Photoactivation of Oxygen-Evolving Enzyme in Dark-Grown Pine Cotyledons: Relationship between Assembly of Photosystem II Proteins and Integration of Manganese and Calcium

Dark-grown cotyledons of pine (Pinus thunbergit) did not exhibitO₂ evolution, but this capability was rapidly activated by illuminationfor a short period (photoactivation). To examine the biochemicalchanges which accompany the process of photoactivation in gymnosperms,a method enabling the preparation of highly active O₂-evolvingphotosystem II (PS II) membranes was applied to light-grown,dark-grown, and photoactivated cotyledons. PS II membranes preparedfrom light-grown cotyledons exhibited high O₂-evolving activity,and contained all the intrinsic proteins as well as the threeextrinsic proteins (32, 23 and 17 kDa) associated with PS II.These membranes were also found to contain 4.4 Mn and 0.83 Ca/PSII reaction center. PS II membranes from dark-grown cotyledonscontained all the intrinsic proteins, but preserved only 32kDa extrinsic protein, and zero Mn and 0.85 Ca/PS II reactioncenter. The two extrinsic proteins (23 and 17 kDa) absent inthe PS II membranes from dark-grown cotyledons were, however,present as mature forms in whole thylakoid membranes from thecorresponding sample. The PS II membranes isolated from photoactivatedcotyledons showed a high activity of O₂ evolution and retainedthe three extrinsic proteins, 5.3 Mn and 1.1 Ca/PS II reactioncenter, respectively. The results indicated that Mn and thetwo extrinsic proteins were tightly integrated in the O₂-evolvingapparatusduring the process of photoactivation but integration of Capreceded the integration of Mn by photoactivation. (Received December 9, 1991; Accepted February 1, 1992)     

Control of Photosynthesis in Wheat by CO2, O2 and Light Intensity

The regulation of photosynthesis in wheat leaves under varyingO₂, CO₂, and light was studied by analyzing certain metabolitepools and enzyme activities. Under high light when the rateof photosynthesis was limited by low intercellular levels ofCO₂ (C₁) there was a high level of ribulose-1,5-bisphosphate(RuBP) (about 100 nmols per mg chlorophyll). As C, increased,there was a parallel decrease in the ratios of RuBP/3-phosphoglycerate(PGA) (from 0.18 to 0.08 under 21% O₂) and triose-phosphate/PGA(from 0.16 to 0.07 under 21% O₂). The results suggest carboxylationis limited at low C_i, and that there is high carboxylation andlimited assimilatory power at high C_i. As photosynthesis increasedwith increasing Jight intensity under atmospheric levels ofCO₂ the ratios of RuBP/PGA and triosephosphate/PGA remainednearly constant (near 0.12 to 0.13) suggesting there may bea coordinate regulation by light of the different phases ofthe cycle. There was increasing activation of ribulose 1,5-bisphosphatecarboxylase oxygenase (Rubisco) and fructose 1,6-bisphosphatase(FBPase) with increasing light intensity. The ways in whichthe light activation of the enzymes Rubisco and FBPase may regulatecarbon metabolism in the cycle are discussed. ¹ Current address: Biological Sciences Center, Desert ResearchInstitute, PO Box 60220, Reno, Nevada 89506, U.S.A. (Received March 24, 1987; Accepted June 23, 1987)     

Influence of Organic-Phosphates on 3-Phosphoglycerate Dependent O2 Evolution in C3 and C4 Mesophyll Chloroplasts

