Roles of a Cryptochrome in Carbon Fixation and Sucrose Metabolism in the Liverwort Marchantia polymorpha.

In vascular plants, cryptochromes acting as blue-light photoreceptors have various functions to adapt plants to the fluctuating light conditions on land, while the roles of cryptochromes in bryophytes have been rarely reported. In this study, we investigated functions of a single-copy ortholog of cryptochrome (MpCRY) in the liverwort Marchantia polymorpha. Knock-out of MpCRY showed that a large number of the mutant plants exhibited asymmetric growth of thalli under blue light. Transcriptome analyses indicated that MpCRY is mainly involved in photosynthesis and sugar metabolism. Further physiological analysis showed that Mpcry mutant exhibited a reduction in CO2 uptake and sucrose metabolism. In addition, exogenous application of sucrose or glucose partially restored the symmetrical growth of the Mpcry mutant thalli. Together, these results suggest that MpCRY is involved in the symmetrical growth of thallus and the regulation of carbon fixation and sucrose metabolism in M. polymorpha.

Structural complexity governs seagrass acclimatization to depth with relevant consequences for meadow production, macrophyte diversity and habitat carbon storage capacity.

Analyses of the integrated seagrass response to depth support the previously documented low plasticity and consistent shade-adapted leaf physiology of a habitat-builder that dominates well-illuminated reef environments. Two structural responses, "canopy-opening" and "below-ground-mass-depletion", govern the photoacclimatory response and facilitate, respectively, light penetration within the canopy and functional adjustments in whole-plant carbon balances. Conversely, "canopy-closing" may also explain dense canopies formed close to the waterline, as they provide shade and photoprotection to a susceptible leaf physiology under high-light. Canopy light attenuation is primarily regulated by the leaf area index (LAI), which is governed by changes in shoot size and density. Shoot density diminishes non-linearly with depth, while shoot size increases to a maximum followed by a decline. The initial increase in shoot size, which resembles a self-thinning response, increases LAI and meadow production in shallow depths. These seagrass structural adjustments have relevant ecological implications. Canopy-thinning allows macrophyte diversity to increase with depth, while seagrass production and carbon storage diminish exponentially, and are maximal only in a shallow coastal fringe. The results support the universality of plant self-thinning, from phytoplankton to complex canopies, likely the consequence of simple physical laws related to light limitation and pigment self-shading within photosynthetic structures and communities.

Modeling the Interplay between Photosynthesis, CO2 Fixation, and the Quinone Pool in a Purple Non-Sulfur Bacterium.

Rhodopseudomonas palustris CGA009 is a purple non-sulfur bacterium that can fix carbon dioxide (CO2) and nitrogen or break down organic compounds for its carbon and nitrogen requirements. Light, inorganic, and organic compounds can all be used for its source of energy. Excess electrons produced during its metabolic processes can be exploited to produce hydrogen gas or biodegradable polyesters. A genome-scale metabolic model of the bacterium was reconstructed to study the interactions between photosynthesis, CO2 fixation, and the redox state of the quinone pool. A comparison of model-predicted flux values with available Metabolic Flux Analysis (MFA) fluxes yielded predicted errors of 5-19% across four different growth substrates. The model predicted the presence of an unidentified sink responsible for the oxidation of excess quinols generated by the TCA cycle. Furthermore, light-dependent energy production was found to be highly dependent on the quinol oxidation rate. Finally, the extent of CO2 fixation was predicted to be dependent on the amount of ATP generated through the electron transport chain, with excess ATP going toward the energy-demanding Calvin-Benson-Bassham (CBB) pathway. Based on this analysis, it is hypothesized that the quinone redox state acts as a feed-forward controller of the CBB pathway, signaling the amount of ATP available.

What is the fate of xylem-transported CO2 in Kranz-type C4 plants?

High concentrations of dissolved inorganic carbon in stems of herbaceous and woody C3 plants exit leaves in the dark. In the light, C3 species use a small portion of xylem-transported CO2 for leaf photosynthesis. However, it is not known if xylem-transported CO2 will exit leaves in the dark or be used for photosynthesis in the light in Kranz-type C4 plants. Cut leaves of Amaranthus hypochondriacus were placed in one of three solutions of [NaH13 CO3 ] dissolved in KCl water to measure the efflux of xylem-transported CO2 exiting the leaf in the dark or rates of assimilation of xylem-transported CO2 * in the light, in real-time, using a tunable diode laser absorption spectroscope. In the dark, the efflux of xylem-transported CO2 increased with increasing rates of transpiration and [13 CO2 *]; however, rates of 13 Cefflux in A. hypochondriacus were lower compared to C3 species. In the light, A. hypochondriacus fixed nearly 75% of the xylem-transported CO2 supplied to the leaf. Kranz anatomy and biochemistry likely influence the efflux of xylem-transported CO2 out of cut leaves of A. hypochondriacus in the dark, as well as the use of xylem-transported CO2 * for photosynthesis in the light. Thus increasing the carbon use efficiency of Kranz-type C4 species over C3 species.

