Biochemical changes in barley plants after excessive supply of copper and manganese

The present study was undertaken to identify changes in some important proteins involved in CO2 fixation (Rubisco, Rubisco activase (RA), Rubisco binding protein (RBP)), NH4+ assimilation (glutamine synthetase (GS) and glutamate synthase (GOGAT)), using immunoblotting, and in the antioxidative defense as a result of Cu or Mn excess in barley leaves (Hordeum vulgare L. cv. Obzor). Activities and isoenzyme patterns of superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and catalase (CAT), as well as the levels of ascorbate (ASC), non-protein sulfhydryl groups, hydrogen peroxide and oxidative damage to proteins were determined. Data were correlated to the accumulation of Cu or Mn in the leaves after 5 days supply of heavy metal (HM) excess in the nutrient solution. In the highest Cu excess (1500 μM), Rubisco LS and SS were reduced considerably whereas under the highest Mn concentrations (18,300 μM) only minor changes in Rubisco subunits were detected. The RBP was diminished under the highest concentrations of both Cu or Mn. The bands of RA changed differently comparing Cu and Mn toxicity. GS decreased and GOGAT was absent under the highest concentration of Cu. At Mn excess Fd-GOGAT diminished whereas GS was not apparently changed. The development of toxicity symptoms corresponded to an accumulation of Cu or Mn in the leaves and to a gradual increase in protein carbonylation, a lower SOD activity and elevated CAT and GPX activities. APX activity was diminished under Mn toxicity and was not changed under Cu excess. Generally, changes in the isoenzyme profiles were similar under both toxicities. An accumulation of H2O2 was observed only at Mn excess. Contrasting changes in the low-molecular antioxidants were detected when comparing both toxicities. Cu excess affected mainly the non-protein SH groups, while Mn influenced the ASC content. Oxidative stress under Cu or Mn toxicity was most probably the consequence of depletion in low-molecular antioxidants as a result of their involvement in detoxification processes and disbalance in antioxidative enzymes. The link between heavy metal accumulation in leaves, leading to different display of oxidative stress, and changes in individual chloroplast proteins is discussed in the article.

Biotic and abiotic controls of nitrogen and phosphorus cycling in Central European forests

The functioning and services of Central European forests are threatened by global change and a loss of biodiversity. Nutrient cycling as a key forest function is affected by biotic drivers (e.g., dominant tree species, understory plants, soil organisms) that interact with abiotic conditions (e.g., climate, soil properties). In contrast to grassland ecosystems, evidence for the relationship of nutrient cycles and biodiversity in forests is scarce because the structural complexity of forests limits experimental control of driving factors. Alternatively, observational studies along gradients in abiotic conditions and biotic properties may elucidate the role of biodiversity for forest nutrient cycles. This thesis aims to improve the understanding of the functional importance of biodiversity for nutrient cycles in forests by analyzing water-bound fluxes of nitrogen (N) and phosphorus (P) along gradients in biodiversity in three regions of Germany. The tested hypotheses included: (1) temperate forest canopies retain atmospheric N and retention increases with increasing plant diversity, (2) N release from organic layers increases with resource availability and population size of decomposers but N leaching decreases along a gradient in plant diversity, (3) P leaching from forest canopies increases with improved P supply from recalcitrant P fractions by a more diverse ectomycorrhizal fungal community. In the canopies of 27 forest stands from three regions, 16 % to 51 % of atmospheric N inputs were retained. Regional differences in N retention likely resulted from different in N availability in the soil. Canopy N retention was greater in coniferous than in beech forests, but this was not the case on loessderived soils. Nitrogen retention increased with increasing tree and shrub diversity which suggested complementary aboveground N uptake. The strength of the diversity effect on canopy N uptake differed among regions and between coniferous and deciduous forests. The N processing in the canopy directly coupled back to N leaching from organic layers in beech forests because throughfall-derived N flushed almost completely through the mull-type organic layers at the 12 studied beech sites. The N release from organic layers increased with stand basal area but was rather low (< 10 % of annual aboveground litterfall) because of a potentially high microbial N immobilization and intensive incorporation of litter into the mineral soil by bioturbation. Soil fauna biomass stimulated N mineralization through trophic interactions with primary producers and soil microorganisms. Both gross and net leaching from organic layers decreased with increasing plant diversity. Especially the diversity but not the cover of herbs increased N uptake. In contrast to N, P was leached from the canopy. Throughfall-derived P was also flushed quickly through the mull-type organic layers and leached P was predominantly immobilized in non directly plant-available P fractions in the mineral soil. Concentrations of plant-available phosphate in mineral soil solution were low and P leaching from the canopy increased with increasing concentrations of the moderately labile P fraction in soil and increasing ectomycorrhiza diversity while leaf C:P ratios decreased. This suggested that tree P supply benefited from complementary mining of diverse mycorrhizal communities for recalcitrant P. Canopy P leaching increased in years with pronounced spring drought which could lead to a deterioration of P supply by an increasing frequency of drought events. This thesis showed that N and P cycling in Central European forests is controlled by a complex interplay of abiotic site conditions with biological processes mediated by various groups of organisms, and that diverse plant communities contribute to tightening the N cycle in Central European forests and that diverse mycorrhizal communities improve the limited P availability. Maintaining forest biodiversity seems essential to ensure forest services in the light of environmental change.