In C₄ plants phosphoenolpyruvate (PEP) of the C₄ cycle may betransported on a chloroplast transporter which also transports3-phosphoglycerate (3-PGA) and triosephosphates. In C₃ plantsPEP is not considered to be effectively transported on the chloroplastphosphate translocator. The influences of certain organic phosphates,having a similar structure to either PEP or triose-phosphates,on 3-PGA dependent O₂ evolution by C₄ (Digitaria sanquinalisL. Scop.) and C₃ (Hordeum vulgare L.) mesophyll chloroplastswere investigated. In the C₄ mesophyll chloroplasts phosphoglycolatewas a competitive inhibitor (K_i = 2.1 mM) of 3-PGA dependentO₂ evolution, and was as effective as previously reported forPEP. 2-Phosphoglycerate was also a competitive inhibitor (K_t= 8.6 mM) of O₂ evolution in the C₄ mesophyll chloroplasts with3-PGA as substrate, while phospholactate was a weak inhibitorand glyphosate had no effect. Neither PEP, phosphoglycolatenor 2-phosphoglycerate were effective inhibitors of 3- PGA dependentO₂ evolution in the C₃ chloroplasts. Phosphohydroxypyruvatewas a competitive inhibitor of 3-PGA dependent O₂2 evolutionin both chloroplast types. The selectivity in inhibition ofO₂ evolution with 3-PGA as substrate suggests that the C₄ mesophyllchloroplasts can recognize certain organic phosphates with thephosphate in the C-2 or C-3 position but that the C₄ mesophyllchloroplasts can only effectively recognize certain organicphosphates with the phosphate in the C-3 position. The resultsalso support the view that 3-PGA and PEP are transported onthe same phosphate translocator in C₄ mesophyll chloroplasts. ¹ Current address: Department of Horticulture, 2001 Fyffe Court,The Ohio State University, Columbus, Ohio 43210-1096. (Received March 24, 1987; Accepted April 16, 1987)     

Model for the Relationships betweenCO2-Concentrating Mechanism, CO2 Fixation, and Glycolate Synthesis during Photosynthesis in Chlamydomonas reinhardtii

We constructed a mathematical model for simulating the relationshipsof extracellular concentration of dissolved inorganic carbon(DIC), the rates of photosynthetic CO₂ fixation and glycolatesynthesis, and the concentrations of intrachloroplast CO₂ andO₂ in Chlamydomonas reinhardtii. When we compared the photosyntheticrates of I0W-CO₂ (air)-grown C. reinhardtii measured experimentallyand the rates simulated with the incubation conditions in themodel, the model was found to function well. The calculatedrates for glycolate synthesis also matched the measured ratesbetween 80 to 200 µM extracellular DIC, found in the presenceof 1 mM aminooxyacetate. The conformity of the calculated ratesto the measured ones of the glycolate synthesis encouraged usto estimate the O₂ concentration at the active site of ribulosebisphosphate carboxylase/oxygenase; the results were 0.36 and0.40 mM at 80 and 200 µM extracellular DIC, respectively.These high concentrations of O₂ were due to stimulation of photosyntheticCO₂ fixation and further O₂ evolution by a CO₂- concentratingmechanism in the low-CO₂-grown cells. These cells were calculatedto consume 43% of ATP formed photosynthetically for CO₂ concentrationat 200 µM extracellular DIC. The model modified to simulatethese relationships in high-CO₂ (3 to 5% CO₂)-grown C. reinhardtiipredicted O₂ concentration in chloroplasts to be 0.36 mM ina 1% CO₂ atmosphere. This high concentration of O₂ caused activeglycolate synthesis at the measured rate in the high-CO₂-growncells even in the presence of 1% CO₂. The comparisons of themeasured and simulated rates of photosynthesis in low- and high-CO₂-grownC. reinhardtii indicated that no matter how the CO₂ accumulatedin the chloroplasts, it increased the O₂ concentration in theorganelles, and consequently enhanced glycolate synthesis. ¹This paper is the twenty-first in a series on glycolate metabolismin Euglena gracilis. (Received March 11, 1987; Accepted August 17, 1987)     

