Lethal DNA Lesions Caused by Direct and Indirect Actions of X rays are Repaired via Different DSB Repair Pathways under Aerobic and Anoxic Conditions.

We examined lethal damages of X rays induced by direct and indirect actions, in terms of double-strand break (DSB) repair susceptibility using two kinds of repair-deficient Chinese hamster ovary (CHO) cell lines. These CHO mutants (51D1 and xrs6) are genetically deficient in one of the two important DNA repair pathways after genotoxic injury [homologous recombination (HR) and non-homologous end binding (NHEJ) pathways, respectively]. The contribution of indirect action on cell killing can be estimated by applying the maximum level of dimethylsulfoxide (DMSO) to get rid of OH radicals. To control the proportion of direct and indirect actions in lethal damage, we irradiated CHO mutant cells under aerobic and anoxic conditions. The contributions of indirect action on HR-defective 51D1 cells were 76% and 57% under aerobic and anoxic conditions, respectively. Interestingly, these percentages were similar to those of the wild-type cells even if the radiosensitivity was different. However, the contributions of indirect action to cell killing on NHEJ-defective xrs6 cells were 52% and 33% under aerobic and anoxic conditions, respectively. Cell killing by indirect action was significantly affected by the oxygen concentration and the DSB repair pathways but was not correlated with radiosensitivity. These results suggest that the lethal damage induced by direct action is mostly repaired by NHEJ repair pathway since killing of NHEJ-defective cells has significantly higher contribution by the direct action. In other words, the HR repair pathway may not effectively repair the DSB by direct action in place of the NHEJ repair pathway. We conclude that the type of DSB produced by direct action is different from that of DSB induced by indirect action.

Symbiosis extended: exchange of photosynthetic O2 and fungal-respired CO2 mutually power metabolism of lichen symbionts.

Lichens are a symbiosis between a fungus and one or more photosynthetic microorganisms that enables the symbionts to thrive in places and conditions they could not compete independently. Exchanges of water and sugars between the symbionts are the established mechanisms that support lichen symbiosis. Herein, we present a new linkage between algal photosynthesis and fungal respiration in lichen Flavoparmelia caperata that extends the physiological nature of symbiotic co-dependent metabolisms, mutually boosting energy conversion rates in both symbionts. Measurements of electron transport by oximetry show that photosynthetic O2 is consumed internally by fungal respiration. At low light intensity, very low levels of O2 are released, while photosynthetic electron transport from water oxidation is normal as shown by intrinsic chlorophyll variable fluorescence yield (period-4 oscillations in flash-induced Fv/Fm). The rate of algal O2 production increases following consecutive series of illumination periods, at low and with limited saturation at high light intensities, in contrast to light saturation in free-living algae. We attribute this effect to arise from the availability of more CO2 produced by fungal respiration of photosynthetically generated sugars. We conclude that the lichen symbionts are metabolically coupled by energy conversion through exchange of terminal electron donors and acceptors used in both photosynthesis and fungal respiration. Algal sugars and O2 are consumed by the fungal symbiont, while fungal delivered CO2 is consumed by the alga.

Unravelling the effects of blue light on aerobic methane emissions from canola.

It is now well documented that plants produce methane (CH4) under aerobic conditions. However, the nature of methane production in plants and all the potential precursors and environmental factors that can be involved in the process are not fully understood. Earlier studies have suggested several chemical compounds, including the amino acid methionine, as precursors of aerobic methane in plants, but none have explored other amino acids as potential precursors or blue light as a driving force of methane emission. We examined the effects of blue light, and the promoter or inhibitor of endogenous ethylene on methane and ethylene emissions, amino acids, and some plant physiological parameters in canola (Brassica napus). Plants were grown under four light conditions: no supplemental blue light, and low, medium, or high blue light, and exposed to three chemical treatments: no chemical application, ethylene promoter (kinetin), or ethylene inhibitor (silver nitrate). Regardless of chemical treatment, blue light significantly increased methane emission, which was accompanied by decreased plant biomass, gas exchange, and flavonoids, but by increased wax, and most amino acids. This study revealed that blue light drives aerobic methane emission from plants by releasing of methyl group from a number of amino acids, and that the methane production in plants may have several pathways.