Stomatal response to blue light in crassulacean acid metabolism plants Kalanchoe pinnata and Kalanchoe daigremontiana.

Blue light (BL) is a fundamental cue for stomatal opening in both C3 and C4 plants. However, it is unknown whether crassulacean acid metabolism (CAM) plants open their stomata in response to BL. We investigated stomatal BL responses in the obligate CAM plants Kalanchoe pinnata and Kalanchoe daigremontiana that characteristically open their stomata at night and close them for part of the day, as contrasted with C3 and C4 plants. Stomata opened in response to weak BL superimposed on background red light in both intact leaves and detached epidermal peels of K. pinnata and K. daigremontiana. BL-dependent stomatal opening was completely inhibited by tautomycin and vanadate, which repress type 1 protein phosphatase and plasma membrane H+-ATPase, respectively. The plasma membrane H+-ATPase activator fusicoccin induced stomatal opening in the dark. Both BL and fusicoccin induced phosphorylation of the guard cell plasma membrane H+-ATPase in K. pinnata. These results indicate that BL-dependent stomatal opening occurs in the obligate CAM plants K. pinnata and K. daigremontiana independently of photosynthetic CO2 assimilation mode.

Enhanced Production of Fatty Acids via Redirection of Carbon Flux in Marine Microalga Tetraselmis sp.

Lipids in microalgae are energy-rich compounds and considered as an attractive feedstock for biodiesel production. To redirect carbon flux from competing pathways to the fatty acid synthesis pathway of Tetraselmis sp., we used three types of chemical inhibitors that can block the starch synthesis pathway or photorespiration, under nitrogen-sufficient and nitrogen-deficient conditions. The starch synthesis pathway in chloroplasts and the cytosol can be inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea and 1,2-cyclohexane diamine tetraacetic acid (CDTA), respectively. Degradation of glycine into ammonia during photorespiration was blocked by aminooxyacetate (AOA) to maintain biomass concentration. Inhibition of starch synthesis pathways in the cytosol by CDTA increased fatty acid productivity by 27% under nitrogen deficiency, whereas the blocking of photorespiration in mitochondria by AOA was increased by 35% under nitrogen-sufficient conditions. The results of this study indicate that blocking starch or photorespiration pathways may redirect the carbon flux to fatty acid synthesis.

A model for the irradiance responses of photosynthesis.

The analysis of the irradiance responses of photosynthetic processes, such as the quantum efficiencies of electron transport by photosystems I and II (PSI and PSII) or the rate of carbon dioxide fixation, is limited by the lack of mechanistically based analytical model for these processes. Starting with a model of P700 redox state, we develop a series of analytical functions which can be used to fit the irradiance responses of the quantum yields for electron transport by PSI and PSII, the irradiance responses of electron transport by PSI and PSII, and even the irradiance response of the fixation rate of carbon dioxide. These functions depend on two or three parameters so they can be fit to typical irradiance response data. We illustrate by example the use of these functions in various applications and discuss further use and development of the basic model described in detail here.

Modeling the diversion of primary carbon flux into secondary metabolism under variable nitrate and light/dark conditions.

In plants, the partitioning of carbon resources between growth and defense is detrimental for their development. From a metabolic viewpoint, growth is mainly related to primary metabolism including protein, amino acid and lipid synthesis, whereas defense is based notably on the biosynthesis of a myriad of secondary metabolites. Environmental factors, such as nitrate fertilization, impact the partitioning of carbon resources between growth and defense. Indeed, experimental data showed that a shortage in the nitrate fertilization resulted in a reduction of the plant growth, whereas some secondary metabolites involved in plant defense, such as phenolic compounds, accumulated. Interestingly, sucrose, a key molecule involved in the transport and partitioning of carbon resources, appeared to be under homeostatic control. Based on the inflow/outflow properties of sucrose homeostatic regulation we propose a global model on how the diversion of the primary carbon flux into the secondary phenolic pathways occurs at low nitrate concentrations. The model can account for the accumulation of starch during the light phase and the sucrose remobilization by starch degradation during the night. Day-length sensing mechanisms for variable light-dark regimes are discussed, showing that growth is proportional to the length of the light phase. The model can describe the complete starch consumption during the night for plants adapted to a certain light/dark regime when grown on sufficient nitrate and can account for an increased accumulation of starch observed under nitrate limitation.

