CO(2) Enhancement of Growth and Photosynthesis in Rice (Oryza sativa) : Modification by Increased Ultraviolet-B Radiation

Two cultivars of rice (Oryza sativa L.) IR-36 and Fujiyama-5 were grown at ambient (360 microbars) and elevated CO₂ (660 microbars) from germination through reproduction in unshaded greenhouses at the Duke University Phytotron. Growth at elevated CO₂ resulted in significant decreases in nighttime respiration and increases in photosynthesis, total biomass, and yield for both cultivars. However, in plants exposed to simultaneous increases in CO₂ and ultraviolet-B (UV-B) radiation, CO₂ enhancement effects on respiration, photosynthesis, and biomass were eliminated in IR-36 and significantly reduced in Fujiyama-5. UV-B radiation simulated a 25% depletion in stratospheric ozone at Durham, North Carolina. Analysis of the response of CO₂ uptake to internal CO₂ concentration at light saturation suggested that, for IR-36, the predominant limitation to photosynthesis with increased UV-B radiation was the capacity for regeneration of ribulose bisphosphate (RuBP), whereas for Fujiyama-5 the primary photosynthetic decrease appeared to be related to a decline in apparent carboxylation efficiency. Changes in the RuBP regeneration limitation in IR-36 were consistent with damage to the photochemical efficiency of photosystem II as estimated from the ratio of variable to maximum chlorophyll fluorescence. Little change in RuBP regeneration and photochemistry was evident in cultivar Fujiyama-5, however. The degree of sensitivity of photochemical reactions with increased UV-B radiation appeared to be related to leaf production of UV-B-absorbing compounds. Fujiyama-5 had a higher concentration of these compounds than IR-36 in all environments, and the production of these compounds in Fujiyama-5 was stimulated by UV-B fluence. Results from this study suggest that in rice alterations in growth or photosynthesis as a result of enhanced CO₂ may be eliminated or reduced if UV-B radiation continues to increase.     

Field Study of the Interaction between Solar Ultraviolet-B Radiation and Drought on Photosynthesis and Growth in Soybean

Soybean, Glycine max (L.) Merr. cv Essex, plants were grown in the field in a 2 × 2 factorial design, under ambient and supplemental levels of ultraviolet-B (UV-B) radiation (supplemental daily dose of 5.1 effective kilojoules per square meter) and were either well-watered or subjected to drought. Soil water potentials were reduced to −2.0 megapascals by the exclusion of natural precipitation in the drought plots and were maintained at approximately −0.5 megapascal by supplemental irrigation in well-watered plots. Plant growth and gas exchange characteristics were affected under both drought and supplemental UV-B radiation. Whole-leaf gas exchange analysis indicated that stomatal limitations on photosynthesis were only significantly affected by the combination of UV-B radiation and drought but substrate (ribulose bisphosphate) regeneration limitations were observed under either stress. The combined effect of both drought and UV-B radiation on photosynthetic gas exchange was a reduction in apparent quantum efficiency and the rapid appearance of biochemical limitations to photosynthesis concomitant with reduced diffusional limitations. However, the combination of stresses did not result in additive effects on total plant growth or seed yield compared to reductions under either stress independently.     

Nitric Oxide Alleviates Oxidative Damage Induced by Enhanced Ultraviolet-B Radiation in Cyanobacterium

