The effects of water-stress on leaf H2 18O enrichment

Summary Water-stress experiments withPhaseolus vulgaris L. were undertaken to determine the transpiration rate dependency of the naturally occurring leaf H₂ ¹⁸O fractionation process. Water-stress leaf H₂ ¹⁸O levels were observed to be unexpectedly higher than controls. Speculations on the cause of this phenomenon are discussed. Since transpiration rate variations should theoretically affect only the rate and not the extent of leaf H₂ ¹⁸O fractionation, the respective time courses for water-stressed and control leaf H₂ ¹⁸O accumulations were compared. Water-stressed leaves displayed a slower rate of isotopic enrichment relative to controls, as was predicted from their reduced transpiration rates. In an absolute sense, however, both control and water-stress leaf H₂ ¹⁸O fractionation rates were markedly greater than projected values from the existing model. Consequently, transpiration rates cannot be derived accurately at present from the observed rates of leaf H₂ ¹⁸O discrimination. Several modifications of the theory are also considered.     

Diurnal changes in the flux of calcium toward meristems and transpiring leaves in tomato and maize plants

Tomato (Lycopersicon esculentum Mill. cv. Moneymaker) and maize (Zea mays L. cv. spec.) plants were supplied with ⁴⁵Ca-labeled nutrient solutions for a period of 8 or 16 h in the dark, in the light, or in a light-dark régime. Plant parts were analyzed for ⁴⁵Ca content. The partitioning of ⁴⁵Ca between mature leaves and meristems was shown to be affected by the presence of light. The transport of ⁴⁵Ca to meristems was higher in a dark period than in a comparable light period. Experiments with excised tomato shoots yielded similar distribution patterns of ⁴⁵Ca over leaves and meristems, thus excluding root pressure as the main driving force for the enhanced import of ⁴⁵Ca into the meristems in the dark. Results are discussed in terms of cation-exchange transport and competition between the various calcium sinks.Abbreviations DM dry material - IAA indole-3-acetic acid - TIBA 2,3,5-trijodbenzoic acid - CEC cation exchange capacityContribution No. 1693 of the Radiation Protection Programme of the Commission of the European Communities     

Controlling the soil moisture environment of transpiring plants

Summary A method of growing plants under constant soil moisture conditions when the transpiration rate is constant is presented. This method uses the principles of unsaturated flow through a porous media to arrive at values for the design variables. If the transpiration rate is not constant, the maximum possible fluctuation in the soil moisture content can be determined.     

Evidence for 18O labeling of photorespiratory CO2 in photoautotrophic cell cultures of higher plants illuminated in the presence of 18O2

The ¹⁸O-enrichment of CO₂ produced in the light or during the post-illumination burst was measured by mass spectrometry when a photoautotrophic cell suspension of Euphorbia characias L. was placed in photorespiratory conditions in the presence of molecular ¹⁸O₂. The only ¹⁸O-labeled species produced was C¹⁸O¹⁶O; no C¹⁸O¹⁸O could be detected. Production of C¹⁸O¹⁶O ceased after addition of two inhibitors of the photosynthetic carbon-oxidation cycle, aminooxyacetate or aminoacetonitrile, and was inhibited by high levels of CO₂. The average enrichment during the post-illumination burst was estimated to be 46 ± 15% of the enrichment of the O₂ present during the preceding light period. Addition of exogenous carbonic anhydrase, by catalyzing the exchange between CO₂ and H₂O, drastically diminished the ¹⁸O-enrichment of the produced CO₂. The very low carbonio-anhydrase level of the photoautotrophic cell suspension probably explains why the ¹⁸O labeling of photorespiratory CO₂ could be observed for the first time. These data allow the establishment of a direct link between O₂ consumption and CO₂ production in the light, and the conclusion that CO₂ produced in the light results, at least partially, from the mitochondrial decarboxylation of the glycine pool synthesized through the photosynthetic carbon-oxidation cycle. Analysis of the C¹⁸O¹⁶O and CO₂ kinetics provides a direct and reliable way to assess in vivo the real contribution of photorespiratory metabolism to CO₂ production in the light.     

Effect of NaCl stress on H2O2 metabolism in rice leaves

The effect of NaCl stress on H₂O₂ metabolismin detached rice leaves was studied. NaCl (200 mM)treatment did not cause the accumulation ofH₂O₂ and resulted in no increase in lipidperoxidation and membrane leakage of leaf tissues. The activities of peroxidase, ascorbate peroxidase,superoxide dismutase, and glutathione reductase wereobserved to be greater in NaCl-stressed rice leavesthan in control leaves. However, glycolate oxidasewas lower in NaCl-treated rice leaves than in thecontrol leaves. There was no difference in catalaseactivity between NaCl and control treatments. Theseresults suggest that some antioxidant enzymes can beactivated in response to oxidative stress induced byNaCl.     