Sensitivity and Fragmentation Calibration of the ROSINA Reflectron-type Time-Of-Flight Mass Spectrometer

The goal of this work was to increase the performance and to calibrate one of the ROSINA sensors, the Reflectron-type Time-Of-Flight mass spectrometer, currently flying aboard the ESA Rosetta spacecraft. Different optimization techniques were applied to both the lab and space models, and a static calibration was performed using different gas species expected to be detected in the vicinity of comet 67P/Churyumov-Gerasimenko. The database thus created was successfully applied to space data, giving consistent results with the other ROSINA sensors.

Computational Comparison of Continuous and Discontinuous Galerkin Time-Stepping Methods for Nonlinear Initial Value Problems

This article centers on the computational performance of the continuous and discontinuous Galerkin time stepping schemes for general first-order initial value problems in R n , with continuous nonlinearities. We briefly review a recent existence result for discrete solutions from [6], and provide a numerical comparison of the two time discretization methods.

Expression of a bacterial serine acetyltransferase in transgenic potato plants leads to increased levels of cysteine and glutathione

The coding sequence of the wild-type, cys-sensitive, cysE gene from Escherichia coli, which encodes an enzyme of the cysteine biosynthetic pathway, namely serine acetyltransferase (SAT, EC 2.3.1.30), was introduced into the genome of potato plants under the control of the cauliflower mosaic virus 35S promoter. In order to target the protein into the chloroplast, cysE was translationally fused to the 5′-signal sequence of rbcS from Arabidopsis thaliana. Transgenic plants showed a high accumulation of the cysE mRNA. The chloroplastic localisation of the E. coli SAT protein was demonstrated by determination of enzymatic activities in enriched organelle fractions. Crude leaf extracts of these plants exhibited up to 20-fold higher SAT activity than those prepared from wild-type plants. The transgenic potato plants expressing the E. coli gene showed not only increased levels of enzyme activity but also exhibited elevated levels of cysteine and glutathione in leaves. Both were up to twofold higher than in control plants. However, the thiol content in tubers of transgenic lines was unaffected. The alterations observed in leaf tissue had no effect on the expression of O-acetylserine(thiol)-lyase, the enzyme which converts O-acetylserine, the product of SAT, to cysteine. Only a minor effect on its enzymatic activity was observed. In conclusion, the results presented here demonstrate the importance of SAT in plant cysteine biosynthesis and show that production of cysteine and related sulfur-containing compounds can be enhanced by metabolic engineering.

Real-time analysis of δ13C- and δD-CH4 in ambient air with laser spectroscopy: method development and first intercomparison results

In situ and simultaneous measurement of the three most abundant isotopologues of methane using mid-infrared laser absorption spectroscopy is demonstrated. A field-deployable, autonomous platform is realized by coupling a compact quantum cascade laser absorption spectrometer (QCLAS) to a preconcentration unit, called trace gas extractor (TREX). This unit enhances CH4 mole fractions by a factor of up to 500 above ambient levels and quantitatively separates interfering trace gases such as N2O and CO2. The analytical precision of the QCLAS isotope measurement on the preconcentrated (750 ppm, parts-per-million, µmole mole−1) methane is 0.1 and 0.5 ‰ for δ13C- and δD-CH4 at 10 min averaging time. Based on repeated measurements of compressed air during a 2-week intercomparison campaign, the repeatability of the TREX–QCLAS was determined to be 0.19 and 1.9 ‰ for δ13C and δD-CH4, respectively. In this intercomparison campaign the new in situ technique is compared to isotope-ratio mass spectrometry (IRMS) based on glass flask and bag sampling and real time CH4 isotope analysis by two commercially available laser spectrometers. Both laser-based analyzers were limited to methane mole fraction and δ13C-CH4 analysis, and only one of them, a cavity ring down spectrometer, was capable to deliver meaningful data for the isotopic composition. After correcting for scale offsets, the average difference between TREX–QCLAS data and bag/flask sampling–IRMS values are within the extended WMO compatibility goals of 0.2 and 5 ‰ for δ13C- and δD-CH4, respectively. This also displays the potential to improve the interlaboratory compatibility based on the analysis of a reference air sample with accurately determined isotopic composition.