玫烟色拟青霉对小菜蛾致病力的时间-剂量-死亡率模型模拟

小菜蛾Plutella xylostella是我国南方十字花科蔬菜上的重要害虫,已对田间常用的化学杀虫剂产生了严重的抗性。为寻找有效的小菜蛾生物防治措施,本实验研究了一株分离自家白蚁的玫烟色拟青霉Paecilomyces fumosoroseus (SCAU-PFCF01)对小菜蛾2～4龄幼虫的致病力。实验采用浸液法,供试浓度为1×10³、1×10⁴、1×10⁵、1×10⁶和1×10⁷个孢子/mL。结果表明：随玫烟色拟青霉孢子浓度的升高,小菜蛾的感病死亡率增加,在浓度为1×10⁷ /mL时,小菜蛾2、3和4龄幼虫的累计死亡率分别为96%、85%和80%。玫烟色拟青霉对小菜蛾各龄幼虫的致病力与供试龄期有关,其感病的敏感顺序为2龄、3龄和4龄。用时间 剂量 死亡率模型（time-dose-mortality model,TDM）对各龄幼虫的致病力数据进行模拟,所建模型均顺利通过Hosmer-Lemeshow拟合异质性检验,表明模型拟合良好,并由模型估计出了该菌株对小菜蛾各龄幼虫的致死剂量与致死时间。2龄幼虫接种后第7天、3龄幼虫接种后第5天、4龄幼虫接种后第4天的LC₅₀估计值分别为1.17×10⁴、1.44×10⁴和5.21×10⁴ /mL,LC₉₀估计值分别为1.98×10⁶、3.82×10⁷和1.29×10⁸ /mL。玫烟色拟青霉对小菜蛾幼虫的致死时间与浓度相关,供试各龄幼虫的LT₅₀值随着孢子悬浮液浓度的增加而递减,在1×10⁵～1×10⁷ /mL的范围内,2龄幼虫的LT₅₀值从3.16天降低到1.72天,3龄幼虫的LT₅₀从3.21天降低到1.83天,4龄幼虫的LT₅₀从3.69天降低到2.04天。即2龄幼虫致死所需的时间最短,其次为3龄幼虫,4龄幼虫致死所需的时间最长。结果显示了该株玫烟色拟青霉在小菜蛾的生物防治中具较强的应用潜力。     

Hydrogen Peroxide-Dependent Oxidation of Flavonoids and Hydroxycinnamic Acid Derivatives in Epidermal and Guard Cells of Tradescantia virginiana L.

Luteolin, kaempferol, quercetin, caffeic acid and ferulic acidwere identified in acid-hydrolyzed epidermal strips of Tradescantiavirginiana using HPLC and spectrophotometry. The amount of flavonoidswas much smaller than that of cinnamic acid derivatives. Morethan 80% of the flavonoids were found in methanol extracts ofepidermal strips. Caffeic acid was found in both methanol extractsand the residues in nearly equal amounts, while more than 80%of the ferulic acid was found in the residues after methanolextraction. These data suggest that most of the ferulic acidand part of the caffeic acid bind to macromolecules as estersin the cell wall and that flavonoids are localized mainly inthe cytoplasm. The localization of esters of hydroxycinnamicacids in cell walls was ascertained by fluorometric analysis.These phenolic compounds were oxidized by H₂O₂ (0.025–1mM) in epidermal and guard cells and the oxidation was inhibitedby KCN and NaN₃: luteolin glycosides were less sensitive toH₂O₂ than quercetin and kaempferol glycosides in flavonoids.Ferulic acid esters were more sensitive to H₂O₂ than caffeicacid esters in hydroxycinnamic acid derivatives. On the basisof these data, the physiological significance of the oxidationof phenolic compounds by H₂O₂ is discussed. (Received October 9, 1987; Accepted February 3, 1988)     

Differential impacts of long-term (CO<Subscript>2</Subscript>) and O<Subscript>3</Subscript> exposure on growth of northern conifer and deciduous tree species

The long-term effects of elevated CO₂ and CO₂+O₃ concentrations on the growth allocation in northern provenances of Norway spruce [Picea abies (L.) Karst.], Scots pine [Pinus sylvestris (L.)] and pubescent birch clones (Betula pubescens Ehrh.) were examined in open-top chambers after a 4-year-long experiment. The total biomass responses of the tree seedlings to increased CO₂ and CO₂+O₃ concentrations were not statistically significant and varied between the provenances and species. The seedlings of northern origin were the least sensitive in their response to treatments. The total biomass of the Norway spruce seedlings slightly decreased in response to CO₂ in three provenances. Scots pine from the local provenance had a slight biomass increase after elevated CO₂+O₃ treatment. The slower-growing birch clone seemed to benefit from elevated CO₂, whereas in the faster-growing clone, reductions in biomass accumulation were seen. The combined CO₂+O₃ treatment reduced the positive effects of elevated CO₂, especially in the slower-growing birches. Observations of significant effects were limited to a few parameters. Carbon dioxide treatment decreased needle dry weight of Norway spruce in one northern provenance. The needle and wood dry weight increased (CO₂ + O₃) in local Scots pine. Significant birch response was limited to increased fine root density (O₃ + CO₂) in the inland clone. The diverse effects of elevated CO₂ and CO₂ +O₃ on seedling growth and biomass provide evidence that exposure of northern trees to the enhanced variable CO₂ and O₃ concentrations of the future will have varied effects on the growth of these species. The direction and magnitude of those effects will differ depending on species and origins.     

