed by global analysis to obtain fluorescence decay-associated spectra. The direct energy transfer from the phycobilisome to photosystem I and energy transfer from photosystem II to photosystem I were modified depending on the light quality, light quantity, and cultivation period. However, the total amount of energy transferred to photosystem I remained constant under the different growth conditions. We discuss the differences in energy-transfer processes under different cultivation and light conditions.
Keywords: Energy transfer; Light adaptation; Phycobilisome; Delayed fluorescence

Spectral properties of a divinyl chlorophyll a harboring mutant of Synechocystis sp. PCC6803 by Md. Rafiqul Islam; Koji Watanabe; Yasuhiro Kashino; Kazuhiko Satoh; Hiroyuki Koike (245-255).
A divinyl chlorophyll (DV-Chl) a harboring mutant of Synechocystis sp. PCC 6803, in which chlorophyll species is replaced from monovinyl(normal)-Chl a to DV-Chl a, was characterized. The efficiency of light utilization for photosynthesis was decreased in the mutant. Absorption spectra at 77 K and their fourth derivative analyses revealed that peaks of each chlorophyll forms were blue-shifted by 1–2 nm, suggesting lowered stability of chlorophylls at their binding sites. This was also true both in PSI and PSII complexes. On the other hand, fluorescence emission spectra measured at 77 K were not different between wild type and the mutant. This indicates that the mode of interaction between chlorophyll and its binding pockets responsible for emitting fluorescence at 77 K is not altered in the mutant. P700 difference spectra of thylakoid membranes and PSI complexes showed that the spectrum in Soret region was red-shifted by 7 nm in the mutant. This is a characteristic feature of DV-Chl a. Microenvironments of iron–sulfur center of a terminal electron acceptor of PSI complex, P430, were practically the same as that of wild type.
Keywords: cvrA ; Divinyl chlorophyll; Divinyl chlorophyllide reductase; Monovinyl chlorophyll; slr1923, Synechocystis sp. PCC 6803

Low-temperature time-resolved spectroscopic study of the major light-harvesting complex of Amphidinium carterae by Václav Šlouf; Marcel Fuciman; Silke Johanning; Eckhard Hofmann; Harry A. Frank; Tomáš Polívka (257-265).
The major light-harvesting complex of Amphidinium (A.) carterae, chlorophyll-a–chlorophyll-c 2–peridinin–protein complex (acpPC), was studied using ultrafast pump-probe spectroscopy at low temperature (60 K). An efficient peridinin–chlorophyll-a energy transfer was observed. The stimulated emission signal monitored in the near-infrared spectral region was stronger when redder part of peridinin pool was excited, indicating that these peridinins have the S1/ICT (intramolecular charge-transfer) state with significant charge-transfer character. This may lead to enhanced energy transfer efficiency from “red” peridinins to chlorophyll-a. Contrary to the water-soluble antenna of A. carterae, peridinin–chlorophyll-a protein, the energy transfer rates in acpPC were slower under low-temperature conditions. This fact underscores the influence of the protein environment on the excited-state dynamics of pigments and/or the specificity of organization of the two pigment–protein complexes.
Keywords: Light-harvesting; Energy transfer; Carotenoid; Dinoflagellates

Spectral and functional studies on siphonaxanthin-type light-harvesting complex of photosystem II from Bryopsis corticulans by Wenda Wang; Xiaochun Qin; Min Sang; Dongqin Chen; Kebin Wang; Rongchen Lin; Congming Lu; Jian-Ren Shen; Tingyun Kuang (267-279).
Carotenoids with conjugated carbonyl groups possess special photophysical properties which have been studied in some water-soluble light-harvesting proteins (Polívka and Sundström, Chem Rev 104:2021–2071, 2004). However, siphonaxanthin-type light-harvesting complexes of photosystem II (LHCII) in siphonous green alga have received fewer studies. In the present study, we determined sequences of genes for several Bryopsis corticulans Lhcbm proteins, which showed that they belong to the group of major LHCII and diverged early from green algae and higher plants. Analysis of pigment composition indicated that this siphonaxanthin-type LHCII contained in total 3 siphonaxanthin and siphonein but no lutein and violaxanthin. In addition, 2 chlorophylls a in higher plant LHCII were replaced by chlorophyll b. These changes led to an increased absorption in green and blue-green light region compared with higher plant LHCII. The binding sites for chlorophylls, siphonaxanthin, and siphonein were suggested based on the structural comparison with that of higher plant LHCII. All of the ligands for the chlorophylls were completely conserved, suggesting that the two chlorophylls b were replaced by chlorophyll a without changing their binding sites in higher plant LHCII. Comparisons of the absorption spectra of isolated siphonaxanthin and siphonein in different organic solutions and the effect of heat treatment suggested that these pigments existed in a low hydrophobic protein environment, leading to an enhancement of light harvesting in the green light region. This low hydrophobic protein environment was maintained by the presence of more serine and threonine residues in B. corticulans LHCII. Finally, esterization of siphonein may also contribute to the enhanced harvesting of green light.
Keywords: Bryopsis corticulans ; Light-harvesting proteins; LHCII; Pigments; Siphonaxanthin; Carotenoids

