Salicylic acid involved in the regulation of nutrient elements uptake and oxidative stress in Vallisneria natans (Lour.) Hara under Pb stress

In this study, the alterations in nutrient elements content, reactive oxygen species level and antioxidant response were studied in leaves of Vallisneria natans (Lour.) Hara exposed to salicylic acid (SA, 10 or 100 μM), or Pb (50 μM) or their combinations for 4 d. No significant alterations in Mn and Ca content were observed but content of Cu, Zn, Fe and P decreased in plants exposed to SA alone. SA application inhibited the uptake of Pb and partially reversed Pb-induced the alterations in Mn, Ca and Fe content in leaves of V. natans exposed to 50 μM Pb. The decreased chlorophyll (a + b) and increased malondialdehyde and O²⁻ and H₂O₂ content were detected in plants exposed to 100 μM SA, 50 μM Pb, 10 μM SA + 50 μM Pb or 100 μM SA + 50 μM Pb. Application SA partially inhibited Pb-induced the increase of malondialdehyde, O²⁻ and H₂O₂ content. 100 μM SA decreased the activity of NADH oxidase and the content of non-protein thiols, carotenoids and ascorbic acid and increased the content of dehydroascorbate in plants treated with or without Pb. SA alone decreased the ascorbate peroxidase activity and increased the catalase and peroxidase activity, while SA application increased catalase activity but had no significant effect on ascorbate peroxidase and peroxidase activity in V. natans exposed to Pb. The results indicate that SA involves in the regulation of Pb uptake, nutrient balance and oxidative stress.     

Investigation of low-level <Superscript>242</Superscript>Pu contamination on nutrition disturbance and oxidative stress in <Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.

Plutonium associated with higher molecular weight molecules is presumed to be poorly mobile and hardly plant available. In our present study, we investigate the uptake and effects of Pu treatments on Solanum tuberosum plants in amended Hoagland medium at concentrations of [²⁴²Pu] = 100 and 500 nm, respectively. We found a direct proof of oxidative stress in the plants caused by these rather low concentrations. For the confirmation of oxidative stress, we explored the production of nitric oxide (NO) and hydrogen peroxide (H₂O₂) by epifluorescence microscopy. Oxidative stress markers like lipid peroxidation and superoxide radicals (O₂ ^??) are monitored through histochemical analysis. The biochemical parameters i.e. chlorophyll and carotenoids are measured as an indicator of cellular damage in the tested plants including the enzymatic parameters such as catalase and glutathione reductase. From our work, we conclude that Pu in low concentration has no significant effects on the uptake of many trace and macroelements. In contrast, the content of O₂ ^?? , malondialdehyde (MDA), and H₂O₂ increases with increasing Pu concentration in the solution, while the opposite effects was found for NO, catalase, and glutathione reductase. These findings prove that even low concentration of Pu regulates ROS production and generate oxidative stress in S. tuberosum L.     

The efficiency of tobacco Bel-W3 and native species for ozone biomonitoring in subtropical climate, as revealed by histo-cytochemical techniques

We aimed to verify whether hydrogen peroxide (H₂O₂) accumulation and cell death are detected early in three bioindicators of ozone (O₃), Nicotiana tabacum ‘Bel-W3’, Ipomoea nil ‘Scarlet O’Hara’ and Psidium guajava ‘Paluma’, and whether environmental factors also affect those microscopic markers. The three species were exposed to chronic levels of O₃ in a subtropical area and a histo-cytochemical technique that combines 3,3′-diaminobenzidine (DAB) with Evans blue staining was used in the assessments. The three species accumulated H₂O₂, but a positive correlation with O₃ concentration was only observed in N. tabacum. A positive correlation between O₃ and cellular death was also observed in N. tabacum. In I. nil and P. guajava, environmental factors were responsible for symptoms at the microscopic level, especially in P. guajava. We conclude that the most appropriate and least appropriate bioindicator plant for O₃ monitoring in the subtropics are N. tabacum ‘Bel-W3’ and P. guajava ‘Paluma’, respectively.     