Integrated aerobic biological-chemical treatment of winery wastewater diluted with urban wastewater. LED-based photocatalysis in the presence of monoperoxysulfate.

The oxidation of Winery Wastewater (WW) by conventional aerobic biological treatment usually leads to inefficient results due to the presence of organic substances, which are recalcitrant or toxic in conventional procedures. This study explores the combination of biological and chemical processes in order to complete the oxidation of biodegradable and non-biodegradable compounds in two sequential steps. Thus, a biological oxidation of a diluted WW is carried out by using the activated sludge process. Activated sludge was gradually acclimated to the Diluted Winery Wastewater (DWW). Some aspects concerning the biological process were evaluated (kinetics of the oxidation and sedimentation of the sludge produced). The biological treatment of the DWW led to a 40-50% of Chemical Oxygen Demand (COD) removal in 8 h, being necessary the application of an additional process. Different chemical processes combining UVA-LEDs radiation, monoperoxysulfate (MPS) and photocatalysts were applied in order to complete the COD depletion and efficient removal of polyphenols content, poorly oxidized in the previous biological step. From the options tested, the combination of UVA, MPS and a novel LaCoO3-TiO2 composite, with double route of MPS decomposition through heterogeneous catalysis and photocatalysis, led to the best results (95% of polyphenol degradation, and additional 60% of COD removal). Initial MPS concentration and pH effect in this process were assessed.

Novel flat-plate photobioreactors for microalgae cultivation with special mixers to promote mixing along the light gradient.

Novel flat-plate photobioreactors (PBRs) with special mixers (type-a, type-b, and type-c) were designed based on increased mixing degree along the light gradient. The hydrodynamic and light regime characteristic of the novel PBRs were investigated through computational fluid dynamics. Compared with the control reactor without mixer, the novel reactors can effectively increase liquid velocity along the light gradient, the frequency of light/dark (L/D) cycles, and the algal growth rates of Chlorella pyrenoidosa. The maximum biomass concentrations in type-a, type-b, and type-c reactors were 42.9% (1.3 g L(-1)), 31.9% (1.2 g L(-1)), and 20.9% (1.1 g L(-1)) higher than that in the control reactor (0.91 g L(-1)), respectively, at an aeration rate of 1.0 vvm. Correlation analysis of algal growth rate with the characteristics of mixing and light regime shows the key factors affecting algal photoautotrophic growth are liquid velocity along the light gradient and L/D cycles rather than the macro-mixing degree.

Effects of elevated [CO2] and low soil moisture on the physiological responses of Mountain Maple (Acer spicatum L.) seedlings to light.

Global climate change is expected to affect how plants respond to their physical and biological environments. In this study, we examined the effects of elevated CO2 ([CO2]) and low soil moisture on the physiological responses of mountain maple (Acer spicatum L.) seedlings to light availability. The seedlings were grown at ambient (392 µmol mol(-1)) and elevated (784 µmol mol(-1)) [CO2], low and high soil moisture (M) regimes, at high light (100%) and low light (30%) in the greenhouse for one growing season. We measured net photosynthesis (A), stomatal conductance (g s), instantaneous water use efficiency (IWUE), maximum rate of carboxylation (V cmax), rate of photosynthetic electron transport (J), triose phosphate utilization (TPU)), leaf respiration (R d), light compensation point (LCP) and mid-day shoot water potential (Ψx). A and g s did not show significant responses to light treatment in seedlings grown at low soil moisture treatment, but the high light significantly decreased the C i/C a in those seedlings. IWUE was significantly higher in the elevated compared with the ambient [CO2], and the effect was greater at high than the low light treatment. LCP did not respond to the soil moisture treatments when seedlings were grown in high light under both [CO2]. The low soil moisture significantly reduced Ψx but had no significant effect on the responses of other physiological traits to light or [CO2]. These results suggest that as the atmospheric [CO2] rises, the physiological performance of mountain maple seedlings in high light environments may be enhanced, particularly when soil moisture conditions are favourable.