Comparison of oligomeric states and polypeptide compositions of fucoxanthin chlorophyll a/c-binding protein complexes among various diatom species by Ryo Nagao; Shuji Takahashi; Takehiro Suzuki; Naoshi Dohmae; Katsuyoshi Nakazato; Tatsuya Tomo (281-288).
Fucoxanthin chlorophyll a/c-binding protein (FCP) is a unique light-harvesting apparatus in diatoms. Several biochemical characteristics of FCP oligomer and trimer from different diatom species have been reported previously. However, the integration of information about molecular organizations and polypeptides of FCP through a comparison among diatoms has not been published. In this study, we used two-dimensional clear-native/SDS-PAGE to compare the oligomeric states and polypeptide compositions of FCP complexes from four diatoms: Chaetoceros gracilis, Thalassiosira pseudonana, Cyclotella meneghiniana, and Phaeodactylum tricornutum. FCP oligomer was found in C. gracilis, T. pseudonana, and C. meneghiniana, but not in P. tricornutum. The oligomerization varied among the three diatoms, although a predominant subunit having similar molecular weight was recovered in each FCP oligomer. These results suggest that the predominant subunit is involved in the formation of high FCP oligomerization in each diatom. In contrast, FCP trimer was found in all the diatoms. The trimerizations were quite similar, whereas the polypeptide compositions were markedly different. On the basis of this information and that from mass spectrometric analyses, the gene products in each FCP complex were identified in T. pseudonana and P. tricornutum. Based on these results, we discuss the role of FCP oligomer and trimer from the four diatoms.
Keywords: CN-PAGE; FCP oligomer; FCP trimer; MS analysis

The energy flux theory 35 years later: formulations and applications by Merope Tsimilli-Michael; Reto J. Strasser (289-320).
Several models have been proposed for the energetic behavior of the photosynthetic apparatus and a variety of experimental techniques are nowadays available to determine parameters that can quantify this behavior. The Energy Flux Theory (EFT) developed by Strasser 35 years ago provides a straightforward way to formulate any possible energetic communication between any complex arrangement of interconnected pigment systems and any energy transduction by these systems. We here revisit the EFT, starting from the basic general definitions and equations and presenting applications in formulating the energy distribution in photosystem (PS) II units with variable connectivity, as originally derived, where certain simplifications were adopted. We then proceed to the derivation of equations for a PSII model of higher complexity, which corresponds, from the formalistic point of view, to the later formulated and now broadly accepted exciton–radical-pair model. We also compare the formulations derived with the EFT with those obtained, by different approaches, in the classic papers on energetic connectivity. Moreover, we apply the EFT for the evaluation of the excitation energy distribution between PSII and PSI and the distinction between state transitions and PSII to PSI excitation energy migration. Our analysis demonstrates that the EFT is a powerful approach for the formulation of any possible model, at any complexity level, even of models that may be proposed in the future, with the advantage that any possible energetic communication or energy transduction can be easily formulated mathematically by trivial algebraic equations. Moreover, the biophysical parameters introduced by the EFT and applicable for any possible model can be linked with obtainable experimental signals, provided that the theoretical resolution of the model does not go beyond the experimental resolution.
Keywords: Energetic connectivity (grouping); Energy distribution; Energy flux; Grouping probability; Photosynthetic unit; Photosystem II models

This study reports on kinetics of the fluorescence decay in a suspension of the alga Scenedesmus quadricauda after actinic illumination. These are monitored as the variable fluorescence signal in the dark following light pulses of variable intensity and duration. The decay reflects the restoration of chlorophyll fluorescence quenching of the photosystem II (PSII) antennas and shows a polyphasic pattern which suggests the involvement of different processes. The overall quenching curve after a fluorescence-saturating pulse (SP) of 250-ms duration, commonly used in pulse amplitude modulation applications as the tool for estimating the maximal fluorescence (F m), has been termed P–O, in which P and O have the same meaning as used in the OJIP induction curve in the light. Deconvolution of this signal shows at least three distinguishable exponential phases with reciprocal rate constants of the order of 10, 102, and 103 ms. The size of the long (>103 ms) and moderate (~102 ms) lasting components relative to the complete quenching signal after an SP increases with the duration of the actinic pulse concomitantly with an increase in the reciprocal rate constants of the fast (~10 ms) and moderate quenching phases. Fluorescence responses upon single turnover flashes of 30-μs duration (STFs) given at discrete times during the P–O quenching were used as tools for identifying the quencher involved in the P–O quenching phase preceding the STF excitation. Results are difficult to interpret in terms of a single-hit two-state trapping mechanism with distinguishable quenching properties of open and closed reaction centers only. They give support for an earlier hypothesis on a double-hit three-state trapping mechanism in which the so-called semi-closed reaction centers of PSII are considered. In these trapping-competent centers the single reduced acceptor pair [PheQ A]1−, depending on the size of photoelectrochemically induced pH effects on the Q B-binding site, functions as an efficient fluorescence quencher.
Keywords: Chlorophyll a fluorescence; Photosystem II; Quenching kinetics; Double-hit model

