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Excess light can have a negative impact on photosynthesis; thus, plants have evolved many different ways to adapt to different light conditions to both optimize energy use and avoid damage caused by excess light. Analysis of the Arabidopsis (Arabidopsis thaliana) mutant snowy cotyledon4 (sco4) revealed a mutation in a chloroplast-targeted protein that shares limited homology with CaaX-type endopeptidases. The SCO4 protein possesses an important function in photosynthesis and development, with point mutations rendering the seedlings and adult plants susceptible to photooxidative stress. The sco4 mutation impairs the acclimation of chloroplasts and their photosystems to excess light, evidenced in a reduction in photosystem I function, decreased linear electron transfer, yet increased nonphotochemical quenching. SCO4 is localized to the chloroplasts, which suggests the existence of an unreported type of protein modification within this organelle. Phylogenetic and yeast complementation analyses of SCO4-like proteins reveal that SCO4 is a member of an unknown group of higher plant-specific proteinases quite distinct from the well-described CaaX-type endopeptidases RAS Converting Enzyme1 (RCE1) and zinc metallopeptidase STE24 and lacks canonical CaaX activity. Therefore, we hypothesize that SCO4 is a novel endopeptidase required for critical protein modifications within chloroplasts, influencing the function of proteins involved in photosynthesis required for tolerance to excess light.

We sought a rapid, non-intrusive, whole-tissue measure of the functional photosystem II (PS II) content in leaves. Summation of electrons, delivered by a single-turnover flash to P700(+) (oxidized PS I primary donor) in continuous background far-red light, gave a parameter S in absorbance units after taking into account an experimentally determined basal electron flux that affects P700 redox kinetics. S was linearly correlated with the functional PS II content measured by the O(2) yield per single-turnover repetitive flash in Arabidopsis thaliana expressing an antisense construct to the PsbO (manganese-stabilizing protein in PS II) proteins of PS II (PsbO mutants). The ratio of S to z(max) (total PS I content in absorbance units) was comparable to the PS II/PS I reaction-center ratio in wild-type A. thaliana and in control Spinacea oleracea. Both S and S/z(max) decreased in photoinhibited spinach leaf discs. The whole-tissue functional PS II content and the PS II/photosystem I (PS I) ratio can be non-intrusively monitored by S and S/z(max), respectively, using a quick P700 absorbance protocol compatible with modern P700 instruments.

The importance of photoprotective strategies for Photosystem (PS) II function under all light regimes was determined from the content of functional PS II, measured by repetitive flash yield of oxygen evolution, in leaves of pea (Pisum sativum L.) grown under 50 (low light), 250 (medium light), and 650 (high light) μmol photons m -2 s -1 . The modulation of PS II functionality in vivo was induced in 1.1% CO 2 after infiltration of leaves with water (control), nigericin (a lipophilic uncoupler) or dithiothreitol (DTT, inhibitor of violaxanthin to zeaxanthin conversion) through the cut petioles of leaves of 22-24 day-old plants. In all nigericin-treated pea leaves, photoinactivation of PS II was greatly increased with increasing photon exposure (mol photons m -2 ). This nigericin-induced increase of photoinactivation of PS II was greater than that induced by DTT in medium- and high-light pea leaves, but comparable in low-light peas. In low-light peas during steady-state photosynthesis, the development of non-photochemical quenching of chlorophyll fluorescence (NPQ, measured as F m /F' m - 1) was lowest, accompanying the highest reduction state of PS II (measured by photochemical quenching, q P ). Further, the susceptibility of PS II to light stress, estimated as (1 - q P )/NPQ [Park et al., Plant Cell Physiol., 36 (1995) 1163-1167], was highest in low-light peas. The decline of chlorophyll fluorescence from maximum (F m ) to steady-state level during induction phase of chlorophyll fluorescence was retarded by nigericin treatment, with a greater inhibitory effect in high- and medium- than low-light pea leaves. From these results, we suggest that the greater susceptibility of low-light peas to light stress than medium- and high-light peas is ascribed to lower capacities for both the utilization of absorbed light by photosynthesis and for non-radiative dissipation of absorbed light as heat.

Detached Pisum sativum leaves were used to study temperature-dependent and temperature-independent effects of supplementary UV-B radiation on chloroplast-associated components and properties. Temperature-independent effects were considered to be due to direct physical damage by the UV-B radiation itself, whereas temperature-dependent damage was thought to be a result of UV-B-triggered chemical reactions. Supplementary UV-B radiation caused a lowering in chlorophyll fluorescence (F v /F m ) at both 2 and 22°C (by about 20 and 2%, respectively) ; it is likely to be caused by direct interaction of UV-B quanta with Photosystem II (PS II), since no additional decrease was seen in F v /F m when leaves irradiated at 2°C were subsequently incubated in darkness for 4 h at 22°C. However, chilling made PS II more prone to UV-B damage. The development of the UV-B effect on the relaxation of the electrochromic shift (t 1/2 ), caused by the trans-thylakoid membrane potential, showed a greater temperature dependency which indicates that UV-B induces chemical reactions that in turn cause the damage. Exposure for 4 h to supplementary UV-B radiation led to a 75% decrease in t 1/2 at 22°C, whereas the effect at 2°C was much smaller (

Supplementary ultraviolet-B radiation (UV-B) caused decreases in mRNA levels for photosynthetic genes and for the chloroplastic defensive enzyme Cu/Zn-superoxide dismutase. The magnitude of these decreases depended on the intensity of the photosynthetically active radiation (PAR): the mRNA levels being considerably higher under high PAR (400-600 /iE m"2 s"1) than under low (150 fiE m~2 s"1). Transcript levels for the non-chloroplastic defence protein chalcone synthase also were PAR-dependent, since an increase of mRNA transcripts occurred under low PAR only. UV-B also caused a PAR-independent increase in the rates of relaxation of the single turnover flashinduced electrochromic shift of thylakoid pigments. The UV-B doses needed were about 10-fold lower than the doses needed to affect photosynthetic components. Chlorophyll content and chlorophyll fluorescence of PSII were unaltered by these doses. We suggest that the lowered sensitivity of mRNA levels under high PAR may be caused by a protective mechanism at the level of DNA or transcription. This mechanism would not be involved in repair of the thylakoid membrane or its lipids. These results also highlight the importance of PAR intensity for the assessing of UV-B effects on plants.

Cyclic electron flux (CEF) around PSI is essential for efficient photosynthesis and aids photoprotection, especially in stressful conditions, but the difficulty in quantifying CEF is non-trivial. The total electron flux through PSI (ETR1) and the linear electron flux (LEFO2) through both photosystems in spinach leaf discs were estimated from the photochemical yield of PSI and the gross oxygen evolution rate, respectively, in CO2-enriched air. ΔFlux=ETR1 - LEFO2 is an upper estimate of CEF. Infiltration of leaf discs with 150μM antimycin A did not affect LEFO2, but decreased ΔFlux 10-fold. ΔFlux was practically negligible below 350μmolphotonsm-2s-1, but increased linearly above it. The following results were obtained at 980μmolphotonsm-2s-1. ΔFlux increased 3-fold as the temperature increased from 5°C to 40°C. It did not decline at high temperature, even when LEFO2 decreased. ΔFlux increased by 80% as the relative water content of leaf discs decreased from 100 to 40%, when LEFO2 decreased 2-fold. The method of using ΔFlux as a non-intrusive upper estimate of steady-state CEF in leaf tissue appears reasonable when photorespiration is suppressed.