Kinetic characteristics and modeling of microalgae Chlorella vulgaris growth and CO2 biofixation considering the coupled effects of light intensity and dissolved inorganic carbon.

Understanding and optimizing the microalgae growth process is an essential prerequisite for effective CO2 capture using microalgae in photobioreactors. In this study, the kinetic characteristics of microalgae Chlorella vulgaris growth in response to light intensity and dissolved inorganic carbon (DIC) concentration were investigated. The greatest values of maximum biomass concentration (Xmax) and maximum specific growth rate (µmax) were obtained as 2.303 g L(-1) and 0.078 h(-1), respectively, at a light intensity of 120 µmol m(-2) s(-1) and DIC concentration of 17 mM. Based on the results, mathematical models describing the coupled effects of light intensity and DIC concentration on microalgae growth and CO2 biofixation are proposed. The models are able to predict the temporal evolution of C. vulgaris growth and CO2 biofixation rates from lag to stationary phases. Verification experiments confirmed that the model predictions agreed well with the experimental results.

High light exposure on seed coat increases lipid accumulation in seeds of castor bean (Ricinus communis L.), a nongreen oilseed crop.

Little was known on how sunlight affects the seed metabolism in nongreen seeds. Castor bean (Ricinus communis L.) is a typical nongreen oilseed crop and its seed oil is an important feedstock in industry. In this study, photosynthetic activity of seed coat tissues of castor bean in natural conditions was evaluated in comparison to shaded conditions. Our results indicate that exposure to high light enhances photosynthetic activity in seed coats and consequently increases oil accumulation. Consistent results were also reached using cultured seeds. High-throughput RNA-Seq analyses further revealed that genes involved in photosynthesis and carbon conversion in both the Calvin-Benson cycle and malate transport were differentially expressed between seeds cultured under light and dark conditions, implying several venues potentially contributing to light-enhanced lipid accumulation such as increased reducing power and CO2 refixation which underlie the overall lipid biosynthesis. This study demonstrated the effects of light exposure on oil accumulation in nongreen oilseeds and greatly expands our understanding of the physiological roles that light may play during seed development in nongreen oilseeds. Essentially, our studies suggest that potential exists to enhance castor oil yield through increasing exposure of the inflorescences to sunlight either by genetically changing the plant architecture (smart canopy) or its growing environment.

Levels of daily light doses under changed day-night cycles regulate temporal segregation of photosynthesis and N2 Fixation in the cyanobacterium Trichodesmium erythraeum IMS101.

While the diazotrophic cyanobacterium Trichodesmium is known to display inverse diurnal performances of photosynthesis and N2 fixation, such a phenomenon has not been well documented under different day-night (L-D) cycles and different levels of light dose exposed to the cells. Here, we show differences in growth, N2 fixation and photosynthetic carbon fixation as well as photochemical performances of Trichodesmium IMS101 grown under 12L:12D, 8L:16D and 16L:8D L-D cycles at 70 µmol photons m-2 s-1 PAR (LL) and 350 µmol photons m-2 s-1 PAR (HL). The specific growth rate was the highest under LL and the lowest under HL under 16L:8D, and it increased under LL and decreased under HL with increased levels of daytime light doses exposed under the different light regimes, respectively. N2 fixation and photosynthetic carbon fixation were affected differentially by changes in the day-night regimes, with the former increasing directly under LL with increased daytime light doses and decreased under HL over growth-saturating light levels. Temporal segregation of N2 fixation from photosynthetic carbon fixation was evidenced under all day-night regimes, showing a time lag between the peak in N2 fixation and dip in carbon fixation. Elongation of light period led to higher N2 fixation rate under LL than under HL, while shortening the light exposure to 8 h delayed the N2 fixation peaking time (at the end of light period) and extended it to night period. Photosynthetic carbon fixation rates and transfer of light photons were always higher under HL than LL, regardless of the day-night cycles. Conclusively, diel performance of N2 fixation possesses functional plasticity, which was regulated by levels of light energy supplies either via changing light levels or length of light exposure.

