Photosystems and photoreceptors in cyanobacterial phototaxis and photophobotaxis.

Cyanobacteria move by gliding motility on surfaces toward the light or away from it. It is as yet unclear how the light direction is sensed on the molecular level. Diverse photoreceptor knockout mutants have a stronger response toward the light than the wild type. Either the light direction is sensed by multiple photoreceptors or by photosystems. In a study on photophobotaxis of the filamentous cyanobacterium Phormidium lacuna, broad spectral sensitivity, inhibition by 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), and a highly sensitive response speaks for photosystems as light direction sensors. Here, it is discussed whether the photosystem theory could hold for phototaxis of other cyanobacteria.

Growth and productivity of Haematococcus pluvialis and Coelastrella saipanensis by photosystem modulation for understanding the heterotrophic nutritional strategy for bioremediation application.

In this study, Haematococcus pluvialis and Coelastrella saipanensis were evaluated for heterotrophic nutrition potential in dairy waste medium by blocking the PSII using DCMU. The study was done by four sets of experiments. In the first set, in the different concentrations DCMU-treatments, 20µL showed pronounced effect in H. pluvialis and C. saipanensis as 89 % and 83% decrease in cells (>30 and > 250 cells/mL) compared to control (536 ± 12.35 × 104 and 1167 ± 15.35 × 104 cells/mL, respectively). Damage to the PS II by DCMU interrupted the growth, which in turn produced a significant drop in the number of cells. In the second round of experiment, growth of algae in various dairy waste concentrations suggest that dairy wastewater (DWW) provides enough nutrients to produce 35.71 % and 64.74 % more cells in H. pluvialis and C. saipanensis, respectively compared to the control. In the third set, high DCMU concentration was added to microalgae cultures in DWW to assess the heterotrophic nutrition potential. Growth in cell number 34.4 ± 19 and 617.46 ± 60.44 cells/mL was recorded in H. pluvialis and C. saipanensis when grown control medium whereas addition of DCMU reduced the cell number to 1.53 ± 0.75 and 55.13 ± 0.75 cells/mL on 15th day, respectively. This shows cells in cultures treated with DCMU reveal that algae can sustain their metabolic activity by utilizing the nutrients of dairy waste inhibiting photosystem. Fourth round of experiments found that microalgae could resume their growth and productivity by adapting to heterotrophic nutritional behaviour when DCMU given in mild dose at different time interval. This study conclude as C. saipanensis grows more readily by absorbing dairy waste nutrients than H. pluvialis. Therefore, C. saipanensis is an excellent choice for wastewater treatment through sustainable environmentally benign process after scale-up investigation. These results provide useful information to advance to molecular study for measuring microalgae's capability for bioremediation application.

Mechanistic insights into the effects of diuron exposure on Alexandrium pacificum.

Diuron (N-(3,4-dichlorophenyl)-N,N­dimethylurea, DCMU), a ureic herbicide, is extensively used in agriculture to boost crop productivity; however, its extensive application culminates in notable environmental pollution, especially in aquatic habitats. Therefore, the present study investigated the effect of diuron on the dinoflagellate Alexandrium pacificum, which is known to induce harmful algal blooms (HAB), and its potential to biodegrade DCMU. Following a four-day DCMU exposure, our results revealed that A. pacificum proficiently assimilated DCMU at concentrations of 0.05 mg/L and 0.1 mg/L in seawater, attaining a complete reduction (100 % efficiency) after 96 h for both concentrations. Moreover, evaluations of paralytic shellfish toxins content indicated that cells subjected to higher DCMU concentrations (0.1 mg/L) exhibited reductions of 73.4 %, 86.7 %, and 75 % in GTX1, GTX4, and NEO, respectively. Exposure to DCMU led to a notable decrease in A. pacificum's photosynthetic efficacy, accompanied by increased levels of reactive oxygen species (ROS) and suppressed cell growth, with a growth inhibition rate of 41.1 % at 72 h. Proteomic investigations pinpointed the diminished expression levels of specific proteins like SxtV and SxtW, linked to paralytic shellfish toxins (PSTs) synthesis, as well as key proteins associated with Photosystem II, namely PsbA, PsbD, PsbO, and PsbU. Conversely, proteins central to the cysteine biosynthesis pathways exhibited enhanced expression. In summary, our results preliminarily resolved the molecular mechanisms underlying the response of A. pacificum to DCMU and revealed that DCMU affected the synthesis of PSTs. Meanwhile, our data suggested that A. pacificum has great potential in scavenging DCMU.

