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The content of cytochrome (cyt) bf complex is the main rate-limiting factor that determines light- and CO2-saturated photosynthetic capacity. A study of the half-life of the cyt f content in leaves was conducted whereby Pisum sativum L. plants, grown in moderately high light (HL), were transferred to low light (LL). The cyt f content in fully-expanded leaves decreased steadily over the 2 weeks after the HL-to-LL transfer, whereas control leaves in HL retained their high contents. The difference between the time courses of HL-to-LL plants and control HL plants represents the time course of loss of cyt f content, with a half-life of 1.7 days, which is >3-fold shorter than that reported for tobacco leaves at constant growth irradiance using an RNA interference approach (Hojka et al. 2014). After transfer to LL (16 h photoperiod), pea plants were re-exposed to HL for 0, 1.5 h or 5 h during the otherwise LL photoperiod, but the cyt f content of fully-expanded leaves declined practically at the same rate regardless of whether HL was re-introduced for 0, 1.5 h or 5 h during each 16 h LL photoperiod. It appears that fully-expanded leaves, having matured under HL, were unable to increase their cyt f content when re-introduced to HL. These findings are relevant to any attempts to maintain a high photosynthetic capacity when the growth irradiance is temporarily decreased by shading or overcast weather.

Pseudomonas syringae pv. tabaci (Pst) is a hemibiotrophic bacterial pathogen responsible for tobacco wildfire disease. Although considerable research has been conducted on the tobacco plant’s tolerance to Pst, the role of light in the responses of the photosystems to Pst infection is poorly understood. This study aimed to elucidate the underlying mechanisms of the reduced photosystem damage in tobacco leaves due to Pst infection under light conditions. Compared to dark conditions, Pst infection under light conditions resulted in less chlorophyll degradation and a smaller decline in photosynthetic function. Although the maximal quantum yield of photosystem II (PSII) and the activity of the photosystem I (PSI) complex decreased as Pst infection progressed, damage to PSI and PSII after infection was reduced under light conditions compared to dark conditions. Pst was 17-fold more abundant in tobacco leaves under dark compared to light conditions at 3 days post-inoculation (dpi). Additionally, hydrogen peroxide (H2O2) accumulated to a high level in tobacco leaves after Pst infection under light conditions; although to a lesser extent, H2O2 accumulation was also significant under dark conditions. Pretreatment with H2O2 alleviated chlorotic lesions and decreased Pst abundance in tobacco leaves at 3 dpi under dark conditions. Methyl viologen (MV) pretreatment had the same effects under light conditions, whereas 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) pretreatment aggravated chlorotic lesions and increased the Pst population. These results indicate that chlorotic symptoms and the size of the bacterial population are each negatively correlated with H2O2 accumulation. In other words, light appears to suppress the Pst population in tobacco leaves through the accumulation of H2O2 during infection.

Salinity is an important abiotic stressor that negatively affects plant growth. In this study, we investigated the physiological and molecular mechanisms underlying moderate and high salt tolerance in diploid (2×) and tetraploid (4×) Robinia pseudoacacia L. Our results showed greater H2O2 accumulation and higher levels of important antioxidative enzymes and non-enzymatic antioxidants in 4× plants compared with 2× plants under salt stress. In addition, 4× leaves maintained a relatively intact structure compared to 2× leaves under a corresponding condition. NaCl treatment didn’t significantly affect the photosynthetic rate, stomatal conductance or leaf intercellular CO2 concentrations in 4× leaves. Moreover, proteins from control and salt treated 2× and 4× leaf chloroplast samples were extracted and separated by two-dimensional gel electrophoresis. A total of 61 spots in 2× (24) and 4× (27) leaves exhibited reproducible and significant changes under salt stress. In addition, 10 proteins overlapped between 2× and 4× plants under salt stress. These identified proteins were grouped into the following 7 functional categories: photosynthetic Calvin-Benson Cycle (26), photosynthetic electron transfer (7), regulation/defense (5), chaperone (3), energy and metabolism (12), redox homeostasis (1) and unknown function (8). This study provides important information of use in the improvement of salt tolerance in plants.

This study resolved correlations between changes in xanthophyll pigments and photosynthetic properties in attached and detached shade-grown avocado (Persea americana) leaves upon sun exposure. Lutein epoxide (Lx) was deepoxidized to lutein (L), increasing the total pool by ƊL over 5 h, whereas violaxanthin (V) conversion to antheraxanthin (A) and zeaxanthin (Z) ceased after 1 h. During subsequent dark or shade recovery, de novo synthesis of L and Z continued, followed by epoxidation of A and Z but not of L. Light-saturated nonphotochemical quenching (NPQ) was strongly and linearly correlated with decreasing [Lx] and increasing [∆L] but showed a biphasic correlation with declining [V] and increasing [A+Z] separated when V deepoxidation ceased. When considering [ƊL+∆Z], the monophasic linear correlation was restored. Photochemical efficiency of photosystem II (PSII) and photosystem (PSI; deduced from the delivery of electrons to PSI in saturating single-turnover flashes) showed a strong correlation in their continuous decline in sunlight and an increase in NPQ capacity. This decrease was also reflected in the initial reduction of the slope of photosynthetic electron transport versus photon flux density. Generally longer, stronger sun exposures enhanced declines in both slope and maximum photosynthetic electron transport rates as well as photochemical efficiency of PSII and PSII/PSI more severely and prevented full recovery. Interestingly, increased NPQ capacity was accompanied by slower relaxation. This was more prominent in detached leaves with closed stomata, indicating that photorespiratory recycling of CO2 provided little photoprotection to avocado shade leaves. Sun exposure of these shade leaves initiates a continuum of photoprotection, beyond full engagement of the Lx and V cycle in the antenna, but ultimately photoinactivated PSII reaction centers.

Orchard yields are a function of the light intercepted, although this linear relation is less tight when greater than 50% of available light is intercepted. Above this level, light exceeds photosynthetic requirements and can expose leaves to photoinhibition. The "asymmetric" peach orchard was designed and planted to obtain in the field six different light intensities at any time of the day and three daily light interception profiles: low (C), middle (E) and high (W). This paper reports on how incoming light energy is managed in attached leaves subjected to several radiative regimes along the day. Combined measurements of gas exchange and chlorophyll fluorescence (quenching analysis) were performed during the day to quantify the absorbed energy used for net photosynthesis and that dissipated by the photoprotective mechanisms. In addition, the amount of inactive PSII after one day of irradiation was calculated via chlorophyll fluorescence determination. Net photosynthesis was linearly related to irradiance up to a saturating point of 1000-1200 μmol photon m-2 s-1. Non-photochemical quenching (NPQ) played the most important role in photoprotection, but its activity was reduced at low irradiance, possibly due to a sub-optimal trans-thylakoid ΔpH. The non-net carboxylative mechanisms (NC) were the main photoprotective mechanisms at middle-low irradiance levels and probably facilitated the establishment of the trans-thylakoid ΔpH needed for full activation of NPQ. At the end of the photosynthetic period the amount of inactive PSII was the highest in W followed by E and C samples. In addition, daily carbon accumulation did not reflect the light interception ranking, with E having the highest, and W the lowest. These findings support the conclusion that under irradiances exceeding the photosynthetic requirements, photoprotection and photoinactivation biochemistries divert energy in the form of carbohydrates and reducing power from tree growth and productivity. The amounts of these losses can be significant.