Electron beam inactivation of selected microbial pathogens and indicator organisms in aerobically and anaerobically digested sewage sludge.

Microbial pathogens in municipal sewage sludges need to be inactivated prior to environmental disposal. The efficacy of high energy (10 MeV) e-beam irradiation to inactivate a variety of selected microbial pathogens and indicator organisms in aerobically and anaerobically digested sewage sludge was evaluated. Both bacterial and viral pathogens and indicator organisms are susceptible to e-beam irradiation. However, as expected there was a significant difference in their respective e-beam irradiation sensitivity. Somatic coliphages, bacterial endospores and enteric viruses were more resistant compared to bacterial pathogens. The current US EPA mandated 10 kGy minimum dose was capable of achieving significant reduction of both bacterial and viral pathogens. Somatic coliphages can be used as a microbial indicator for monitoring e-beam processes in terms of pathogen inactivation in sewage sludges.

The field-dependence of the solid-state photo-CIDNP effect in two states of heliobacterial reaction centers.

The solid-state photo-CIDNP (photochemically induced dynamic nuclear polarization) effect is studied in photosynthetic reaction centers of Heliobacillus mobilis at different magnetic fields by (13)C MAS (magic-angle spinning) NMR spectroscopy. Two active states of heliobacterial reaction centers are probed: an anaerobic preparation of heliochromatophores ("Braunstoff", German for "brown substance") as well as a preparation of cells after exposure to oxygen ("Grünstoff", "green substance"). Braunstoff shows significant increase of enhanced absorptive (positive) signals toward lower magnetic fields, which is interpreted in terms of an enhanced differential relaxation (DR) mechanism. In Grünstoff, the signals remain emissive (negative) at two fields, confirming that the influence of the DR mechanism is comparably low.

Effects of light, food availability and temperature stress on the function of photosystem II and photosystem I of coral symbionts.

BACKGROUND: Reef corals are heterotrophic coelenterates that achieve high productivity through their photosynthetic dinoflagellate symbionts. Excessive seawater temperature destabilises this symbiosis and causes corals to "bleach," lowering their photosynthetic capacity. Bleaching poses a serious threat to the persistence of coral reefs on a global scale. Despite expanding research on the causes of bleaching, the mechanisms remain a subject of debate. METHODOLOGY/PRINCIPAL FINDINGS: This study determined how light and food availability modulate the effects of temperature stress on photosynthesis in two reef coral species. We quantified the activities of Photosystem II, Photosystem I and whole chain electron transport under combinations of normal and stressful growth temperatures, moderate and high light levels and the presence or absence of feeding of the coral hosts. Our results show that PS1 function is comparatively robust against temperature stress in both species, whereas PS2 and whole chain electron transport are susceptible to temperature stress. In the symbiotic dinoflagellates of Stylophora pistillata the contents of chlorophyll and major photosynthetic complexes were primarily affected by food availability. In Turbinaria reniformis growth temperature was the dominant influence on the contents of the photosynthetic complexes. In both species feeding the host significantly protected photosynthetic function from high temperature stress. CONCLUSIONS/SIGNIFICANCE: Our findings support the photoinhibition model of coral bleaching and demonstrate that PS1 is not a major site for thermal damage during bleaching events. Feeding mitigates bleaching in two scleractinian corals, so that reef responses to temperature stresses will likely be influenced by the coinciding availabilities of prey for the host.

The photosynthetic apparatus and photoinduced electron transfer in the aerobic phototrophic bacteria Roseicyclus mahoneyensis and Porphyrobacter meromictius.