Probing the Influence of Novel Organometallic Copper(II) Complexes on Spinach PSII Photochemistry Using OJIP Fluorescence Transient Measurements.

Modern agricultural cultivation relies heavily on genetically modified plants that survive after exposure to herbicides that kill weeds. Despite this biotechnology, there is a growing need for new sustainable, environmentally friendly, and biodegradable herbicides. We developed a novel [CuL2]Br2 complex (L = bis{4H-1,3,5-triazino[2,1-b]benzothiazole-2-amine,4-(2-imidazole) that is active on PSII by inhibiting photosynthetic oxygen evolution on the micromolar level. [CuL2]Br2 reduces the FV of PSII fluorescence. Artificial electron donors do not rescind the effect of [CuL2]Br2. The inhibitory mechanism of [CuL2]Br2 remains unclear. To explore this mechanism, we investigated the effect of [CuL2]Br2 in the presence/absence of the well-studied inhibitor DCMU on PSII-containing membranes by OJIP Chl fluorescence transient measurements. [CuL2]Br2 has two effects on Chl fluorescence transients: (1) a substantial decrease of the Chl fluorescence intensity throughout the entire kinetics, and (2) an auxiliary "diuron-like" effect. The initial decrease dominates and is observed both with and without DCMU. In contrast, the "diuron-like" effect is small and is observed only without DCMU. We propose that [CuL2]Br2 has two binding sites for PSII with different affinities. At the high-affinity site, [CuL2]Br2 produces effects similar to PSII reaction center inhibition, while at the low-affinity site, [CuL2]Br2 produces effects identical to those of DCMU. These results are compared with other PSII-specific classes of herbicides.

Unexpected growth promotion of Chlorella sacchrarophila triggered by herbicides DCMU.

The ecotoxicological effects of herbicide contamination on the autotrophic growth of microalgae in aquatic environments have been major concerns. However, little is known about the influence of herbicides on the mixotrophic growth of microalgae. This study investigated the ecotoxicological effect of 3-(3,4-dichlorophenyl)-1,1-dimethyl-urea (DCMU) on the mixotrophic growth of Chlorella sacchrarophila FACHB 4. Results showed that C. sacchrarophila in mixotrophy was more resistant to DCMU than in photoautotrophy. Moreover, a low content of DCMU (20-80 µg·L-1) promoted the mixotrophic growth of C. sacchrarophila, and the promotion effect was obviously enhanced with the increase in light intensity. The chlorophyll content and glucose absorption rate of C. sacchrarophila were found to increase after incubation with DCMU for 24 h. Transcriptome analyses revealed that the mechanism of DCMU to promote the mixotrophic growth of C. sacchrarophila was probably through accelerating glucose uptake and utilization, which was accomplished by reducing photodamage and increasing the chlorophyll content of C. sacchrarophila. This study not only revealed an unexpected bloom of mixotrophic microalgae triggered by herbicides, but it also shed new light on an effective and low-cost strategy to improve the microalgae productivity for utilization.

Measurement of Fluorescence for Monitoring Algal Growth and Health.

Measuring fluorescence is a noninvasive, inexpensive, and quick way of determining biomass concentration and health of the algae. Fluorescence is generally positively correlated with chlorophyll a and can as such be used as a proxy for biomass. In addition, the proportion variable fluorescence of maximal fluorescence is a measure of photochemical efficiency, which is affected by stress in a very early stage and can as such be used as a proxy for algal health.

Noninvasive determination of toxic stress biomarkers by high-throughput screening of photoautotrophic cell suspension cultures with multicolor fluorescence imaging.