Plants use a small part of the total absorbed light energy for net carboxylation, while the remaining amount is dissipated via alternative pathways involving thermal processes, fluorescence and non- carboxylation photochemistry in order to limit the formation of reactive oxygen species (ROS) and other photooxidative risks. The commonly used analysis of the Photosystem II (PSII) fluorescence signals gives qualitative information about absorbed light energy management by plants, but it is difficult to appreciate the relative contribution of each pathway in energy partitioning. This study reports the application of quenching partitioning through a chlorophyll fluorescence approach performed on peach leaves subjected to three different light intensities for four durations of exposure in absence of recovery from photo-damage. This methodology was compared with the P700 redox kinetic method for determining the functional PSII fraction in leaves. In the absence of recovery processes the active PSII concentration decayed with an increase in photon exposure (the product of irradiance and the time of exposure), following an exponential pattern according to the reciprocity law. The photoprotective thermal dissipation ( ̊NPQ ) was proportional to irradiance up to 30 min of photoin- hibitory treatment. Afterwards ̊NPQ was limited by the increasing competition for the absorbed energy re-emitted by the inactive PSII ( ̊NF ). ̊NF increased with the photon exposure dissipating up to 70% of the total incoming energy. The energy funnelled to photochemistry ( ̊PSII ) decreased with increasing exposure time or intensity, becoming zero after 120 min of photoinhibitory treatment at the maximum irradiance (2100 ␮mol photon m−2 s−1 ). The relation between the fraction of energy dissipated by the inactive PSII (derived from the quenching partitioning) and the inactive PSII fraction (measured with the P700 redox kinetic method) was linear. The quenching partitioning through light-modulated chlorophyll fluorescence is a useful tool to anal- yse plant energy management and gives also a reasonable estimation of the active PSII fraction. This methodology can easily be used in the field as measurements are rapid, non-destructive and detection devices are portable.

Low temperature (LT) is one of the major factors that limit crop production and reduce yield. To better understand the cold-tolerance mechanism in the plantains, a sensitive cultivar Williams (Musa acuminata AAA cv. Williams) and a tolerant cultivar Cachaco (Musa paradisiaca ABB cv. Dajiao) were used. LT resulted in increased malondialdehyde (MDA) content, elevated contents of hydrogen peroxide (H2O2) and superoxide radical (O 2 ·− ), and decreased photochemical efficiency (Fv/Fm) and net photosynthetic rate (P N), but cv. Cachaco showed better LT tolerance than cv. Williams. After LT treatment for 120 h, total scavenging capability (DPPH· scavenging capability) in Williams showed a significant decrease but no significant alternations was found in Cachaco. Ascorbate peroxidase (APX) and peroxidase (POD) displayed a significant increase but superoxide dismutase (SOD) showed no significant alternations and catalase (CAT) showed a significant decrease in Cachaco after 120 h of LT treatment. All the four antioxidant enzymes above showed a significant decrease in Williams after 120 h of LT treatment. Our results suggest that higher activities of APX, POD, SOD, and DPPH· scavenging capability to a certain extent can be used to explain the higher cold tolerance in the plantain, which would provide a theoretical guidance for bananas production and screening cold-resistant variety.

• We studied how different color lights cause gradients of photoinhibition within a leaf, to attempt to resolve the controversy of whether photon absorption by chlorophyll or by manganese (Mn) is the primary cause of photoinhibition, as suggested by the excess-energy hypothesis or the two-step hypothesis, respectively. • Lincomycin-treated leaf discs were photoinhibited by white, blue, green or red light. Combining a microfiber fluorometer, a fiber-thinning technique and a micro-manipulator enabled us to measure the chlorophyll fluorescence signals within a leaf. Photoinhibition gradients were also compared with results from various conventional fluorometers to estimate their depth of signal detection. • The severity of photoinhibition was 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 correlated with the chlorophyll and the Mn absorption spectrums, respectively. These results cannot be explained by either hypothesis alone. • These data strongly suggest that both the excess-energy and the two-step mechanisms occur in photoinhibition, and fluorometers with red or blue measuring light give overestimated or underestimated F(v)/F(m) values of photoinhibited leaves compared with the whole tissue average, respectively; that is, they measured deeper or shallower leaf tissue, respectively.

Photosystem II (PS II) is photoinactivated during photosynthesis, requiring repair to maintain full function during the day. What is the mechanism(s) of the initial events that lead to photoinactivation of PS II? Two hypotheses have been put forward. The 'excess-energy hypothesis' states that excess energy absorbed by chlorophyll (Chl), neither utilized in photosynthesis nor dissipated harmlessly in non-photochemical quenching, leads to PS II photoinactivation; the 'Mn hypothesis' (also termed the two-step hypothesis) states that light absorption by the Mn cluster in PS II is the primary effect that leads to dissociation of Mn, followed by damage to the reaction centre by light absorption by Chl. Observations from various studies support one or the other hypothesis, but each hypothesis alone cannot explain all the observations. We propose that both mechanisms operate in the leaf, with the relative contribution from each mechanism depending on growth conditions or plant species. Indeed, in a single system, namely, the interior of a leaf, we could observe one or the other mechanism at work, depending on the location within the tissue. There is no reason to expect the two mechanisms to be mutually exclusive.

Plants adjust the relative sizes of PSII and PSI antennae in response to the spectral composition of weak light favouring either photosystem by processes known as state transitions (ST), attributed to a discrete antenna migration involving phosphorylation of light-harvesting chlorophyll-protein complexes in PSII. Here for the first time we monitored the extent and dynamics of ST in leaves from estimates of optical absorption cross-section (relative PSII antenna size; aPSII). These estimates were obtained from in situ measurements of functional absorption cross-section (σPSII) and maximum photochemical efficiency of PSII (φPSII); i.e. aPSII = σPSII/φPSII (Kolber et al. 1998) and other parameters from a light induced fluorescence transient (LIFT) device (Osmond et al. 2017). The fast repetition rate (FRR) QA flash protocol of this instrument monitors chlorophyll fluorescence yields with reduced QA irrespective of the redox state of plastoquinone (PQ), as well as during strong ~1 s white light pulses that fully reduce the PQ pool. Fitting this transient with the FRR model monitors kinetics of PSII → PQ, PQ → PSI, and the redox state of the PQ pool in the ‘PQ pool control loop’ that underpins ST, with a time resolution of a few seconds. All LIFT/FRR criteria confirmed the absence of ST in antenna mutant chlorina-f2 of barley and asLhcb2–12 of Arabidopsis, as well as STN7 kinase mutants stn7 and stn7/8. In contrast, wild-type barley and Arabidopsis genotypes Col, npq1, npq4, OEpsbs, pgr5 bkg and pgr5, showed normal ST. However, the extent of ST (and by implication the size of the phosphorylated LHCII pool participating in ST) deduced from changes in aʹPSII and other parameters with reduced QA range up to 35%. Estimates from strong WL pulses in the same assay were only ~10%. The larger estimates of ST from the QA flash are discussed in the context of contemporary dynamic structural models of ST involving formation and participation of PSII and PSI megacomplexes in an ‘energetically connected lake’ of phosphorylated LHCII trimers (Grieco et al. 2015). Despite the absence of ST, asLhcb2-12 displays normal wild-type modulation of electron transport rate (ETR) and the PQ pool during ST assays, reflecting compensatory changes in antenna LHCIIs in this genotype. Impaired LHCII phosphorylation in stn7 and stn7/8 accelerates ETR from PSII →PQ, over-reducing the PQ pool and abolishing the yield difference between the QA flash and WL pulse, with implications for photochemical and thermal phases of the O-J-I-P transient.

Joan Mary (Jan) Anderson pioneered the investigation of the molecular organisation of the plant thylakoid membrane, making seminal discoveries that laid the foundations for the current understanding of photosynthesis. She grew up in Queenstown, New Zealand, obtaining a BSc and MSc at the University of Otago in Dunedin. After completing her PhD at the University of California, she embarked on a glittering career at the Commonwealth Scientific and Industrial Research Organisation (CSIRO) and then Australian National University (ANU) in Canberra. Not only a gifted experimentalist, Jan was a creative thinker, not afraid to put her insightful and prophetic hypotheses into the public domain. Her many notable achievements include establishing the details and the physiological significance of lateral heterogeneity in the distribution of the two photosystems between stacked and unstacked thylakoid membranes and the dynamic changes in the extent of stacking that occur in response to changes in the light environment. Her investigations brought her into collaboration with prominent researchers throughout the world. Recognised with many honours as a leading scientist in Australia, international recognition included Lifetime Achievement Award from the International Society of Photosynthesis Research, and Honorary Fellowships at Universities in the UK and USA.