Temporal Gene Expression of the Cyanobacterium Arthrospira in Response to Gamma Rays.

The edible cyanobacterium Arthrospira is resistant to ionising radiation. The cellular mechanisms underlying this radiation resistance are, however, still largely unknown. Therefore, additional molecular analysis was performed to investigate how these cells can escape from, protect against, or repair the radiation damage. Arthrospira cells were shortly exposed to different doses of 60Co gamma rays and the dynamic response was investigated by monitoring its gene expression and cell physiology at different time points after irradiation. The results revealed a fast switch from an active growth state to a kind of 'survival modus' during which the cells put photosynthesis, carbon and nitrogen assimilation on hold and activate pathways for cellular protection, detoxification, and repair. The higher the radiation dose, the more pronounced this global emergency response is expressed. Genes repressed during early response, suggested a reduction of photosystem II and I activity and reduced tricarboxylic acid (TCA) and Calvin-Benson-Bassham (CBB) cycles, combined with an activation of the pentose phosphate pathway (PPP). For reactive oxygen species detoxification and restoration of the redox balance in Arthrospira cells, the results suggested a powerful contribution of the antioxidant molecule glutathione. The repair mechanisms of Arthrospira cells that were immediately switched on, involve mainly proteases for damaged protein removal, single strand DNA repair and restriction modification systems, while recA was not induced. Additionally, the exposed cells showed significant increased expression of arh genes, coding for a novel group of protein of unknown function, also seen in our previous irradiation studies. This observation confirms our hypothesis that arh genes are key elements in radiation resistance of Arthrospira, requiring further investigation. This study provides new insights into phasic response and the cellular pathways involved in the radiation resistance of microbial cells, in particularly for photosynthetic organisms as the cyanobacterium Arthrospira.

Induction of Photosynthetic Carbon Fixation in Anoxia Relies on Hydrogenase Activity and Proton-Gradient Regulation-Like1-Mediated Cyclic Electron Flow in Chlamydomonas reinhardtii.

The model green microalga Chlamydomonas reinhardtii is frequently subject to periods of dark and anoxia in its natural environment. Here, by resorting to mutants defective in the maturation of the chloroplastic oxygen-sensitive hydrogenases or in Proton-Gradient Regulation-Like1 (PGRL1)-dependent cyclic electron flow around photosystem I (PSI-CEF), we demonstrate the sequential contribution of these alternative electron flows (AEFs) in the reactivation of photosynthetic carbon fixation during a shift from dark anoxia to light. At light onset, hydrogenase activity sustains a linear electron flow from photosystem II, which is followed by a transient PSI-CEF in the wild type. By promoting ATP synthesis without net generation of photosynthetic reductants, the two AEF are critical for restoration of the capacity for carbon dioxide fixation in the light. Our data also suggest that the decrease in hydrogen evolution with time of illumination might be due to competition for reduced ferredoxins between ferredoxin-NADP(+) oxidoreductase and hydrogenases, rather than due to the sensitivity of hydrogenase activity to oxygen. Finally, the absence of the two alternative pathways in a double mutant pgrl1 hydrogenase maturation factor G-2 is detrimental for photosynthesis and growth and cannot be compensated by any other AEF or anoxic metabolic responses. This highlights the role of hydrogenase activity and PSI-CEF in the ecological success of microalgae in low-oxygen environments.

Proteomic analysis of Physcomitrella patens treated with 12-oxo-phytodienoic acid, an important oxylipin in plants.

12-Oxo-phytodienoic acid (OPDA) is biosynthesized in the octadecanoid pathway and is considered to be a signaling molecule in plants. In Physcomitrella patens, OPDA is induced by bacterial infection and mechanical stress and is known to suppress growth; however, the functional mechanism of OPDA signaling remains elusive. In this study, we performed a proteomic analysis of P. patens treated with OPDA and found that the expression of 82 proteins was significantly altered, with approximately 80% of these proteins being downregulated by OPDA. The identified proteins were mainly categorized as being involved in photosynthesis, metabolism, and protein synthesis, and most of the proteins that were upregulated by OPDA are involved in light-dependent reactions, suggesting that OPDA regulates a function in chloroplasts. Additionally, OPDA induced the expression of an allene oxide cyclase (PpAOC1) in the octadecanoid pathway, demonstrating positive feedback regulation by OPDA in P. patens.