Development of Epidermal Cell Wall Peroxidases along the Mung Bean Hypocotyl: Possible Involvement in the Cell Wall Stiffening Process

Goldberg, R., Liberman, M., Mathieu, C, Pierron, M. and Catesson,A. M. 1987. Development of epidermal cell wall peroxidases alongthe mung bean hypocotyl: possible involvement in the cell wallstiffening process.—J. exp. Bot. 38: 1378–1390. Ultrastructural investigation showed that in the epidermis ofmung bean hypocotyls, cell wall peroxidatic activities couldbe detected mainly below the maximal elongation zone. In theepidermis the peroxidatic activities were preferentially locatedin the radial cell walls. Cell wall peroxidases were then isolatedfrom epidermal strips and further characterized. The possiblepresence of a H₂O₂-generating system in the epidermis of mungbean hypocotyls was also investigated. When whole segments wereprocessed for electron microscopy, H₂O₂ could be detected cytochemicallyin the cell walls with the CeCl₃ technique. A positive reactionwas obtained in the same location when specimens were incubatedin a 3-3'-diaminobenzidine medium for peroxidases in which H₂O₂was replaced by its possible precursors (NADH or NAD + malate).However, isolated epidermal cell walls could not generate H₂O₂at the expense of NADH although they were able to oxidize thereduced nicotinamide-adenine-dinucleotide. The possible relationshipsbetween peroxidase activities, H₂O₂, and Ca²⁺ ions are discussedwith respect to their involvement in the cell wall stiffeningprocess. Key words: Epidermis, cell wall, elongation, peroxidases     

Oxidation of Flavonols by Hydrogen Peroxide in Epidermal and Guard Cells of Vicia faba L.

Epidermal strips of Vicia faba were found to contain kaempferoland quercetin glycosides. These flavonols were oxidized by H₂O₂and oxidation was inhibited by KCN (3.5 nM). Quercetin glycosideswere more sensitive to H₂O₂ than kaempferol glycosides. Oxidationcould be detected in epidermal strips even at 30 µM H₂O₂.Flavonol oxidation by H₂O₂ was observed in both guard and epidermalcells. In guard cells, oxidation appeared as the bleaching ofabsorption bands of flavonols. Epidermal cells could be roughlydivided into two types on the basis of their absorption characteristicsin the UV-light region. In one type, only flavonol oxidationwas observed; in the other, both flavonol and 3,4-dihydroxyphenylalanine(DOPA) oxidation were observed. Oxidation of flavonols and DOPAby H₂O₂ was also observed in cell-free extracts of the epidermalstrips, even at 10µ H₂O₂. Oxidation was inhibited by 1mM KCN, suggesting the participation of peroxidase in the reactions.The data obtained in this study indicate the cellular specificdistribution of phenolic compounds in the epidermis and thepossibility of their oxidation by H₂O₂ generated in epidermaland guard cells. (Received August 24, 1987; Accepted January 21, 1988)     

Elevated CO2 reduces O3 flux and O3-induced yield losses in soybeans: possible implications for elevated CO2 studies