Photoinactivation of Photosystem II (PS II) during photosynthesis can lead to the loss of photochemical efficiency and decrease in crop yield. Plants have evolved various photoprotective strategies to ameliorate photoinactivation of PS II. Non-leaf organs of cotton also contribute to carbon gain, but it is not clear how they photoprotect themselves. This study investigated the photoprotective mechanisms in the leaf, bract, main stem and capsule wall of cotton. Our results suggested that the bract mainly relies on high activities of antioxidative enzymes and high ΔpH- and xanthophyll-regulated thermal dissipation (ΦNPQ) for photoprotection. The main stem preferentially dissipated its absorbed light energy via light-regulated as well as light-independent non-photochemical quenching, aided by the moderately high activities of antioxidative enzymes. The capsule wall was less able to remove reactive oxygen species due to lower activities of antioxidative enzymes, and less able to dissipate energy via heat due to its lower ΦNPQ. Its main photoprotective mechanisms seem to be (a) direct quenching of the energy by abundant carotenoids and (b) light-independent constitutive thermal dissipation via Φf,D. The green organs of cotton have different ways to use or dissipate energy.

We quantified the photosynthetic cyclic electron flux (CEF) around Photosystem I as the difference between the total electron flux through PS I (ETR1) and the linear electron flux through both photosystems. Both measurements were made in the whole tissue of spinach leaf discs illuminated in the same geometry and in CO2-enriched air to suppress photorespiration. (1) CEF was negligibly small below 300 umol photons m−2s−1. Above this irradiance, CEF increased approximately linearly up to the highest irradiance used (1,900 umol photons m−2s−1). (2) CEF at a fixed irradiance of 980 μmol m−2s−1 increased by a factor of almost 3 as the temperature was increased from 5 °C to 40 °C. It did not decline, even when the linear electron flux decreased at high temperatures. (3) Antimycin A, at a high concentration, decreased CEF to about 10% of the control value without affecting the linear electron flux. This method appears to be reliable for quantifying CEF non-intrusively. By contrast, estimation of the linear electron flux from chlorophyll fluorescence over-estimated CEF in the above treatments.

Gradients of photoinhibition within a leaf caused by different color lights were studied to get more insight into the controversy whether photon absorption by chlorophyll or Mn is the primary cause of photoinhibition, suggested by the excess-energy hypothesis or the Mn (two-step) hypothesis, respectively. We photoinhibited lincomycin-treated leaf-discs with white, blue, green or red light. A combination of a micro-fiber fluorometer, a fiber-thinning technique and a micro-manipulator enabled us to measure the chlorophyll fluorescence signals within a leaf. Gradients of photoinhibition were also compared with results from various conventional fluorometers to estimate their depth of signal detection. The photoinhibition was more severe in the descending order of blue, red and green light near the adaxial surface, and in the descending order of blue, green and red light in the deeper tissue, which is correlated with the absorption spectrum of chlorophyll and Mn, respectively. These results cannot be explained by either hypothesis alone and strongly suggest that both mechanisms occur in photoinhibition. Fv/Fm values of photoinhibited leaves estimated with the conventional fluorometers were different from the whole tissue. This is because the depths, in which these systems detect fluorescence signals, differ depending on the wavelengths of measuring beam and detector.

Arabidopsis thaliana leaves were examined in short-term (1 h) and long-term (10 h) irradiance experiments involving growth, saturating and excess light. Changes in photosynthetic and chlorophyll fluorescence parameters and in populations of functional photosystem II (PSII) centers were independently measured. Xanthophyll pigments, 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU)-binding sites, the amounts of D1 protein, and the rates of D1 protein synthesis were determined. These comprehensive studies revealed that under growth or light-saturating conditions, photosynthetic parameters remained largely unaltered. Photoprotection occurred at light saturation indicated by a dark-reversible increase in non-photochemical quenching accompanied by a 5-fold increase in antheraxanthin and zeaxanthin. No consistent change in the concentrations of functional PSII centers, DCMU-binding sites, or D1 protein pool size occurred. D1 protein synthesis was rapid. In excess irradiance, quantum yield of O2 evolution and the efficiency of PSII were reduced, associated with a 15- to 20-fold increase in antheraxanthin and zeaxanthin and a sustained increase in nonphotochemical quenching. A decrease in functional PSII center concentration occurred, followed by a decline in the concentration of D1 protein; the latter, however, was not matched by a decrease in DCMU-binding sites. In the most extreme treatments, DCMU-binding site concentration remained 2 times greater than the concentration of D1 protein recognized by antibodies. D1 protein synthesis rates remained unaltered at excess irradiances.

Photosynthetic membrane sacs (thylakoids) of plants form granal stacks interconnected by non-stacked thylakoids, thereby being able to fine-tune (i) photosynthesis, (ii) photoprotection and (iii) acclimation to the environment. Growth in low light leads to the formation of large grana, which sometimes contain as many as 160 thylakoids. The net surface charge of thylakoid membranes is negative, even in low-light-grown plants; so an attractive force is required to overcome the electrostatic repulsion. The theoretical van der Waals attraction is, however, at least 20-fold too small to play the role. We determined the enthalpy change, in the spontaneous stacking of previously unstacked thylakoids in the dark on addition of Mg 2+ , to be zero or marginally positive (endothermic). The Gibbs free-energy change for the spontaneous process is necessarily negative, a requirement that can be met only by an increase in entropy for an endothermic process. We conclude that the dominant attractive force in thylakoid stacking is entropy-driven. Several mechanisms for increasing entropy upon stacking of thylakoid membranes in the dark, particularly in low-light plants, are discussed. In the light, which drives the chloroplast far away from equilibrium, granal stacking accelerates non-cyclic photophosphorylation, possibly enhancing the rate at which entropy is produced.

Long-term acclimation of shade versus sun plants modulates the composition, function and structural organization of the architecture of the thylakoid membrane network. Significantly, these changes in the macroscopic structural organization of shade and sun plant chloroplasts during long-term acclimation are also mimicked following rapid transitions in irradiance: reversible ultrastructural changes in the entire thylakoid membrane network increase the number of grana per chloroplast, but decrease the number of stacked thylakoids per granum in seconds to minutes in leaves. It is proposed that these dynamic changes depend on reversible macro-reorganization of some light-harvesting complex IIb and photosystem II supracomplexes within the plant thylakoid network owing to differential phosphorylation cycles and other biochemical changes known to ensure flexibility in photosynthetic functionin vivo.Some lingering grana enigmas remain: elucidation of the mechanisms involved in the dynamic architecture of the thylakoid membrane network under fluctuating irradiance and its implications for function merit extensive further studies.

Photosystem (PS) II is the multisubunit complex which uses light energy to split water, providing the reducing equivalents needed for photosynthesis. The complex is susceptible to damage from environmental stresses such as excess excitation energy and high temperature. This research investigated the in vivo photosynthetic consequences of impairments to PSII in Arabidopsis thaliana (ecotype Columbia) expressing an antisense construct to the PsbO proteins of PSII. Transgenic lines were obtained with between 25 and 60% of wild-type (WT) total PsbO protein content, with the PsbO1 isoform being more strongly reduced than PsbO2. These changes coincided with a decrease in functional PSII content. Low PsbO (less than 50% WT) plants grew more slowly and had lower chlorophyll content per leaf area. There was no change in content per unit area of cytochrome b6f, ATP synthase, or Rubisco, whereas PSI decreased in proportion to the reduction in chlorophyll content. The irradiance response of photosynthetic oxygen evolution showed that low PsbO plants had a reduced quantum yield, but matched the oxygen evolution rates of WT plants at saturating irradiance. It is suggested that these plants had a smaller pool of PSII centres, which are inefficiently connected to antenna pigments resulting in reduced photochemical efficiency.