Effect of monochromatic illumination on lipid accumulation of Nannochloropsis gaditana under continuous cultivation.

Although nitrogen starvation is frequently used to increase lipid contents in microalgae, it has a negative effect on cellular growth. Since light supply is essential for photosynthetic organisms, the effects of cultivation under monochromatic illumination on the growth and lipid contents of Nannochloropsis gaditana were assessed. Continuous cultivation under blue and red light conditions improved the productivity and physical properties for biodiesel from this microalga. FAME yield was twofold higher under red light than under normal white light (21.12% vs 11.35%), with no significant difference in growth rates. Blue and red light increased photosynthetic oxygen evolution, carbon fixation and nutrient uptake. In total, more significant physiological changes were observed under red than under blue light. These results show that red light illumination may be useful for enhancing lipid production by N. gaditana, with the increased photosynthetic reducing equivalents induced by red light which could be deposited as lipids and carbohydrates.

Light driven CO2 fixation by using cyanobacterial photosystem I and NADPH-dependent formate dehydrogenase.

The ultimate goal of this research is to construct a new direct CO2 fixation system using photosystems in living algae. Here, we report light-driven formate production from CO2 by using cyanobacterial photosystem I (PS I). Formate, a chemical hydrogen carrier and important industrial material, can be produced from CO2 by using the reducing power and the catalytic function of formate dehydrogenase (FDH). We created a bacterial FDH mutant that experimentally switched the cofactor specificity from NADH to NADPH, and combined it with an in vitro-reconstituted cyanobacterial light-driven NADPH production system consisting of PS I, ferredoxin (Fd), and ferredoxin-NADP(+)-reductase (FNR). Consequently, light-dependent formate production under a CO2 atmosphere was successfully achieved. In addition, we introduced the NADPH-dependent FDH mutant into heterocysts of the cyanobacterium Anabaena sp. PCC 7120 and demonstrated an increased formate concentration in the cells. These results provide a new possibility for photo-biological CO2 fixation.

Phosphorylation of BLUS1 kinase by phototropins is a primary step in stomatal opening.

Opening of stomata in the plant facilitates photosynthetic CO2 fixation and transpiration. Blue-light perception by phototropins (phot1, phot2) activates the plasma membrane H(+)-ATPase, causing stomata to open. Here we describe a regulator that connects these components, a Ser/Thr protein kinase, BLUS1 (BLUE LIGHT SIGNALING1), which mediates a primary step for phototropin signalling in guard cells. blus1 mutants identified by infrared thermography result in a loss of blue light-dependent stomatal opening. BLUS1 encodes a protein kinase that is directly phosphorylated by phot1 in vitro and in vivo at Ser-348 within its C-terminus. Both phosphorylation of Ser-348 and BLUS1 kinase activity are essential for activation of the H(+)-ATPase. blus1 mutants show lower stomatal conductance and CO2 assimilation than wild-type plants under decreased ambient CO2. Together, our analyses demonstrate that BLUS1 functions as a phototropin substrate and primary regulator of stomatal control to enhance photosynthetic CO2 assimilation under natural light conditions.

Chlorophyll fluorescence kinetics, photosynthetic activity, and pigment composition of blue-shade and half-shade leaves as compared to sun and shade leaves of different trees.

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

Mutate Chlorella sp. by nuclear irradiation to fix high concentrations of CO2.

To improve biomass productivity and CO2 fixation of microalgae under 15% (v/v) CO2 of flue gas, Chlorella species were mutated by nuclear irradiation and domesticated with high concentrations of CO2. The biomass yield of Chlorella pyrenoidosa mutated using 500 Gy of (60)Co γ irradiation increased by 53.1% (to 1.12 g L(-1)) under air bubbling. The mutants were domesticated with gradually increased high concentrations of CO2 [from 0.038% (v/v) to 15% (v/v)], which increased the biomass yield to 2.41 g L(-1). When light transmission and culture mixing in photo-bioreactors were enhanced at 15% (v/v) CO2, the peak growth rate of the domesticated mutant (named Chlorella PY-ZU1) was increased to 0.68 g L(-1) d(-1). When the ratio of gas flow rate (L min(-1)) to 1L of microalgae culture was 0.011, the peak CO2 fixation rate and the efficiency of Chlorella PY-ZU1 were 1.54 g L(-1) d(-1) and 32.7%, respectively.