Multivariate Patterns of Antioxidative and Photoprotective Defence Compounds in Spruce Needles at Two Central European Forest Sites of Different Elevation

In this work we measured a set of antioxidative and photoprotective compounds (chlorophylls, carotenoids, tocopherol, ascorbate and glutathione), which were suggested previously as stress markers in conifer needles, at two spruce forest sites at different elevation in Saxony, Germany. Most variables differed significantly between current and 1-year-old needles, but only the content of the xanthophyll cycle per mg total chlorophyll and the oxidation state of glutathione were significantly different between the sites. We applied principal component analysis (PCA) to address the question if underlying accumulated variables are similar to the ones found in spruce needles across Alpine elevation profiles and/or for pines in Mediterranean ecosystems. Four principal components (accumulated variables, PC) representing 68% of the total variance of the dataset were extracted. PC 1 encompassed total chlorophyll, lutein, and β-carotene contents, PC 2 combined the epoxidation state of xanthophylls, ascorbate content and redox state, and glutathione content, PC 3 represented the content of xanthophylls and the redox state of glutathione, and PC 4 encompassed the content of α-carotene and the epoxidation state of xanthophylls. Only PC 3 was significantly different between sites. The PCA structure shows many similarities to corresponding findings in studies on spruce in mountain forests in the Alps and pines in Mediterranean systems. This corroborates the interpretation of PCs as indicative for underlying physiological processes. However, separation of the two investigated sites by PCs was in the present case study not superior to the separation by single input variables.     

Development of enzyme-linked immunosorbent assay (ELISA) for glutathione S-transferase (GST-S) protein in the intertidal copepod Tigriopus japonicus and its application for environmental monitoring

To utilize the GST-S protein as a useful biomarker for environmental contamination, we developed a polyclonal antibody-based enzyme-linked immunosorbent assay (ELISA) in the intertidal copepod Tigriopus japonicus. Two polyclonal antibodies, TJ-GST-S1 and TJ-GST-S2, were raised against two TJ-GST-S synthetic peptides. Also a recombinant TJ-GST-S protein was purified as a standard for ELISA development. Each polyclonal antibody was tested by Western blot analysis and indirect ELISA. Of two polyclonal antibodies, TJ-GST-S2 ELISA was further employed due to its wide range of detection and the limit of specificity compared to those of TJ-GST-S1 ELISA system. After exposure to 4 metals (Ag, As, Cd, and Cu) to T. japonicus, the amount of TJ-GST-S protein was significantly elevated in a concentration-dependent manner. Also, TJ-GST-S protein was upregulated at relative high concentrations of B[α]P, PCB, and TBT. In this paper, we suggest that T. japonicas ELISA for TJ-GST-S2 is useful as a potential indicator system for marine contaminants.     

Effect of municipal waste water effluent upon the expression of Glutathione S-transferase isoenzymes of brine shrimp Artemia

Multiple isoenzymes of the detoxification enzyme family Glutathione S-transferase are expressed in the brine shrimp Artemia. The number of the major ones detected in crude extract by means of chromatofocusing varied between three and four, depending on the age. Two isoenzymes, one alkaline and one neutral (with corresponding isoelectric points of 8.5 and 7.2) appear to be dominant in all three developmental stages studied, (24, 48, and 72 h after hatching). Culturing Artemia for 48 h after hatching, in artificial sea water prepared by municipal wastewater effluent resulted to significant alterations of the isoenzyme profile. In comparison to organisms cultured for the same period of time in artificial sea water prepared by filtered tap water, the expression of the alkaline isoenzyme decreased by 62% while that of the neutral isoenzyme increased by 58%. Furthermore, the enzyme activity of the major isoenzyme of the acidic area increased by more than two folds. It is worth mentioning that although the specific activity of the total enzyme in the whole body homogenate was elevated, no statistically significant alteration of the Km value was observed. These findings suggest that study of the isoenzyme profile of Glutathione S-transferase may offer high sensitivity in detecting environmental pollution and needs to be further investigated.     