Soybeans were grown for three seasons in open-top field chambersto determine (1) whether elevated CO₂ (360 versus 700 µmolmol^–1) alleviates some of the yield loss due to pollutantO₃, (2) whether the partial stomatal closure resulting fromchronic O₃ exposure (charcoal-filtered air versus 1.5 ambientconcentrations) is a cause or result of decreased photosynthesis,and (3) possible implications of CO₂/O₃ interactions to climatechange studies using elevated CO₂. Leaf conductance was reducedby elevated CO₂, regardless of O₃ level, or by exposure to O₃alone. As.a result of these effects on conductance, high CO₂reduced estimated midday O₃ flux into the leaf by an averageof 50% in charcoal-filtered air and 35% in the high O₃ treatment.However, while exposure to O₃ reduced seed yields by 41% atambient CO₂ levels, the yield reduction was completely amelioratedby elevated CO₂. The threshold midday O₃ flux for yield lossappears to be 20–30 nmol m^–2 s^–1 in this study.Although elevated CO₂ increased total biomass production, itdid not increase seed yields. A/C_i curves show a large reductionin the stomatal limitation to photosynthesis due to elevatedCO₂ but no effect of O₃. These data demonstrate that (1) reducedconductance due to O₃ is the result, and not the cause, of reducedphotosynthesis, (2) 700 µmol mol^–1 CO₂ can completelyameliorate yield losses due to O₃ within the limits of theseexperiments, and (3) some reports of increased yields underelevated CO₂ treatments may, at least in part, reflect the ameliorationof unrecognized suppression of yield by O₃ or other stresses. Key words: Stomatal limitation, elevated CO₂, O₃ flux, Glycine max, yield suppression     

Impact of elevated atmospheric CO2 and O3 on gas exchange and chlorophyll content in spring wheat (Triticum aestivum L.)

Stands of spring wheat grown in open-top chambers (OTCs) wereused to assess the individual and interactive effects of season-longexposure to elevated atmospheric carbon dioxide (CO₂ and ozone(O₃) on the photosynthetic and gas exchange properties of leavesof differing age and position within the canopy. The observedeffects were related to estimated ozone fluxes to individualleaves. Foliar chlorophyll content was unaffected by elevatedCO₂ but photosynthesis under saturating irradiances was increasedby up to 100% at 680 µmol mol^–1 CO₂ relative tothe ambient CO₂ control; instantaneous water use efficiencywas improved by a combination of increased photosynthesis andreduced transpiration. Exposure to a seasonal mean O₃ concentration(7 h d^–1) of 84 nmol mol^–1 under ambient CO₂ acceleratedleaf senescence following full expansion, at which time chlorophyllcontent was unaffected. Stomatal regulation of pollutant uptakewas limited since estimated O₃ fluxes to individual leaves werenot reduced by elevated atmospheric CO₂, A common feature ofO₃-treated leaves under ambient CO₂ was an initial stimulationof photosynthesis and stomatal conductance for up to 4 d and10 d, respectively, after full leaf expansion, but thereafterboth variables declined rapidly. The O₃-induced decline in chlorophyllcontent was less rapid under elevated CO₂ and photosynthesiswas increased relative to the ambient CO₂ treatment. A/C_i analysessuggested that an increase in the amount of in vivo active RuBisCOmay be involved in mitigating O₃-induced damage to leaves. Theresults obtained suggest that elevated atmospheric CO₂ has animportant role in restricting the damaging effects of O₃ onphotosynthetic activity during the vegetative growth of springwheat, and that additional direct effects on reproductive developmentwere responsible for the substantial reductions in grain yieldobtained at final harvest, against which elevated CO₂ providedlittle or no protection. Key words: Elevated CO₂ and O₃, gas exchange, O₃ flux, stomata, chlorophyll, Triticum aestivum     

Pollutant particles enhanced H2O2 production from NAD(P)H oxidase and mitochondria in human pulmonary artery endothelial cells