BACKGROUND: With increasing pollution, herbicide application and interest in plant phenotyping, sensors capturing early responses to toxic stress are demanded for screening susceptible or resistant plant varieties. Standard toxicity tests on plants are laborious, demanding in terms of space and material, and the measurement of growth-inhibition based endpoints takes relatively long time. The aim of this work was to explore the potential of photoautotrophic cell suspension cultures for high-throughput early toxicity screening based on imaging techniques. The investigation of the universal potential of fluorescence imaging methods involved testing of three toxicants with different modes of action (DCMU, glyphosate and chromium). RESULTS: The increased pace of testing was achieved by using non-destructive imaging methods-multicolor fluorescence (MCF) and chlorophyll fluorescence (ChlF). These methods detected the negative effects of the toxicants earlier than it was reflected in plant growth inhibition (decrease in leaf area and final dry weight). Moreover, more subtle and transient effects not resulting in growth inhibition could be detected by fluorescence. The pace and sensitivity of stress detection was further enhanced by using photoautotrophic cell suspension cultures. These reacted sooner, more pronouncedly and to lower concentrations of the tested toxicants than the plants. Toxicant-specific stress signatures were observed as a combination of MCF and ChlF parameters and timing of the response. Principal component analysis was found to be useful for reduction of the collected multidimensional data sets to a few informative parameters allowing comparison of the toxicant signatures. CONCLUSIONS: Photoautotrophic cell suspension cultures have proved to be useful for rapid high-throughput screening of toxic stress and display a potential for employment as an alternative to tests on whole plants. The MCF and ChlF methods are capable of distinguishing early stress signatures of at least three different modes of action.

Symbiont Identity Influences Patterns of Symbiosis Establishment, Host Growth, and Asexual Reproduction in a Model Cnidarian-Dinoflagellate Symbiosis.

The genus Symbiodinium is physiologically diverse and so may differentially influence symbiosis establishment and function. To explore this, we inoculated aposymbiotic individuals of the sea anemone Exaiptasia pallida (commonly referred to as "Aiptasia"), a model for coral symbiosis, with one of five Symbiodinium species or types (S. microadriaticum, S. minutum, phylotype C3, S. trenchii, or S. voratum). The spatial pattern of colonization was monitored over time via confocal microscopy, and various physiological parameters were measured to assess symbiosis functionality. Anemones rapidly formed a symbiosis with the homologous symbiont, S. minutum, but struggled or failed to form a long-lasting symbiosis with Symbiodinium C3 or S. voratum, respectively. Symbiodinium microadriaticum and S. trenchii were successful but reached their peak density two weeks after S. minutum. The spatial pattern of colonization was identical for all Symbiodinium taxa that were ultimately successful, starting in the oral disk and progressing to the tentacles, before invading the column and, finally, the pedal disk. In all cases, proliferation through the anemone's tentacles was patchy, suggesting that symbionts were being expelled into the gastrovascular cavity and re-phagocytosed by the host. However, the timing of these various spatial events differed between the different Symbiodinium taxa. Furthermore, S. microadriaticum and S. trenchii were less beneficial to the host, as indicated by lower rates of photosynthesis, anemone growth, and pedal laceration. This study enhances our understanding of the link between symbiont identity and the performance of the overall symbiosis, which is important for understanding the potential establishment and persistence of novel host-symbiont pairings. Importantly, we also provide a baseline for further studies on this topic with the globally adopted "Aiptasia" model system.

H2 production pathways in nutrient-replete mixotrophic Chlamydomonas cultures under low light. Response to the commentary article "On the pathways feeding the H2 production process in nutrient-replete, hypoxic conditions," by Alberto Scoma and Szilvia Z. Tóth.