Bioaccumulation and function analysis of glutathione S-transferase isoforms in Manila clam Ruditapes philippinarum exposed to different kinds of PAHs

This study analyzed the function of different glutathione S-transferase(GST) isoforms and detoxification metabolism responses in Manila clam, Ruditapes philippinarum, exposed to 4 kinds of polycyclic aromatic hydrocarbons(PAHs) single, and their mixtures for 15 days under laboratory conditions. 13 kinds of GSTs in R. philippinarum were classified, and the results of tissue distribution indicated that 12 kinds of GSTs(except GST sigma 3) expressed most in digestive glands. We detected the m RNA e...     

Antioxidant enzyme activities of <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> in response to nonylphenols and degradation of nonylphenols by <Emphasis Type="Italic">M. aeruginosa</Emphasis>

The aim of this study was to examine the effects of chemical nonylphenols (NPs) on the antioxidant system of Microcystis aeruginosa strains. The degradation and sorption of NPs by M. aeruginosa were also evaluated. High concentrations of NPs (1 and 2 mg/l) were found to cause increases in superoxidase dismutase (SOD) and glutathione-S-transferase (GST) activities and in glutathione (GSH) levels. These results suggest that toxic stress manifested by elevated SOD and GST levels and GSH contents may be responsible for the toxicity of NPs to M. aeruginosa and that the algal cells could improve their antioxidant and detoxification ability through the enhancement of enzymatic and nonenzymatic prevention substances. The observed elevations in GSH levels and GST activities were relatively higher than those in SOD activities, indicating that GSH and GST contributed more in eliminating toxic effects than SOD. Low concentrations of NPs (0.05–0.2 mg/l) enhanced cell growth and decreased GST activity in algal cells of M. aeruginosa, suggesting that NPs may have acted as a protecting factor, such as an antioxidant. The larger portion of the NPs (>60%) disappeared after 12 days of incubation, indicating the strong ability of M. aeruginosa to degrade the moderate persistent NP compounds. The sorption ratio of M. aeruginosa after a 12-day exposure to low nominal concentrations of NPs (0.02–0.5 mg/l) was relatively high (>30%). The fact that M. aeruginosa effectively resisted the toxic effects of NPs and strongly degraded these pollutants indicate that M. aeruginosa cells have a strong ability to adapt to variations in environmental conditions and that low and moderate concentrations of organic compounds may favor its survival. Further studies are needed to provide detailed information on the fate of persistent organic pollutants and the survival of algae and to determine the possible role of organic pollutants in the occurrence of water blooms in eutrophic lakes.     

Cadmium-induced toxicity reversal by zinc in Ceratophyllum demersum L. (a free floating aquatic macrophyte) together with exogenous supplements of amino- and organic acids

This paper analyzes the detoxification mechanisms adopted by amino- and organic acids to alleviate Cd toxicity. In addition, with our published data on Zn–Cd interactions, the influence of Zn (200 μM) supplements on the detoxification mechanisms of amino- and organic acids have also been studied. The experimental studies on metal uptake, lipid peroxidation levels, estimation of reduced and oxidized glutathione levels as well as γ-glutamylcysteine synthetase activity in amino acid supplemented Cd treatments indicated glutathione-mediated detoxification system, which was also enhanced by Zn (200 μM) supplements. However Zn did not aid in glutathione synthesis, but maintained the ratio of reduced and oxidized forms. The supplementation of organic acids to Cd treatments indicated detoxification through the mechanism of chelation. Zn seemed to be less influential on organic acids-mediated detoxification mechanism as compared to amino acid mediated detoxification system.     