More intense, more frequent, and longer heat waves are expected in the future due to global warming, which could have dramatic ecological impacts. However, few studies have involved invasive species. The aims of this study were to examine the effect of extreme heating (40/35 degrees C for 30 d) on the growth and photosynthesis of an alien invasive species Wedelia trilobata and its indigenous congener (Wedelia chinensis) in South China, and to determine the development of this invasive species and its potential adaptive mechanism. In comparison with W. chinensis, W. trilobata suffered less inhibition of the relative growth rate (RGR) and biomass production due to high temperature, which was consistent with the changes of photosystem II (PSII) activity and net photosynthetic rate (P(n)). High temperature caused a partial inhibition of PSII, but the adverse effect was more severe in W. chinensis. Measurement of the minimum fluorescence (F(o)) versus temperature curves showed that W. trilobata had a higher inflexion temperature of F(o) (T(i)), indicating greater thermostability of the photosynthetic apparatus. Moreover, comparisons of absorbed light energy partitioning revealed that W. trilobata increased xanthophyll-dependent thermal dissipation (Phi(NPQ)) under high temperature, while retaining the higher fraction of absorbed light allocated to photochemistry (Phi(PSII)) relative to W. chinensis. The results suggest that the invasive W. trilobata has a high thermostability of its photosynthetic apparatus and an effective regulating mechanism in energy partitioning of PSII complexes to minimize potential damage and to retain greater capability for carbon assimilation. These factors confer greater heat stress tolerance compared with the native species. Therefore, the invasive W. trilobata may become more aggressive with the increasingly extreme heat climates.

Alex Hope was for many years a Foundation Professor of Biology in Biophysics and later Emeritus Professor at The Flinders University of South Australia. Throughout his career he strove to understand the energetics of plant cells, and devoted the latter two-thirds of his research career to the study of photosynthesis. His earlier, highly successful, research had focused on electrical properties and ionic relations of plant cells. The change of research direction, however, was only an apparent one, since a continuing theme was the role of electrochemical potential gradients in energy capture and conversion. His life work was ‘‘driven by electricity’’ (Hope 2002, 2004, 2006). Alex was born in Launceston (in Tasmania), the first Australian city to have hydroelectricity as early as 1895. Electricity was seemingly ‘‘in the air’’, as several members of his family appear to have made their careers based on electricity. Alex studied at The University of Tasmania, majoring in physics. During his Honours degree year in 1949, he chose to investigate the electric fields in and around plant roots and shoots, suggested by his supervisor Alexander Leicester McAulay as possibly contributing to developmental forces in plant growth. McAulay, Professor of Physics from 1926 to 1959 (and son of a professor of mathematics and physics), was almost certainly Australia’s first biophysicist, having pioneered the study of mutations caused in yeast by ultraviolet radiation. Decades later, Alex was instrumental in establishing in the Australian Society for Biophysics a prize for innovative biophysics to honour the memory of McAulay, helped by Alex’s generous personal donation in support of that prize. Reluctantly, Alex let the Australian Society for Biophysics extend the name of the prize to The McAulay–Hope Prize for Original Biophysics. In his PhD work (1950–1952), Alex continued under the supervision of McAulay to investigate the mechanism by which nutrient minerals entered plant roots, with financial support from an appointment as a Temporary Research Officer in the Commonwealth Scientific and Industrial Research Organisation (CSIRO) for the final 2 years. The CSIRO Division of Food Preservation and Transport, as it was then called, had a Plant Physiology Unit headed jointly by the influential (later Sir) R. N. Robertson (see Robertson 1992) and F. V. Mercer for the study of salt absorption by plant cells. Alex was appointed on the understanding that he would be available for future employment in the Division! W. S. Chow (&) Division of Plant Science, Research School of Biology, College of Medicine, Biology and Environment, The Australian National University, Canberra, ACT 0200, Australia e-mail: Fred.Chow@anu.edu.au

Expression of the genes for carotenoid biosynthesis (crt) is dependent on light, but little is known about the underlying mechanism of light sensing and signalling in the cyanobacterium Synechocystis sp. PCC 6803 (hereafter, Synechocystis). In the present study, we investigated the light-induced increase in the transcript levels of Synechocystiscrt genes, including phytoene synthase (crtB), phytoene desaturase (crtP), ζ-carotene desaturase (crtQ), and β-carotene hydroxylase (crtR), during a dark-to-light transition period. During the dark-to-light shift, the increase in the crt transcript levels was not affected by mutations in cyanobacterial photoreceptors, such as phytochromes (cph1, cph2 and cph3) and a cryptochrome-type photoreceptor (ccry), or respiratory electron transport components NDH and Cyd/CtaI. However, treatment with photosynthetic electron transport inhibitors significantly diminished the accumulation of crt gene transcripts. Therefore, the light induction of the Synechocystis crt gene expression is most likely mediated by photosynthetic electron transport rather than by cyanobacterial photoreceptors during the dark-to-light transition.

Under severe water stress, leaf wilting is quite general in higher plants. This passive movement can reduce the energy load on a leaf. This paper reports an experimental test of the hypothesis that leaf wilting movement has a protective function that mitigates against photoinhibition of photosynthesis in the field. The experiments exposed cotton (Gossypium hirsutum L.) to two water regimes: water-stressed and well-watered. Leaf wilting movement occurred in water-stressed plants as the water potential decreased to −4.1 MPa, reducing light interception but maintaining comparable quantum yields of photosystem II (PS II; Yield for short) and the proportion of total PS II centers that were open (qP). Predrawn F v/F m (potential quantum yield of PS II) as an indicator of overnight recovery of PS II from photoinhibition was higher than or similar to that in well-watered plants. Compared with water-stressed cotton leaves for which wilting movement was permitted, water-stressed cotton leaves restrained from such movement had significantly increased leaf temperature and instantaneous CO2 assimilation rates in the short term, but reduced Yield, qP, and F v/F m. In the long term, predrawn F v/F m and CO2 assimilation capacity were reduced in water-stressed leaves restrained from wilting movement. These results suggest that, under water stress, leaf wilting movement could reduce the incident light on leaves and their heat load, alleviate damage to the photosynthetic apparatus due to photoinhibition, and maintain considerable carbon assimilation capacity in the long term despite a partial loss of instantaneous carbon assimilation in the short term.

The multiple roles of light-harvesting chlorophyll a/b-protein complexes in the structure and function of Arabidopsis chloroplasts were investigated using two chlorophyll b-less mutants grown under metal halide lamps with a significant far-red component. In ch1-3, all six light-harvesting proteins of photosystem (PS) II were greatly decreased; in ch1-3lhcb5, Lhcb5 was completely absent while the other five proteins were further decreased. The thylakoids of ch1-3 were less negatively-charged than the wild type, and those of ch1-3lhcb5 were even less so. Despite the expected weaker electrostatic repulsion, however, thylakoids in leaves of the mutants were not well stacked, an effect we attribute to lower van der Waals attraction, lower electrostatic attraction between opposite charges, and the absence or instability of PSII supercomplexes and peripheral light-harvesting trimers. The quantum yield of oxygen evolution in leaves decreased from 0.109 (wild type) to 0.087 (ch1-3) and 0.081 (ch1-3lhcb5) O(2) (photon absorbed)(-1); we attribute this decrease to an excessive spillover from PSII to PSI, a limited PSII antenna, and increased light-independent thermal dissipation in PSII in the mutants. Destabilization of the donor side of PSII, indicated by slower electron donation to the redox-active tyrosine Y(Z)(*) in ch1-3, probably enhanced PSII susceptibility to photoinactivation, increased the non-functional PSII complexes in vivo, and further inactivated PSII complexes in vitro. The evolution of chlorophyll b-containing chloroplasts seems to fine-tune oxygenic photosynthesis.