Since photosystem II (PS II) performs the demanding function of water oxidation using light energy, it is susceptible to photoinactivation during photosynthesis. The time course of photoinactivation of PS II yields useful information about the process. Depending on how PS II function is assayed, however, the time course seems to differ. Here, we revisit this problem by using two additional assays: (1) the quantum yield of oxygen evolution in limiting, continuous light and (2) the flash-induced cumulative delivery of PS II electrons to the oxidized primary donor (P700(+)) in PS I measured as a 'P700 kinetics area'. The P700 kinetics area is based on the fact that the two photosystems function in series: when P700 is completely photo-oxidized by a flash added to continuous far-red light, electrons delivered from PS II to PS I by the flash tend to re-reduce P700(+) transiently to an extent depending on the PS II functionality, while the far-red light photo-oxidizes P700 back to the steady-state concentration. The quantum yield of oxygen evolution in limiting, continuous light indeed decreased in a way that deviated from a single-negative exponential. However, measurement of the quantum yield of oxygen in limiting light may be complicated by changes in mitochondrial respiration between darkness and limiting light. Similarly, an assay based on chlorophyll fluorescence may be complicated by the varying depth in leaf tissue from which the signal is detected after progressive photoinactivation of PS II. On the other hand, the P700 kinetics area appears to be a reasonable assay, which is a measure of functional PS II in the whole leaf tissue and independent of changes in mitochondrial respiration. The P700 kinetics area decreased in a single-negative exponential fashion during progressive photoinactivation of PS II in a number of plant species, at least at functional PS II contents ≥6 % of the initial value, in agreement with the conclusion of Sarvikas et al. (Photosynth Res 103:7-17, 2010). That is, the single-negative-exponential time course does not provide evidence for photoprotection of functional PS II complexes by photoinactivated, connected neighbours.

Given its unique function in light-induced water oxidation and its susceptibility to photoinactivation during photosynthesis, photosystem II (PS II) is often the focus of studies of photosynthetic structure and function, particularly in environmental stress conditions. Here we review four approaches for quantifying or monitoring PS II functionality or the stoichiometry of the two photosystems in leaf segments, scrutinizing the approximations in each approach. (1) Chlorophyll fluorescence parameters are convenient to derive, but the information-rich signal suffers from the localized nature of its detection in leaf tissue. (2) The gross O(2) yield per single-turnover flash in CO(2)-enriched air is a more direct measurement of the functional content, assuming that each functional PS II evolves one O(2) molecule after four flashes. However, the gross O(2) yield per single-turnover flash (multiplied by four) could over-estimate the content of functional PS II if mitochondrial respiration is lower in flash illumination than in darkness. (3) The cumulative delivery of electrons from PS II to P700(+) (oxidized primary donor in PS I) after a flash is added to steady background far-red light is a whole-tissue measurement, such that a single linear correlation with functional PS II applies to leaves of all plant species investigated so far. However, the magnitude obtained in a simple analysis (with the signal normalized to the maximum photo-oxidizable P700 signal), which should equal the ratio of PS II to PS I centers, was too small to match the independently-obtained photosystem stoichiometry. Further, an under-estimation of functional PS II content could occur if some electrons were intercepted before reaching PS I. (4) The electrochromic signal from leaf segments appears to reliably quantify the photosystem stoichiometry, either by progressively photoinactivating PS II or suppressing PS I via photo-oxidation of a known fraction of the P700 with steady far-red light. Together, these approaches have the potential for quantitatively probing PS II in vivo in leaf segments, with prospects for application of the latter two approaches in the field.

Artificial photosynthesis in the broadest sense encompasses human attempts to convert sunlight, water and carbon dioxide into carbohydrates, other organic compounds and oxygen, to split water into hydrogen and oxygen using sunlight, or to convert sunlight directly into electricity. Although largely still in its infancy, artificial photosynthesis is being researched on various fronts. Moderately-high-efficiency silicon photovoltaic generators are the most established and durable devices, but their efficiencies are still very much below thermodynamic limitation; thus, further improvements are feasible. Of great concern is how to lower the costs of manufacturing silicon photovoltaic devices. Dye-sensitized photovoltaic cells, when designed to absorb a broad spectrum of solar radiation, seem to offer great promise, especially in terms of lower manufacturing costs. The field of photon-harvesting for non-biological hydrogen production has advanced steadily, ever since the discovery of light-induced splitting of water into hydrogen and oxygen at a semiconductor electrode (Fujishima A, Honda K, Nature 238:37–38, 1972). A deficiency that needs to be addressed, however, is the limited absorption of the solar spectrum by a semiconductor. A tandem device with two semiconductor electrodes, absorbing complementary portions of the solar spectrum and working in series, partially overcomes the poor absorption, in a way that is analogous to the “Z Scheme” of natural photosynthesis in which two photosystems operate in series. Photoconversion efficiency, however, has to be further improved. Artificial reaction centers capable of light-induced charge separation have been constructed, based on porphyrin-fullerene complexes, ruthenium complexes or synthetic proteins holding redox cofactors. The longer-term application of such artificial reaction centers is their coupling to chemical/biochemical reactions that produce useful products/devices. Another spin-off from research on artificial reaction centers is the basic understanding of the mechanisms of forward and back charge transfers that can be gained from a comparison of natural and artificial reaction centers. Perhaps the most ambitious goal in artificial photosynthesis is the in vitro making of carbon-based end products, especially foodstuffs. Such a supplementation of the natural process may be necessitated by a gradual loss of agricultural land, a shortage of fresh water and a need to feed an increasing world population. Artificial photosynthesis for food production may present one of the greatest challenges to future scientists and technologists.

Non-foliar green organs are recognized as important carbon sources after leaves. However, the contribution of each organ to total yield has not been comprehensively studied in relation to the time-course of changes in surface area and photosynthetic activity of different organs at different growth stages. We studied the contribution of leaves, main stem, bracts and capsule wall in cotton by measuring their time-course of surface area development, O(2) evolution capacity and photosynthetic enzyme activity. Because of the early senescence of leaves, non-foliar organs increased their surface area up to 38.2% of total at late growth stage. Bracts and capsule wall showed less ontogenetic decrease in O(2) evolution capacity per area and photosynthetic enzyme activity than leaves at the late growth stage. The total capacity for O(2) evolution of stalks and bolls (bracts plus capsule wall) was 12.7 and 23.7% (total ca. 36.4%), respectively, as estimated by multiplying their surface area by their O(2) evolution capacity per area. We also kept the bolls (from 15 days after anthesis) or main stem (at the early full bolling stage) in darkness for comparison with non-darkened controls. Darkening the bolls and main stem reduced the boll weight by 24.1 and 9%, respectively, and the seed weight by 35.9 and 16.3%, respectively. We conclude that non-foliar organs significantly contribute to the yield at the late growth stage.