To study the role of nitric oxide (NO) on enhanced ultraviolet-B (UV-B) radiation (280–320 nm)-induced damage of Cyanobacterium, the growth, pigment content, and antioxidative activity of Spirulina platensis-794 cells were investigated under enhanced UV-B radiation and under different chemical treatments with or without UV-B radiation for 6 h. The changes in chlorophyll-a, malondialdehyde content, and biomass confirmed that 0.5 mM sodium nitroprusside (SNP), a donor of nitric oxide (NO), could markedly alleviate the damage caused by enhanced UV-B. Specifically, the biomass and the chlorophyll-a content in S. platensis-794 cells decreased 40% and 42%, respectively under enhanced UV-B stress alone, but they only decreased 10% and 18% in the cells treated with UV-B irradiation and 0.5 mM SNP. Further experiments suggested that NO treatment significantly increased the activities of superoxide dismutase (SOD) and catalase (CAT), and decreased the accumulation of O ₂ ⁻ in enhanced UV-B-irradiated cells. SOD and CAT activity increased 0.95- and 6.73-fold, respectively. The accumulation of reduced glutathione (GSH) increased during treatment with 0.5 mM SNP in normal S. platensis cells, but SNP treatment could inhibit the increase of GSH in enhanced UV-B-stressed S. platensis cells. Thus, these results suggest that NO can strongly alleviate oxidative damage caused by UV-B stress by increasing the activities of SOD, peroxidase, CAT, and the accumulation of GSH, and by eliminating O ₂ ⁻ in S. platensis-794 cells. In addition, the difference of NO origin between plants and cyanobacteria are discussed.     

Decreased Ultraviolet-B Radiation Alters the Vertical Biomass Distribution in Cocksfoot

A greenhouse experiment was conducted to investigate whether small differences in UV-B irradiance would lead to changes in competition between cocksfoot (Dactylis glomerata L. cv. Athos) and white clover (Trifolium repens L. cv. Mervi). Plants were grown in greenhouses covered with different thicknesses of UV-transmittant plexi (3 and 5 mm) resulting in 82 % and 88 % of ambient UV-B radiation. Aboveground biomass was harvested at 4-week intervals and the vertical distribution of biomass, leaf thickness and specific leaf area were determined. Tillering, stubble and root biomass and crop height were also measured. There was only one significant effect: at 88 % of ambient UV-B radiation a larger fraction of the biomass was present in the lower layers and a smaller fraction was present in the upper layers.     

Inactivation of Cyanobacterial Nitrogenase After Exposure to Ultraviolet-B Radiation

Exposure of the N₂-fixing cyanobacterium Anabaena BT2 to ultraviolet-B radiation (2.5 W m⁻²) for 30 min resulted in complete loss of nitrogenase activity but 100% cell killing occurred only after a 90-min exposure. Inactivation of nitrogenase activity was not specific to Anabaena BT2; other species also showed a similar effect. The time required for 100% killing and inactivation of nitrogenase activity differed in various species, and this difference may be ascribed to the presence of different levels of UV-B protection mechanisms in individual species. Inhibition of nitrogenase activity was immediate, since exposure of cultures to UV-B for as little as 5 min elicited some inhibition of activity. The activity of UV-B-inhibited nitrogenase did not recover upon transfer of exposed cells to fluorescent light, suggesting that the inhibition may be due to specific inactivation of the enzyme. By employment of inhibitors of protein synthesis and PS-II activity, it was demonstrated that restoration of nitrogenase activity in a UV-B-treated culture occurred by fresh synthesis of nitrogenase polypeptide. Our findings suggest that estimation of nitrogenase activity in diazotrophic species may be used as a marker enzyme for assessing the impact of UV-B radiation. Received: 13 June 2002 / Accepted: 22 July 2002     

Influence of UV-B Supplemental Radiation on Growth and Pigment Content in Suaeda Maritima L.

In a field experiment with a mangrove species Suaeda maritima L. grown under ambient and supplementary UV-B radiation corresponding to 20 % ozone depletion, changes in growth and contents of photosynthetic and UV-absorbing pigments were determined. Supplemental UV-B irradiation for 9 d significantly reduced the growth and concentration of photosynthetic pigments. However, anthocyanin and flavonoid contents were significantly increased in UV-treated plants and which could be reduce the UV-B penetration and damage to the underlying tissues.     