Chlorophyll a fluorescence induction (FI) kinetics, in the microseconds to the second range, reflects the overall performance of the photosynthetic apparatus. In this paper, we have developed a novel FI model, using a rule-based kinetic Monte Carlo method, which incorporates not only structural and kinetic information on PSII, but also a simplified photosystem I. This model has allowed us to successfully simulate the FI under normal or different treatment conditions, i.e., with different levels of measuring light, under 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea treatment, under 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone treatment, and under methyl viologen treatment. Further, using this model, we have systematically studied the mechanistic basis and factors influencing the FI kinetics. The results of our simulations suggest that (1) the J step is caused by the two-electron gate at the Q B site; (2) the I step is caused by the rate limitation of the plastoquinol re-oxidation in the plastoquinone pool. This new model provides a framework for exploring impacts of modifying not only kinetic but also structural parameters on the FI kinetics.
Keywords: Chlorophyll fluorescence induction; Kinetic model; Photosynthesis; Rule-based kinetic Monte Carlo methods; Systems biology

The chlorophyll (Chl) fluorescence induction kinetics, net photosynthetic CO2 fixation rates P N, and composition of photosynthetic pigments of differently light exposed leaves of several trees were comparatively measured to determine the differences in photosynthetic activity and pigment adaptation of leaves. The functional measurements were carried out with sun, half-shade and shade leaves of seven different trees species. These were: Acer platanoides L., Ginkgo biloba L., Fagus sylvatica L., Platanus x acerifolia Willd., Populus nigra L., Quercus robur L., Tilia cordata Mill. In three cases (beech, ginkgo, and oak), we compared the Chl fluorescence kinetics and photosynthetic rates of blue-shade leaves of the north tree crown receiving only blue sky light but no direct sunlight with that of sun leaves. In these cases, we also determined in detail the pigment composition of all four leaf types. In addition, we determined the quantum irradiance and spectral irradiance of direct sunlight, blue skylight as well as the irradiance in half shade and full shade. The results indicate that sun leaves possess significantly higher mean values for the net CO2 fixation rates P N (7.8–10.7 μmol CO2 m−2 s−1 leaf area) and the Chl fluorescence ratio R Fd (3.85–4.46) as compared to shade leaves (mean P N of 2.6–3.8 μmol CO2 m−2 s−1 leaf area.; mean R Fd of 1.94–2.56). Sun leaves also exhibit higher mean values for the pigment ratio Chl a/b (3.14–3.31) and considerably lower values for the weight ratio total chlorophylls to total carotenoids, (a + b)/(x + c), (4.07–4.25) as compared to shade leaves (Chl a/b 2.62–2.72) and (a + b)/(x + c) of 5.18–5.54. Blue-shade and half-shade leaves have an intermediate position between sun and shade leaves in all investigated parameters including the ratio F v/F o (maximum quantum yield of PS2 photochemistry) and are significantly different from sun and shade leaves but could not be differentiated from each other. The mean values of the Chl fluorescence decrease ratio R Fd of blue-shade and half-shade leaves fit well into the strong linear correlation with the net photosynthetic rates P N of sun and shade leaves, thus unequivocally indicating that the determination of the Chl fluorescence decrease ratio R Fd is a fast and indirect measurement of the photosynthetic activity of leaves. The investigations clearly demonstrate that the photosynthetic capacity and pigment composition of leaves and chloroplasts strongly depend on the amounts and quality of light received by the leaves.
Keywords: Blue skylight spectrum; Chlorophyll fluorescence decrease ratio R Fd ; Light adaptation; Net photosynthetic CO2 fixation; Spectral irradiance of half-shade and full shade

It has been shown that removal of manganese from the water-oxidizing complex (WOC) of photosystem II (PSII) leads to flash-induced oxygen consumption (FIOC) which is activated by low concentration of Mn2+ (Yanykin et al., Biochim Biophys Acta 1797:516–523, 2010). In the present work, we examined the effect of transition and non-transition divalent metal ions on FIOC in Mn-depleted PSII (apo-WOC-PSII) preparations. It was shown that only Mn2+ ions are able to activate FIOC while other transition metal ions (Fe2+, V2+ and Cr2+) capable of electron donation to the apo-WOC-PSII suppressed the photoconsumption of O2. Co2+ ions with a high redox potential (E 0 for Co2+/Co3+ is 1.8 V) showed no effect. Non-transition metal ions Ca2+ by Mg2+ did not stimulate FIOC. However, Ca2+ (in contrast to Mg2+) showed an additional activation effect in the presence of exogenic Mn2+. The Ca2+ effect depended on the concentration of both Mn2+ and Ca2+. The Ca effect was only observed when: (1) the activation of FIOC induced by Mn2+ did not reach its maximum, (2) the concentration of Ca2+ did not exceed 40 μM; at higher concentrations Ca2+ inhibited the Mn2+-activated O2 photoconsumption. Replacement of Ca2+ by Mg2+ led to a suppression of Mn2+-activated O2 photoconsumption; while, addition of Ca2+ resulted in elimination of the Mg2+ inhibitory effect and activation of FIOC. Thus, only Mn2+ and Ca2+ (which are constituents of the WOC) have specific effects of activation of FIOC in apo-WOC-PSII preparations. Possible reactions involving Mn2+ and Ca2+ which could lead to the activation of FIOC in the apo-WOC-PSII are discussed.
Keywords: Photosystem II; Oxygen photoconsumption; Manganese; Reactive oxygen species