The enrichment of18O in leaf water under natural conditions

Summary The enrichment of¹⁸O in the water of transpiring leaves under natural conditions is described. In the first series two, later three species at the same location and at the same time are compared (birchBetula pubescens L., oakQuercus robur Ehrh., larchLarix decidua Mill., and sprucePicea abies Karst). All four show parallel enrichment properties. In addition two beeches (Fagus silvatica L.) were observed, one at Jülich, the other at the Solling hills. They show a similar¹⁸O/¹⁶O ratio fluctuation in their leaves during the time of observation. Three plant communities (beech forest B1, spruce forest F1, and meadow W, sites of the Solling-Project, German Research Foundation, part of International Biological Program, at the Solling hills) show a daily course of the¹⁸O enrichment comparable to each other.     

Components of apoplastic ascorbate use in Betula pendula leaves exposed to CO2 and O3 enrichment

Here, the aim was to estimate loads imposed on the apoplastic ascorbate (ASC) pool by enzymatic and nonenzymatic reactions in Betula pendula exposed to doubled CO2 and O3 concentrations in open-top chambers. Leaf apoplastic extracts were analysed for peroxidase and oxidase activities in vitro, using different substrates. Partial loads in vivo were deduced using measured kinetic constants and substituted-enzyme catalysis approaches. Ascorbate use in O3 scavenging was calculated using measured stomatal conductances and ASC concentrations. Under elevated O3, stomatal conductance and O3 uptake were higher. O3 fluxes to the plasmalemma were levelled off by higher apoplastic ASC concentrations. The effect of CO2 enrichment on ASC concentrations under elevated O3 was minor. Under ambient O3, the ascending hierarchy of ASC users was: peroxidases, O3 scavenging, oxidases, coniferyl alcohol re-reduction. Under elevated O3, ASC use in O3 scavenging was higher than by oxidases. The redox state of ASC was not depressed by O3; there was no leaf injury. The cell wall/plasmalemma/cytosol system in birch had sufficient capacity to maintain ASC redox status in the apoplast, without necessity to restrict O3 uptake by stomatal closure.     

Fatty acid hydroperoxides and H2O2 in the execution of hypersensitive cell death in tobacco leaves

We initially compared lipid peroxidation profiles in tobacco (Nicotiana tabacum) leaves during different cell death events. An upstream oxylipin assay was used to discriminate reactive oxygen species (ROS)-mediated lipid peroxidation from 9- and 13-lipoxygenase (LOX)-dependent lipid peroxidation. Free radical-mediated membrane peroxidation was measured during H(2)O(2)-dependent cell death in leaves of catalase-deficient plants. Taking advantage of these transgenic plants, we demonstrate that, under light conditions, H(2)O(2) plays an essential role in the execution of cell death triggered by an elicitor, cryptogein, which provokes a similar ROS-mediated lipid peroxidation. Under dark conditions, however, cell death induction by cryptogein was independent of H(2)O(2) and accompanied by products of the 9-LOX pathway. In the hypersensitive response induced by the avirulent pathogen Pseudomonas syringae pv syringae, both 9-LOX and oxidative processes operated concurrently, with ROS-mediated lipid peroxidation prevailing in the light. Our results demonstrate, therefore, the tight interplay between H(2)O(2) and lipid hydroperoxides and underscore the importance of light during the hypersensitive response.     

The H2 18O enrichment in the leaf water of tropic trees: Comparison of species from the tropical rain forest and the semi-arid region in Brazil

Summary The H₂ ¹⁸O enrichment,, in the water of leaves from four Brazilian trees, was studied. In all trees the leaf water showed a periodic variation in, with a maximum in the early afternoon and a minimum around 6 a.m. In general was found to be either higher or lower than the stationary enrichment which is supposed to depend only on the relative atmospheric humidity. This effect is due to the slow response of the system to variations of the humidity. For a special case, where steady-state conditions could be anticipated, the kinetic enrichment was obtained to 20 ± 3, which agrees with theoretical predictions.     

The contribution of land photosynthesis to the stationary enrichment of18O in the atmosphere

Summary The stationary enrichment of¹⁸O in the earth's atmosphere (Dole-effect) does not result exclusively from respiration processes, but also from photosynthesis in transpiring plants. Using the global distribution patterns of relative humidity, photosynthetic activity and H₂ ¹⁸O concentration in precipitation water, the contribution of land photosynthesis to the Dole-effect is estimated to 8 .     

On the Mechanism of Action of H2O2 in the Cellular Stress

We propose a hypothesis according to which the reactive and reduced species of oxygen could be the intracellular inducers of the stress (or “heat-shock”) response. This hypothesis is based on the following observations on Drosphila cells: -a) the return to normoxia after 24 h anaerobiosis is suficient to induce the synthesis of the ‘heat shock’ proteins without elevation of temperature together with a rapid increase of O₂ consumption; -b) hydrogen peroxide introduced in the culture medium induces the early transcrip-tional activation of the ‘heat shock’ genes (maximal after 5 minutes); -c) hydrogen peroxide added to cellular extracts in vitro (thus acting as an intracellular metabolite) activates instantaneously the binding capacity of a ‘heat shock’ factor to a DNA ‘heat shock’ regulatory element. Thus, hydrogen peroxide, and possibly other reactive reduced species ofoxygen, could trigger the onset of the stress (or ‘heat shock’) response.     