This paper reports an experimental test of the hypothesis that cotton and soybean differing in leaf movement have distinct strategies to perform photosynthesis under drought. Cotton and soybean were exposed to two water regimes: drought stressed and well watered. Drought-stressed cotton and soybean had lower maximum CO2 assimilation rates than well-watered (control) plants. Drought reduced the light saturation point and photorespiration of both species – especially in soybean. Area-based leaf nitrogen decreased in drought-stressed soybean but increased in drought-stressed cotton. Drought decreased PSII quantum yield (ΦPSII) in soybean leaves, but increased ΦPSII in cotton leaves. Drought induced an increase in light absorbed by the PSII antennae that is dissipated thermally via ΔpH- and xanthophylls-regulated processes in soybean leaves, but a decrease in cotton leaves. Soybean leaves appeared to have greater cyclic electron flow (CEF) around PSI than cotton leaves and drought further increased CEF in soybean leaves. In contrast, CEF slightly decreased in cotton under drought. These results suggest that the difference in leaf movement between cotton and soybean leaves gives rise to different strategies to perform photosynthesis and to contrasting photoprotective mechanisms for utilisation or dissipation of excess light energy. We suggest that soybean preferentially uses light-regulated non-photochemical energy dissipation, which may have been enhanced by the higher CEF in drought-stressed leaves. In contrast, cotton appears to rely on enhanced electron transport flux for light energy utilisation under drought, for example, in enhanced nitrogen assimilation.

Leaf anatomy of C3 plants is mainly regulated by a systemic irradiance signal. Since the anatomical features of C4 plants are different from that of C3 plants, we investigated whether the systemic irradiance signal regulates leaf anatomical structure and photosynthetic performance in sorghum (Sorghum bicolor), a C4 plant. Compared with growth under ambient conditions (A), no significant changes in anatomical structure were observed in newly developed leaves by shading young leaves alone (YS). Shading mature leaves (MS) or whole plants (S), on the other hand, caused shade-leaf anatomy in newly developed leaves. By contrast, chloroplast ultrastructure in developing leaves depended only on their local light conditions. Functionally, shading young leaves alone had little effect on their net photosynthetic capacity and stomatal conductance, but shading mature leaves or whole plants significantly decreased these two parameters in newly developed leaves. Specifically, the net photosynthetic rate in newly developed leaves exhibited a positive linear correlation with that of mature leaves, as did stomatal conductance. In MS and S treatments, newly developed leaves exhibited severe photoinhibition under high light. By contrast, newly developed leaves in A and YS treatments were more resistant to high light relative to those in MS- and S-treated seedlings. We suggest that (1) leaf anatomical structure, photosynthetic capacity, and high-light tolerance in newly developed sorghum leaves were regulated by a systemic irradiance signal from mature leaves; and (2) chloroplast ultrastructure only weakly influenced the development of photosynthetic capacity and high-light tolerance. The potential significance of the regulation by a systemic irradiance signal is discussed.

Trandescantia albiflora (Kunth) was grown under two different light quality regimes of comparable light quantity: in red + far-red light absorbed mainly by photosystem I(PSI light) and yellow light absorbed mainly by photosystem II (PSII light). The composition, function and ultrastructure of chloroplasts, and photoinhibition of photosynthesis in the two types of leaves were compared. In contrast to regulation by light quantity (Chow et al. 1991. Physiol. Plant. 81: 175-182), light quality exerted an effect on the composition of pigment complexes, function and structure of chloroplasts in Tradescantia: PSII light-grown leaves had higher Chi a/b ratios, higher PSI concentrations, lower PSII/PSI reaction centre ratios and less extensive thylakoid stacking than PSI light-grown leaves. Light quality triggered modulations of chloroplast components, leading to a variation of photosynthetic characteristics. A larger proportion of primary quinone acceptor (Q(A)) in PSI light-grown leaves was chemically reduced at any given irradiance. It was also observed that the quantum yield of PSII photochemistry was lower in PSI light-grown leaves. PSI light-grown leaves were more sensitive to photoinihibition and recovery was slower compared to PSII light-grown leaves, showing that the PSII reaction centre in PSI light-grown leaves was more easily impaired by photoinhibition. The increase in susceptibility of leaves to photoinhibition following blockage of chloroplast-encoded protein synthesis was greater in PSII light-grown leaves, showing that these leaves normally have a greater capacity for PSII repair. Inhibition of zeaxanthin formation by dithiothreitol slightly increased sensitivity to photoinhibition in both PSI and PSII light-grown leaves.

Wild-type and an atrazine-resistant biotype of Brassica napus, in which a glycine is substituted for the serine-264 of the D1protein, were grown over a wide range of constant irradiances in a growth cabinet. In the absence of serine-264, the function of photosystem II (PSII) was changed as reflected by changes in chlorophyll fluorescence parameters and in photosynthetic oxygen-evolving activity. The photochemical quenching coefficient was lower, showing that a larger proportion of the primary quinone acceptor is reduced at all irradiances. At low actinic irradiances, the nonphotochemical quenching coefficient was higher, showing a greater tendency for heat emission. Decreased rates of light-limited photosynthesis (quantum yield) and lower oxygen yields per single-turnover flash were also observed. These changes were observed even when the plants had been grown under low irradiances, indicating that the changes in PSII function are direct and not consequences of photoinhibition. In spite of the lowered PSII efficiency under light-limiting conditions, the light-saturated photosynthesis rate of the atrazine-resistant mutant was similar to that of the wild type. An enhanced susceptibility to photoinhibition was observed for the atrazine-resistant biotype compared to the wild type when plants were grown under high and intermediate, but not low, irradiance. We conclude that the replacement of serine by glycine in the D1 protein has a direct effect on PSII function, which in turn causes increased photoinhibitory damage and increased rates of turnover of the D1 protein. Both the intrinsic lowering of light-limited photosynthetic efficiency and the increased sensitivity to photoinhibition probably contribute to reduced crop yields in the field, to different extents, depending on growth conditions.

The productivity of fruit trees is a linear function of the light intercepted, although the relationship is less tight when greater than 50% of available light is intercepted. This paper investigates the management of light energy in peach using the measurement of whole-tree light interception and gas exchange, along with the absorbed energy partitioning at the leaf level by concurrent measurements of gas exchange and chlorophyll fluorescence. These measurements were performed on trees of a custom-built ‘asymmetric’ orchard. Whole-tree gas exchange for north–south, vertical canopies (C) was similar to that for canopies intercepting the highest irradiance in the morning hours (W), but trees receiving the highest irradiance in the afternoon (E) had the highest net photosynthesis and transpiration while maintaining a water use efficiency (WUE) comparable to the other treatments. In the W trees, 29% and 8% more photosystems were damaged than in C and E trees, respectively. The quenching partitioning revealed that the non-photochemical quenching (NPQ) played the most important role in excess energy dissipation, but it was not fully active at low irradiance, possibly due to a sub-optimal trans-thylakoid DpH. The non-net carboxylative mechanisms (NC) appeared to be the main photoprotective mechanisms at low irradiance levels and, probably, they could facilitate the establishment of a trans-thylakoid DpH more appropriate for NPQ. These findings support the conclusion that irradiance impinging on leaves may be excessive and can cause photodamage, whose repair requires energy in the form of carbohydrates that are thereby diverted from tree growth and productivity.

Ten anthocyanin components have been detected in roots of purple sweet potato (Ipomoea batatas Lam.) by high-performance liquid chromatography coupled to diode array detection and electrospray ionization tandem mass spectrometry. All the anthocyanins were exclusively cyanidins or peonidin 3-sophoroside-5-glucosides and their acylated derivatives. The total anthocyanin content in purple sweet potato powder obtained by solid-phase extraction was 66 mg g(-1). A strong capacity of purple sweet potato anthocyanins (PSPA) to scavenge reactive oxygen species (superoxide, hydroxyl radical) and the stable 1,1-diphenyl-2-picrylhydrazyl organic free radical was found in vitro using the electron spin resonance technique. To determine the functional roles of anthocyanins in leaves in vivo, for the first time, supplemental anthocyanins were infiltrated into leaves of Arabidopsis thaliana double mutant of the ecotype Landsberg erecta (tt3tt4) deficient in anthocyanin biosynthesis. Chlorophyll fluorescence imaging showed that anthocyanins significantly ameliorated the inactivation of photosystems II during prolonged high-light (1300 micromol m(-2) s(-1)) exposure. Comet assay of DNA revealed an obvious role of supplemental PSPA in alleviating DNA damage by high light in leaves. Our results suggest that anthocyanins could function in vitro and in vivo to alleviate the direct or indirect oxidative damage of the photosynthetic apparatus and DNA in plants caused by high-light stress.