Ozone affects pollen viability and NAD(P)H oxidase release from Ambrosia artemisiifolia pollen

Air pollution is frequently proposed as a cause of the increased incidence of allergy in industrialised countries. We investigated the impact of ozone (O₃) on reactive oxygen species (ROS) and allergen content of ragweed pollen (Ambrosia artemisiifolia). Pollen was exposed to acute O₃ fumigation, with analysis of pollen viability, ROS and nitric oxide (NO) content, activity of nicotinamide adenine dinucleotide phosphate (NAD[P]H) oxidase, and expression of major allergens. There was decreased pollen viability after O₃ fumigation, which indicates damage to the pollen membrane system, although the ROS and NO contents were not changed or were only slightly induced, respectively. Ozone exposure induced a significant enhancement of the ROS-generating enzyme NAD(P)H oxidase. The expression of the allergen Amb a 1 was not affected by O₃, determined from the mRNA levels of the major allergens. We conclude that O₃ can increase ragweed pollen allergenicity through stimulation of ROS-generating NAD(P)H oxidase.     

The use of experimental data and their uncertainty for assessing ozone photochemistry in the Eastern Iberian Peninsula

Observation-based methods are useful tools to explore the sensitivity of ozone concentrations to precursor controls. With the aim of assessing the ozone precursor sensitivity in two locations: Paterna (suburban) and Villar del Arzobispo (rural) of the Turia river basin in the east of Spain, the photochemical indicator O₃/NO_y and the Extent-of-Reaction (EOR) parameter have been calculated from field measurements. In Paterna, the O₃/NO_y ratio varied from 0 to 13 with an average value of 5.1 (SD 3.2), whereas the averaged value for the EOR was 0.43 (SD 0.14). In Villar del Arzobispo, the O₃/NO_y ratio changed from 5 to 30 with a mean value of 13.6 (SD 4.7) and the EOR gave an averaged value of 0.72 (SD 0.11). The results show two different patterns of ozone production as a function of the location. The suburban area shows a VOC-sensitive regime whereas the rural one shows a transition regime close to NO_x-sensitive conditions. No seasonal differences in these regimes are observed along the monitoring campaigns. Finally, an analysis of the influence of the measurement quality of NO_y, NO_x and O₃ on the uncertainty of the O₃/NO_y ratio and the EOR was performed showing that the uncertainty of O₃/NO_y is not dependent on either its value or the individual values of O₃ and NO_y but just on the quality of O₃ and NO_y measurements. The maximum uncertainty is 26% as long as the combined uncertainties of O₃ and NO_y remain below the 7.5%. The case of the EOR is different and its uncertainty depends on both the value of the EOR parameter and the individual concentration values of NO_y and NO_x. The uncertainty of the EOR estimation can be very high (>200%) if the combined uncertainties of both NO_y and NO_x are high (>7.5%), or especially, if u(NO_y) and u(NO_x) differ considerably from each other (>3.5%).     

Enhanced ozone strongly reduces carbon sink strength of adult beech (Fagus sylvatica) - Resume from the free-air fumigation study at Kranzberg Forest

Ground-level ozone (O₃) has gained awareness as an agent of climate change. In this respect, key results are comprehended from a unique 8-year free-air O₃-fumigation experiment, conducted on adult beech (Fagus sylvatica) at Kranzberg Forest (Germany). A novel canopy O₃ exposure methodology was employed that allowed whole-tree assessment in situ under twice-ambient O₃ levels. Elevated O₃ significantly weakened the C sink strength of the tree-soil system as evidenced by lowered photosynthesis and 44% reduction in whole-stem growth, but increased soil respiration. Associated effects in leaves and roots at the gene, cell and organ level varied from year to year, with drought being a crucial determinant of O₃ responsiveness. Regarding adult individuals of a late-successional tree species, empirical proof is provided first time in relation to recent modelling predictions that enhanced ground-level O₃ can substantially mitigate the C sequestration of forests in view of climate change.     

Apoplastic ascorbate contributes to the differential ozone sensitivity in two varieties of winter wheat under fully open-air field conditions

We studied leaf apoplastic ascorbates in relation to ozone (O₃) sensitivity in two winter wheat (Triticum aestivum L.) varieties: Yangfumai 2 (Y2) and Yangmai 16 (Y16). The plants were exposed to elevated O₃ concentration 27% higher than the ambient O₃ concentration in a fully open-air field from tillering stage until final maturity. The less sensitive variety Y16 had higher concentration of reduced ascorbate in the apoplast and leaf tissue by 33.5% and 12.0%, respectively, than those in the more sensitive variety Y2, whereas no varietal difference was detected in the decline of reduced ascorbate concentration in response to elevated O₃. No effects of O₃ or variety were detected in either oxidized ascorbate or the redox state of ascorbate in the apoplast and leaf tissue. The lower ascorbate concentrations in both apoplast and leaf tissue should have contributed to the higher O₃ sensitivity in variety Y2.     