H2O2 activates CD11b/CD18-dependent cell adhesion

Treatment of monoblastoid U-937 cells with low concentrations of H2O2 caused adhesion of the cells to plastic. The H2O2 induced adhesion was rapid with a t1/2 of congruent to 6 min and was optimally stimulated by 100 microM H2O2 with an ED50 of congruent to 50 microM. The response to H2O2 closely resembled the adhesive response of U-937 cells to phorbol esters in its time dependency, requirement for extracellular Mg2+ and inhibition by cytochalasin B as well as inhibition by monoclonal antibodies against the leucocyte adhesion molecules CD11b and CD18. Phorbol ester treatment of U-937 cells stimulated the phosphorylation of at least three endogenous substrates, pp28, pp34 and pp43, of which pp28 and pp43 also responded to H2O2-treatment with increased 32P-incorporation. The results suggest that H2O2 might be a physiological modulator of leucocyte adhesion, possibly operating by activating protein kinase C.     

Response of young barley plants to CO2 enrichment

Barley (Hordeum vulgare L. cv. Digger) was grown for 22 d inenclosed chambers with a CO₂ enrichment of 35, 155, 400 or 675µmol CO₂ mol1. CO₂ enrichment increased photosyntheticcapacity in the plants grown at either of the two highest levelsof pCO₂. A CO₂ enrichment of 675µmol CO₂ caused a significantincrement of shoot dry weight, whereas no changes were observedin fresh weight, chlorophyll or protein levels. At a light intensityof 860µmol m^–2s^–1 CO₂ enrichment caused photosyntheticcapacity to increase by 250%, whereas no effect was observedat 80 µmol m^–2 s^–1. Over time, photosynthesisdecreased by 70% independent of CO₂. A time-dependent increasein the level of extractable fructose was observed whereas totalextractable carbohydrate only changed slightly. Key words: Carbohydrates, CO₂ enrichment, Hordeum vulgare, photosynthesis, respiration     

Estimation of an effective soil water potential at the root surface of transpiring plants

Abstract. A simple method is described for estimating an average of 'effective' soil water potential at the root surface for transpiring plants. The method is based on measurements of leaf water potential and leaf conductance to water vapour in stressed plants and in well-watered controls, and uses the simple Ohm's law analogue for water flow in the soil-plant system. The technique is applied to data for field-grown apple trees and to previously published data for wheat and cowpea.     

Evidence against a direct antimicrobial role of H2O2 in the infection of plants by Erwinia chrysanthemi

We have investigated the role of bacterial resistance to oxidative stress in pathogenesis. The oxyR gene from the pathogenic bacterium Erwinia chrysanthemi has been characterized. It is closely related to that found in Escherichia coli (88% overall amino acid identity). An E. chrysanthemi oxyR mutant strain was constructed by marker exchange. After induction with a sublethal dose of H2O2, this mutant was more sensitive to H2O2 and showed reduced levels of catalase and glutathione reductase activities, compared with the wild type. The oxyR mutant was unable to form individual colonies on agar plates unless catalase was added exogenously. However, it retained full virulence in potato tubers and tobacco leaves. These results suggest that the host-produced H2O2 has no direct antimicrobial effect on the interaction of E. chrysanthemi with the two plant species.     

TNF-alpha and H2O2 induce IL-18 and IL-18R beta expression in cardiomyocytes via NF-kappa B activation

Myocardial ischemia/reperfusion is characterized by oxidative stress and induction of proinflammatory cytokines. Interleukin (IL)-18, a member of the IL-1 family, acts as a proinflammatory cytokine, and is induced during various immune and inflammatory disorders. Therefore, in the present study we investigated whether IL-18 expression is regulated by cytokines and oxidative stress in cardiomyocytes. TNF-alpha induced rapid and sustained activation of NF-kappaB whereas H(2)O(2) induced delayed and transient activation. Both TNF-alpha and H(2)O(2) induced IL-18 mRNA and precursor protein in cardiomyocytes, and IL-18 release into culture supernatants. However, only TNF-alpha led to sustained expression. Expression of IL-18Rbeta, but not alpha, was induced by both agonists. TNF-alpha and H(2)O(2) induced delayed expression of IL-18 BP. Pretreatment with PDTC attenuated TNF-alpha and H(2)O(2) induced IL-18 and IL-18Rbeta, but not basal expression of IL-18Ralpha. These results indicate that adult cardiomyocytes express IL-18 and its receptors, and proinflammatory cytokines and oxidative stress regulate their expression via activation of NF-kappaB. Presence of both ligand and receptors suggests IL-18 impacts myocardial biology through an autocrine pathway.