Alleviation of Ultraviolet-B Radiation-Induced Growth Inhibition of Green Gram by Triadimefon

Supplementary UV-B (12.2 kJ m⁻² d⁻¹ UV-B_BE) provided to Vigna radiata for 2 h d⁻¹ suppressed the length of root, shoot and whole plants, number of leaves, total leaf area, leaf area index, specific leaf mass, fresh and dry mass of leaves and shoot, relative growth rate and net productivity. In unstressed green gram plants (10 kJ m⁻² d⁻¹ UV-B_BE), triadimefon (TRIAD) (20 mg dm⁻³) enhanced growth in all parameters over control. The growth promoting effect of TRIAD enabled the UV-B impacted plants to overcome the growth inhibitions to varying degrees indicating its protective potential against UV-B stress. This revised version was published online in July 2006 with corrections to the Cover Date.     

Flux Imbalance Analysis and the Sensitivity of Cellular Growth to Changes in Metabolite Pools

Stoichiometric models of metabolism, such as flux balance analysis (FBA), are classically applied to predicting steady state rates - or fluxes - of metabolic reactions in genome-scale metabolic networks. Here we revisit the central assumption of FBA, i.e. that intracellular metabolites are at steady state, and show that deviations from flux balance (i.e. flux imbalances) are informative of some features of in vivo metabolite concentrations. Mathematically, the sensitivity of FBA to these flux imbalances is captured by a native feature of linear optimization, the dual problem, and its corresponding variables, known as shadow prices. First, using recently published data on chemostat growth of Saccharomyces cerevisae under different nutrient limitations, we show that shadow prices anticorrelate with experimentally measured degrees of growth limitation of intracellular metabolites. We next hypothesize that metabolites which are limiting for growth (and thus have very negative shadow price) cannot vary dramatically in an uncontrolled way, and must respond rapidly to perturbations. Using a collection of published datasets monitoring the time-dependent metabolomic response of Escherichia coli to carbon and nitrogen perturbations, we test this hypothesis and find that metabolites with negative shadow price indeed show lower temporal variation following a perturbation than metabolites with zero shadow price. Finally, we illustrate the broader applicability of flux imbalance analysis to other constraint-based methods. In particular, we explore the biological significance of shadow prices in a constraint-based method for integrating gene expression data with a stoichiometric model. In this case, shadow prices point to metabolites that should rise or drop in concentration in order to increase consistency between flux predictions and gene expression data. In general, these results suggest that the sensitivity of metabolic optima to violations of the steady state constraints carries biologically significant information on the processes that control intracellular metabolites in the cell.     

Protection of Photosynthesis against Ultraviolet-B Radiation by Carotenoids in Transformants of the Cyanobacterium Synechococcus PCC7942

The cyanobacterium Synechococcus PCC7942 was transformed with various carotenogenic genes, and the resulting transformants either accumulated higher amounts of β-carotene and zeaxanthin or showed a shift in the carotenoid pattern toward the formation of zeaxanthin. These transformants were exposed to ultraviolet-B (UV-B) radiation, and the degradation of phycobilins, the inactivation of photosynthetic oxygen evolution, and the activity of photosystem II were determined. In the genetically modified cells, the influence on destruction of phycobilins was negligible. However, protection of photosynthetic reactions against UV-B damage was observed and was dependent on the carotenoid concentrations in the different transformants. Furthermore, it was shown that endogenous zeaxanthin is more effective than β-carotene. Our results suggest that carotenoids exert their protective function as antioxidants to inactivate UV-B-induced radicals in the photosynthetic membrane.     

Lysozyme Modification by the Fenton Reaction and Gamma Radiation

A comparative study was performed on lysozyme modification after exposure to Fenton reagent (Fe(II)/H 2 O 2 ) or hydroxyl radicals produced by &#110 radiation. The conditions were adjusted to obtain, with both systems, a 50% loss of activity of the modified ensemble. &#110 radiation modified almost all types of amino acid residues in the enzyme, with little specificity. The modification order was Tyr > Met=Cys > Lys > Ile+Leu > Gly > Pro=Phe>Thr+Ala>Trp=Ser>Arg>Asp+Glu, with 42 mol of modified residues per initial mole of native enzyme. In contrast, when the enzyme was exposed to the Fenton reaction, only some types of amino acids were modified. Furthermore, a smaller number of residues (13.5) were damaged per initial mole of enzyme. The order of the modified residues was Tyr > Cys > Trp > Met >His > Ile+Leu > Val > Arg. These results demonstrate that the modifications elicited by these two free radical sources follow different mechanisms. An intramolecular free radical chain reaction is proposed to play a dominant role in the oxidative modification of the protein promoted by &#110 radiation.     

