LIGHT‐INDUCED RESPONSES OF OXYGEN PHOTOREDUCTION,REACTIVE OXYGEN SPECIES PRODUCTION AND SCAVENGING IN TWO DIATOM SPECIES1

Diatoms are frequently exposed to high light (HL) levels, which can result in photoinhibition and damage to PSII. Many microalgae can photoreduce oxygen using the Mehler reaction driven by PSI, which could protect PSII. The ability of Nitzschia epithemioides Grunow and Thalassiosira pseudonana Hasle et Heimdal grown at 50 and 300 μmol photons · m^?2 · s^?1 to photoreduce oxygen was examined by mass spectrometric measurements of ¹⁸O₂. Both species exhibited significant rates of oxygen photoreduction at saturating light levels, with cells grown in HL exhibiting higher rates. HL‐grown T. pseudonana had maximum rates of oxygen photoreduction five times greater than N. epithemoides, with 49% of electrons transported through PSII being used to reduce oxygen. Exposure to excess light (1,000 μmol photons · m^?2 · s^?1) produced similar decreases in the operating quantum efficiency of PSII (F_q′/F_m′) of low light (LL)‐ and HL‐grown N. epithemoides, whereas HL‐grown T. pseudonana exhibited much smaller decreases in F_q′/F_m′ than LL‐grown cells. HL‐grown T. pseudonana and N. epithemioides exhibited greater superoxide and hydrogen peroxide production, higher activities (in T. pseudonana) of superoxide dismutase (SOD) and ascorbate peroxidase (APX), and increased expression of three SOD‐ and one APX‐encoding genes after 60 min of excess light compared to LL‐grown cells. These responses provide a mechanism that contributes to the photoprotection of PSII against photodamage.     

Scavenging of reactive oxygen species by N‐substituted indole‐2 and 3‐carboxamides

Indole‐2 and 3‐carboxamides (IDs) are proposed to be selective cyclooxygenase inhibitors. Since cyclooxygenase‐1 may be involved in reactive oxygen species (ROS) production, we hypothesize that these indole derivatives have antioxidative properties. We have employed chemiluminescence (CL) and electron spin resonance (ESR) spin trapping to examine this hypothesis. We report here the results of a study of reactivity of 10 selected indole derivatives towards ROS. The following generators of ROS were applied: potassium superoxide (KO₂) as a source of superoxide radicals (O₂^·?), the Fenton reaction (Co‐EDTA/H₂O₂) for hydroxyl radicals (HO^·), and a mixture of alkaline aqueous H₂O₂ and acetonitrile for singlet oxygen (¹O₂). Hydroxyl radicals were detected as 5,5‐dimethyl‐1‐pyrroline‐N‐oxide (DMPO) spin adduct, whereas 2,2,6,6‐tetramethyl‐piperidine (TEMP) was used as a detector of ¹O₂. Using the Fenton reaction, 0.5 mmol/L IDs were found to inhibit DMPO‐?H radical formation in the range 7–37%. Furthermore the tested compounds containing the thiazolyl group also inhibited the ¹O₂‐dependent TEMPO radical, generated in the acetonitrile + H₂O₂ system. About 20% inhibition was obtained in the presence of 0.5 mmol/L IDs. 1 mmol/L IDs caused an approximately 13–70% decrease in the CL sum from the O₂^·? generating system (1 mmol/L). The aim of this paper is to evaluate these indole derivatives as antioxidants and their abilities to scavenge ROS. Copyright © 2004 John Wiley & Sons, Ltd.     

Does growth under elevated CO2 moderate photoacclimation in rice?