水体中3种磺胺类药物对斑马鱼的生态毒性效应

采用斑马鱼作为模式动物,针对斑马鱼的代谢能力,进行3种磺胺类药物(磺胺甲恶唑,磺胺嘧啶,磺胺二甲嘧啶)的暴露实验。实验选取谷胱甘肽s-转移酶(GST)和丙二醛(MDA)含量作为评判斑马鱼代谢能力的指标。在实验过程中,随着暴露时间的改变,GST活性和MDA含量都呈现出不同的变化。在实验前3 d,GST酶活性在不同浓度下都呈现出显著增长。但是MDA的含量在第1天达到顶峰,随后都呈现出下降趋势。由结果可得,MDA含量指标对磺胺类药物的毒性更加敏感,其指示作用更优于GST活性指标。同时,实验结果中可以看出,在磺胺嘧啶作用下的GST酶活性和MDA含量都呈现更加明显的变化,因此实验推断磺胺嘧啶会对水环境具有更潜在的危害。     

Response of antioxidant enzymes in Nicotiana tabacum clones during phytoextraction of heavy metals

Background, aim, and scope  Tobacco, Nicotiana tabacum, is a widely used model plant for growth on heavy-metal-contaminated sites. Its high biomass and deep rooting system make it interesting for phytoextraction. In the present study, we investigated the antioxidative activities and glutathione-dependent enzymes of different tobacco clones optimized for better Cd and Zn accumulation in order to characterize their performance in the field. Main features  The improved heavy metal resistance also makes the investigated tobacco clones interesting for understanding the plant defense enzyme system in general. Freshly harvested plant material (N. tabacum leaves) was used to investigate the antioxidative cascade in plants grown on heavy metal contaminated sites with and without amendments of different ammonium nitrate and ammonium sulfate fertilizers. Materials and methods  Plants were grown on heavily polluted soils in north-east Switzerland. Leaves were harvested at the field site and directly deep frozen in liquid N₂. Studies were concentrated on the antioxidative enzymes of the Halliwell–Asada cycle, and spectrophotometric measurements of catalase (CAT, EC 1.11.1.6), ascorbate peroxidase (APX, EC 1.11.1.11), superoxide dismutase (SOD, EC 1.15.1.1), glutathione peroxidase (GPX, EC 1.11.1.9), glutathione reductase (GR, EC 1.6.4.2), glutathione S-transferase (GST, EC 2.5.1.18) were performed. Results and discussion  We tried to explain the relationship between fertilizer amendments and the activity of the enzymatic defense systems. When tobacco (N. tabacum) plants originating from different mutants were grown under field conditions with varying fertilizer application, the uptake of cadmium and zinc from soil increased with increasing biomass. Depending on Cd and Zn uptake, several antioxidant enzymes showed significantly different activities. Whereas SOD and CAT were usually elevated, several other enzymes, and isoforms of GST were strongly inhibited. Conclusions  Heavy metal uptake represents severe stress to plants, and specific antioxidative enzymes are induced at the cost of more general reactions of the Halliwell–Asada cycle. In well-supplied plants, the glutathione level remains more or less unchanged. The lack of certain glutathione S-transferases upon exposure to heavy metals might be problematic in cases when organic pollutants coincide with heavy metal pollution. When planning phytoremediation of sites, mixed pollution scenarios have to be foreseen and plants should be selected according to both, their stress resistance and hyperaccumulative capacity.     

Spray drift of pesticides and stream macroinvertebrates: experimental evidence of impacts and effectiveness of mitigation measures

Impoverished stream communities in agricultural landscapes have been associated with pesticide contamination, but conclusive evidence of causality is rare. We address this deficiency by adopting an experimental approach to investigate the effects of the insecticides cypermethrin and chlorpyrifos on benthic macroinvertebrates. Three treatments were established and a combination of biomarker, bioassay and biomonitoring approaches was employed to investigate, individual, population and community-level effects. Animals deployed during pesticide application had altered enzyme activity, depressed feeding rate and reduced survival, but these effects were only observed where pesticide was sprayed to the stream edge. There were no clear pesticide-related effects on macroinvertebrate community structure or on the population densities of individual species. Hence, short-term pesticide exposure did cause individual-level effects in stream macroinvertebrates, but these were not translated to effects at the population or community-level and were effectively mitigated by the adoption of a no-spray buffer zone.