Protective Effect of Rare Earth Against Oxidative Stress Under Ultraviolet-B Radiation

The effects of lanthanum (III) (La(III)) in protecting soybean leaves against oxidative stress induced by ultraviolet-B (UV-B) radiation were investigated. The increase in contents of hydrogen peroxide (H₂O₂) and superoxide () due to UV-B radiation suggested oxidative stress. The increase in the content of malondialdehyde (MDA) and the decrease in the index of unsaturated fatty acid (IUFA) indicated oxidative damage on cell membrane induced by UV-B radiation. La(III) partially reversed UV-B-radiation-induced damage of plant growth. The reduction in the contents of H₂O₂, , and MDA and increase in the content of IUFA, compared with UV-B treatment, also indicated that La(III) alleviated the oxidative damage induced by UV-B radiation. The increase in the activities of superoxide dismutase and peroxidase and the contents of ascorbate, carotinoids, and flavonoids were observed in soybean leaves with La(III) + UV-B treatment, compared with UV-B treatment. Our data suggested that La(III) could protect soybean plants from UV-B-radiation-induced oxidative stress by reacting with reactive oxygen species directly or by improving the defense system of plants.     

Influence of Ultraviolet-B Radiation on Peroxidase Activity of Allium schoenoprasum Leaves

An approximately 7 % difference in biologically effective ultraviolet-B (UV-B) radiation did not significantly influence leaf length or leaf peroxidase activity of chives (Allium schoenoprasum L.). However, correlation and regression analyses with different climatic parameters revealed that increased UV-B radiation enhanced ascorbate peroxidase activity in chive leaves whereas guaiacol peroxidase was inhibited. This revised version was published online in July 2006 with corrections to the Cover Date.     

Inhibition of Pigments and Phycocolloid in a Marine Red Alga Gracilaria Edulis by Ultraviolet-B Radiation

Vegetative fragments of the subtidal macroalga Gracilaria edulis (Gmel.) Silva cultured under field conditions at Thonithurai were subjected in the laboratory to UV-B radiation (280 – 320 nm). UV-B inhibited the accumulation of chlorophyll and phycobiliproteins, and lowered agar yield (23 to 43%) and its gel strength (22 to 36%) under 12 to 72 h exposure. The longer the exposure to UV-B radiation the more significant impact on pigments and phycocolloid properties of Gracilaria edulis was observed.     

Effects of Ultraviolet-B Radiation Stress on the Abscisic Acid Status of Rumex patientia Leaves

To determine if increased abscisic acid (ABA) levels are associated with ultraviolet-B (UV-B) radiation (288–320 nm) stress, Rumex patientia L. plants were exposed to visible light with high or low (control) UV-B irradiance and then assayed for ABA by gas chromatography. Though leaf growth was inhibited by the UV-B irradiation, no differences in levels of free ABA were found between leaves from the two treatments after 1, 3 or 5 days of exposure. Unlike the situation in most environmental stresses, increased ABA levels are not associated with UV-B radiation stress. Further, photolysis or isomerization are not likely explanations for the absence of ABA increase. Calculations indicate that if ABA in the leaf were fully exposed to solar UV-B radiation only about 6.7% would be photolyzed by existing daily radiation at 40°N latitude, and 11.3% by radiation resulting from a 40% decrease in atmospheric ozone. The quantum yield for isomerization of cistrans-ABA by radiation in the 288–312 nm waveband was determined. Existing daily solar UV-B irradiation is capable of isomerizing ABA in solution to an equilibrium mixture of 50% cisjrans and 50%irans, trans-ABA., However, because of radiation attenuation in the leaf, it is estimated that only a small fraction of the cistrans-A.BA. would be isomerized.     