Acclimation of plant photosynthesis to light irradiance (photoacclimation) involves adjustments in levels of pigments and proteins and larger scale changes in leaf morphology. To investigate the impact of rising atmospheric CO₂ on crop physiology, we hypothesize that elevated CO₂ interacts with photoacclimation in rice (Oryza sativa). Rice was grown under high light (HL: 700 µmol m^?2 s^?1), low light (LL: 200 µmol m^?2 s^?1), ambient CO₂ (400 µl l^?1) and elevated CO₂ (1000 µl l^?1). Leaf six was measured throughout. Obscuring meristem tissue during development did not alter leaf thickness indicating that mature leaves are responsible for sensing light during photoacclimation. Elevated CO₂ raised growth chamber photosynthesis and increased tiller formation at both light levels, while it increased leaf length under LL but not under HL. Elevated CO₂ always resulted in increased leaf growth rate and tiller production. Changes in leaf thickness, leaf area, Rubisco content, stem and leaf starch, sucrose and fructose content were all dominated by irradiance and unaffected by CO₂. However, stomata responded differently; they were significantly smaller in LL grown plants compared to HL but this effect was significantly suppressed under elevated CO₂. Stomatal density was lower under LL, but this required elevated CO₂ and the magnitude was adaxial or abaxial surface‐dependent. We conclude that photoacclimation in rice involves a systemic signal. Furthermore, extra carbohydrate produced under elevated CO₂ is utilized in enhancing leaf and tiller growth and does not enhance or inhibit any feature of photoacclimation with the exception of stomatal morphology.     

Influence of knockout of <Emphasis Type="Italic">At4g20990</Emphasis> gene encoding α-CA4 on photosystem II light-harvesting antenna in plants grown under different light intensities and day lengths

Effect of knockout of the At4g20990 gene encoding α-carbonic anhydrase 4 (α-CA4) in Arabidopsis thaliana in plants grown in low light (LL, 80 μmol photons m^?2 s^?1) or in high light (HL, 400 μmol photons m^?2 s^?1) under long (LD, 16 h) or short (SD, 8 h) day length was studied. In α-CA4 knockout plants, under all studied conditions, the non-photochemical quenching was lower; the decrease was more pronounced under HL. This pointed to α-CA4 implication in the processes leading to energy dissipation in PSII antenna. In this context the content of major antenna proteins Lhcb1 and Lhcb2 was lower in α-CA4 knockouts than in wild-type (WT) plants under all growth conditions. The expression level of lhcb2 gene was also lower in mutants grown under LD, LL and HL in comparison to WT. At the same time, this level was higher in mutants grown under SD, LL and it was the same under SD, HL. Overall, the data showed that the knockout of the At4g20990 gene affected both the contents of proteins of PSII light-harvesting complex and the expression level of genes encoding these proteins, with peculiarities dependent on day length. These data together with the fact of a decrease of non-photochemical quenching of leaf chlorophyll a fluorescence in α-CA4-mut as compared with that in WT plants implied that α-CA4 participates in acclimation of photosynthetic apparatus to light intensity, possibly playing important role in the photoprotection. The role of this CA can be especially important in plants growing under high illumination conditions.     

Early burst of reactive oxygen species positively regulates resistance of eggplant against bacterial wilt

Reactive oxygen species (ROS) play a crucial role in the early response to plant biotic and abiotic stresses. In this study, bacterial wilt‐resistant and wilt‐susceptible eggplants were inoculated with Ralstonia solanacearum and the ROS content was analysed. The result revealed an increased accumulation of hydrogen peroxide (H₂O₂) and superoxide (O₂^?) in resistant and susceptible eggplant roots after R. solanacearum inoculation. H₂O₂ and O₂^? accumulation increased earlier in the inoculated resistant eggplant root than in the inoculated susceptible eggplant root. Real‐time polymerase chain reaction results revealed that respiratory burst oxidase homologue (Rboh) A, RbohB, RbohF and PR1 expression levels increased in inoculated resistant eggplant roots at an early stage (0–60 h postinoculation) and were at higher expression levels than those in susceptible eggplant roots. Ascorbate peroxidase, peroxidase and catalase activities were higher in inoculated resistant eggplant roots than in susceptible eggplant roots at the early stage. Hence, an early ROS burst positively regulates bacterial wilt resistance in eggplant.     