This introductory article is the first of four short articles from the Tribute to Alex Hope Symposium held at the 2008 Australian Society for Biophysics meeting in Canberra, Australia, as a tribute to Professor Alex Hope, who died in July last year. As well as briefly introducing the other three articles by three former PhD students, it will also be a biographical memoir of Alex Hope.

The literature and our present examinations indicate that the intra-leaf light absorption profile is in most cases steeper than the photosynthetic capacity profile. In strong white light, therefore, the quantum yield of photosynthesis would be lower in the upper chloroplasts, located near the illuminated surface, than that in the lower chloroplasts. Because green light can penetrate further into the leaf than red or blue light, in strong white light, any additional green light absorbed by the lower chloroplasts would increase leaf photosynthesis to a greater extent than would additional red or blue light. Based on the assessment of effects of the additional monochromatic light on leaf photosynthesis, we developed the differential quantum yield method that quantifies efficiency of any monochromatic light in white light. Application of this method to sunflower leaves clearly showed that, in moderate to strong white light, green light drove photosynthesis more effectively than red light. The green leaf should have a considerable volume of chloroplasts to accommodate the inefficient carboxylation enzyme, Rubisco, and deliver appropriate light to all the chloroplasts. By using chlorophylls that absorb green light weakly, modifying mesophyll structure and adjusting the Rubisco/chlorophyll ratio, the leaf appears to satisfy two somewhat conflicting requirements: to increase the absorptance of photosynthetically active radiation, and to drive photosynthesis efficiently in all the chloroplasts. We also discuss some serious problems that are caused by neglecting these intra-leaf profiles when estimating whole leaf electron transport rates and assessing photoinhibition by fluorescence techniques.

Grana are not essential for photosynthesis, yet they are ubiquitous in higher plants and in the recently evolved Charaphyta algae; hence grana role and its need is still an intriguing enigma. This article discusses how the grana provide integrated and multifaceted functional advantages, by facilitating mechanisms that fine-tune the dynamics of the photosynthetic apparatus, with particular implications for photosystem II (PSII). This dynamic flexibility of photosynthetic membranes is advantageous in plants responding to ever-changing environmental conditions, from darkness or limiting light to saturating light and sustained or intermittent high light. The thylakoid dynamics are brought about by structural and organizational changes at the level of the overall height and number of granal stacks per chloroplast, molecular dynamics within the membrane itself, the partition gap between appressed membranes within stacks, the aqueous lumen encased by the continuous thylakoid membrane network, and even the stroma bathing the thylakoids. The structural and organizational changes of grana stacks in turn are driven by physicochemical forces, including entropy, at work in the chloroplast. In response to light, attractive van der Waals interactions and screening of electrostatic repulsion between appressed grana thylakoids across the partition gap and most probably direct protein interactions across the granal lumen (PSII extrinsic proteins OEEp-OEEp, particularly PsbQ-PsbQ) contribute to the integrity of grana stacks. We propose that both the light-induced contraction of the partition gap and the granal lumen elicit maximisation of entropy in the chloroplast stroma, thereby enhancing carbon fixation and chloroplast protein synthesizing capacity. This spatiotemporal dynamic flexibility in the structure and function of active and inactive PSIIs within grana stacks in higher plant chloroplasts is vital for the optimization of photosynthesis under a wide range of environmental and developmental conditions. © 2008 Springer Science+Business Media B.V., Australian Research Council for financial support through Discovery Project Grant DP0664719.

This study investigated the roles of sucrose synthase (SUS) in developing seeds and siliques of Arabidopsis thaliana. Enzyme activity assays showed that SUS activity was highest in developing whole siliques and young rosette leaves compared with other tissues including mature leaves, stems, and flowers. Surprisingly, quantitative PCR analyses revealed little correlation between SUS activity and transcript expression, which indicated the importance of examining the role of SUS at the protein level. Therefore, immunolocalization was performed over a developmental time course to determine the previously unreported cellular localization of SUS in Arabidopsis seed and silique tissues. At 3 d and 10 d after flowering (daf), SUS protein localized to the silique wall, seed coat, funiculus, and endosperm. By 13 daf, SUS protein was detected in the embryo and aleurone layer, but was absent from the seed coat and funiculus. Starch grains were also present in the seed coat at 3 and 10 daf, but were absent at 13 daf. Co-localization of SUS protein and starch grains in the seed coat at 3 and 10 daf indicates that SUS may be involved in temporary starch deposition during the early stages of seed development, whilst in the later stages SUS metabolizes sucrose in the embryo and cotyledon. Within the silique wall, SUS localized specifically to the companion cells, indicating that SUS activity may be required to provide energy for phloem transport activities in the silique wall. The results highlight the diverse roles that SUS may play during the development of silique and seed in Arabidopsis.

Background Pseudomonas syringae pv. tabaci (Pst), which is the pathogen responsible for tobacco wildfire disease, has received considerable attention in recent years. The objective of this study was to clarify the responses of photosystem I (PSI) and photosystem II (PSII) to Pst infection in tobacco leaves. Results The net photosynthetic rate (Pn) and carboxylation efficiency (CE) were inhibited by Pst infection. The normalized relative variable fluorescence at the K step (Wk) and the relative variable fluorescence at the J step (VJ) increased while the maximal quantum yield of PSII (Fv/Fm) and the density of QA-reducing PSII reaction centers per cross section (RC/CSm) decreased, indicating that the reaction centers, and the donor and acceptor sides of PSII were all severely damaged after Pst infection. The PSI activity decreased as the infection progressed. Furthermore, we observed a considerable overall degradation of PsbO, D1, PsaA proteins and an over-accumulation of reactive oxygen species (ROS). Conclusions Photoinhibition and photoinhibition-like damage were observed under light and dark conditions, respectively, after Pst infection of tobacco leaves. The damage was greater in the dark. ROS over-accumulation was not the primary cause of the photoinhibition and photoinhibition-like damage. The PsbO, D1 and PsaA proteins appear to be the targets during Pst infection under light and dark conditions. Electronic supplementary material The online version of this article (doi:10.1186/s12870-016-0723-6) contains supplementary material, which is available to authorized users.

The stoichiometry of Photosystem II (PSII) to Photosystem I (PSI) reaction centres in spinach leaf segments was determined by two methods, each capable of being applied to monitor the presence of both photosystems in a given sample. One method was based on a fast electrochromic (EC) signal, which in the millisecond time scale represents a change in the delocalized electric potential difference across the thylakoid membrane resulting from charge separation in both photosystems. This method was applied to leaf segments, thus avoiding any potential artefacts associated with the isolation of thylakoid membranes. Two variations of this method, suppressing PSII activity by prior photoinactivation (in spinach and poplar leaf segments) or suppressing PSI by photo-oxidation of P700 (the chlorophyll dimer in PSI) with background far-red light (in spinach, poplar and cucumber leaf segments), each gave the separate contribution of each photosystem to the fast EC signal; the PSII/PSI stoichiometry obtained by this method was in the range 1.5–1.9 for the three plant species, and 1.5–1.8 for spinach in particular. A second method, based on electron paramagnetic resonance (EPR), gave values in a comparable range of 1.7–2.1 for spinach. A third method, which consisted of separately determining the content of functional PSII in leaf segments by the oxygen yield per single turnover-flash and that of PSI by photo-oxidation of P700 in thylakoids isolated from the corresponding leaves, gave a PSII/PSI stoichiometry (1.5–1.7) that was consistent with the above values. It is concluded that the ratio of PSII to PSI reaction centres is considerably higher than unity in typical higher plants, in contrast to a surprisingly low PSII/PSI ratio of 0.88, determined by EPR, that was reported for spinach grown in a cabinet under far-red-deficient light in Sweden [Danielsson et al. (2004) Biochim. Biophys. Acta 1608: 53–61]. We suggest that the low PSII/PSI ratio in the Swedish spinach, grown in far-red-deficient light with a lower PSII content, is not due to greater accuracy of the EPR method of measurement, as suggested by the authors, but is rather due to the growth conditions.