Crystal structure of the Psb28 accessory factor of Thermosynechococcus elongatus photosystem II at 2.3 Å by Wojciech Bialek; Songjia Wen; Franck Michoux; Martina Beckova; Josef Komenda; James W. Murray; Peter J. Nixon (375-383).
Members of the Psb28 family of proteins are accessory factors implicated in the assembly and repair of the photosystem II complex. We present here the crystal structure of the Psb28 protein (Tlr0493) found in the thermophilic cyanobacterium Thermosynechococcus elongatus at a resolution of 2.3 Å. Overall the crystal structure of the Psb28 monomer is similar to the solution structures of C-terminally His-tagged Psb28-1 from Synechocystis sp. PCC 6803 obtained previously by nuclear magnetic resonance spectroscopy. One new aspect is that Escherichia coli-expressed T. elongatus Psb28 is able to form dimers in solution and packs as a dimer of dimers in the crystal. Analysis of wild type and mutant strains of Synechocystis 6803 by blue native-polyacrylamide gel electrophoresis suggests that Psb28-1, the closest homologue to T. elongatus Psb28 in this organism, also exists as an oligomer in vivo, most likely a dimer. In line with the prediction based on the crystal structure of T. elongatus Psb28, the addition of a 3× Flag-tag to the C-terminus of Synechocystis 6803 Psb28-1 interferes with the accumulation of the Psb28-1 oligomer in vivo. In contrast, the more distantly related Psb28-2 protein found in Synechocystis 6803 lacks the residues that stabilize dimer formation in the T. elongatus Psb28 crystal and is detected as a monomer in vivo. Overall our data suggest that the dimer interface in the Psb28 crystal might be physiologically relevant.
Keywords: Assembly factor; Psb28; Psb28-2; X-ray crystallography; Photosystem II; Cyanobacteria

Production of reactive oxygen species in decoupled, Ca2+-depleted PSII and their use in assigning a function to chloride on both sides of PSII by Boris K. Semin; Lira N. Davletshina; Kirill N. Timofeev; Il’ya I. Ivanov; Andrei B. Rubin; Michael Seibert (385-399).
Extraction of Ca2+ from the oxygen-evolving complex of photosystem II (PSII) in the absence of a chelator inhibits O2 evolution without significant inhibition of the light-dependent reduction of the exogenous electron acceptor, 2,6-dichlorophenolindophenol (DCPIP) on the reducing side of PSII. The phenomenon is known as “the decoupling effect” (Semin et al. Photosynth Res 98:235–249, 2008). Extraction of Cl from Ca2+-depleted membranes (PSII[–Ca]) suppresses the reduction of DCPIP. In the current study we investigated the nature of the oxidized substrate and the nature of the product(s) of the substrate oxidation. After elimination of all other possible donors, water was identified as the substrate. Generation of reactive oxygen species HO, H2O2, and O 2 ·− , as possible products of water oxidation in PSII(–Ca) membranes was examined. During the investigation of O 2 ·− production in PSII(–Ca) samples, we found that (i) O 2 ·− is formed on the acceptor side of PSII due to the reduction of O2; (ii) depletion of Cl does not inhibit water oxidation, but (iii) Cl depletion does decrease the efficiency of the reduction of exogenous electron acceptors. In the absence of Cl under aerobic conditions, electron transport is diverted from reducing exogenous acceptors to reducing O2, thereby increasing the rate of O 2 ·− generation. From these observations we conclude that the product of water oxidation is H2O2 and that Cl anions are not involved in the oxidation of water to H2O2 in decoupled PSII(–Ca) membranes. These results also indicate that Cl anions are not directly involved in water oxidation by the Mn cluster in the native PSII membranes, but possibly provide access for H2O molecules to the Mn4CaO5 cluster and/or facilitate the release of H+ ions into the lumenal space.
Keywords: Photosystem II; Superoxide anion radical; Hydrogen peroxide; Chloride; Decoupling; Oxygen-evolving complex

Efficiency of photosynthetic water oxidation at ambient and depleted levels of inorganic carbon by Dmitriy Shevela; Birgit Nöring; Sergey Koroidov; Tatiana Shutova; Göran Samuelsson; Johannes Messinger (401-412).
Over 40 years ago, Joliot et al. (Photochem Photobiol 10:309–329, 1969) designed and employed an elegant and highly sensitive electrochemical technique capable of measuring O2 evolved by photosystem II (PSII) in response to trains of single turn-over light flashes. The measurement and analysis of flash-induced oxygen evolution patterns (FIOPs) has since proven to be a powerful method for probing the turnover efficiency of PSII. Stemler et al. (Proc Natl Acad Sci USA 71(12):4679–4683, 1974), in Govindjee’s lab, were the first to study the effect of “bicarbonate” on FIOPs by adding the competitive inhibitor acetate. Here, we extend this earlier work by performing FIOPs experiments at various, strictly controlled inorganic carbon (Ci) levels without addition of any inhibitors. For this, we placed a Joliot-type bare platinum electrode inside a N2-filled glove-box (containing 10–20 ppm CO2) and reduced the Ci concentration simply by washing the samples in Ci-depleted media. FIOPs of spinach thylakoids were recorded either at 20-times reduced levels of Ci or at ambient Ci conditions (390 ppm CO2). Numerical analysis of the FIOPs within an extended Kok model reveals that under Ci-depleted conditions the miss probability is discernibly larger (by 2–3 %) than at ambient conditions, and that the addition of 5 mM HCO3 to the Ci-depleted thylakoids largely restores the original miss parameter. Since a “mild” Ci-depletion procedure was employed, we discuss our data with respect to a possible function of free or weakly bound HCO3 at the water-splitting side of PSII.
Keywords: Flash-induced oxygen evolution patterns; S states; An extended Kok model; Hydrogen carbonate (bicarbonate); Photosynthetic water oxidation