Particulate matter (PM) induces oxidative stress and cardiovascular adverse health effects, but the mechanistic link between the two is unclear. We hypothesized that PM enhanced oxidative stress in vascular endothelial cells and investigated the enzymatic sources of reactive oxygen species and their effects on mitogen-activated protein kinase (MAPK) activation and vasoconstriction. We measured the production of extracellular H₂O₂, activation of extracellular signal-regulated kinases1/2 (ERK1/2) and p38 MAPKs in human pulmonary artery endothelial cells (HPAEC) treated with urban particles (UP; SRM1648), and assessed the effects of H₂O₂ on vasoconstriction in pulmonary artery ring and isolated perfused lung. Within minutes after UP treatment, HPAEC increased H₂O₂ production that could be inhibited by diphenyleneiodonium (DPI), apocynin (APO), and sodium azide (NaN₃). The water-soluble fraction of UP as well as its two transition metal components, Cu and V, also stimulated H₂O₂ production. NaN₃ inhibited H₂O₂ production stimulated by Cu and V, whereas DPI and APO inhibited only Cu-stimulated H₂O₂ production. Inhibitors of other H₂O₂-producing enzymes, including N-methyl-L-argnine, indomethacin, allopurinol, cimetidine, rotenone, and antimycin, had no effects. DPI but not NaN₃ attenuated UP-induced pulmonary vasoconstriction and phosphorylation of ERK1/2 and p38 MAPKs. Knockdown of p47phox gene expression by small interfering RNA attenuated UP-induced H₂O₂ production and phosphorylation of ERK1/2 and p38 MAPKs. Intravascular administration of H₂O₂ generated by glucose oxidase increased pulmonary artery pressure. We conclude that UP induce oxidative stress in vascular endothelial cells by activating NAD(P)H oxidase and the mitochondria. The endothelial oxidative stress may be an important mechanism for PM-induced acute cardiovascular health effects. mitogen-activated protein kinase; extracellular signal-regulated kinase; p38; vasoconstriction     

Resistance of Photosynthesis to Hydrogen Peroxide in Algae

The effects of H₂O₂ on the photosynthetic fixation of CO₂ andon thiol-modulated enzymes involved in the photosynthetic reductionof carbon in algae were studied in a comparison with those inchloroplasts isolated from spinach leaves. In both systems,H₂O₂-scavenging enzymes were inhibited by addition of 0.1 mMNaN₃ 1 h prior to the addition of H₂O₂. A concentration (10^-4M) of H₂O₂ caused strong inhibition of the CO₂ fixation by intactspinach chloroplasts, as observed by Kaiser [(1976) Biochim.Biophys. Acta 440: 476], but not that by Euglena and Chlamydomonascells. The same results were also obtained with cells of thecyanobacteria Synechococcus PCC 7942 and Synechocystis PCC 6803in the presence of 1 mM hydroxylamine. These results indicatethat algal photosynthesis is rather resistant to H₂O₂. The insusceptibilityto H₂O₂ of thiolmodulated enzymes, namely, fructose-1,6-bisphosphatase,NADP-glyceraldehyde-3-phosphate dehydrogenase, and ribulose-5-phosphatekinase, was also observed in the chloroplasts of Euglena andChlamydomonas and in cyanobacterial cells. It seems likely thatthe resistance of photosynthesis to H₂O₂ is due in part to theinsusceptibility of the algal thiol-modulated enzymes to H₂O₂. (Received April 22, 1995; Accepted June 29, 1995)     

Requirement of Manganese for Electron Donation of Hydrogen Peroxide in Photosystem II Reaction Center Complex

Mn²⁺ was required for the electron donating reaction from H₂O₂,but not for that from diphenylcarbazide (DPC), in the PS IIreaction center complex which was prepared from spinach chloroplastsby Triton X-100 extraction. The reaction center complex showeda high activity of 2,6-dichloroindophenol (DCIP) photoreductionin the presence of DPC, but a low activity with H₂O₂. The H₂O₂-supportedDCIP photoreduction was suppressed by EDTA and enhanced by asmall amount of Mn²⁺. Ca²⁺ and Mg²⁺ could not replace Mn²⁺.The activation by Mn²⁺ and its binding showed two binding sitesof Mn²⁺ in the reaction center complex, with high (1.5?10⁷ M^–1)and low (1 ? 10⁶ M^–1) binding constants. (Received November 8, 1986; Accepted April 10, 1987)     