Light damages photosynthetic machinery, primarily photosystem II (PSII) and it results in photoinhibition. A new photodamage model, the two-step photodamage model, suggests that photodamage to PSII initially occurs at the oxygen evolving complex (OEC) by light energy absorbed by manganese and that the PSII reaction center is subsequently damaged by light energy absorbed by photosynthetic pigments due to the limitation of electrons to the PSII reaction center. However, it is still uncertain whether this model is applicable to photodamage to PSII under visible light as manganese absorbs visible light only weakly. In the present study, we identified the initial site of photodamage to PSII upon illumination of visible light using PSII membrane fragments isolated from spinach leaves. When PSII samples were exposed to visible light in the presence of an exogenous electron acceptor, both PSII total activity and the PSII reaction centre activity declined due to photodamage. The supplemental addition of an electron donor to the PSII reaction centre alleviated the decline of the reaction centre activity but not the PSII total activity upon the light exposure. Our results demonstrate that visible light damages OEC prior to photodamage to the PSII reaction center, consistent with two-step photodamage model.

To better understand the adaptation mechanisms of the photosynthetic apparatus of cotton plants to water deficit conditions, the influence of water deficit on photosynthesis, chlorophyll a fluorescence and the activities of antioxidant systems were determined simultaneously in Gossypium hirsutum L. cv. Xinluzao 45 (upland cotton) and Gossypium barbadense L. cv. Xinhai 21 (pima cotton). Water deficit decreased photosynthesis in both cotton species, but did not decrease chlorophyll content or induce any sustained photoinhibition in either cotton species. Water deficit increased ETR/4 − AG, where ETR/4 estimates the linear photosynthetic electron flux and AG is the gross rate of carbon assimilation. The increase in ETR/4 − AG, which represents an increase in photorespiration and alternative electron fluxes, was particularly pronounced in Xinluzao 45. In Xinluzao 45, water deficit increased the activities of antioxidative enzymes, as well as the contents of reactive oxygen species (ROS), which are related to the Mehler reaction. In contrast, moderate water deficit particularly increased non-photochemical quenching (NPQ) in Xinhai 21. Our results suggest that Xinluzao 45 relied on enhanced electron transport such as photorespiration and the Mehler reaction to dissipate excess light energy under mild and moderate water deficit. Xinhai 21 used enhanced photorespiration for light energy utilisation under mild water deficit but, when subjected to moderate water deficit, possessed a high capacity for dissipating excess light energy via heat dissipation.

To explore replacement control of the invasive weed Ipomoea cairica, we studied the competitive effects of two valuable natives, Pueraria lobata and Paederia scandens, on growth and photosynthetic characteristics of I. cairica, in pot and field experiments. When I. cairica was planted in pots with P. lobata or P. scandens, its total biomass decreased by 68.7% and 45.8% and its stem length by 33.3% and 34.1%, respectively. The two natives depressed growth of the weed by their strong effects on its photosynthetic characteristics, including suppression of leaf biomass and the abundance of the CO2-fixing enzyme RUBISCO. The field experiment demonstrated that sowing seeds of P. lobata or P. scandens in plots where the weed had been largely cleared produced 11.8-fold or 2.5-fold as much leaf biomass of the two natives, respectively, as the weed. Replacement control by valuable native species is potentially a feasible and sustainable means of suppressing I. cairica.

Photosynthesis is always accompanied by light induced damage to the Photosystem II (PSII) which is compensated by its subsequent repair. Photoinhibition of PSII is a complex process, balancing between photoinactivation, protective and repair mechanisms. Current understanding of photoinactivation is limited with competing hypotheses where the photosensitiser is either photosynthetic pigments or the Mn4CaO5 cluster itself, with little consensus on the mechanisms and consequences of PSII photoinactivation. The mechanism of photoinactivation should be reflected in the action spectrum of PSII photoinactivation, but there is a great diversity of the action spectra reported thus far. The only consensus is that PSII photoinactivation is greatest in the UV region of the electromagnetic spectrum. In this review, the authors revisit the methods, technical constraints and the different action spectra of PSII photoinactivation reported to date and compare them against the diverse mechanisms proposed. Upon critical examination of the reported action spectra, a hybrid mechanism of photoinactivation, sensitised by both photosynthetic pigments and the Mn4CaO5 appears to be the most plausible rationalisation.

Several auxin herbicides, such as 2, 4-D and dicamba, have been used to eradicate an exotic invasive weed Ipomoea cairica in subtropical China, but restraining the re-explosion of this weed is still a challenge. Since ethylene is one of the major intermediate functioning products during the eradication process, we explored the possibility, mechanism and efficiency of using ethephon which can release ethylene to control Ipomoea cairica. The results of the pot experiment showed that 7.2 g /L ethephon could totally kill Ipomoea cairica including the stems and roots. The water culture experiment indicated that ethephon released an abundance of ethylene directly in leaves and caused increases in electrolyte leakage, 1-aminocyclopropane-1-carboxylic acid (ACC), abscisic acid (ABA) and H2O2 and decreases in chlorophyll content and photosynthetic activity, finally leading to the death of Ipomoea cairica. The field experiment showed that the theoretical effective concentration of ethephon for controlling Ipomoea cairica (weed control efficacy, WCE = 98%) was 4.06 g/L and the half inhibitory concentration (I50) was 0.56 g/L. More than 50% of the accompanying species were insensitive to the phytotoxicity of ethephon. Therefore, ethephon is an excellent alternative herbicide for controlling Ipomoea cairica.

Molecular analyses of plants have revealed a number of genes whose expression changes in response to high light (HL), including the H2O2 scavenger, ASCORBATE PEROXIDASE 2 (APX2). We carried out a screen in Arabidopsis thaliana for lesions that alter HL-induced expression of APX2 to identify components in abiotic stress signalling pathways. High light was used as it can be instantaneously applied or removed and accurately measured. We identified a number of alx mutations causing altered APX2 expression. Here we describe the gain-of-function mutant, alx8, which has constitutively higher APX2 expression and higher levels of foliar abscisic acid (ABA) than wild type. In fact, exogenous ABA increased APX2 expression and the APX2 promoter contains ABA response elements. Furthermore, we have shown that HL stress increases ABA in wild-type plants, implicating ABA in the regulation of HL-inducible genes. The alx8 mutant is drought tolerant, exhibits improved water-use efficiency and a number of drought-tolerance genes are upregulated. Additionally, alx8 demonstrates the complexity of ABA-dependent and ABA-independent transcriptional networks as some components in both pathways are upregulated in alx8. This study provides evidence for common steps in drought and HL stress response pathways.

Photoinactivation of Photosystem II (PS II), the light-induced loss of ability to evolve oxygen, inevitably occurs under any light environment in nature, counteracted by repair. Under certain conditions, the extent of photoinactivation of PS II depends on the photon exposure (light dosage, x), rather than the irradiance or duration of illumination per se, thus obeying the law of reciprocity of irradiance and duration of illumination, namely, that equal photon exposure produces an equal effect. If the probability of photoinactivation (p) of PS II is directly proportional to an increment in photon exposure (p = kDeltax, where k is the probability per unit photon exposure), it can be deduced that the number of active PS II complexes decreases exponentially as a function of photon exposure: N = Noexp(-kx). Further, since a photon exposure is usually achieved by varying the illumination time (t) at constant irradiance (I), N = Noexp(-kI t), i.e., N decreases exponentially with time, with a rate coefficient of photoinactivation kI, where the product kI is obviously directly proportional to I. Given that N = Noexp(-kx), the quantum yield of photoinactivation of PS II can be defined as -dN/dx = kN, which varies with the number of active PS II complexes remaining. Typically, the quantum yield of photoinactivation of PS II is ca. 0.1micromol PS II per mol photons at low photon exposure when repair is inhibited. That is, when about 10(7) photons have been received by leaf tissue, one PS II complex is inactivated. Some species such as grapevine have a much lower quantum yield of photoinactivation of PS II, even at a chilling temperature. Examination of the longer-term time course of photoinactivation of PS II in capsicum leaves reveals that the decrease in N deviates from a single-exponential decay when the majority of the PS II complexes are inactivated in the absence of repair. This can be attributed to the formation of strong quenchers in severely-photoinactivated PS II complexes, able to dissipate excitation energy efficiently and to protect the remaining active neighbours against damage by light.