To explore the mechanisms underlying water regulation in clonal plants and its effects on carbon assimilation under water stress, we studied the responses of water status, gas exchange and abscisic acid (ABA) contents to water stress in leaves of pairs of strawberry ramets that consist of mother and daughter ramets. There was a greater decrease in photosynthetic rates (P(n)) and stomatal conductance (G(s)) in the disconnected mother ramets than the connected mother ramets upon exposure to water stress, indicating that water stress in mother ramets was alleviated by water translocation from the well-watered daughter ramets. Conversely, the connected mother ramets displayed enhanced symptoms of water stress when the connected daughter ramets were exposed to water deficit. The mother ramets had lower water potential (psi(w)) due to their stronger osmotic adjustment than in well-watered daughter ramets; this resulted in water flow from the connected daughter ramets to mother ramets, thus alleviating water stress of mother ramets. During soil drying, there was a striking increase in ABA concentrations in leaves of the disconnected mother ramets, whereas leaf bulk ABA was much lower in the connected and water-stressed mother ramets than that in the drought-affected mother ramets in the disconnected group. In this study, though G(s) was linearly correlated with leaf bulk ABA and psi(w), G(s) in water-stressed mother ramets in disconnected group exhibited less sensitivity to the variation in leaf bulk ABA and psi(w) than that in connected and water-stressed mother ramets. Taken together, these results indicate that: (1) the flux of water translocation between the connected ramets is determined by a water potential gradient; (2) water translocation between connected ramets helps to keep sensitivity of G(s) to ABA and psi(w) in drought-affected ramets, thereby benefit to effectively maintain the homeostasis of leaf water status and (3) the improvements in P(n) in water-stressed ramets due to water translocation from well-watered ramets suggest the advantages of physiological integration in clonal plants in environments with heterogeneous water distribution.

For plants, light is an indispensable resource. However, it also causes a loss of photosynthetic activity associated with photoinactivation of photosystem II (PSII). In studies of the mechanism of this photoinactivation, there are two conflicting hypotheses at present. One is that excess energy received by leaves, being neither utilized by photosynthesis nor dissipated safely in non-photochemical quenching, causes the photoinactivation. The other involves a two-step mechanism in which excitation of Mn by photons is the primary cause. In the former hypothesis, photoinactivation of PSII should not occur in low light that provides little excess energy, but in the latter hypothesis it should. Therefore, we tested these two hypotheses in different irradiances. We used a system that can measure the fraction of functional PSII complexes under natural conditions and over a long period in intact leaves, which were attached to a plant treated with lincomycin taken up via the roots. The leaves were photoinactivated in low, medium or high light (30, 60 or 950 micromol m(-2) s(-1)) with white, blue, green or red light-emitting diode arrays. Our results showed that the extent of photoinactivation per photon exposure was higher in high light than in low light, consistent with the abundance of excess energy. However, photoinactivation did occur in low light with little excess energy, and blue light caused the greatest extent of photoinactivation followed by white, green and red light in this order, an order that can be predicted from the Mn absorbance spectrum. These results suggest that both mechanisms occur in the photoinactivation process.

Methyl viologen (MV) is a well-known electron mediator that works on the acceptor side of photosystem I. We investigated the little-known, MV-induced inhibition of linear electron flow through photosystem II (PS II) in spinach-leaf discs. Even a low [MV] decreased the (1) average, light-adapted photochemical efficiency of PS II traps, (2) oxidation state of the primary quinone acceptor Q(A) in PS II during illumination, (3) photochemical efficiency of light-adapted open PS II traps, (4) fraction of absorbed light energy dissipated constitutively in a light-independent manner or as chlorophyll (Chl) a fluorescence emission, (5) Chl a fluorescence yield corresponding to dark-adapted open reaction-center traps (F (o)) and closed reaction-center traps (F (m)), and (6) half-time for re-oxidation of Q (A) (-) in PS II after a single-turnover flash. These effects suggest that the presence of MV accelerates various "downhill" electron-transfer steps in PS II. Therefore, when using the MV to quantify cyclic electron flow, the inhibitory effect of MV on PS II should be taken into account.

Cyclic electron flow (CEF) around photosystem I has a role in avoiding photoinhibition of photosystem II (PSII), which occurs under conditions in which the rate of photodamage to PSII exceeds the rate of its repair. However, the molecular mechanism underlying how CEF contributes to photoprotection is not yet well understood. We examined the effect of impairment of CEF and thermal energy dissipation (qE) on photoinhibition using CEF (pgr5) and qE (npq1 and npq4) mutants of Arabidopsis (Arabidopsis thaliana) exposed to strong light. Impairment of CEF by mutation of pgr5 suppressed qE and accelerated photoinhibition. We found that impairment of qE, by mutations of pgr5, npq1, and npq4, caused inhibition of the repair of photodamaged PSII at the step of the de novo synthesis of the D1 protein. In the presence of the chloroplast protein synthesis inhibitor chloramphenicol, impairment of CEF, but not impairment of qE, accelerated photoinhibition, and a similar effect was obtained when leaves were infiltrated with the protonophore nigericin. These results suggest that CEF-dependent generation of ΔpH across the thylakoid membrane helps to alleviate photoinhibition by at least two different photoprotection mechanisms: one is linked to qE generation and prevents the inhibition of the repair of photodamaged PSII at the step of protein synthesis, and the other is independent of qE and suppresses photodamage to PSII.

Tradescantia albiflora (Kunth), a trailing ground species naturally occurring in deep shade in rainforests, has an unusual photosynthetic acclimation profile for growth irradiance. Although capable of increasing its capacity for electron transport, photophosphorylation and carbon fixation when grown in full sunlight, Tradescantia has constant chlorophyll alb ratios, photosystem reaction centre stoichiometry and pigment-protein composition at all growth irradiances (Chow et al. 1991. Physiol. Plant. 81: 175–182). To gain an insight into the compensatory strategies which allow Tradescantia to grow in both high and low lights, plants were grown under shade cloth (100 to 1.4% relative growth irradiance) and leaf and chloroplast attributes were compared. While shade Tradescantia chloroplasts had three times more chlorophyll per chloroplast and twice the length of thylakoid membranes compared to plants grown in full sunlight, the ratios of appressed to nonappressed thylakoid membranes were constant. The average net surface charge density of destacked thylakoids was the same for plants grown at moderate and low-irradiance, consistent with their similar stacking profiles. Tradescantia plants grown in direct sunlight had 10-times more fresh and dry weight per plant compared to plants grown in shade, despite a lower photosynthetic capacity on a leaf area basis with partial photoinhibition. We conclude that having a light-harvesting apparatus permanently locked into the “shade-plant mode” does not necessarily prevent a plant from thriving in high light. Analyses of leaf growth at different irradiances provide a partial explanation of the manner in which Tradescantia compensates for very low photosynthetic capacity per unit leaf in sunlight.