BACKGROUND: A recent Commentary article entitled "On the pathways feeding the H2 production process in nutrient-replete, hypoxic conditions" by Dr. Scoma and Dr. Tóth, Biotechnology for Biofuels (2017), opened a very interesting debate about the H2 production photosynthetic-linked pathways occurring in Chlamydomonas cultures grown in acetate-containing media and incubated under hypoxia/anoxia conditions. This Commentary article mainly focused on the results of our previous article "Low oxygen levels contribute to improve photohydrogen production in mixotrophic non-stressed Chlamydomonas cultures," by Jurado-Oller et al., Biotechnology for Biofuels (7, 2015; 8:149). MAIN BODY: Here, we review some previous knowledge about the H2 production pathways linked to photosynthesis in Chlamydomonas, especially focusing on the role of the PSII-dependent and -independent pathways in acetate-containing nutrient-replete cultures. The potential contributions of these pathways to H2 production under anoxia/hypoxia are discussed. CONCLUSION: Despite the fact that the PSII inhibitor DCMU is broadly used to discern between the two different photosynthetic pathways operating under H2 production conditions, its use may lead to distinctive conclusions depending on the growth conditions. The different potential sources of reductive power needed for the PSII-independent H2 production in mixotrophic nutrient-replete cultures are a matter of debate and conclusive evidences are still missing.

Carbon starvation induces lipid degradation via autophagy in the model alga Micrasterias.

Autophagy is regarded as crucial intracellular process in plant development but also in intracellular stress response. It is known to be controlled by the energy level of the cell and consequently can be triggered by energy deprivation. In this study carbon starvation evoked in different ways was investigated in the freshwater algae model system Micrasterias denticulata (Streptophyta) which is closely related to higher plants. Cells exposed to the photosynthesis inhibiting herbicide DCMU, to the glycolysis inhibitor 2-Deoxy-d-glucose and to complete darkness over up to 9 weeks for preventing metabolism downstream of glucose supply, were investigated by means of Nile red staining and analyses in CLSM, and TEM after cryo-preparation. Our results show that lipid bodies containing both neutral and polar lipids are evenly distributed inside the chloroplast in control cells. During carbon starvation they are displaced into the cytoplasm and are either degraded via autophagy and/or excreted from the cell. Upon discharge from the chloroplast lipid bodies become engulfed by double membranes probably deriving from the ER, thus forming autophagosomes which later fuse with vacuoles. Coincidently indications for autophagy of other organelles and cytoplasmic portions were found during starvation and particularly in DCMU treated cells the number of starch grains decreased and pyrenoids disintegrated. Additionally our molecular data provide first evidence for the existence of a single ATG8 isoform in Micrasterias. ATG8 is known as main regulator of both bulk and selective autophagy in eucaryotes. Our study indicates that lipid degradation during carbon starvation is achieved via "classical" autophagy in the alga Micrasterias. This process has so far only been very rarely observed in plant cells and seems to allow recruitment of lipids for energy supply on the one hand and elimination of unusable or toxicated lipids on the other hand.

Light-triggered selective nitration of PsbO1 in isolated Arabidopsis thylakoid membranes is inhibited by photosynthetic electron transport inhibitors.

PsbO1 is exclusively nitrated when isolated thylakoid membranes are incubated in a buffer bubbled with nitrogen dioxide (NO2) containing NO2 and nitrite. NO2 is the primary intermediate for this selective nitration. Isolated thylakoid membranes were incubated in NO2-bubbled buffer at 25°C in the light or dark. Protein analysis confirmed the selective nitration of PsbO1. Illumination was found to be essential in PsbO1 nitration. A nitration mechanism whereby nitratable tyrosine residues of PsbO1 are, prior to nitration, selectively photo-oxidized by photosynthetic electron transport to tyrosyl radicals to combine with NO2 to form 3-nitrotyrosine was hypothesized. We tested the electron transport inhibitors 3-(3,4-dichlorophenyl)-1,1- dimethylurea, sodium azide, and 1,5-diphenylcarbazide and found distinct inhibition of nitration of PsbO1. We also propose a possible nitration mechanism.

Assessing the Impact of Photosynthetic Sugars on the Arabidopsis Circadian Clock.

Circadian clocks drive 24 h biological rhythms to optimize physiology and development in response to the rotation of the planet. In plants, photosynthesis is modulated by the circadian clock and contributes to daily rhythms in cellular metabolism. In addition to light and temperature, sugar produced from photosynthesis acts as a zeitgeber to contribute to setting of the plant circadian clock. Here, we describe methods to manipulate photosynthetic output and sugar availability in Arabidopsis seedlings. These protocols have been applied to investigate the effects on the Arabidopsis circadian network, but are easily adaptable to other processes in plants.