Photosynthetic electron transfer has been examined in whole cells, isolated membranes and in partially purified reaction centers (RCs) of Roseicyclus mahoneyensis, strain ML6 and Porphyrobacter meromictius, strain ML31, two species of obligate aerobic anoxygenic phototrophic bacteria. Photochemical activity in strain ML31 was observed aerobically, but the photosynthetic apparatus was not functional under anaerobic conditions. In strain ML6 low levels of photochemistry were measured anaerobically, possibly due to incomplete reduction of the primary electron acceptor (Q(A)) prior to light excitation, however, electron transfer occurred optimally under low oxygen conditions. Photoinduced electron transfer involves a soluble cytochrome c in both strains, and an additional reaction center (RC)-bound cytochrome c in ML6. The redox properties of the primary electron donor (P) and Q(A) of ML31 are similar to those previously determined for other aerobic phototrophs, with midpoint redox potentials of +463 mV and -25 mV, respectively. Strain ML6 showed a very narrow range of ambient redox potentials appropriate for photosynthesis, with midpoint redox potentials of +415 mV for P and +94 mV for Q(A). Cytoplasm soluble and photosynthetic complex bound cytochromes were characterized in terms of apparent molecular mass. Fluorescence excitation spectra revealed that abundant carotenoids not intimately associated with the RC are not involved in photosynthetic energy conservation.

A heme oxygenase isoform is essential for aerobic growth in the cyanobacterium Synechocystis sp. PCC 6803: modes of differential operation of two isoforms/enzymes to adapt to low oxygen environments in cyanobacteria.

Heme oxygenase (HO) catalyzes the oxygen-dependent cleavage of heme to produce biliverdin IXα in phycobilin biosynthesis. In the genome of the cyanobacterium Synechocystis sp. PCC 6803 there are two genes, ho1 (sll1184) and ho2 (sll1875), encoding HO isoforms. Reverse transcription-PCR indicated that ho1 is constitutively expressed, and ho2 is induced under micro-oxic conditions. A mutant lacking ho1 (Δho1) failed to grow under aerobic conditions while it did grow at a significantly slower rate than the wild type under anaerobic (micro-oxic) conditions. When micro-oxically grown Δho1 was incubated under aerobic conditions, the cells underwent chlorosis with a significant decrease in phycocyanin accompanied by anomalous accumulation of protoporphyrin IX. These results suggested that HO1 is essential for aerobic growth as the sole HO and is dispensable under micro-oxic conditions. A mutant lacking ho2 (Δho2) grew under both aerobic and micro-oxic conditions like the wild type at low light intensity (50 µmol(photon) m⁻² s⁻¹). At higher light intensity (120 µmol(photon) m⁻² s⁻¹) the Δho2 mutant showed significant growth retardation under micro-oxic conditions. It is suggested that HO2 operates as a dominant HO under high light and micro-oxic environments and acts as an accessory HO at low light intensity. Constitutive expression of HO2 in a neutral site of the chromosome restored aerobic growth of Δho1, suggesting that HO2 has an activity high enough to substitute for HO1 under aerobic conditions. The differential operation of two isoforms/enzymes in cyanobacterial tetrapyrrole biosynthesis to adapt to low oxygen environments is discussed, including three other reactions.

The effect of lignin photodegradation on decomposability of Calamagrostis epigeios grass litter.

The common grass Calamagrostis epigeions produces a large amount of dead biomass, which remain above the soil surface for many months. In this study, we determined how exposure of dead biomass above the soil affects its subsequent decomposition in soil. Collected dead standing biomass was divided in two parts, the first one (initial litter) was stored in a dark, dry place. The other part was placed in litterbags in the field. The litterbags were located in soil, on the soil surface, or hanging in the air without contact with soil but exposed to the sun and rain. After 1 year of field exposure, litter mass loss and C and N content were measured, and changes in litter chemistry were explored using NMR and thermochemolysis-GC-MS. The potential decomposability of the litter was quantified by burying the litter from the litterbags and the initial litter in soil microcosms and measuring soil respiration. Soil respiration was greater with litter that had been hanging in air than with all other kinds of litter. These finding could not be explained by changes in litter mass or C:N ratio. NMR indicated a decrease in polysaccharides relative to lignin in litter that was buried in soil but not in litter that was placed on soil surface or that was hanging in the air. Thermochemolysis indicated that the syringyl units of the litter lignin were decomposed when the litter was exposed to light. We postulate that photochemical decay of lignin increase decomposability of dead standing biomass.

Energy starved Candidatus Pelagibacter ubique substitutes light-mediated ATP production for endogenous carbon respiration.