NADPH oxidase is involved in H2O2-induced differentiation of human promyelocytic leukaemia HL-60 cells

The expression and activity of NADPH oxidase increase when HL‐60 cells are induced into terminally differentiated cells. However, the function of NADPH oxidase in differentiation is not well elucidated. With 150–500 μM H₂O₂ inducing differentiation of HL‐60 cells, we measured phagocytosis of latex beads and investigated cell electrophoresis. Two inhibitors of NADPH oxidase, DPI (diphenyleneiodonium) and APO (apocynin), blocked the differentiation potential of cells induced by 200 μM H₂O₂. However, H₂O₂ stimulated the generation of intracellular superoxide (O₂ ^{? ?}), which decreased in the presence of the two inhibitors. DPI also inhibited H₂O₂‐induced ERK (extracellular‐signal‐regulated kinase) activation, as detected by Western blotting. Furthermore, PD98059, the inhibitor of the ERK pathway, inhibited the differentiation of HL‐60 cells induced by H₂O₂. This shows that H₂O₂ can activate NADPH oxidase, leading to O₂ ^{? ?} production, followed by ERK activation and ultimately resulting in the differentiation of HL‐60 cells. The data indicate that NADPH oxidase is an important cell signal regulating cell differentiation.     

Can the progressive increase of C4 bundle sheath leakiness at low PFD be explained by incomplete suppression of photorespiration?

The ability to concentrate CO₂ around Rubisco allows C₄ crops to suppress photorespiration. However, as phosphoenolpyruvate regeneration requires ATP, the energetic efficiency of the C₄ pathway at low photosynthetic flux densities (PFD) becomes a balancing act between primary fixation and concentration of CO₂ in mesophyll (M) cells, and CO₂ reduction in bundle sheath (BS) cells. At low PFD, retro‐diffusion of CO₂ from BS cells, relative to the rate of bicarbonate fixation in M cells (termed leakiness φ), is known to increase. This paper investigates whether this increase in ? could be explained by incomplete inhibition of photorespiration. The PFD response of φ was measured at various O₂ partial pressures in young Zea mays plants grown at 250 (LL) and 750 µmol m^?2 s^?1 PFD (HL). φ increased at low PFD and was positively correlated with O₂ partial pressure. Low PFD during growth caused BS conductance and interveinal distance to be lower in the LL plants, compared to the HL plants, which correlated with lower φ. Model analysis showed that incomplete inhibition of photorespiration, especially in the HL plants, and an increase in the relative contribution of mitochondrial respiration at low PFD could explain the observed increases in φ.     

AUTOTROPHIC GROWTH AND PHOTOACCLIMATION IN KARLODINIUM MICRUM (DINOPHYCEAE) AND STOREATULA MAJOR (CRYPTOPHYCEAE)1

We compared autotrophic growth of the dinoflagellate Karlodinium micrum (Leadbeater et Dodge) and the cryptophyte Storeatula major (Butcher ex Hill) at a range of growth irradiances (E_g). Our goal was to determine the physiological bases for differences in growth–irradiance relationships between these species. Maximum autotrophic growth rates of K. micrum and S. major were 0.5 and 1.5 div.·d^?1, respectively. Growth rates were positively correlated with C‐specific photosynthetic performance (PP^C, g C·g C^?1·h^?1) (r²=0.72). Cultures were grouped as light‐limited (LL) and high‐light (HL) treatments to allow interspecific comparisons of physiological properties that underlie the growth–irradiance relationships. Interspecific differences in the C‐specific light absorption rate (E_a^C, mol photons·g C^?1·h^?1) were observed only among HL acclimated cultures, and the realized quantum yield of C fixation (φ_C(real.), mol C·mol photons^?1) did not differ significantly between species in either LL or HL treatments. The proportion of fixed C that was incorporated into new biomass was lower in K. micrum than S. major at each E_g, reflecting lower growth efficiency in K. micrum. Photoacclimation to HL in K. micrum involved a significant loss of cellular photosynthetic capacity (P_max^cell), whereas in S. major, P_max^cell was significantly higher in HL acclimated cells. We conclude that growth rate differences between K. micrum and S. major under LL conditions relate primarily to cell metabolism processes (i.e. growth efficiency) and that reduced chloroplast function, reflected in PP^C and photosynthesis–irradiance curve acclimation in K. micrum, is also important under HL conditions.     