Nondestructive evaluation of photosynthesis by delayed luminescence in Arabidopsis in Petri dishes.

Nondestructive evaluation of photosynthesis is a valuable tool in the field and laboratory. Delayed luminescence (DL) can reflect charge recombination through the backflow of electrons. However, DL detection has not yet been adapted for whole plants in Petri dishes. To compensate for differences in DL decay between sibling Arabidopsis plants grown under the same conditions, we developed a time-sequential double measurement method. Using this method, we examined the influence of photosynthetic electron flow inhibitors, and differences in the DL decay curves were categorized by considering the initial and late phases of the decay curves, as well as their intermediate slopes. The appearance of concavity and convexity in DL curves in Arabidopsis was different from unicellular algae, suggesting complexity in the photosynthetic machinery of higher plants. This detection method should be invaluable for evaluating photosynthetic defects in higher plants under sterile conditions without interrupting plant culture.

Low oxygen levels contribute to improve photohydrogen production in mixotrophic non-stressed Chlamydomonas cultures.

BACKGROUND: Currently, hydrogen fuel is derived mainly from fossil fuels, but there is an increasing interest in clean and sustainable technologies for hydrogen production. In this context, the ability of some photosynthetic microorganisms, particularly cyanobacteria and microalgae, to produce hydrogen is a promising alternative for renewable, clean-energy production. Among a diverse array of photosynthetic microorganisms able to produce hydrogen, the green algae Chlamydomonas reinhardtii is the model organism widely used to study hydrogen production. Despite the well-known fact that acetate-containing medium enhances hydrogen production in this algae, little is known about the precise role of acetate during this process. RESULTS: We have examined several physiological aspects related to acetate assimilation in the context of hydrogen production metabolism. Measurements of oxygen and CO2 levels, acetate uptake, and cell growth were performed under different light conditions, and oxygenic regimes. We show that oxygen and light intensity levels control acetate assimilation and modulate hydrogen production. We also demonstrate that the determination of the contribution of the PSII-dependent hydrogen production pathway in mixotrophic cultures, using the photosynthetic inhibitor DCMU, can lead to dissimilar results when used under various oxygenic regimes. The level of inhibition of DCMU in hydrogen production under low light seems to be linked to the acetate uptake rates. Moreover, we highlight the importance of releasing the hydrogen partial pressure to avoid an inherent inhibitory factor on the hydrogen production. CONCLUSION: Low levels of oxygen allow for low acetate uptake rates, and paradoxically, lead to efficient and sustained production of hydrogen. Our data suggest that acetate plays an important role in the hydrogen production process, during non-stressed conditions, other than establishing anaerobiosis, and independent of starch accumulation. Potential metabolic pathways involved in hydrogen production in mixotrophic cultures are discussed. Mixotrophic nutrient-replete cultures under low light are shown to be an alternative for the simultaneous production of hydrogen and biomass.

Plastoquinone redox state modifies plant response to pathogen.

The role of PQ (plastoquinione) redox state in establishment of response to pathogen infection (Botrytis cinerea) was tested along the regulation of main antioxidative enzymes (superoxide dismutase - SOD, catalase - CAT) and photochemistry of PSII (photosystem II) in Mesembryanthemum crystallinum plants performing C3 and CAM (Crassulacean acid metabolism) carbon metabolism. The redox state of PQ was modified by two inhibitors of photosynthetic electron transport resulting in a more oxidised (3-(3,4-dichlorophenyl)-1,1-dimethylurea; DCMU) or reduced (2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone; DBMIB) PQ redox state simulating darkness and high light conditions, respectively. Irrespective of the type of treatment (mock inoculation or pathogen inoculation) SOD activity depended on the PQ pool. Our results suggest that regarding changes in infection-induced CAT activity, plants developed response that is vital for hypersensitive-like (HR-like) response establishment only when PQ pool generated signal was similar to that in light presence (DBMIB pre-treatment). When PQ pool generated signal was similar to darkness, CAT activity response remained stress-independent, similarly to SOD. Fluorescence parameters of PSII, Qp (photochemical quenching coefficient) and NPQ (non-photochemical quenching) were affected only in the tissues treated with DCMU in stress-independent manner. We suggest that in case of abiotic and biotic stresses signals emerging from PQ pool indirectly orchestrate plant response and carbon metabolism affects this regulatory pathway.