Imidazolium or guanidinium/layered manganese (III, IV) oxide hybrid as a promising structural model for the water-oxidizing complex of Photosystem II for artificial photosynthetic systems by Mohammad Mahdi Najafpour; Mahmoud Amouzadeh Tabrizi; Behzad Haghighi; Julian J. Eaton-Rye; Robert Carpentier; Suleyman I. Allakhverdiev (413-421).
Photosystem II is responsible for the light-driven biological water-splitting system in oxygenic photosynthesis and contains a cluster of one calcium and four manganese ions at its water-oxidizing complex. This cluster may serve as a model for the design of artificial or biomimetic systems capable of splitting water into oxygen and hydrogen. In this study, we consider the ability of manganese oxide monosheets to self-assemble with organic compounds. Layered structures of manganese oxide, including guanidinium and imidazolium groups, were synthesized and characterized by scanning electron microscopy, transmission electron microscopy, X-ray diffraction spectrometry, and atomic absorption spectroscopy. The compounds can be considered as new structural models for the water-oxidizing complex of Photosystem II. The overvoltage of water oxidation for the compounds in these conditions at pH = 6.3 is ~0.6 V. These compounds may represent the first step to synthesize a hybrid of guanidinium or imidazole together with manganese as a biomimetic system for the water-oxidizing complex of Photosystem II.
Keywords: Artificial photosynthesis; Photosystem II; Manganese; Nano-layered manganese oxide; Imidazolium; Guanidinium

Gold or silver deposited on layered manganese oxide: a functional model for the water-oxidizing complex in photosystem II by Mohammad Mahdi Najafpour; Fahimeh Rahimi; Davood Jafarian Sedigh; Robert Carpentier; Julian J. Eaton-Rye; Jian-Ren Shen; Suleyman I. Allakhverdiev (423-429).
In this report, gold or silver deposited on layered manganese oxide has been synthesized by a simple method and characterized by scanning electron microscopy, transmission electron microscopy, X-ray diffraction spectrometry, atomic absorption spectroscopy, and energy-dispersive X-ray mapping. The gold deposited on layered manganese oxide showed efficient catalytic activity toward water oxidation in the presence of cerium(IV) ammonium nitrate. The properties associated with this compound suggest it is a functional model for the water-oxidizing complex in photosystem II.
Keywords: Gold; Nano-sized manganese oxide; Oxygen; Silver; Water oxidation

Chl fluorescence induction (FI) was recorded in sunflower leaves pre-adapted to darkness or low preferentially PSI light, or inhibited by DCMU. For analysis the FI curves were plotted against the cumulative number of excitations quenched by PSII, n q, calculated as the cumulative complementary area above the FI curve. In the +DCMU leaves n q was <1 per PSII, suggesting pre-reduction of Q A during the dark pre-exposure. A strongly sigmoidal FI curve was constructed by complementing (shifting) the recorded FI curves to n q = 1 excitation per PSII. The full FI curve in +DCMU leaves was well fitted by a model assuming PSII antennae are excitonically connected in domains of four PSII. This result, obtained by gradually reducing Q A in PSII with pre-blocked Q B (by DCMU or PQH2), differs from that obtained by gradually blocking the Q B site (by increasing DCMU or PQH2 level) in leaves during (quasi)steady-state e transport (Oja and Laisk, Photosynth Res 114, 15–28, 2012). Explanations are discussed. Donor side quenching was characterized by comparison of the total n q in one and the same dark-adapted leaf, which apparently increased with increasing PFD during FI. An explanation for the donor side quenching is proposed, based on electron transfer from excited P680* to oxidized tyrosine Z (TyrZox). At high PFDs the donor side quenching at the J inflection of FI is due mainly to photochemical quenching by TyrZox. This quenching remains active for subsequent photons while TyrZ remains oxidized, following charge transfer to Q A. During further induction this quenching disappears as soon as PQ and Q A become reduced, charge separation becomes impossible and TyrZ is reduced by the water oxidizing complex.
Keywords: Leaf; Fluorescence induction; Photosystem II; Electron transport