Chloroplasts in plants and some green algae contain a continuous thylakoid membrane system that is structurally differentiated into stacked granal membranes interconnected by unstacked thylakoids, the stromal lamellae. Experiments were conducted to test the hypothesis that the thermodynamic tendency to increase entropy in chloroplasts contributes to thylakoid stacking to form grana. We show that the addition of bovine serum albumin or dextran, two very different water-soluble macromolecules, to a suspension of envelope-free chloroplasts with initially unstacked thylakoids induced thylakoid stacking. This novel restacking of thylakoids occurred spontaneously, accompanied by lateral segregation of PSII from PSI, thereby mimicking the natural situation. We suggest that such granal formation, induced by the macromolecules, is partly explained as a means of generating more volume for the diffusion of macromolecules in a crowded stromal environment, i.e., greater entropy overall. This mechanism may be relevant in vivo where the stroma has a very high concentration of enzymes of carbon metabolism, and where high metabolic fluxes are required.

Changes in the extent of P700 oxidation (P700 + ) were investigated after chilling of barley, rice, pumpkin, and cucumber leaf segments at 4°C for 1 h under light with various photon flux densities. At 50 μmol photons m -2 s -1 , the decrease in P700 + was observed only in cucumber, but at 150 μmol photons m -2 s -1 , it was found in all plants except barley, revealing their expected chilling sensitivities. However, the decrease in P700 + by this short-term chilling was reversible in the presence of 3-(3',4'-dichlorophenyl)-1,1-dimethylurea or methyl viologen, and it did not show any causal relationship with the decrease in the electron transfer rate nor with the down-regulation of photosystem II through the accumulation of zeaxanthin and the development of non-photochemical quenching. These results led to the suggestion that photosystem I (PSI) acceptor side limitation is a prerequisite for the decrease of P700. Furthermore, PSI acceptor side limitation could be mainly due to limitation of electron-sink pathways such as CO 2 assimilation and ascorbate-glutathione cycle, because treatment with glycolaldehyde which inhibits the former pathway, and with KCN which inhibits both pathways, decreased P700 + by 20-30% in barley leaves after chilling in the light.

Cucumber leaf discs were illuminated at room-temperature with far-red light to photo-oxidise P700, the chlorophyll dimer in Photosystem (PS) I. The post-illumination kinetics of P700(+) re-reduction were studied in the presence of inhibitors or cofactors of photosynthetic electron transport. The re-reduction kinetics of P700(+) were well fitted as the sum of three exponentials, each with its amplitude and rate coefficient, and an initial flux (at the instant of turning off far-red light) given as the product of the two. Each initial flux is assumed equal to a steady state flux during far-red illumination. The fast phase of re-reduction, with rate coefficient k (1) approximately 10 s(-1), was completely abolished by a saturating concentration of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU); it is attributed to electron flow to P700(+) from PS II, which was stimulated to some extent by far-red light. The intermediate phase, with rate coefficient k (1) approximately 1 s(-1), was only partly diminished by methyl viologen (MV) which diverts electron flow to oxygen. The intermediate phase is attributed to electron donation from reduced ferredoxin to the intersystem pool; reduced ferredoxin could be formed: (1) directly by electron donation on the acceptor of PS I; and/or (2) indirectly by stromal reductants, in line with only a partial inhibition of the intermediate phase by MV. Duroquinol enhanced the intermediate phase in the presence of DCMU, presumably through its interaction with thylakoid membrane components leading to the partial reduction of plastoquinone. The slow phase of P700(+) re-reduction, with rate coefficient k (1) approximately 0.1 s(-1), was unaffected by DCMU and only slightly affected by MV; it could be associated with electron donation to either: (1) the intersystem chain by stromal reductants catalysed by NAD(P)H dehydrogenase slowly; or (2) plastocyanin/P700(+) by ascorbate diffusing across the thylakoid membrane to the lumen. It is concluded that a post-illumination analysis of the fluxes to P700(+) can be used to probe the pathways of electron flow to PS I in steady state illumination.

Growth and photosynthesis of grapevine ( Vitis vinifera L.) planted on two sloping cool climate vineyards were measured during the early growth season. At both vineyards, a small difference in mean minimum air temperature (1‐ 3 ∞ C) between two microsites accumulated over time, producing differences in shoot growth rate. The growth rates of the warmer (upper) microsite were 34‐63% higher than the cooler (lower) site. Photosynthesis measurements of both east and west canopy sides revealed that the difference in carbon gain between the warmer and cooler microsites was due to low temperatures restricting the photosynthetic contribution of east-facing leaves. East-facing leaves at the warmer microsite experienced less time at suboptimal temperature while being exposed to high irradiance, contributing to an average 10% greater net carbon gain compared to the east-facing leaves at the cooler microsite. This chilling-induced reduction in photosynthesis was not due to net photo-inhibition. Further analysis revealed that CO 2 - and light-saturated photosynthesis of grapevines was restricted by stomatal closure from 15 to 25 ∞ C and by a limitation of RuBP regeneration and/or end-product limitation from 5 to 15 ∞ C. Changes in photosynthetic carboxylation efficiency implied that Rubisco activity may also play a regulatory role at all temperatures. This restriction of total photosynthetic carbon gain is proposed to be a major contributor to the temperature dependence of growth rate at both vineyards during the early season growth period.

Photoinactivation of photosystem II (PSII) and energy dissipation at low leaf temperatures were investigated in leaves of glasshouse-grown grapevine (Vitis vinifera L. cv. Riesling). At low temperatures (< 15 degrees C), photosynthetic rates of CO(2) assimilation were reduced. However, despite a significant increase in the amount of light excessive to that required by photosynthesis, grapevine leaves maintained high intrinsic quantum efficiencies of PSII (F(v)/F(m)) and were highly resistant to photoinactivation compared to other species. Non-photochemical energy dissipation involving xanthophylls and fast D1 repair were the main protective processes reducing the 'gross' rate of photoinactivation and the 'net' rate of photoinactivation, respectively. We developed an improved method of energy dissipation analysis that revealed up to 75% of absorbed light is dissipated thermally via pH- and xanthophyll-mediated non-photochemical quenching at low temperatures (5-15 degrees C) and moderate (800 micro mol quanta m(-2) s(-1)) light. Up to 20% of the energy flux contributing to electron transport was dissipated via photorespiration when taking into account temperature-dependent mesophyll conductance; however, this flux used in photorespiration was only a relatively small amount of the total absorbed light energy. Photoreduction of O(2) at photosystem I (PSI) and subsequent superoxide detoxification (water-water cycle) was more sensitive to inhibition by low temperature than photorespiration. Therefore the water-water cycle represents a negligibly small energy sink below 15 degrees C, irrespective of mesophyll conductance.

We propose a simplified alternative method for quantifying the partitioning of excitation energy between photochemistry, fluorescence and thermal dissipation. This alternative technique uses existing well-defined quantum efficiencies such as Phi(PS II), leaving no 'excess' efficiency unaccounted for, effectively separates regulated and constitutive thermal dissipation processes, does not require the use of F(o) and F'(o) measurements and gives very similar results to the method proposed by Kramer et al. [(2004) Photosynth Res 79: 209-218]. We demonstrate the use of the technique using chlorophyll fluorescence measurements in grapevine leaves and observe a high dependence on thermal dissipation processes (up to 75%) at both high light and low temperature.