Expression and distribution of a vacuolar aquaporin in young and mature leaf tissues of Brassica napus in relation to water fluxes

Recently, it has been shown that water fluxes across biological membranes occur not only through the lipid bilayer but also through specialized water-conducting proteins, the so called aquaporins. In the present study, we investigated in young and mature leaves of Brassica napus L. the expression and localization of a vacuolar aquaporin homologous to radish γ-tonoplast intrinsic protein/vacuolar-membrane integral protein of 23 kDa (TIP/VM 23). In-situ hybridization showed that these tonoplast aquaporins are highly expressed not only in developing but also in mature leaves, which export photosynthates. No substantial differences could be observed between different tissues of young and mature leaves. However, independent of the developmental stage, an immunohistochemical approach revealed that the vacuolar membrane of bundle-sheath cells contained more protein cross-reacting with antibodies raised against radish γ-TIP/VM 23 than the mesophyll cells. The lowest labeling was detected in phloem cells. We compared these results with the distribution of plasma-membrane aquaporins cross-reacting with antibodies detecting a domain conserved among members of the plasma-membrane intrinsic protein 1 (PIP1) subfamily. We observed the same picture as for the vacuolar aquaporins. Furthermore, a high density of gold particles labeling proteins of the PIP1 group could be observed in plasmalemmasomes of the vascular parenchyma. Our results indicate that γ-TIP/VM 23 and PIP1 homologous proteins show a similar expression pattern. Based on these results it is tempting to speculate that bundle-sheath cells play an important role in facilitating water fluxes between the apoplastic and symplastic compartments in close proximity to the vascular tissue.

Redistribution of cobalt and nickel in detached wheat shoots: effects of steam-girdling and of cobalt and nickel supply

Detached wheat shoots (ear with peduncle and flag leaf) were incubated for 4 d in a solution containing 1 mM RbCl and 1 mM SrCl2 as well as 10, 40 or 160 µM NiCl2 and CoCl2. The phloem of some plants was interrupted by steam-girdling the stem below the ear to distinguish between xylem and phloem transport. The phloem-immobile Sr flowed mainly to the leaf lamina and to the glumes via the xylem. The Sr transport was not sensitive to steam-girdling. In contrast, the phloem-mobile Rb accumulated during the incubation time mainly in the stem and the leaf sheath. The Rb transport to the grains was impaired by steam-girdling as well as by elevated Ni and Co concentrations in the incubation solution indicating that Rb was transported via the phloem to the maturing grains and that this transport was affected by the heavy metals. Ni was removed more efficiently from the xylem in the peduncle than Co (but far less efficiently than Rb). It became evident that the two heavy metals can also be transferred from the xylem to the phloem in the stem of wheat and reach the maturing grains via the phloem.

Children's Perceptions of Male and Female Athletes as Presented in Sports Illustrated for Kids

Despite gains made by Title IX in the past 36 years, including increased female participation in high school and collegiate sport, there is evidence that gender equity in sport is not fully achieved. Researchers target the media because they tend to shape social values and disseminate information to the masses (Kane, 1978, in Fink & Kensicki, 2002). As sports become more pervasive, framing theory has become particularly relevant. The purpose of this study is to build on the Hardin et al. (2002) study by examining the relationship among media sports coverage, gender equity in sport and the perceptions young sports fans begin to form about gender and sport based on media consumption. The researcher hypothesized that since women face discrimination in sport starting from the time that they choose to participate, children will perceive male athletes and their sports as more legitimate. Additionally, the media play a major role in shaping the views of audiences, so the way that they represent male and female athletes, including juxtaposing them, may have an impact on children. The researcher conducted a content analysis of 24 Sports Illustrated for Kids issues from 1996 to 1999 and 24 issues from 2006 to 2007. The researcher analyzed the content of photographs (N=3219) and of headlines (N=762) by using the definitions determined by Hardin et al. (2002). We found that there is a disproportionate amount of coverage devoted to male athletes and that the discrepancy between media representation between men and women in sport has grown since the mid-1990s. This study also includes a focus group conducted with three children from a community swimming program in a northeastern town and found that those children were acutely aware of the differences between men and women in sport based on the discussion. The researcher does not attempt to find a causal relationship between these children’s perceptions and the way media represents them, but rather uses the focus group to complement the content analysis. As children become sports consumers in later life, future research exploring the relationship between children’s perceptions and the media’s representations need to be done before causality and the significance of media effects are determined.