The chloroplast must rapidly and precisely adjust photosynthetic ATP and NADPH output to meet changing metabolic demands imposed by fluctuating environmental conditions. Cyclic electron flow (CEF) around photosystem I is thought to contribute to this adjustment by providing ATP in excess of that supplied by linear electron low, balancing chloroplast energy budget when relative demand for ATP is high. We assessed the kinetics and energy production of CEF activation in Chlamydomonas reinhardtii under rapid changes of organic and inorganic carbon availability. Comparisons of transient electric field and chlorophyll fluorescence measurements indicated CEF was activated under conditions where ATP demand is expected to be high, consistent with a role in balancing the cellular ATP/NADPH budget under fluctuating environmental or metabolic conditions. CEF activation was not correlated with antenna state transitions, both in wild-type and the state transition mutant stt7-9, suggesting that CEF is rapidly regulated by allosteric or redox modulators. Comparing the CEF under ambient and high CO2 conditions suggests an increase in required energy output of approximately 1ATP/CO2 fixed, nearly sufficient to power proposed mechanistic models for the carbon-concentrating mechanism. Additionally, we see three-fold higher CEF rates in cells under steady-state conditions than cells under similar conditions with inhibited photosystem II, and up to five times higher in cells with severe depletion of inorganic carbon, implying that CEF has larger energetic capacity than predicted from some previous work.
Keywords: Photosynthesis; Chlamydomonas; State transitions; Cyclic electron flow; Stt7 ; Carbon-concentrating mechanism; Electrochromic shift

The field-dependence of the solid-state photo-CIDNP effect in two states of heliobacterial reaction centers by Smitha Surendran Thamarath; A. Alia; Esha Roy; Karthick Babu Sai Sankar Gupta; John H. Golbeck; Jörg Matysik (461-469).
The solid-state photo-CIDNP (photochemically induced dynamic nuclear polarization) effect is studied in photosynthetic reaction centers of Heliobacillus mobilis at different magnetic fields by 13C MAS (magic-angle spinning) NMR spectroscopy. Two active states of heliobacterial reaction centers are probed: an anaerobic preparation of heliochromatophores (“Braunstoff”, German for “brown substance”) as well as a preparation of cells after exposure to oxygen (“Grünstoff”, “green substance”). Braunstoff shows significant increase of enhanced absorptive (positive) signals toward lower magnetic fields, which is interpreted in terms of an enhanced differential relaxation (DR) mechanism. In Grünstoff, the signals remain emissive (negative) at two fields, confirming that the influence of the DR mechanism is comparably low.
Keywords: Heliobacteria; Electron transfer; Photo-CIDNP; Solid-state NMR; Magnetic field effect

Continuous ECS-indicated recording of the proton-motive charge flux in leaves by Christof Klughammer; Katharina Siebke; Ulrich Schreiber (471-487).
Technical features and examples of application of a special emitter–detector module for highly sensitive measurements of the electrochromic pigment absorbance shift (ECS) via dual-wavelength (550–520 nm) transmittance changes (P515) are described. This device, which has been introduced as an accessory of the standard, commercially available Dual-PAM-100 measuring system, not only allows steady-state assessment of the proton motive force (pmf) and its partitioning into ΔpH and ΔΨ components, but also continuous recording of the overall charge flux driven by photosynthetic light reactions. The new approach employs a double-modulation technique to derive a continuous signal from the light/dark modulation amplitude of the P515 signal. This new, continuously measured signal primarily reflects the rate of proton efflux via the ATP synthase, which under quasi-stationary conditions corresponds to the overall rate of proton influx driven by coupled electron transport. Simultaneous measurements of charge flux and CO2 uptake as a function of light intensity indicated a close to linear relationship in the light-limited range. A linear relationship between these two signals was also found for different internal CO2 concentrations, except for very low CO2, where the rate of charge flux distinctly exceeded the rate of CO2 uptake. Parallel oscillations in CO2 uptake and charge flux were induced by high CO2 and O2. The new device may contribute to the elucidation of complex regulatory mechanisms in intact leaves.
Keywords: CO2 gas exchange; DIRK method; Dual-PAM-100; Electrochromic absorbance shift; Photosynthetic electron transport; P515

Dibromothymoquinone (DBMIB) has been used as a specific inhibitor of plastoquinol oxidation at the Q0 binding site of the cytochrome b6f complex for 40 years. It is thought to suppress electron transfer between photosystem (PS) II and I, as well as cyclic electron transfer around PSI. However, DBMIB has also been reported to act as a quencher of chlorophyll excited states. In this study, we have re-evaluated the effects of DBMIB on chlorophyll excited states and PSII photochemistry. The results show that DBMIB significantly quenches the chlorophyll excited states of PSII antenna even at low concentration (from 0.1 μM), lowering the effective excitation rate of the actinic light. It also acts as a potent PSII electron acceptor retarding the reduction of the plastoquinone pool with almost maximal potency at 2 μM. Altogether, these results suggest that experiments using DBMIB can easily be misinterpreted and stress on the importance of taking into account all these side effects that occur in the same range of DBMIB concentration used for inhibition of plastoquinol oxidation (1 μM).
Keywords: Electron transport; Photosystem II; DBMIB; Fluorescence quenching; Thermoluminescence