紫外光源及太阳UV-B辐射的模拟实验

介绍了用于UV-B生物学效应研究的几种常见光源,对如何利用选择性薄膜获得理想的UV-B波段的光谱,进而能较真实模拟平流层O3耗损所导致的近地表面太阳UV-B辐射的增强,以及在野外进行UV-B模拟研究时,紫外荧光灯管的排列方式等进行了分析讨论。介绍并讨论了当前UV-B辐射实验的各种方法,并就各实验设计的注意事项和安全操作等提出了建议。     

Dna Radiation Damage and its Modification by Metallothionein

Thiol compounds have long been known to protect living cells against the harmful effects of ionizing radiation. Maetallothionein is a naturally occurring low molecular weight polypeptide rich in cysteine residues and may be useful in protection against low-level radiation effects.

Radiation damage to DNA and its nucleotide components and the radioprotective effect of metallothionein have been studied in model chemical systems and compared to its effect on cells. Metallothionein acts both as a free radical scavenger and a reductant, and its radioprotective effectiveness has been studied as a function of dose, drug concentration, and in the presence and absence of oxygen. It is more effective in protecting against sugar-phosphate damage under hypoxic conditions. The chemical modification is greater than that of cell killing as measured by the loss of colony-forming ability. Dose reduction factors greater than two are observed for DNA radioprotection, but the values in cells are much lower. These findings will be discussed in terms of the molecular mechanisms and their implications.     

Dna Radiation Damage and its Modification by Metallothionein

Thiol compounds have long been known to protect living cells against the harmful effects of ionizing radiation. Maetallothionein is a naturally occurring low molecular weight polypeptide rich in cysteine residues and may be useful in protection against low-level radiation effects.

Radiation damage to DNA and its nucleotide components and the radioprotective effect of metallothionein have been studied in model chemical systems and compared to its effect on cells. Metallothionein acts both as a free radical scavenger and a reductant, and its radioprotective effectiveness has been studied as a function of dose, drug concentration, and in the presence and absence of oxygen. It is more effective in protecting against sugar-phosphate damage under hypoxic conditions. The chemical modification is greater than that of cell killing as measured by the loss of colony-forming ability. Dose reduction factors greater than two are observed for DNA radioprotection, but the values in cells are much lower. These findings will be discussed in terms of the molecular mechanisms and their implications.     

Radiation Sensitivity of GL261 Murine Glioma Model and Enhanced Radiation Response by Flavopiridol

Response of a solid tumor to radiation treatment depends, in part, on the intrinsic radiosensitivity of tumor cells, the proliferation rate of tumor cells between radiation treatments and the hypoxic state of the tumor cells. A successful radiosensitizing agent would target S-phase cells and hypoxia. Recently, we demonstrated the anti-tumor effects of flavopiridol in the GL261 murine glioma model might involve 1) recruitment of tumor cells to S-phase (Newcomb et al., Cell Cycle 2004; 3:230-234) and 2) an anti-angiogenic effect on the tumor vasculature by downregulation of hypoxia-inducible factor -1? (HIF-1?) (Newcomb et al., Neuro-Oncology 2005; 7:225-235). Given that flavopiridol has demonstrated radiosensitizing activity in several murine tumor models, we tested whether it would enhance the response of GL261 tumors to radiation. In the present study, we evaluated the intrinsic radiation sensitivity of the GL261 glioma model using the tumor control/cure dose of radiation assay (TCD50). We found that a single dose of 65 Gy (CI 57.1-73.1) was required to cure 50% of the tumors locally. Using the tumor growth delay assay, fractionated radiation (5 fractions of 5 Gy over 10 days) combined with flavopiridol (5 mg/kg) given three times weekly for 3 cycles produced a significant growth delay. Our results indicate that the GL261 murine glioma model mimics the radioresistance encountered in human gliomas, and thus should prove useful in identifying promising new investigational radiosensitizers for use in the treatment of glioma patients.