Effects of Exogenous γ-Aminobutyric Acid (GABA) on Photosynthesis and Antioxidant System in Pepper (Capsicum annuum L.) Seedlings Under Low Light Stress

The effects of γ-aminobutyric acid (GABA) treatment on parameters of photosynthesis and antioxidant defense system were measured in pepper (Capsicum annuum L.) leaves under low-light (LL) stress. Seedlings exposed to LL stress showed increased chlorophyll content as well as decreased net photosynthetic rate (P _n), stomatal conductance (g _s), maximum quantum yield of PSII (F _v/F _m), actual PSII photochemical efficiency (Φ_PSII), electron transport rates and photochemical quenching coefficient (q _p). However, almost all the photosynthetic parameters above were enhanced markedly in seedlings treated with GABA under LL stress. Moreover, LL stress increased malondialdehyde (MDA) content, superoxide anion radical (O₂ ^·?) and hydrogen peroxide (H₂O₂) production. GABA-treated, LL-stressed seedlings exhibited lower MDA, O₂ ^·? and H₂O₂ production, and showed an activated antioxidant defense system, including increased activities of superoxide dismutase, catalase, ascorbate peroxidase, glutathione peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase, glutathione reductase, ascorbate and glutathione. Moreover, seedlings subjected to LL stress showed increased endogenous GABA levels, and the level was further improved by application of exogenous GABA. These results suggest that GABA mitigates the LL-induced stress via regulating the antioxidant defense system and maintaining a high level of photochemical efficiency in pepper seedlings.

     

Reactive oxygen species are involved in regulation of pollen wall cytomechanics

Production and scavenging of reactive oxygen species (ROS) in somatic plant cells is developmentally regulated and plays an important role in the modification of cell wall mechanical properties. Here we show that H₂O₂ and the hydroxyl radical (^?OH) can regulate germination of tobacco pollen by modifying the mechanical properties of the pollen intine (inner layer of the pollen wall). Pollen germination was affected by addition of exogenous H₂O₂, ^?OH, and by antioxidants scavenging endogenous ROS: superoxide dismutase, superoxide dismutase/catalase mimic Mn‐5,10,15,20‐tetrakis(1‐methyl‐4‐pyridyl)21H, 23H‐porphin, or a spin‐trap α‐(4‐pyridyl‐1‐oxide)‐N‐tert‐butylnitrone, which eliminates ^?OH. The inhibiting concentrations of exogenous H₂O₂ and ^?OH did not decrease pollen viability, but influenced the mechanical properties of the wall. The latter were estimated by studying the resistance of pollen to hypo‐osmotic shock. ^?OH caused excess loosening of the intine all over the surface of the pollen grain, disrupting polar growth induction. In contrast, H₂O₂, as well as partial removal of endogenous ^?OH, over‐tightened the wall, impeding pollen tube emergence. Feruloyl esterase (FAE) was used as a tool to examine whether H₂O₂‐inducible inter‐polymer cross‐linking is involved in the intine tightening. FAE treatment caused loosening of the intine and stimulated pollen germination and pollen tube growth, revealing ferulate cross‐links in the intine. Taken together, the data suggest that pollen intine properties can be regulated differentially by ROS. ^?OH is involved in local loosening of the intine in the germination pore region, while H₂O₂ is necessary for intine strengthening in the rest of the wall through oxidative coupling of feruloyl polysaccharides.     

Reactive oxygen species (ROS) reduce the expression of BRAK/CXCL14 in human head and neck squamous cell carcinoma cells

Abstract

The present study investigated the effects of oxidative stress induced by reactive oxygen species (ROS), such as hydrogen peroxide (H₂O₂) and hydroxyl radical (HO^?), on the expression of both BRAK , which is also known as non-ELR motif angiostatic CXC chemokine ligand 14 (CXCL14), in head and neck squamous cell carcinoma (HNSCC) cells. When HNSCC cells were cultured in the presence of ROS, the expression of BRAK was significantly decreased whereas that of IL-8 was increased. Interestingly, the effects on the expression of both genes in HNSCC cells were much greater with HO^? than with H₂O₂. The effects of ROS on both BRAK and IL-8 expression were attenuated by pre-treatment with N-acetyl-L-cysteine (NAC), epidermal growth factor receptor (EGFR), and mitogen-activated protein kinase (MAPK) inhibitors. These results indicate that oxidative stress induced by H₂O₂ or HO^? stimulates angiogenesis and tumuor progression by altering the gene expression of BRAK and IL-8 via the EGFR/MEK/ERK pathway in human HNSCC cells.     