We have previously investigated the response mechanisms of photosystem II complexes from spinach to strong UV and visible irradiations (Wei et al J Photochem Photobiol B 104:118–125, 2011). In this work, we extend our study to the effects of strong light on the unusual cyanobacterium Acaryochloris marina, which is able to use chlorophyll d (Chl d) to harvest solar energy at a longer wavelength (740 nm). We found that ultraviolet (UV) or high level of visible and near-far red light is harmful to A. marina. Treatment with strong white light (1,200 μmol quanta m−2 s−1) caused a parallel decrease in PSII oxygen evolution of intact cells and in extracted pigments Chl d, zeaxanthin, and α-carotene analyzed by high-performance liquid chromatography, with severe loss after 6 h. When cells were irradiated with 700 nm of light (100 μmol quanta m−2 s−1) there was also bleaching of Chl d and loss of photosynthetic activity. Interestingly, UVB radiation (138 μmol quanta m−2 s−1) caused a loss of photosynthetic activity without reduction in Chl d. Excess absorption of light by Chl d (visible or 700 nm) causes a reduction in photosynthesis and loss of pigments in light harvesting and photoprotection, likely by photoinhibition and inactivation of photosystem II, while inhibition of photosynthesis by UVB radiation may occur by release of Mn ion(s) in Mn4CaO5 center in photosystem II.
Keywords: Acaryochloris marina ; Photoinhibition; UV irradiation; Chlorophyll d ; High-performance liquid chromatography

Expression of a highly active catalase VktA in the cyanobacterium Synechococcus elongatus PCC 7942 alleviates the photoinhibition of photosystem II by Haruhiko Jimbo; Akiko Noda; Hidenori Hayashi; Takanori Nagano; Isao Yumoto; Yoshitake Orikasa; Hidetoshi Okuyama; Yoshitaka Nishiyama (509-515).
The repair of photosystem II (PSII) after photodamage is particularly sensitive to reactive oxygen species—such as H2O2, which is abundantly produced during the photoinhibition of PSII. In the present study, we generated a transformant of the cyanobacterium Synechococcus elongatus PCC 7942 that expressed a highly active catalase, VktA, which is derived from a facultatively psychrophilic bacterium Vibrio rumoiensis, and examined the effect of expression of VktA on the photoinhibition of PSII. The activity of PSII in transformed cells declined much more slowly than in wild-type cells when cells were exposed to strong light in the presence of H2O2. However, the rate of photodamage to PSII, as monitored in the presence of chloramphenicol, was the same in the two lines of cells, suggesting that the repair of PSII was protected by the expression of VktA. The de novo synthesis of the D1 protein, which is required for the repair of PSII, was activated in transformed cells under the same stress conditions. Similar protection of the repair of PSII in transformed cells was also observed under strong light at a relatively low temperature. Thus, the expression of the highly active catalase mitigates photoinhibition of PSII by protecting protein synthesis against damage by H2O2 with subsequent enhancement of the repair of PSII.
Keywords: Catalase; Oxidative stress; Photoinhibition; Photosystem II; Protein synthesis

Using radioactively labelled amino acids to investigate repair of photoinactivated photosystem II (PS II) gives only a relative rate of repair, while using chlorophyll fluorescence parameters yields a repair rate coefficient for an undefined, variable location within the leaf tissue. Here, we report on a whole-tissue determination of the rate coefficient of photoinactivation k i , and that of repair k r in cotton leaf discs. The method assays functional PS II via a P700 kinetics area associated with PS I, as induced by a single-turnover, saturating flash superimposed on continuous background far-red light. The P700 kinetics area, directly proportional to the oxygen yield per single-turnover, saturating flash, was used to obtain both k i and k r . The value of k i , directly proportional to irradiance, was slightly higher when CO2 diffusion into the abaxial surface (richer in stomata) was blocked by contact with water. The value of k r , sizable in darkness, changed in the light depending on which surface was blocked by contact with water. When the abaxial surface was blocked, k r first peaked at moderate irradiance and then decreased at high irradiance. When the adaxial surface was blocked, k r first increased at low irradiance, then plateaued, before increasing markedly at high irradiance. At the highest irradiance, k r differed by an order of magnitude between the two orientations, attributable to different extents of oxidative stress affecting repair (Nishiyama et al., EMBO J 20: 5587–5594, 2001). The method is a whole-tissue, convenient determination of the rate coefficient of photoinactivation k i and that of repair k r .
Keywords: Chlorophyll fluorescence; P700; Photosystem II; Photoinactivation of photosystem II; Repair of photosystem II