Electron transfers in the photosynthetic electron transport chain including the cytochrome (cyt) bf and Photosystem (PS) I complexes were studied in leaves of several plant species by measuring flash-induced absorbancy changes at specific wavelengths. The electrochromic signal (ECS), indicative of a trans-thylakoid membrane electric field, consisted of a fast phase arising from charge separation in both photosystems, and a slow rise usually interpreted as charge transfer in the cyt bf complex (part of the Q-cycle). The amplitude of the slow phase of the ECS was frequently greater than could be accounted for by the withdrawal of an electron from cyt bf via plastocyanin (PC) by oxidised P700 in PS I. The ‘extra’ slow ECS, variable depending on the number of turnovers and plant species, can be attributed to a variable operation of proton-pumping activity of the cyt bf complex. The redox kinetics of cyt f and b were obtained by deconvolution of the signals at three or four wavelengths. Rates of cyt b reduction were very high, and never the same as the onset kinetics of the slow ECS. The cyt f signal suggests that a fraction of the oxidised cyt f was re-reduced only slowly in the time of 5 s between consecutive flashes. Leaf discs in far-red light were given single-turnover flashes to measure the rates of P700ox reduction and reoxidation. To simulate the redox kinetics of the ECS, cyt f, cyt b and P700 it was assumed that a Q-cycle normally operated in bf complexes; reasonable values for the appropriate rate coefficients, and for the equilibrium constants for the cyt f/PC and P700/PC reactions were chosen. Close similarity of the observed data with those predicted from the simulation was obtained for cyt b, P700 (far-red light experiments) and the ECS, but not for cyt f. The results contribute to an understanding of photosynthetic electron transfers in vivo.

During photosynthesis, photoinactivation and repair of photosystem II (PSII) occur simultaneously, resulting in a net loss of functional PSII under a given irradiance. This study determines the rate coefficients for the partial processes, allowing the calculation of the partial rates at any concentration of functional/non-functional PSII. The rate coefficient of photoinactivation was obtained from the onset of photoinactivation of PSII in leaf segments of Capsicum annuum L. in the absence of repair, and was in turn used to obtain the rate coefficient (k r ) of repair of PSII when repair was occurring. The value of k r was found to be near maximum at an irradiance as low as 29μmol photons m -2 s -1 and peaked at or somewhat above the growth irradiance; however, it declined on further increasing the irradiance, possibly due to oxidative stress. The value of k r was considerably decreased by elevating the CO 2 to about 1%, particularly at low irradiance, probably due to acidification of the stroma to a pH outside the range that is optimal for protein synthesis. The method of determining k r is convenient to apply, not relying on radiolabelling and pulse-chase experiments.

Photosynthesis, the natural process that yields food, fuel and fibre, spans physical and biological sciences, spatially from atomic scales to the global and temporally from electronic transitions to the evolutionary time frame. Photosynthesis is highly efficient in its primary energy capture, but much less so in terms of conversion to crop yield. The natural photosynthetic system provides fertile ground for exploring and dissecting partial processes that may be mimicked inartificial systems for human needs, perhaps with improved efficiency. Future developments are limited only by the imagination.

Light-chilling stress, the combination of low-light illumination and low temperature, preferentially inactivated photosystem I (PSI) of cucumber (Cucumis sativus L.) leaves, resulting in the photoinhibition of photosynthesis. The extent of PSI photoinhibition, determined in vivo by monitoring absorption changes around 810 nm (induced by far-red light), was closely correlated with the redox state of the PSII electron acceptor Q(A), measured as the chlorophyll fluorescence parameter, 1-qP, where qP is a photochemical quenching coefficient. In contrast, the decrease in the far-red-induced leaf absorptance signal was not well correlated with the limited fragmentation of the PsaA/B gene products in the PSI reaction center after the light-chilling stress. Amongst various enzymes involved in the photooxidative damage such as superoxide dismutase (SOD), ascorbate peroxidase, and NAD(P)H dehydrogenase, only SOD was inhibited by light-chilling treatment. Further, an approximately 3-fold increase in the leaf content of H(2)O(2), a potent inhibitor of Cu/Zn-SOD, was observed after light-chilling stress. From these results, we suggest that Cu/Zn-SOD is the primary target of the light-chilling stress, followed by subsequent inactivation of PSI by reactive oxygen species.

Photoinactivation of photosystem II (PSII), the light–induced loss of ability to evolve oxygen, is an inevitable event during normal photosynthesis, exacerbated by saturating light but counteracted by repair via new protein synthesis. The photoinactivation of PSII is dependent on the dosage of light: in the absence of repair, typically one PSII is photoinactivated per 107photons, although the exact quantum yield of photoinactivation is modulated by a number of factors, and decreases as fewer active PSII targets are available. PSII complexes initially appear to be photoinactivated independently; however, when less than 30% functional PSII complexes remain, they seem to be protected by strongly dissipative PSII reaction centres in several plant species examined so far, a mechanism which we term ‘inactive PSII–mediated quenching‘. This mechanism appears to require a pH gradient across the photosynthetic membrane for its optimal operation. The residual fraction of functional PSII complexes may, in turn, aid in the recovery of photoinactivated PSII complexes when conditions become less severe. This mechanism may be important for the photosynthetic apparatus in extreme environments such as those experienced by over–wintering evergreen plants, desert plants exposed to drought and full sunlight and shade plants in sustained sunlight.

Dinoflagellates from the genus Symbiodinium form symbiotic relationships with many marine invertebrates, including reef-building corals. Symbiodinium is genetically diverse, and acquiring suitable Symbiodinium phylotypes is crucial for the host to survive in habitat environments, such as high-light conditions. The sensitivity of Symbiodinium to high light differs among Symbiodinium phylotypes, but the mechanism that controls light sensitivity has not yet been fully resolved. In the present study using high-light-tolerant and -sensitive Symbiodinium phylotypes, we examined what determines sensitivity to high light. In growth experiments under different light intensities, Symbiodinium CS-164 (clade B1) and CCMP2459 (clade B2) were identified as high-light-tolerant and -sensitive phylotypes, respectively. Measurements of the maximum quantum yield of photosystem II (PSII) and the maximum photosynthetic oxygen production rate after high-light exposure demonstrated that CCMP2459 is more sensitive to photoinhibition of PSII than CS-164, and tends to lose maximum photosynthetic activity faster. Measurement of photodamage to PSII under light of different wavelength ranges demonstrated that PSII in both Symbiodinium phylotypes was significantly more sensitive to photodamage under shorter wavelength regions of light spectra (

Most chloroplasts undergo changes in composition, function and structure in response to growth irradiance. However, Tradescantia albiflora, a facultative shade plant, is unable to modulate its light-harvesting components and has the same Chl a/Chl b ratios and number of functional PS II and PS I reaction centres on a Chl basis at all growth irradiances. With increasing growth irradiance, Tradescantia leaves have the same relative amount of chlorophyll-proteins of PS II and PS I, but increased xanthophyll cycle components and more zeaxanthin formation under high light. Despite high-light leaves having enhanced xanthophyll cycle content, all Tradescantia leaves acclimated to varying growth irradiances have similar non-photochemical quenching. These data strongly suggest that not all of the zeaxanthin formed under high light is necessarily non-covalently bound to major and minor light-harvesting proteins of both photosystems, but free zeaxanthin may be associated with LHC II and LHC I or located in the lipid bilayer. Under the unusual circumstances in light-acclimated Tradescantia where the numbers of functional PS II and PS I reaction centres and their antenna size are unaltered during growth under different irradiances, the extents of PS II photoinactivation by high irradiances are comparable. This is due to the extent of PS II photoinactivation being a light dosage effect that depends on the input (photon exposure, antenna size) and output (photosynthetic capacity, non-radiative dissipation) parameters, which in Tradescantia are not greatly varied by changes in growth irradiance.