Hormesis depends upon the life-stage and duration of exposure: Examples for a pesticide and a nanomaterial.

Tests to assess toxic effects on the reproduction of adult C. elegans after 72h exposure for two chemicals, (3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU)), also known as diuron, and silver nanoparticles (Ag NPs) indicated potential, although not significant hormesis. Follow up toxicity tests comparing the potential hormesis concentrations with controls at high replication confirmed that the stimulatory effect was repeatable and also statistically significant within the test. To understand the relevance of the hormesis effects for overall population fitness, full life-cycle toxicity tests were conducted for each chemical. When nematodes were exposed to DCMU over the full life-span, the hormesis effect for reproduction seen in short-term tests was no longer evident. Further at the putative hormesis concentrations, a negative effect of DCMU on time to maturation was also seen. For the Ag NPs, the EC50 for effects on reproduction in the life-cycle exposure was substantially lower than in the short-term test, the EC50s estimated by a three parameter log logistic model being 2.9mg/L and 0.75mg/L, respectively. This suggests that the level of toxicity for Ag NPs for C. elegans reproduction is dependant on the life stage exposed and possibly the duration of the exposure. Further, in the longer duration exposures, hormesis effects on reproduction seen in the short-term exposures were no longer apparent. Instead, all concentrations reduced both overall brood size and life-span. These results for both chemicals suggest that the hormesis observed for a single endpoint in short-term exposure may be the result of a temporary reallocation of resources between traits that are not sustained over the full life-time. Such reallocation is consistent with energy budget theories for organisms subject to toxic stress.

The effect of inhibitors on photosynthetic electron transport chain in canola leaf discs / Efeito de inibidores na cadeia de transporte de elétrons fotossintética em discos foliares de canola

The mechanisms of photosynthetic electron transport can be elucidated by inhibition of electron flow through the use of specific substances that, when combined with the chlorophyll chlorophyll a fluorescence emission was measured to investigate the effect of several inhibitors of the photosynthetic electron transport chain in canola leaf discs. Leaf discs were incubated in the dark for 2 hours in different solutions: (a) water ­ control; (b) DCMU at 500 µM; (c) phenanthroline at 10 mM; and (d) hydroxylamine at 10, 50, 100 and 200 mM. Similar effects were observed between DCMU and phenanthroline, the initial fluorescence value was altered, but not the maximum fluorescence, with the disappearance of the IP phase. Hydroxylamine interacted and inhibited the oxygen evolving complex and caused an imbalance between the rate of QA reduction by photosystem II and the rate of QA oxidation by photosystem I.

Os mecanismos de transporte de elétrons podem ser elucidados pela inibição do fluxo de elétrons pelo uso de substâncias específicas que, juntamente com a técnica de fluorescência da clorofila, torna-se uma ferramenta importante para detalhar a cadeia de transporte de elétrons. Neste trabalho, a emissão da fluorescência da clorofila foi mensurada para investigar o efeito dos diferentes inibidores da cadeia de transporte de elétrons fotossintéticos de discos foliares de canola. Os discos foliares foram incubados no escuro durante 02 h em diferentes soluções: (a) água - controle, (b) 500 uM de DCMU, (c) 10 mM de fenantrolina, e (d) 10, 50, 100 e 200 mM de hidroxilamina. Foram observados efeitos similares entre DCMU e fenantrolina, o valor da fluorescência inicial foi alterado, contudo a fluorescência máxima não se alterou, havendo o desaparecimento da fase de IP. Hidroxilamina interagiu e inibiu o complexo de evolução de oxigênio e causou desequilíbrio entre a taxa de redução QA pelo fotossistema II e a taxa de oxidação QA pelo fotossistema I.

PsbS is required for systemic acquired acclimation and post-excess-light-stress optimization of chlorophyll fluorescence decay times in Arabidopsis.