Expanding Learning Time in Schools: Considering the Challenges of Implementation and the Potential Impact

Considering the broader context of school reform that is seeking education strategies that might deliver substantial impact, this article examines four questions related to the policy and practice of expanding learning time: (a) why do educators find the standard American school calendar insufficient to meet students’ educational needs, especially those of disadvantaged students? (b) how do educators implement a longer day and/or year, addressing concerns about both educational quality and costs? (c) what does research report about outcomes of expanding time in schools? and (d) what are the future prospects for increasing the number of expanded-time schools? The paper examines these questions by considering research, policy, and practice at the national level and, throughout, by drawing upon additional evidence from Massachusetts, one of the leading states in the expanded-time movement. In considering the latter two questions, the article explores the knowns and unknowns related to expanded learning time and offers suggestions for further research.

Impact of Liquid Swine Manure Application and Cover Crops on Ground Water Quality

The primary objective of this project was to determine the impact of appropriate rates of swine manure applications to corn and soybeans based on nitrogen and phosphorus requirements of crops, soil phosphorus accumulation, and the potential of nitrate and phosphorus leaching to groundwater. Another purpose of this long-term experimental study was to develop and recommend appropriate manure and nutrient management practices to producers to minimize the water contamination potential and enhance the use of swine manure as inorganic fertilizer. A third component of this study was to determine the potential effects of rye as a cover crop to reduce nitrate loss to shallow ground water.

Dinoflagellate cyst assemblages, oxygen isotope records of foraminifera, and alkenone and Mg/Ca-based SST estimates for the time interval 3.40-3.18 Ma (Late Pliocene) from five North Atlantic and Caribbean sediment cores

The early Late Pliocene (3.6 to ~3.0 million years ago) is the last extended interval in Earth's history when atmospheric CO2 concentrations were comparable to today's and global climate was warmer. Yet a severe global glaciation during marine isotope stage (MIS) M2 interrupted this phase of global warmth ~3.30 million years ago, and is seen as a premature attempt of the climate system to establish an ice-age world. Our geochemical and palynological records from five marine sediment cores along a Caribbean to eastern North Atlantic transect show that increased Pacific-to-Atlantic flow via the Central American Seaway weakened the North Atlantic Current (NAC) and attendant northward heat transport prior to MIS M2. The consequent cooling of the northern high latitude oceans permitted expansion of the Greenland ice sheet during MIS M2, despite near-modern atmospheric CO2 concentrations. Before and after MIS M2, heat transport via the NAC was crucial in maintaining warm climates comparable to those predicted for the end of this century.

Stable isotope record of benthic and planktonic foraminifera in sediment cores of ODP Leg 177, Southern Ocean

While onboard ship during Leg 177, we used variations in sediment physical properties (mainly percent color reflectance) in conjunction with biomagnetostratigraphy to correlate among sites and predict the position of marine isotope stages (MISs) (e.g., see fig. F11 in Shipboard Scientific Party, 1999, p. 45). Our working assumption was that physical properties of Leg 177 sediments are controlled mainly by variations in carbonate content. Previous studies of Southern Ocean sediment cores have shown that carbonate concentrations are relatively high during interglacial stages and low during glacial stages at sites located within the Polar Frontal Zone (PFZ). Today, the PFZ marks a lithologic boundary in underlying sediment separating calcareous oozes to the north and silica-rich facies to the south (Hays et al., 1976). Although there is debate whether the position of the "physical" PFZ actually moved during glacial-interglacial cycles (Charles and Fairbanks, 1990; Matsumoto et al., 2001), the "biochemical" PFZ, as expressed by the CaCO3/opal boundary in sediments, certainly migrated north during glacials and south during interglacials. This gave rise to lithologic variations that are useful for stratigraphic correlation. At Leg 177 sites located north of the PFZ and at sublysoclinal depths, we expected the same pattern of carbonate variation because cores in the Atlantic basin are marked by increased carbonate dissolution during glacial periods and increased preservation during interglacials (Crowley, 1985).

(Table 1) Uranium and Thorium concentrations, activity ratios and calcium carbonate content of DSDP Site 54-424

The geochemical behaviour of uranium and thorium in metalliferous sediments and hydrothermal deposits has been widely studied and the main results have been summarised by Boström and Rydell. These isotopes may be used to clarify how the metal-rich solutions are introduced into sediment cover and seawater. Using radiochemistry followed by alpha spectrometry, we have measured uranium concentrations as high as several hundred p.p.m., which must clearly be associated with ocean ridge thermal activity, in sediments interbedded between the basaltic basement and the green hydrothermal mud at DSDP Site 424. These high uranium concentrations indicate the path followed by the hydrothermal fluid which, debouching at the sediment-water interface, formed the green mud.