Photosynthetic electron transport and specific photoprotective responses in wheat leaves under drought stress by Marek Zivcak; Marian Brestic; Zuzana Balatova; Petra Drevenakova; Katarina Olsovska; Hazem M. Kalaji; Xinghong Yang; Suleyman I. Allakhverdiev (529-546).
The photosynthetic responses of wheat (Triticum aestivum L.) leaves to different levels of drought stress were analyzed in potted plants cultivated in growth chamber under moderate light. Low-to-medium drought stress was induced by limiting irrigation, maintaining 20 % of soil water holding capacity for 14 days followed by 3 days without water supply to induce severe stress. Measurements of CO2 exchange and photosystem II (PSII) yield (by chlorophyll fluorescence) were followed by simultaneous measurements of yield of PSI (by P700 absorbance changes) and that of PSII. Drought stress gradually decreased PSII electron transport, but the capacity for nonphotochemical quenching increased more slowly until there was a large decrease in leaf relative water content (where the photosynthetic rate had decreased by half or more). We identified a substantial part of PSII electron transport, which was not used by carbon assimilation or by photorespiration, which clearly indicates activities of alternative electron sinks. Decreasing the fraction of light absorbed by PSII and increasing the fraction absorbed by PSI with increasing drought stress (rather than assuming equal absorption by the two photosystems) support a proposed function of PSI cyclic electron flow to generate a proton-motive force to activate nonphotochemical dissipation of energy, and it is consistent with the observed accumulation of oxidized P700 which causes a decrease in PSI electron acceptors. Our results support the roles of alternative electron sinks (either from PSII or PSI) and cyclic electron flow in photoprotection of PSII and PSI in drought stress conditions. In future studies on plant stress, analyses of the partitioning of absorbed energy between photosystems are needed for interpreting flux through linear electron flow, PSI cyclic electron flow, along with alternative electron sinks.
Keywords: Drought stress; Wheat; Photosynthetic electron transport; Cyclic electron transport around PSI; Photosystem stoichiometry; Chlorophyll fluorescence; Alternative electron sinks

Age-dependent changes in the functions and compositions of photosynthetic complexes in the thylakoid membranes of Arabidopsis thaliana by Krishna Nath; Bong-Kwan Phee; Suyeong Jeong; Sun Yi Lee; Yoshio Tateno; Suleyman I. Allakhverdiev; Choon-Hwan Lee; Hong Gil Nam (547-556).
Photosynthetic complexes in the thylakoid membrane of plant leaves primarily function as energy-harvesting machinery during the growth period. However, leaves undergo developmental and functional transitions along aging and, at the senescence stage, these complexes become major sources for nutrients to be remobilized to other organs such as developing seeds. Here, we investigated age-dependent changes in the functions and compositions of photosynthetic complexes during natural leaf senescence in Arabidopsis thaliana. We found that Chl a/b ratios decreased during the natural leaf senescence along with decrease of the total chlorophyll content. The photosynthetic parameters measured by the chlorophyll fluorescence, photochemical efficiency (F v/F m) of photosystem II, non-photochemical quenching, and the electron transfer rate, showed a differential decline in the senescing part of the leaves. The CO2 assimilation rate and the activity of PSI activity measured from whole senescing leaves remained relatively intact until 28 days of leaf age but declined sharply thereafter. Examination of the behaviors of the individual components in the photosynthetic complex showed that the components on the whole are decreased, but again showed differential decline during leaf senescence. Notably, D1, a PSII reaction center protein, was almost not present but PsaA/B, a PSI reaction center protein is still remained at the senescence stage. Taken together, our results indicate that the compositions and structures of the photosynthetic complexes are differentially utilized at different stages of leaf, but the most dramatic change was observed at the senescence stage, possibly to comply with the physiological states of the senescence process.
Keywords: Arabidopsis thaliana ; Chl contents and Chl a/b ratio; Developmental stage and senescence; Nutrient mobilization; Photosynthetic performance; Photosynthetic complexes and their components

Activation of interspecies-hybrid Rubisco enzymes to assess different models for the Rubisco–Rubisco activase interaction by Rebekka M. Wachter; Michael E. Salvucci; A. Elizabete Carmo-Silva; Csengele Barta; Todor Genkov; Robert J. Spreitzer (557-566).
Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is prone to inactivation from non-productive binding of sugar-phosphates. Reactivation of Rubisco requires conformational remodeling by a specific chaperone, Rubisco activase. Rubisco activase from tobacco and other plants in the family Solanaceae is an inefficient activator of Rubisco from non-Solanaceae plants and from the green alga Chlamydomonas reinhardtii. To determine if the Rubisco small subunit plays a role in the interaction with Rubisco activase, a hybrid Rubisco (SSNT) composed of tobacco small subunits and Chlamydomonas large subunits was constructed. The SSNT hybrid, like other hybrid Rubiscos containing plant small subunits, supported photoautotrophic growth in Chlamydomonas, but growth in air was much slower than for cells containing wild-type Rubisco. The kinetic properties of the SSNT hybrid Rubisco were similar to the wild-type enzyme, indicating that the poor growth in air was probably caused by disruption of pyrenoid formation and the consequent impairment of the CO2concentrating mechanism. Recombinant Rubisco activase from Arabidopsis activated the SSNT hybrid Rubisco and hybrid Rubiscos containing spinach and Arabidopsis small subunits at rates similar to the rates with wild-type Rubisco. However, none of the hybrid Rubiscos was activated by tobacco Rubisco activase. That replacement of Chlamydomonas small subunits with plant small subunits does not affect the species-specific interaction between Rubisco and Rubisco activase suggests that the association is not dominated by the small subunits that surround the Rubisco central solvent channel. Therefore, the geometry of a side-on binding mode is more consistent with the data than a top-on or ring-stacking binding mode.
Keywords: CO2 fixation; Molecular chaperone; Protein–protein interaction; Conformational remodeling; Chlamydomonas ; Chloroplast