Spinach grown in light which preferentially excites Photosystem II (PSII-light) had increased amounts of PS I reaction centres on a chlorophyll (Chl) basis, a higher Chl a /Chl b ratio but less extensive thylakoid stacking, when compared with growth in light preferentially favouring excitation of PS I (PSI-light). Surprisingly, despite the marked difference in the extent of thylakoid stacking, the overall surface charge densities of thylakoid membranes were similar for plants grown in PSII- and PSI-light. The amounts of PS II reaction centres, cytochrome f and adenosine triphosphate synthase were also similar on a Chl basis for the two growth light qualities. Thylakoids of the Chl- b -less barley mutant had a smaller magnitude of surface charge density, although the extent of thylakoid stacking (Goodchild, D.J., Highkin, H.R. and Boardman, N.K. (1966) Exp. Cell Res. 43, 684–688) was much smaller, compared with wild-type barley. The results are interpreted in terms of the possible mechanisms governing the heterogeneous lateral distribution of the two photosystems. They suggest that PS I is laterally segregated to non-appressed thylakoid membrane domains because of the excess negative charges it carries on the outer membrane surface, whereas PS II is mainly sequestered into the appressed regions because of an association with its Chl a/b -proteins which appear to contribute more to van der Waals attraction than hitherto recognized.

The photosynthetic acclimation of Tradescantia albiflora (Kunth), a trailing ground species naturally occurring in the deep shade of rainforests, was studied in relation to growth irradiance (glasshouse; direct light and 1 to 4 layers of shade cloth, giving 100 to 1.4% relative growth irradiance). Contrary to other irradiance studies of higher plants grown in natural habitats or controlled light environments, the chlorophyll a/b ratios of Tradescantia leaves were low (∼2.2) and constant. Acclimation to growth irradiance caused no changes in the relative amounts of specific Chl-proteins or the numbers of photosystem I (PSI) and PSII reaction centres on a chlorophyll basis, indicating that the light-harvesting antenna sizes of PSII and PSI, as well as the photosystem stoichiometry, were independent of growth irradiance. However, the amount of cytochrome f and ATP synthase on a chlorophyll basis increased with increasing the relative growth irradiance from 1.4 to 35%, showing acclimation of electron transport and photophosphorylation capacity. The photosynthetic capacity and ribulose 1, 5-bisphosphate carboxylase (EC 4.1.1.39) activity also increased with increase of the growth irradiance to 35%. Beyond that, the inflexible PSII/PSI stoichiometry and shade-type photosystem II/light-harvesting units in Tradescaniia are a disadvantage for long-term exposure to high irradiance since the leaves are more prone to photoinhibition.

Pre-illumination of cucumber leaf discs at 4 degrees C with low-irradiance white light (i) led to a marked decrease in the extent of photo-oxidation of P700 (the special chlorophyll pair in the PSI reaction center) in actinic light at room temperature and (ii) hastened the post-illumination re-reduction of P700+. Quantifying the linear, cyclic and stroma-sourced electron fluxes to P700+ in two actinic light regimes, we found that there was no increase in cyclic or linear electron fluxes to account for these changes. Rather, we observed a decrease in the maximum extent of P700 photo-oxidation assayed by a strong flash superimposed on continuous, background light of wavelength 723 nm, which we interpret to represent a loss of stable charge separation in PSI due to enhanced charge recombination as a result of the pre-illumination treatment. The funneling of electrons towards fewer non-damaged PSI complexes could explain the hastened post-illumination re-reduction of P700+, aided by a slight increase in a stroma-sourced electron flux after prolonged pre-illumination at 4 degrees C. Quantifying the separate fluxes to P700+ helps to elucidate the effects of chilling of cucumber leaf discs in the light and the reasons for the hastened post-illumination re-reduction of P700+.

Linear and cyclic electron fluxes through Photosystem I in 1% CO(2) were quantified in spinach leaf tissue under severe water stress. Using actinic light with a peak at 697 nm for preferential light absorption by Photosystem I while also stimulating Photosystem II to improve redox poising, the cyclic electron flux after 60 s of illumination was a substantial proportion (33-44%) of the total electron flux through PSI at irradiances up to ~1,070 micromol photons m(-2) s(-1). At the maximum irradiance, the cyclic electron flux changed little with the progressive water loss from leaf tissue up to ~60%; by contrast, the linear electron flux was approximately halved. A reason for this differential effect of water stress on the capacity for cyclic and linear electron flow could be the increased crowding of soluble proteins in the stroma due to chloroplast shrinkage. Indeed the confinement of soluble proteins to a smaller chloroplast volume was indicated by cryo-scanning electron microscopy. It is known that the diffusion coefficient of large proteins is decreased when the background concentration of small proteins is raised; by contrast, the diffusion coefficient of small proteins is not affected by increasing the concentration of a large protein (Muramatsu and Minton in Proc Natl Acad Sci USA 85:2984-2988, 1988). Therefore, we suggest that linear electron flow, being coupled to the Calvin-Benson cycle, is limited by the diffusion of large macromolecules, especially the ribulose 1, 5-bisphosphate carboxylase/oxygenase complex. By contrast, cyclic electron flow, involving relatively small macromolecules such as ferredoxin, is less susceptible to inhibition by crowding in the stroma.

Assaying the number of functional PSII complexes by the oxygen yield from leaf tissue per saturating, single-turnover flash, assuming that each functional PSII evolves one oxygen molecule after four flashes, is one of the most direct methods but time-consuming. The ratio of variable to maximum Chl fluorescence yield (F(v)/F(m)) in leaves can be correlated with the oxygen yield per flash during a progressive loss of PSII activity associated with high-light stress and is rapid and non-intrusive, but suffers from being representative of chloroplasts near the measured leaf surface; consequently, the exact correlation depends on the internal leaf structure and on which leaf surface is being measured. Our results show that the average F(v)/F(m) of the adaxial and abaxial surfaces has a reasonable linear correlation with the oxygen yield per flash after varied extents of photoinactivation of PSII. However, we obtained an even better linear correlation between (1) the integrated, transient electron flow (Sigma) to P700+, the dimeric Chl cation in PSI, after superimposing a single-turnover flash on steady background far-red light and (2) the relative oxygen yield per flash. Leaves of C3 and C4 plants, woody and herbaceous species, wild-type and a Chl-b-less mutant, and monocot and dicot plants gave a single straight line, which seems to be a universal relation for predicting the relative oxygen yield per flash from Sigma. Measurement of Sigma is non-intrusive, representative of the whole leaf tissue, rapid and applicable to attached leaves; it may even be applicable in the field.

The stoichiometry of photosystem II to photosystem I reaction centres in spinach leaf segments was determined by two methods, each capable of monitoring both photosystems in a given sample. One method, based on the fast electrochromic (EC) signal, was applied to leaf segments, thereby avoiding potential artefacts associated with the isolation of thylakoids. Two variations of the EC method were used, either suppression of PSII activity by prior photoinactivation or suppression PSI by photo- oxidation of P700, gave the separate contribution of each photosystem to the fast EC signal. The PSII/PSI stoichiometries obtained by the EC methods ranged from 1.5 to 1.8 for spinach, and 1.5 to 1.9 for two other plant species. A second method, based on electron paramagnetic resonance (EPR), gave comparable values of 1.7–2.1 for spinach. A third method consisting of separate determination of the contents of functional PSII by oxygen yield per single turnover flash and of P700 gave a PSII/PSI stiochiometry consistent with above values. We conclude that the content of functional PSII is greater than that of PSI, and PSII/PSI reaction centre ratios considerably higher than unity in typical higher plants.