Systemic acquired acclimation (SAA) is an important light acclimatory mechanism that depends on the global adjustments of non-photochemical quenching and chloroplast retrograde signaling. As the exact regulation of these processes is not known, we measured time-resolved fluorescence of chlorophyll a in Arabidopsis thaliana leaves exposed to excess light, in leaves undergoing SAA, and in leaves after excess light episode. We compare the behavior induced in wild-type plants with null mutant of non-photochemical quenching (npq4-1). The wild type rosettes exhibit a small reduction of fluorescence decay times in leaves directly exposed to excess light and in leaves undergoing SAA in ambient low light. However in npq4-1 exposition to excess light results in much faster fluorescence decay, which is insensitive to excitation power. At the same time npq4-1 leaves undergoing SAA displayed intermediate fluorescence decay. The npq4-1 plants also lost the ability to optimize florescence decay, and thus chlorophyll a dynamics up to 2 h after excess light episode. The fluorescence decay dynamics in both WT and npq4-1 can be described by a set of 3 maximum decay times. Based on the results, we concluded that functional PsbS is required for optimization of absorbed photon fate and optimal light acclimatory responses such as SAA or after excess light stress.

Light-dependent expression of flg22-induced defense genes in Arabidopsis.

Chloroplasts have been reported to generate retrograde immune signals that activate defense gene expression in the nucleus. However, the roles of light and photosynthesis in plant immunity remain largely elusive. In this study, we evaluated the effects of light on the expression of defense genes induced by flg22, a peptide derived from bacterial flagellins which acts as a potent elicitor in plants. Whole-transcriptome analysis of flg22-treated Arabidopsis thaliana seedlings under light and dark conditions for 30 min revealed that a number of (30%) genes strongly induced by flg22 (>4.0) require light for their rapid expression, whereas flg22-repressed genes include a significant number of genes that are down-regulated by light. Furthermore, light is responsible for the flg22-induced accumulation of salicylic acid (SA), indicating that light is indispensable for basal defense responses in plants. To elucidate the role of photosynthesis in defense, we further examined flg22-induced defense gene expression in the presence of specific inhibitors of photosynthetic electron transport: 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) and 2,5-dibromo-3-methyl-6-isopropyl-benzoquinone (DBMIB). Light-dependent expression of defense genes was largely suppressed by DBMIB, but only partially suppressed by DCMU. These findings suggest that photosynthetic electron flow plays a role in controlling the light-dependent expression of flg22-inducible defense genes.

Modification of the pheophytin redox potential in Thermosynechococcus elongatus Photosystem II with PsbA3 as D1.

In Photosystem II (PSII) of the cyanobacterium Thermosynechococcus elongatus, glutamate 130 in the high-light variant of the D1-subunit (PsbA3) was changed to glutamine in a strain lacking the two other genes for D1, psbA1 and psbA2. The resulting PSII (PsbA3/Glu130Gln) was compared with those from the "native" high-light (PsbA3-PSII) and low-light (PsbA1-PSII) variants, which differ by 21 amino acid including Glu130Gln. H-bonding from D1-Glu130Gln to the primary electron acceptor, PheophytinD1 (PheoD1), is known to affect the Em of the PheoD1/PheoD1(-) couple. The Gln130 mutation here had little effect on water splitting, charge accumulation and photosensitivity but did slow down S2QA(-) charge recombination and up-shift the thermoluminescence while increasing its yield. These changes were consistent with a ≈-30mV shift of the PheoD1/PheoD1(-)Em, similar to earlier single site-mutation results from other species and double the ≈-17mV shift seen for PsbA1-PSII versus PsbA3-PSII. This is attributed to the influence of the other 20 amino-acids that differ in PsbA3. A computational model for simulating S2QA(-) recombination matched the experimental trend: the S2QA(-) recombination rate in PsbA1-PSII differed only slightly from that in PsbA3-PSII, while in Glu130-PsbA3-PSII there was a more pronounced slowdown of the radical pair decay. The simulation predicted a major effect of the PheoD1/PheoD1(-) potential on (1)O2 yield (~60% in PsbA1-PSII, ~20% in PsbA3-PSII and ~7% in Gln130-PsbA3-PSII), reflecting differential sensitivities to high light.