Previous studies have demonstrated that Candidatus Pelagibacter ubique, a member of the SAR11 clade, constitutively expresses proteorhodopsin (PR) proteins that can function as light-dependent proton pumps. However, exposure to light did not significantly improve the growth rate or final cell densities of SAR11 isolates in a wide range of conditions. Thus, the ecophysiological role of PR in SAR11 remained unresolved. We investigated a range of cellular properties and here show that light causes dramatic changes in physiology and gene expression in Cand. P. ubique cells that are starved for carbon, but provides little or no advantage during active growth on organic carbon substrates. During logarithmic growth there was no difference in oxygen consumption by cells in light versus dark. Energy starved cells respired endogenous carbon in the dark, becoming spheres that approached the minimum predicted size for cells, and produced abundant pili. In the light, energy starved cells maintained size, ATP content, and higher substrate transport rates, and differentially expressed nearly 10% of their genome. These findings show that PR is a vital adaptation that supports Cand. P. ubique metabolism during carbon starvation, a condition that is likely to occur in the extreme conditions of ocean environments.

Modeling the light- and redox-dependent interaction of PpsR/AppA in Rhodobacter sphaeroides.

Facultative photosynthetic bacteria switch their energy generation mechanism from respiration to photosynthesis depending on oxygen tension and light. Part of this transition is mediated by the aerobic transcriptional repressor PpsR. In Rhodobacter sphaeroides, the repressive action of PpsR is antagonized by the redox- and blue-light-sensitive flavoprotein AppA which results in a unique phenotype: the repression of photosynthesis genes at intermediate oxygen levels and high light intensity, which is believed to reduce the risk of photooxidative stress. To analyze the underlying mechanism we developed a simple mathematical model based on the AppA-dependent reduction of a disulfide bond in PpsR and the light-sensitive complex formation between the reduced forms of AppA and PpsR. A steady-state analysis shows that high light repression can indeed occur at intermediate oxygen levels if PpsR is reduced on a faster timescale than AppA and if the electron transfer from AppA to PpsR is effectively irreversible. The model further predicts that if AppA copy numbers exceed those of PpsR by at least a factor of two, the transition from aerobic to anaerobic growth mode can occur via a bistable regime. We provide necessary conditions for the emergence of bistability and discuss possible experimental verifications.

Global methane emission estimates from ultraviolet irradiation of terrestrial plant foliage.

SUMMARY: *Several studies have reported in situ methane (CH(4)) emissions from vegetation foliage, but there remains considerable debate about its significance as a global source. Here, we report a study that evaluates the role of ultraviolet (UV) radiation-driven CH(4) emissions from foliar pectin as a global CH(4) source. *We combine a relationship for spectrally weighted CH(4) production from pectin with a global UV irradiation climatology model, satellite-derived leaf area index (LAI) and air temperature data to estimate the potential global CH(4) emissions from vegetation foliage. *Our results suggest that global foliar CH(4) emissions from UV-irradiated pectin could account for 0.2-1.0 Tg yr(-1), of which 60% is from tropical latitudes, corresponding to < 0.2% of total CH(4) sources. *Our estimate is one to two orders of magnitude lower than previous estimates of global foliar CH(4) emissions. Recent studies have reported that pectin is not the only molecular source of UV-driven CH(4) emissions and that other environmental stresses may also generate CH(4). Consequently, further evaluation of such mechanisms of CH(4) generation is needed to confirm the contribution of foliage to the global CH(4) budget.

Photorespiratory 2-phosphoglycolate metabolism and photoreduction of O2 cooperate in high-light acclimation of Synechocystis sp. strain PCC 6803.