In studies of the mechanism of the photoinactivation of photosystem II (PSII), there are presently two conflicting hypotheses. One hypothesis is that excess energy received by leaves, neither utilized by photosynthesis nor safely dissipated in non-photochemical quenching, causes photoinactivation. The other involves a two-step mechanism in which excitation of Mn by photons is the primary cause of photoinactivation. In the former hypothesis, photoinactivation of PSII should not occur under low light with no excess energy conditions, but in the latter photoinactivation should occur. Therefore, we tested these two hypotheses under low light conditions. The leaves were photoinhibited in low light (30 μmol m−2 s−1) or in high light (950 μmol m−2 s−1) with blue, white, green or red LED lamps. The results showed that there was a difference in the extent of photoinactivation per photon exposure between low- and high-light conditions, and that photoinactivation did occur under low light with little excess energy condition. These results suggest that both mechanisms occur in the photoinactivation process.

Assaying functional Photosystem II (PSII) complexes by flash-induced oxygen yield from leaf tissue after photoinhibition is most direct but time-consuming, while measurement of chlorophyll fluorescence in leaves is only representative of chloroplasts near the leaf surface. To circumvent these deficiencies, we obtained an excellent linear correlation between (a) the integrated, transient electron flow (Σ) to PS I, after superimposing a single-turnover flash on steady far-red light, and (b) the relative oxygen yield per flash during progressive photoinactivation of Photosystem II. Leaves of C3 and C4 plants, woody and herbaceous species, wild type and a chlorophyll b-less mutant, and monocot and dicot plants gave a single linear correlation, which seems to be a universal relation for predicting the relative oxygen yield per flash from Σ.

Pea plants (Pisum sativum L., cv. Greenfeast) were exposed to supplementary UV-B light (up to 8 days) starting on the 17th day after sowing. The effects of this exposure on photosynthesis and the content and activities of some chloroplast components of the mature leaves of these plants were studied. (i) The total chorophyll content of pea leaves was approximately 40% of that in the control leaves on the 8th day of UV-B exposure. Chlorophyll a levels decreased to a greater extent than the content of chlorophyll b. The decrease in carotenoids paralleled the decrease in chlorophyll b. (ii) On a chlorophyll basis, the contents of Photosystem I and cytochrome f were stable, whereas Photosystem II, ATP hydrolysis by the ATP synthase and the maximum ribulose-1,5-bisphosphate carboxylase (Rubisco) activity decreased by 55, 47 and 80%, respectively, when compared with the controls at the end of the 8-day illumination period. (iii) On a leaf-area basis, Photosystem I and cytochrome f content decreased by 58%, Photosystem II by 80%, ATP hydrolysis by 80%, and Rubisco activity by 90%, when compared with the controls. The in vivo activation of Rubisco was markedly increased in UV-B-treated pea leaves. The underlying mechanisms for these results are discussed.

The efficiency of photosynthetic electron transport depends on the coordinated interaction of photosystem II (PSII) and photosystem I (PSI) in the electron-transport chain. Each photosystem contains distinct pigment-protein complexes that harvest light from different regions of the visible spectrum. The light energy is utilized in an endergonic electron-transport reaction at each photosystem. Recent evidence has shown a large variability in the PSII/PSI stoichiometry in plants grown under different environmental irradiance conditions. Results in this work are consistent with the notion of a dynamic, rather than static, thylakoid membrane in which the stoichiometry of the two photosystems is adjusted and optimized in response to different light quality conditions. Direct evidence is provided that photosystem stoichiometry adjustments in chloroplasts are a compensation strategy designed to correct unbalanced absorption of light by the two photosystems. Such adjustments allow the plant to maintain a high quantum efficiency of photosynthesis under diverse light quality conditions and constitute acclimation that confers to plants a significant evolutionary advantage over that of a fixed photosystem stoichiometry in thylakoid membranes.

Peas were grown in controlled environments (12h white fluorescent light. ∼47 μmol photons m-2 s 1/12 dark, 25 °C), using (1) 15-min far-red illumination at the end of each photoperiod (brief FR) to simulate the increase in the far-red/red ratio near the end of the day, and (2) high levels of supplementary far-red light (red:far-red ratio=0.04) during the entire photoperiod (long-term FR) to simulate extreme shade conditions under a plant canopy. Brief FR illumination led to marked morphological effects attributable to phytochrome regulation, namely, an increase in internodal length, but a decrease in leaflet area, chloroplast size and chlorophyll content per chloroplast compared with the control. Significantly, brief FR illumination had little or no effect on the amounts of the major chloroplast components (ribulose 1.5-biphosphate carboxylase, adenosine triphosphate synthase, cytochrome b/f complex and Photosystem II) relative to chlorophyll or Photosystem I, and the leaf photosynthetic capacities per unit chlorophyll were similar. In contrast, supplementing high levels of far-red light during the entire photoperiod not only led to the phytochrome effects above, but there was also a marked increase in leaf photosynthetic capacity per unit chlorophyll. due to increased amounts of the major chloroplast components relative to chlorophyll or Photosystem I. We hypothesize that supplementary far-red light, absorbed by Photosystem I, induced an increase in the major chloroplast components by a photosynthetic feedback mechanism. In fully greened leaves, we propose that the two photosystems themselves, rather than phytochrome, may be the predominent sensors of light quantity in triggering modulations of the stoichiometries of chloroplast components, which in turn lead to varying photosynthetic capacities.

Phosphatidylglycerol (PG), containing the unique fatty acid Delta3, trans-16:1-hexadecenoic acid, is a minor but ubiquitous lipid component of thylakoid membranes of chloroplasts and cyanobacteria. We investigated its role in electron transfers and structural organization of Photosystem II (PSII) by treating Arabidopsis thaliana thylakoids with phospholipase A(2) to decrease the PG content. Phospholipase A(2) treatment of thylakoids (a) inhibited electron transfer from the primary quinone acceptor Q(A) to the secondary quinone acceptor Q(B), (b) retarded electron transfer from the manganese cluster to the redox-active tyrosine Z, (c) decreased the extent of flash-induced oxidation of tyrosine Z and dark-stable tyrosine D in parallel, and (d) inhibited PSII reaction centres such that electron flow to silicomolybdate in continuous light was inhibited. In addition, phospholipase A(2) treatment of thylakoids caused the partial dissociation of (a) PSII supercomplexes into PSII dimers that do not have the complete light-harvesting complex of PSII (LHCII); (b) PSII dimers into monomers; and (c) trimers of LHCII into monomers. Thus, removal of PG by phospholipase A(2) brings about profound structural changes in PSII, leading to inhibition/retardation of electron transfer on the donor side, in the reaction centre, and on the acceptor side. Our results broaden the simple view of the predominant effect being on the Q(B)-binding site.