Cross-talk between salicylic acid and NaCl-generated reactive oxygen species and nitric oxide in tomato during acclimation to high salinity

Hydrogen peroxide (H₂O₂) and nitric oxide (NO) generated by salicylic acid (SA) are considered to be functional links of cross‐tolerance to various stressors. SA‐stimulated pre‐adaptation state was beneficial in the acclimation to subsequent salt stress in tomato (Solanum lycopersicum cv. Rio Fuego). At the whole‐plant level, SA‐induced massive H₂O₂ accumulation only at high concentrations (10^?3–10^?2M), which later caused the death of plants. The excess accumulation of H₂O₂ as compared with plants exposed to 100 mM NaCl was not associated with salt stress response after SA pre‐treatments. In the root tips, 10^?3–10^?2M SA triggered the production of reactive oxygen species (ROS) and NO with a concomitant decline in the cell viability. Sublethal concentrations of SA, however, decreased the effect of salt stress on ROS and NO production in the root apex. The attenuation of oxidative stress because of high salinity occurred not only in pre‐adapted plants but also at cell level. When protoplasts prepared from control leaves were exposed to SA in the presence of 100 mM NaCl, the production of NO and ROS was much lower and the viability of the cells was higher than in salt‐treated samples. This suggests that, the cross‐talk of signalling pathways induced by SA and high salinity may occur at the level of ROS and NO production. Abscisic acid (ABA), polyamines and 1‐aminocyclopropane‐1‐carboxylic acid, the compounds accumulating in pre‐treated plants, enhanced the diphenylene iodonium‐sensitive ROS and NO levels, but, in contrast to others, ABA and putrescine preserved the viability of protoplasts.     

Acclimation to low light by C4 maize: implications for bundle sheath leakiness

C4 plants have a biochemical carbon concentrating mechanism (CCM) that increases CO₂ concentration around ribulose bisphosphate carboxylase oxygenase (Rubisco) in the bundle sheath (BS). Under limiting light, the activity of the CCM generally decreases, causing an increase in leakiness, (Φ), the ratio of CO₂ retrodiffusing from the BS relative to C4 carboxylation processes. Maize plants were grown under high and low light regimes (respectively HL, 600 versus LL, 100 μE m^?2 s^?1). Short‐term acclimation of Φ was compared from isotopic discrimination (Δ), gas exchange and photochemistry. Direct measurement of respiration in the light, and ATP production rate (J_ATP), allowed us use a novel approach to derive Φ, compared with the conventional fitting of measured and predicted Δ. HL grown plants responded to decreasing light intensities with the well‐documented increase in Φ. Conversely, LL plants showed a constant Φ, which has not been observed previously. We explain the pattern by two contrasting acclimation strategies: HL plants maintained a high CCM activity at LL, resulting in high CO₂ overcycling and increased Φ; LL plants acclimated by down‐regulating the CCM, effectively optimizing scarce ATP supply. This surprising plasticity may limit the impact of Φ‐dependent carbon losses in leaves becoming shaded within developing canopies.     

Multi-mechanisms are involved in reactive oxygen species regulation of mTORC1 signaling

The mammalian target of rapamycin complex 1(mTORC1) integrates diverse signals to control cell growth, proliferation, survival, and metabolism. Role of reactive oxygen species (ROS) on mTORC1 signaling remains obscure and mechanisms through which ROS modulate mTORC1 are not known. We demonstrate that low doses ROS exposure stimulate mTORC1 while high concentrations or long-term ROS treatment decrease mTORC1 activity in vivo and in a variety of cell lines. The dose/time needed for inhibition or activation are cell type-dependent. In HEK293 cells hydrogen peroxide (H₂O₂) stimulates phosphorylation of AMP-activated kinase (AMPK) (T172) and Raptor (S792), enhances association of activated AMPK with Raptor. Furthermore, AMPK inhibitor compound c inhibits H₂O₂-induced Raptor (S792) phosphorylation and reverses H₂O₂-induced de-phosphorylation of mTORC1 downstream targets p70-S6K1 (T389), S6 (S235/236) and 4E-BP1 (T37/46). H₂O₂ also stimulates association of endogenous protein phosphatase 2A catalytic subunit (PP2Ac) with p70-S6K1. Like compound c, inhibitor of PP2A, okadaic acid partially reverses inactivation of mTORC1 substrates induced by H₂O₂. Moreover, inhibition of PP2A and AMPK partially rescued cells from H₂O₂-induced cell death. High doses of H₂O₂ inhibit while low doses of H₂O₂ activate mTORC1 both in TSC2^?/? P53^?/? and TSC2^+/+ P53^?/? MEFs. These data suggest that PP2A and AMPK-mediated phosphorylation of Raptor mediate H₂O₂-induced inhibition of mTORC1 signaling.     