In cyanobacteria, photorespiratory 2-phosphoglycolate (2PG) metabolism is mediated by three different routes, including one route involving the glycine decarboxylase complex (Gcv). It has been suggested that, in addition to conversion of 2PG into non-toxic intermediates, this pathway is important for acclimation to high-light. The photoreduction of O(2) (Mehler reaction), which is mediated by two flavoproteins Flv1 and Flv3 in cyanobacteria, dissipates excess reductants under high-light by the four electron-reduction of oxygen to water. Single and double mutants defective in these processes were constructed to investigate the relation between photorespiratory 2PG-metabolism and the photoreduction of O(2) in the cyanobacterium Synechocystis sp. PCC 6803. The single mutants Deltaflv1, Deltaflv3, and DeltagcvT, as well as the double mutant Deltaflv1/DeltagcvT, were completely segregated but not the double mutant Deltaflv3/DeltagcvT, suggesting that the T-protein subunit of the Gcv (GcvT) and Flv3 proteins cooperate in an essential process. This assumption is supported by the following results: (1) The mutant Deltaflv3/DeltagcvT showed a considerable longer lag phase and sometimes bleached after shifts from slow (low light, air CO(2)) to rapid (standard light, 5% CO(2)) growing conditions. (2) Photoinhibition experiments indicated a decreased ability of the mutant Deltaflv3/DeltagcvT to cope with high-light. (3) Fluorescence measurements showed that the photosynthetic electron chain is reduced in this mutant. Our data suggest that the photorespiratory 2PG-metabolism and the photoreduction of O(2), particularly that catalyzed by Flv3, cooperate during acclimation to high-light stress in cyanobacteria.

Is the redox state of the ci heme of the cytochrome b6f complex dependent on the occupation and structure of the Qi site and vice versa?

Oxidoreductases of the cytochrome bc(1)/b(6)f family transfer electrons from a liposoluble quinol to a soluble acceptor protein and contribute to the formation of a transmembrane electrochemical potential. The crystal structure of cyt b(6)f has revealed the presence in the Q(i) site of an atypical c-type heme, heme c(i). Surprisingly, the protein does not provide any axial ligand to the iron of this heme, and its surrounding structure suggests it can be accessed by exogenous ligand. In this work we describe a mutagenesis approach aimed at characterizing the c(i) heme and its interaction with the Q(i) site environment. We engineered a mutant of Chlamydomonas reinhardtii in which Phe(40) from subunit IV was substituted by a tyrosine. This results in a dramatic slowing down of the reoxidation of the b hemes under single flash excitation, suggesting hindered accessibility of the heme to its quinone substrate. This modified accessibility likely originates from the ligation of the heme iron by the phenol(ate) side chain introduced by the mutation. Indeed, it also results in a marked downshift of the c(i) heme midpoint potential (from +100 mV to -200 mV at pH 7). Yet the overall turnover rate of the mutant cytochrome b(6)f complex under continuous illumination was found similar to the wild type one, both in vitro and in vivo. We propose that, in the mutant, a change in the ligation state of the heme upon its reduction could act as a redox switch that would control the accessibility of the substrate to the heme and trigger the catalysis.

Nonmicrobial aerobic methane emission from poplar shoot cultures under low-light conditions.

The aerobic formation of methane in plants has been reported previously, but has been questioned by a number of researchers. Recently, isotopic evidence demonstrated that ultraviolet irradiation and heating lead to photochemical or thermal aerobic methane formation mainly from plant pectin in the absence of microbial methane production. However, the origin of aerobic methane formation from plant material observed under low temperature and low-light/dark conditions is still unclear. Here we show that Grey poplar (Populus × canescens, syn. Populus tremula × Populus alba) plants derived from cell cultures under sterile conditions released 13C-labeled methane under low-light conditions after feeding the plants with 13CO2. Molecular biological analysis proved the absence of any microbial contamination with known methanogenic microorganisms and ruled out the possibility that methane emission from our poplar shoot cultures under aerobic low-light/dark and ambient temperature conditions could be of microbial origin. The CH4 release rates in our experiment were in the range of 0.16-0.7 ng g-1 DW h-1, adding evidence to the growing opinion that the quantitative role of aerobic methane emissions from plants in the global methane budget, at least from cold temperate or boreal regions, is only of minor importance.

Loss of the mitochondrial nucleoid protein, Abf2p, destabilizes repetitive DNA in the yeast mitochondrial genome.

Loss of Abf2p, an abundant mitochondrial nucleoid-associated protein, results in increased mitochondrial frameshifts and direct-repeat mediated deletions but has no effect on the rate of mitochondrial point mutations. The instability of repeated sequences in this strain may be linked to the loss of mitochondrial DNA in abf2-Delta strains.