The Relationship Between Growth and Oxygen Uptake in Hypoxic Rice Seedlings

Atwell, B. J. and Green way, H. 1987. The relationship betweengrowth and oxygen uptake in hypoxic rice seedlings.—J.exp. Bot. 38: 454–465. Rice seedlings (Oryza saliva L.) were grown in the dark forup to 4 d in solutions containing various concentrations ofO₂. Compared with seedlings grown at 0·250 mol O₂ m^–3,the dry weight of the growing seedling was 14% lower at 0·110mol O₂ m^–3 and 60% lower at 0 mol O₂ m^–3. Decreasesin fresh weight were similar but not identical to decreasesin dry weight, possibly because leaf growth was suppressed evenabove 0·110 mol O₂ m^–3. Oxygen deficiency inhibitedroot growth more severely than coleoptile growth. Coleoptiles from seedlings grown in aerated solution were exposedto an atmosphere of pure N₂ for 30 min. Anoxia caused a declinein ATP content and energy charge, suggestive of decreased oxidativephosphorylation. It is not clear whether the decline in oxidativephosphorylation was solely responsible for impaired growth inhypoxia. In seedlings growing at O₂ concentrations less than 0·110mol O₂ m^–3, significant amounts of ethanol were synthesized.The rate of O₂ uptake decreased markedly below 0·06 molO₂ m^–3; this was presumably near the external O₂ concentrationat which oxidative phosphorylation became limited by the supplyof O₂. The stage of development of the seedlings appeared toinfluence O₂ uptake, possibly through changes in conductanceof the tissue to O₂. Uncouplers were used to confirm that thecritical O₂ concentration was dependent on O₂ diffusion ratherthan enzyme kinetics. Impaired growth above 0·110 molO₂ m^–3 may have been due to a decreased activity of oxygenasesof relatively low affinity for O₂, which in turn altered cellmetabolism. Key words: Growth, oxygen uptake, rice seedlings, hypoxia     

Synthesis of Isozymes of Superoxide Dismutase in Maize Leaves in Response to O3, SO2 and Elevated O2

Matters, G. L. and Scandalios, J. G. 1987. Synthesis of isozymesof superoxide dismutase in maize leaves in response to O₃ SO₂and elevated O₂.—J. exp. Bot 38: 842–852. The activities of the enzymes superoxide dismutase (SOD) andcatalase were determined in maize leaves treated with O₃or SO₂for8 h, or with elevated levels of oxygen for up to 96 h. NeitherO₃nor SO₂significantly increased the levels of superoxide dismutaseor catalase activity. However, after 72 h in an atmosphere containing90% oxygen, superoxide dismutase activity was increased, butnot the activities of catalase, ascorbate pcroxidase, and malatedehydrogenase. Immunological analysis showed that amounts ofthe cytosolic superoxide dismutase isozymes, SOD-2 and SOD-4,were increased by the elevated oxygen but not the chroloplast(SOD-1) or mitochondrial (SOD-3) isozymes. Immunoprecipitationof translation products of leaf polysomes indicated that thehigher levels of SOD-2 and SOD-4 were due to increased amountsof polysome-bound mRNA coding for these proteins. The specificresponse of SOD-2 and SOD-4 to 90% oxygen treatments contrastswith the increase in all SOD isozymes in maize leaves treatedwith the herbicide paraquat. Key words: Air pollutants, maize, oxidative stress, oxygen, superoxide dismutase     

Organic Acid Metabolism in Cellular Organelles of Vigna cylindrica (Mitorisasage) Cotyledons

In starchy cotyledons of Vigna cylindrica (L.) Skeels (Mitorisasage)during seed germination, the enzymes of the glyoxylate cyclewere located in the matrix of mitochondria. Glyoxysomes wereabsent. The glyoxylate cycle in the mitochondria supplies organicacids to the tricarboxylic acid cycle. In mitochondria, isocitratelyase activity was much higher than malate synthase activity.Part of the glyoxylate thus produced in mitochondria may benonenzymatically converted to formate by H₂O₂ and the formatethen converted to CO₂ by peroxidase or by formic dehydrogenase.The activity of superoxide dismutase, which supplies H₂O₂, washigher in mitochondria than in peroxisomes. The remaining glyoxylatein mitochondria is possibly converted to glycine by alanine-glyoxylateaminotransferase or transported to peroxisomes which lackedisocitrate lyase activity but had high malate synthase activity.In peroxisomes, glyoxylate may be also produced from urate,as is suggested by the fairly high activities of uricase, allantoinaseand allantoicase. Judging from the enzyme distribution, Vignaperoxisomes should be capable of producing malate, oxalacetate,citrate, isocitrate and a-ketoglutarate. ¹Present address: Department of Horticulture, College of Agricultureand Animal Science, Yeugnam University, Gyeongsan 632, Korea. (Received May 27, 1987; Accepted October 7, 1987)     