When light absorption by a plant exceeds its capacity for light utilization, photosynthetic light harvesting is rapidly downregulated by photoprotective thermal dissipation, which is measured as nonphotochemical quenching of chlorophyll fluorescence (NPQ). To address the involvement of specific xanthophyll pigments in NPQ, we have analyzed mutants affecting xanthophyll metabolism in Arabidopsis thaliana. An npq1 lut2 double mutant was constructed, which lacks both zeaxanthin and lutein due to defects in the violaxanthin de-epoxidase and lycopene ∈-cyclase genes. The npq1 lut2 strain had normal Photosystem II efficiency and nearly wild-type concentrations of functional Photosystem II reaction centers, but the rapidly reversible component of NPQ was completely inhibited. Despite the defects in xanthophyll composition and NPQ, the npq1 lut2 mutant exhibited a remarkable ability to tolerate high light.

Leaf discs of Capsicum annuum L. were illuminated in air enriched with 1% CO 2 in the absence or presence of lincomycin, an inhibitor of chloroplast-encoded protein synthesis. The loss of functional photosystem (PS) II complexes with increase in cumulative light dose (photon exposure), assessed by the O 2 yield per single-turnover flash, was greater in leaves of plants grown in low light than those in high light; it was also exacerbated in the presence of lincomycin. A single exponential decay can describe the relationship between the loss of functional PSII and increase in cumulative photon exposure. From this relationship we obtained both the maximum quantum yield of photoinactivation of PSII at limiting photon exposures and the coefficient k, interpreted as the probability of photoinactivation of PSII per unit photon exposure. Parallel measurements of chlorophyll fluorescence after light treatment showed that 1/F o - 1/F m was linearly correlated with the functionality of PSII, where F o and F m are the chlorophyll fluorescence yields corresponding to open and closed PSII reaction centers, respectively. Using 1/F o - 1/F m as a convenient indicator of PSII functionality, it was found that PSII is present in excess; only after the loss of about 40% functional PSII complexes did PSII begin to limit photosynthetic capacity in capsicum leaves.

We present a unifying mechanism for photoinhibition based on current obsevations from in vivo studies rather than from in vitro studies with isolated thylakoids or PS II membranes. In vitro studies have limited relevance for in vivo photoinhibition because very high light is used with photon exposures rarely encountered in nature, and most of the multiple, interacting, protective strategies of PS II regulation in living cells are not functional. It is now established that the photoinactivation of Photosystem II in vivo is a probability and light-dosage event which depends on the photons absorbed and not the irradiance per se. As the reciprocity law is obeyed and target theory analysis strongly suggests that only one photon is required, we propose that a single dominant molecular mechanism occurs in vivo with one photon inactivating PS II under limiting, saturating or sustained high light. Two mechanisms have been proposed for photoinhibition under high light, acceptor-side and donor-side photoinhibition [see Aro et al. (1994) Biochim Biophys Acta 1143: 113–134], and another mechanism for very low light, the low-light syndrome [Keren et al. (1995) J Biol Chem 270: 806–814]. Based on the exciton-radical pair equilibrium model of exciton dynamics, we propose a unifying mechanism for the photoinactivation of PS II in vivo under steady-state photosynthesis that depends on the generation and maintenance of increased concentrations of the primary radical pair, P680+Pheo−, and the different ways charge recombination is regulated under varying environmental conditions [Anderson et al. (1997) Physiol Plant 100: 214–223]. We suggest that the primary cause of damage to D1 protein is P680+, rather than singlet O2 formed from triplet P680, or other reactive oxygen species.

According to the Second Law of Thermodynamics, an overall increase of entropy contributes to the driving force for any physicochemical process, but entropy has seldom been investigated in biological systems. Here, for the first time, we apply Isothermal Titration Calorimetry (ITC) to investigate the Mg(2+)-induced spontaneous stacking of photosynthetic membranes isolated from spinach leaves. After subtracting a large endothermic interaction of MgCl₂ with membranes, unrelated to stacking, we demonstrate that the enthalpy change (heat change at constant pressure) is zero or marginally positive or negative. This first direct experimental evidence strongly suggests that an entropy increase significantly drives membrane stacking in this ordered biological structure. Possible mechanisms for the entropy increase include: (i) the attraction between discrete oppositely-charged areas, releasing counterions; (ii) the release of loosely-bound water molecules from the inter-membrane gap; (iii) the increased orientational freedom of previously-aligned water dipoles; and (iv) the lateral rearrangement of membrane components.

Utilization of absorbed light energy by photosystem (PS) II for O2 evolution depends on the light-harvesting antenna size, but the role of antenna size in the photoinactivation of PSII seems controversial. To address this controversy, pea (Pisum sativum L.) plants were grown in low (50 [mu]mol m-2 s-1) or high (650 [mu]mol m-2 s-1) light. The doubled functional antenna size of PSII in low light allows each PSII to utilize twice as many photons at given flash light energies for O2 evolution. The application of a target theory to depict the photon dose dependency of PSII photoinactivation measured by repetitive-flash O2 yield and the ratio of variable to maximal chlorophyll fluorescence indicates that photoinactivation of PSII is probably a single-hit process in which repair or photoprotective mechanisms are only slightly involved. Furthermore, the exacerbation of photoinactivation of PSII with greater antenna size under anaerobic conditions strongly indicates that photoinactivation of PSII depends on antenna size.

Photosystem II plays a central role not only in energy transduction, but also in monitoring the molecular redox mechanisms involved in signal transduction for acclimation to environmental stresses. Central to the regulation of photosystem II (PSII) function as a light-driven molecular machine in higher plant leaves, is an inevitable photoinactivation of one PSII after 10 6 -10 7 photons have been delivered to the leaf, although the act of photoinactivation per se requires only one photon. PSII function in acclimated pea leaves shows a reciprocity between irradiance and the time of illumination, demonstrating that the photoinactivation of PSII is a light dosage effect, depending on the number of photons absorbed rather than the rate of photon absorption. Hence, PSII photoinactivation will occur at low as well as high irradiance. There is a heterogeneity of PSII functional stability, possibly with less stable PSII monomers being located in grana margins and more stable PSII dimers in appressed granal domains. Matching the inevitable photoinactivation of PSII, green plants have an intrinsic capacity for D1 protein synthesis to restore PSII function which is saturated at very low light. Photoinhibition of PSII in vivo is often a photoprotective strategy rather than a damaging process.

Tobacco transgenics with decreased amounts of the FeS apoprotein were generated using an antisense RNA construct targeted against the nuclear-encoded Rieske FeS protein of cytochrome bf complex [Price et al. (1995) Aust J Plant Physiol 22: 285–297]. FeS phenotypes ranging from intermediate to low were obtained which had 69% and 26% of the Rieske FeS protein of wild type. Similar reductions in the other subunits of cytochrome bf complex, cytochrome f, cytochrome b563and the 17 kDa subunit, were demonstrated in the thylakoids of intermediate and low FeS phenotypes. Confirmation that the levels of assembled cytochrome bf in leaves matched the levels of the FeS protein was demonstrated by laser flash-induced redox absorbance changes in leaves, with the extents of cytochrome f oxidation and cytochrome b563reduction being equivalent to the decreased amounts of the subunits in isolated thylakoids of the antisense plants. Despite greatly enhanced photochemical reduction of QAand the plastoquinone pool in the antisense plants, light acclimation of the FeS phenotypes to irradiance did not occur. Furthermore, the state 1–state 2 transitions were identical in wild type and antisense plants. Our results suggest that neither QAnor the plastoquinone pool acts alone in either the redox control of gene expression or the regulation of light energy distribution between the photosystems. We suggest rather that reduced plastoquinone acting at the inner Qpsite of cytochrome bf complex is involved in molecular redox signalling.