Hydrogen peroxide is the major oxidant product of xanthine oxidase

Xanthine oxidase (XO) is a critical source of reactive oxygen species (ROS) in inflammatory disease. Focus, however, has centered almost exclusively on XO-derived superoxide (O₂^??), whereas direct H₂O₂ production from XO has been less well investigated. Therefore, we examined the relative quantities of O₂^?? and H₂O₂ produced by XO under a range (1–21%) of O₂ tensions. At O₂ concentrations between 10 and 21%, H₂O₂ accounted for ～75% of ROS production. As O₂ concentrations were lowered, there was a concentration-dependent increase in H₂O₂ formation, accounting for 90% of ROS production at 1% O₂. Alterations in pH between 5.5 and 7.4 did not affect the relative proportions of H₂O₂ and O₂^?? formation. Immobilization of XO, by binding to heparin–Sepharose, further enhanced relative H₂O₂ production by ～30%, under both normoxic and hypoxic conditions. Furthermore, XO bound to glycosaminoglycans on the apical surface of bovine aortic endothelial cells demonstrated a similar ROS production profile. These data establish H₂O₂ as the dominant (70–95%) reactive product produced by XO under clinically relevant conditions and emphasize the importance of H₂O₂ as a critical factor when examining the contributory roles of XO-catalyzed ROS in inflammatory processes as well as cellular signaling.     

Cadmium-induced subcellular accumulation of O2·− and H2O2 in pea leaves

Cadmium is a toxic metal that produces disturbances in plant antioxidant defences giving rise to oxidative stress. The effect of this metal on H₂O₂ and O₂^·? production was studied in leaves from pea plants growth for 2 weeks with 50 µm Cd, by histochemistry with diaminobenzidine (DAB) and nitroblue tetrazolium (NBT), respectively. The subcellular localization of these reactive oxygen species (ROS) was studied by cytochemistry with CeCl₃ and Mn/DAB staining for H₂O₂ and O₂^·?, respectively, followed by electron microscopy observation. In leaves from pea plants grown with 50 µm CdCl₂ a rise of six times in the H₂O₂ content took place in comparison with control plants, and the accumulation of H₂O₂ was observed mainly in the plasma membrane of transfer, mesophyll and epidermal cells, as well as in the tonoplast of bundle sheath cells. In mesophyll cells a small accumulation of H₂O₂ was observed in mitochondria and peroxisomes. Experiments with inhibitors suggested that the main source of H₂O₂ could be a NADPH oxidase. The subcellular localization of O₂^·? production was demonstrated in the tonoplast of bundle sheath cells, and plasma membrane from mesophyll cells. The Cd‐induced production of the ROS, H₂O₂ and O₂^·?, could be attributed to the phytotoxic effect of Cd, but lower levels of ROS could function as signal molecules in the induction of defence genes against Cd toxicity. Treatment of leaves from Cd‐grown plants with different effectors and inhibitors showed that ROS production was regulated by different processes involving protein phosphatases, Ca²⁺ channels, and cGMP.     