Toxic effects of imidazolium-based ionic liquids on Caenorhabditis elegans: The role of reactive oxygen species

By using Caenorhabditis elegans (C. elegans) as a model animal, the present work is aimed to evaluate the acute toxicity of imidazolium-based bromide Ionic Liquids (ILs), and to elucidate the underlying mechanisms involved. Firstly, 24-h median lethal concentration (LC₅₀) for eight ILs with different alkyl chain lengths and one or two methyl groups in the imidazolium ring were determined to be in a range of 0.09–6.64 mg mL⁻¹. Four ILs were selected to investigate the toxic mechanisms. Mortality, levels of reactive oxygen species (ROS), lipofuscin accumulation and expression of superoxide dismutase 3 in C. elegans were determined after exposed to ILs at sub-lethal concentrations for 12 h. A significant increase in the levels of these biomarkers was observed in accordance with the results of 12-h lethality assay. The addition of 0.5% dimethyl sulfoxide, which acts as a radical scavenger, remarkably rescued the lethality of C. elegans and significantly decreased the ROS level in C. elegans. Our results suggest that ROS play an important role in IL-induced toxicity in C. elegans.     

Effect of 3 years' free-air exposure to elevated ozone on mature Norway spruce (Picea abies (L.) Karst.) needle epicuticular wax physicochemical characteristics

We examined the effect of ozone (O₃) on Norway spruce (Picea abies) needle epicuticular wax over three seasons at the Kranzberg Ozone Fumigation Experiment. Exposure to 2× ambient O₃ ranged from 64.5 to 74.2 μl O₃ l⁻¹ h AOT40, and 117.1 to 123.2 nl O₃ l⁻¹ 4th highest daily maximum 8-h average O₃ concentration. The proportion of current-year needle surface covered by wax tubes, tube aggregates, and plates decreased (P = 0.011) under 2× O₃. Epistomatal chambers had increased deposits of amorphous wax. Proportion of secondary alcohols varied due to year (P = 0.004) and O₃ treatment (P = 0.029). Secondary alcohols were reduced by 9.1% under 2× O₃. Exposure to 2× O₃ increased (P = 0.037) proportions of fatty acids by 29%. Opposing trends in secondary alcohols and fatty acids indicate a direct action of O₃ on wax biosynthesis. These results demonstrate O₃-induced changes in biologically important needle surface characteristics of 50-year-old field-grown trees.     

Screening of Cd tolerant genotypes and isolation of metallothionein genes in alfalfa (Medicago sativa L.)

In order to evaluate Cd tolerance in wide-ranging sources of alfalfa (Medicago sativa) and to identify Cd tolerant genotypes which may potentially be useful for restoring Cd-contaminated environments, thirty-six accessions of alfalfa were screened under hydroponic culture. Our results showed that the relative root growth rate varied from 0.48 to 1.0, which indicated that different alfalfa accessions had various responses to Cd stress. The candidate fragments derived from differentially expressed metallothionein (MT) genes were cloned from leaves of two Cd tolerant genotypes, YE and LZ. DNA sequence and the deduced protein sequence showed that MsMT2a and MsMT2b had high similarity to those in leguminous plants. DDRT-PCR analysis showed that MsMT2a expressed in both YE and LZ plants under control and Cd stress treatment, but MsMT2b only expressed under Cd stress treatment. This suggested that MsMT2a was universally expressed in leaves of alfalfa but expression of MsMT2b was Cadmium (Cd) inducible.     

Catalase and ascorbate peroxidase—representative H<Subscript>2</Subscript>O<Subscript>2</Subscript>-detoxifying heme enzymes in plants

Plants have to counteract unavoidable stress-caused anomalies such as oxidative stress to sustain their lives and serve heterotrophic organisms including humans. Among major enzymatic antioxidants, catalase (CAT; EC 1.11.1.6) and ascorbate peroxidase (APX; EC 1.11.1.11) are representative heme enzymes meant for metabolizing stress-provoked reactive oxygen species (ROS; such as H₂O₂) and controlling their potential impacts on cellular metabolism and functions. CAT mainly occurs in peroxisomes and catalyzes the dismutation reaction without requiring any reductant; whereas, APX has a higher affinity for H₂O₂ and utilizes ascorbate (AsA) as specific electron donor for the reduction of H₂O₂ into H₂O in organelles including chloroplasts, cytosol, mitochondria, and peroxisomes. Literature is extensive on the glutathione-associated H₂O₂-metabolizing systems in plants. However, discussion is meager or scattered in the literature available on the biochemical and genomic characterization as well as techniques for the assays of CAT and APX and their modulation in plants under abiotic stresses. This paper aims (a) to introduce oxidative stress-causative factors and highlights their relationship with abiotic stresses in plants; (b) to overview structure, occurrence, and significance of CAT and APX in plants; (c) to summarize the principles of current technologies used to assay CAT and APX in plants; (d) to appraise available literature on the modulation of CAT and APX in plants under major abiotic stresses; and finally, (e) to consider a brief cross-talk on the CAT and APX, and this also highlights the aspects unexplored so far.     