The variation of the rate of cyclic electron transport around Photosystem I (PS I) during photosynthetic induction was investigated by illuminating dark-adapted spinach leaf discs with red + far-red actinic light for a varied duration, followed by abruptly turning off the light. The post-illumination re-reduction kinetics of P700+, the oxidized form of the photoactive chlorophyll of the reaction centre of PS I (normalized to the total P700 content), was well described by the sum of three negative exponential terms. The analysis gave a light-induced total electron flux from which the linear electron flux through PS II and PS I could be subtracted, yielding a cyclic electron flux. Our results show that the cyclic electron flux was small in the very early phase of photosynthetic induction, rose to a maximum at about 30 s of illumination, and declined subsequently to

Arabidopsis plants in which the major trimeric light harvesting complex (LHCIIb) is eliminated by antisense expression still exhibit the typical macrostructure of photosystem II in the granal membranes. Here the detailed analysis of the composition and the functional state of the light harvesting antennae of both photosystem I and II of these plants is presented. Two new populations of trimers were found, both functional in energy transfer to the PSII reaction center, a homotrimer of CP26 and a heterotrimer of CP26 and Lhcb3. These trimers possess characteristic features thought to be specific for the native LHCIIb trimers they are replacing: the long wavelength form of lutein and at least one extra chlorophyll b, but they were less stable. A new population of loosely bound LHCI was also found, contributing to an increased antenna size for photosystem I, which may in part compensate for the loss of the phosphorylated LHCIIb that can associate with this photosystem. Thus, the loss of LHCIIb has triggered concerted compensatory responses in the composition of antennae of both photosystems. These responses clearly show the importance of LHCIIb in the structure and assembly of the photosynthetic membrane and illustrate the extreme plasticity at the level of the composition of the light harvesting system.

Photosystem II (PSII) complexes, which split water into oxygen, protons and electrons in photosynthesis, require light but are also inactivated by it. Recovery of PSII from photoinactivation requires de novo protein synthesis. PSII in capsicum leaf segments were photoinactivated in the absence of chloroplast-encoded protein synthesis. At large photon exposures and despite the absence of repair, a residual fraction of PSII remained functional, being ca 0.08-0.2 depending on the ease of gas exchange in the tissue. This study revealed that the residual functional PSII was photoprotected by both (1) reaction-center quenching of excitation energy by photoinactivated PSII even when little or no PSII activity was permitted, and (2) antenna quenching, which was dependent on a trans-thylakoid pH gradient sustained mainly by linear electron transport and facilitated by the residual functional PSII complexes themselves. Significantly, little or no contribution to photoprotection of PSII was observed from cyclic electron flow around PSI. Further, the small residual functional PSII population was critical for recovery of the photoinactivated PSII complexes. Thus, photoinactivated and residual functional PSII complexes in leaves play a mutually beneficial role in each other's ultimate survival.

The formation of grana in chloroplasts of higher plants is examined in terms of the subtle interplay of physicochemical forces of attraction and repulsion. The attractive forces between two adjacent membranes comprise (1) van der Waals attraction that depends on the abundance and type of atoms in each membrane, on the distance between the membranes and on the dielectric constant, (2) depletion attraction that generates local order by granal stacking at the expense of greater disorder (i.e. entropy) in the stroma, and (3) an electrostatic attraction of opposite charges located on adjacent membranes. The repulsive forces comprise (1) electrostatic repulsion due to the net negative charge on the outer surface of thylakoid membranes, (2) hydration repulsion that operates at small separations between thylakoid membranes due to layers of bound water molecules, and (3) steric hindrance due to bulky protrusions of Photosystem I (PSI) and ATP synthase into the stroma. In addition, specific interactions may occur, but they await experimental demonstration. Although grana are not essential for photosynthesis, they are ubiquitous in higher plants. Grana may have been selected during evolution for the functional advantages that they confer on higher plants. The functional consequences of grana stacking include (1) enhancement of light capture through a vastly increased area-to-volume ratio and connectivity of several PSIIs with large functional antenna size, (2) the ability to control the lateral separation of PSI from PSII and, therefore, the balanced distribution of excitation energy between two photosystems working in series, (3) the reversible fine-tuning of energy distribution between the photosystems by State 1-State 2 transitions, (4) the ability to regulate light-harvesting via controlled thermal dissipation of excess excitation energy, detected as non-photochemical quenching, (5) dynamic flexibility in the light reactions mediated by a granal structure in response to regulation by a trans-thylakoid pH gradient, (6) delaying the premature degradation of D1 and D2 reaction-centre protein(s) in PSII by harbouring photoinactived PSIIs in appressed granal domains, (7) enhancement of the rate of non-cyclic synthesis of adenosine triphosphate (ATP) as well as the regulation of non-cyclic vs. cyclic ATP synthesis, and (8) the potential increase of photosynthetic capacity for a given composition of chloroplast constituents in full sunlight, concomitantly with enhancement of photochemical efficiency in canopy shade. Hence chloroplast ultrastructure and function are intimately intertwined.

In 27 C4 grasses grown under adequate or deficient nitrogen (N) supplies, N-use efficiency at the photosynthetic (assimilation rate per unit leaf N) and whole-plant (dry mass per total leaf N) level was greater in NADP-malic enzyme (ME) than NAD-ME species. This was due to lower N content in NADP-ME than NAD-ME leaves because neither assimilation rates nor plant dry mass differed significantly between the two C4 subtypes. Relative to NAD-ME, NADP-ME leaves had greater in vivo (assimilation rate per Rubisco catalytic sites) and in vitro Rubisco turnover rates (k cat; 3.8 versus 5.7 s−1 at 25°C). The two parameters were linearly related. In 2 NAD-ME (Panicum miliaceum and Panicum coloratum) and 2 NADP-ME (Sorghum bicolor and Cenchrus ciliaris) grasses, 30% of leaf N was allocated to thylakoids and 5% to 9% to amino acids and nitrate. Soluble protein represented a smaller fraction of leaf N in NADP-ME (41%) than in NAD-ME (53%) leaves, of which Rubisco accounted for one-seventh. Soluble protein averaged 7 and 10 g (mmol chlorophyll)−1 in NADP-ME and NAD-ME leaves, respectively. The majority (65%) of leaf N and chlorophyll was found in the mesophyll of NADP-ME and bundle sheath of NAD-ME leaves. The mesophyll-bundle sheath distribution of functional thylakoid complexes (photosystems I and II and cytochrome f) varied among species, with a tendency to be mostly located in the mesophyll. In conclusion, superior N-use efficiency of NADP-ME relative to NAD-ME grasses was achieved with less leaf N, soluble protein, and Rubisco having a faster k cat.

Photoinactivation of Photosystem II (PS II) is unavoidable in oxygenic photosynthesis, but the photodamage is counteracted by an elaborate repair process without which the photosynthetic apparatus would soon perish. Photoinactivation of PS II depends on the light dosage; the quantum yield of photoinactivation is such that practically the entire population of PS II would be photoinactivated during the course of a sunny day if repair were inhibited. An agent predominantly and inadvertently responsible for the photoinactivation of PS II is likely to be P680+, the strongest oxidant in photosynthesis needed for the oxidation of water molecules. Amelioration of photoinactivation of PS II occurs via many strategies, including one mechanism that appears to sustain activity in a small population of PS II during prolonged high-light stress. The majority of PS II complexes, however, have to be repaired following their photoinactivation. The repair process, culminating in the biosynthesis and insertion of a new copy of the D1 protein in a re-assembled PS II reaction center, consists of many steps. These include: (1) ‘triggering’ of the D1 protein, leading to monomerization and partial disassembly of the PS II dimer complex in granal appressed membranes in higher-plant chloroplasts; (2) migration of the PS II core monomer to stroma-exposed thylakoids; (3) D1 proteolysis catalyzed, for example, first by the proteases DegP2 followed by FtsHl, or directly by FtsH2; (4) targeting of the ribosome/nascent D1 chain complex to the thylakoid membrane; (5) translation elongation of D1 and insertion of new D1 into a D1-depleted PS II; (6) ligation of cofactors in the PS II reaction center; (7) termination of translation and carboxyl-terminal processing of D1; (8) post-translational assembly of PS II monomers; and (9) migration of re-assembled PS II monomers to granal membranes, where functional PS II dimers are formed. In this way, the functionality of PS II is maintained despite the inevitability of photoinactivation.