Activation of Electron Donation from Hydrogen Peroxide by Manganese in Non-oxygen evolving Photosystem II Particles

Analysis of the activation of H₂O₂-supported 2,6-dichloroindophenol(DCIP) photoreduction by MnCl₂ showed two Mn²⁺-binding sitesin non-oxygen evolving PS II particles, with large (0.4IM) andsmall (0.04 µm) K_m values for Mn²⁺. Photoreduction throughthe high-affinity Mn²⁺.-binding site was inhibited by treatmentwith H₂O₂. (Received April 20, 1987; Accepted July 13, 1987)     

Photosynthesis and photorespiration in soybean [Glycine max (L.) Merr.] chronically exposed to elevated carbon dioxide and ozone

The effects of elevated carbon dioxide (CO₂ and ozone (O₃) onsoybean (Glycine max (L.) Merr.] photosynthesis and photorespiration-relatedparameters were determined periodically during the growing seasonby measurements of gas exchange, photorespiratory enzyme activitiesand amino acid levels. Plants were treated in open-top fieldchambers from emergence to harvest maturity with seasonal meanconcentrations of either 364 or 726 µmol mol^–1 CO₂in combination with either 19 or 73 nmol mol^–1 O₃ (12h daily averages). On average at growth CO₂ concentrations,net photosynthesis (A) increased 56% and photorespiration decreased36% in terminal mainstem leaves with CO₂ enrichment. Net photosynthesisand photorespiration were suppressed 30% and 41%, respectively,by elevated O₃ during late reproductive growth in the ambientCO₂ treatment, but not in the elevated CO₂ treatment. The ratioof photorespiration to A at growth CO₂ was decreased 61% byelevated CO₂ There was no statistically significant effect ofelevated O₃ on the ratio of photorespiration to A. Activitiesof glycolate oxidase, hydroxypyruvate reductase and catalasewere decreased 10–25% by elevated CO₂ and by 46–66%by elevated O₃ at late reproductive growth. The treatments hadno significant effect on total amino acid or glycine levels,although serine concentration was lower in the elevated CO₂and O₃ treatments at several sampling dates. The inhibitoryeffects of elevated O₃ on photorespiration-related parameterswere generally commensurate with the O₃-induced decline in A.The results suggest that elevated CO₂ could promote productivityboth through increased photoassimilation and suppressed photorespiration. Key words: Photorespiration, CO₂-enrichment, ozone, climate change, air pollution     

大气臭氧浓度升高对水稻叶片膜脂过氧化及保护酶活性的影响

采用开顶式气室（open top chambers,OTCs）装置,以水稻品种“3694繁”(Oryza sativa L.)为材料,研究3种处理：过滤大气（CF,O₃浓度约为20 nl·L^-1）、环境大气（NF,O₃浓度约为40 nl·L^-1）和高浓度O₃（EO,O₃浓度约为75 nl·L^-1）下叶片可溶性蛋白质含量、膜脂过氧化程度与主要保护酶活性的变化.结果表明：过滤大气与环境大气处理之间各个指标无显著差异.与CF处理相比,高浓度O₃处理条件下水稻叶片中可溶性蛋白含量显著下降,而过氧化氢（H₂O₂）和抗坏血酸（ASA）含量显著增加;超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、过氧化物酶（POD）活性升高,最大升高幅度分别为93.7%、39.9%和312.4%;抗坏血酸过氧化物酶（APX）活性呈先升高后下降趋势; 不同O₃处理条件下叶片中丙二醛（MDA)含量无显著变化.表明抗氧化系统可有效抑制水稻叶片中的O₃胁迫引起的膜质过氧化作用,说明水稻品种“3694繁”对O₃胁迫有一定抗性.