Biphasic regulation of H2O2 on angiogenesis implicated NADPH oxidase

ROS (reactive oxygen species) take an important signalling role in angiogenesis. Although there are several ways to produce ROS in cells, multicomponent non‐phagocytic NADPH oxidase is an important source of ROS that contribute to angiogenesis. In the present work, we examined the effects of H₂O₂ on angiogenesis including proliferation and migration in HUVECs (human umbilical vein endothelial cells), new vessel formation in chicken embryo CAM (chorioallantoic membrane) and endothelial cell apoptosis, which is closely related to anti‐angiogenesis. Our results showed that H₂O₂ dose‐dependently increased the generation of O₂ ^? (superoxide anion) in HUVECs, which was suppressed by DPI (diphenylene iodonium) and APO (apocynin), two inhibitors of NADPH oxidase. H₂O₂ at low concentrations (10 µM) stimulated cell proliferation and migration, but at higher concentrations, inhibited both. Similarly, H₂O₂ at 4 nmol/cm² strongly induced new vessel formation in CAM, while it suppressed at high concentrations (higher than 4 nmol/cm²). Also, H₂O₂ (200～500 µM) could stimulate apoptosis in HUVECs. All the effects of H₂O₂ on angiogenesis could be suppressed by NADPH oxidase inhibitors, which suggests that NADPH oxidase acts downstream of H₂O₂ to produce O₂ ^? and then to regulate angiogenesis. In summary, our results suggest that H₂O₂ as well as O₂ ^? mediated by NADPH oxidase have biphasic effects on angiogenesis in vitro and in vivo.     

Protective effects of complementary Ca2+ on low-light-induced oxidative damage in tall fescue

Low-light (LL) intensity is a primary abiotic stressor that negatively influences turf grass quality. In the present experiment, we studied the effect of exogenous Ca²⁺ (0, 10, 50, 100, and 200 mM) on the antioxidant system, the accumulation of MDA and proline, the content of photosynthetic pigments in plant leaves in order to investigate whether exogenous Ca²⁺ treatment improves LL tolerance in tall fescue (Festuca arundinacea Schreb.). We have found that LL significantly reduced a number of growth parameters (plant height, leaf width, leaf fresh weight, root fresh weight, leaf dry weight, and root dry weight), chlorophyll (Chl) a and Chl b contents, and carotenoid (Car) levels, while considerably enhancing electrolyte leakage (EL), MDA accumulation, calcium (Ca²⁺) concentration, and generation of reactive oxygen species (ROS), such as hydrogen peroxide (H₂O₂) and superoxide radical (O ₂ ^·? ). Moreover, LL significantly induced the activities of antioxidant enzymes, such as peroxidase (POD) and catalase (CAT), and slightly increased the activity of superoxide dismutase (SOD) in tall fescue leaves. In contrast, POD and SOD activities declined considerably while CAT activity significantly increased in plant roots under LL stress. The application of 50 mM Ca²⁺ significantly improved the aforementioned growth parameters, the content of photosynthetic pigments, and further enhanced the activities of POD, SOD, and CAT, but decreased electrolyte leakage and MDA and H₂O₂ levels in the leaves and roots of tall fescue under LL stress. These results suggest that Ca²⁺ is likely involved in a resistance to LL by regulating antioxidant enzyme action in tall fescue leaves and roots.     

Impact of irradiance on the C allocation in the coastal marine diatom Skeletonema marinoi Sarno and Zingone

Elemental stoichiometry and organic composition were investigated in an Adriatic strain of Skeletonema marinoi, cultured at 25 [low light (LL)] and 250 [high light (HL)]µmol photon m^?2 s^?1. Inorganic carbon acquisition, fixation and allocation, and silicic acid and orthophosphate uptake were also studied. The C : P ratio was below the Redfield ratio, especially at LL. In HL cells, N quota was halved, C quota was similar, silica quota was lower, growth rate and long‐term net primary productivity were almost doubled, relative to LL cells. The HL : LL cell quota ratios were 6 for lipid, 0.5 for protein and 0.4 for carbohydrate. Phosphoenolpyruvate carboxylase (PEPc) and glutamine synthetase (GS) activities were unaffected by the growth irradiance; phosphoenolpyruvate carboxykinase (PEPck) was 2.5‐fold more active in LL cells. This suggests that in S. marinoi, C₄ photosynthesis is unlikely, PEPc is anaplerotic and PEPck may be involved in the conversion of lipid C to carbohydrates, especially in LL cells. Because about 50% of the cost for the production of an HL cell is caused by lipid biosynthesis, we propose that the preferential allocation of C to lipid at HL takes advantage of the relatively high volume‐based energy content of lipids, in an organism that reduces its size at each vegetative cell division.