Ozone exposure of field-grown winter wheat affects soil mesofauna in the rhizosphere

A 2-year open-top chamber experiment with field-grown winter wheat (Triticum aestivum L. cv. Astron) was conducted to examine the effects of ozone on plant growth and selected groups of soil mesofauna in the rhizosphere. From May through June in each year, plants were exposed to two levels of O₃: non-filtered (NF) ambient air or NF+ 40 ppb O₃ (NF+). During O₃ exposure, soil sampling was performed at two dates according to different plant growth stages. O₃ exposure reduced above- and below-ground plant biomass in the first year, but had little effect in the second year. The individual density of enchytraeids, collembolans and soil mites decreased significantly in the rhizosphere of plants exposed to NF+ in both years. Differences were highest around anthesis, i.e. when plants are physiologically most active. The results suggest that elevated O₃ concentrations may influence the dynamic of decomposition processes and the turnover of nutrients.     

Oxidative stress and arsenic toxicity: Role of NADPH oxidases

The effect of arsenic (25 and 50 μM As for 1 and 5 d) was analysed in wild type (WT) and Arabidopsis thaliana (L.) Heynh plants deficient in NADPH oxidase C (AtrbohC). The content of H₂O₂ and malondialdehyde (MDA) increased with the As concentration, while the opposite effect was found for NO in WT and AtrbohC plants. The As treatment reduced catalase and increased glutathione reductase activities to the same extent in WT and AtrbohC plants, although the induction of all SOD isoforms (mainly CuZn–SODs) was observed in WT plants, the opposite effects being found in AtrbohC plants. Glycolate oxidase (H₂O₂ producers) considerably increased with the concentration and time of treatment with As in WT and AtrbohC mutants. Arsenic induced the uptake and translocation of P, S, Cu, Zn, and Fe in WT plants, while in AtrbohC plants the opposite trend was noted and the uptake of As became considerably lower than in WT plants. These results suggest that As causes oxidative stress by inducing glycolate oxidase, while NADPH oxidase does not appear to participate in ROS overproduction but could be critical in regulating antioxidant defences as well as the transport and translocation of As and macro/micronutrients.     

Decrease in catalase activity of Folsomia candida fed a Bt rice diet

Here we report the effects of three Bt-rice varieties and their non-Bt conventional isolines on biological traits including survival, reproduction, and the activities of three antioxidant enzymes superoxide dismutase, catalase and peroxidase, in the Collembolan, Folsomia candida. The reproduction was significantly lower when fed Kemingdao and Huahui1 than those feeding on their non-GM near-isogenic varieties Xiushui and Minghui63 respectively, this can be explained by the differences of plant compositions depended on variety of rice. The catalase activity of F. candida was significantly lower when fed the Bt-rice variety Kemingdao compared to the near-isogenic non-Bt-rice variety Xiushui. This suggests that some Bt-rice varieties may impose environmental stress to collembolans. We emphasize that changes in activity of antioxidant enzymes of non-target organisms are important in understanding the ecological consequences for organisms inhabiting transgenic Bt-rice plantations.     

Photosynthesis, chloroplast pigments, and antioxidants in Pinus canariensis under free-air ozone fumigation

High O₃ levels, driving uptake and challenging defense, prevail on the Canary Islands, being associated with the hot and dry summers of the Mediterranean-type climate. Pinus canariensis is an endemic conifer species that forms forests across these islands. We investigated the effects of ozone on photosynthesis and biochemical parameters of P. canariensis seedlings exposed to free-air O₃ fumigation at Kranzberg Forest, Germany, where ambient O₃ levels were similar to those at forest sites in the Canary Islands. The twice-ambient O₃ regime (2xO₃) neither caused visible injury-like chlorotic or necrotic spots in the needles nor significantly affected violaxanthin, antheraxanthin and zeaxanthin levels and the de-epoxidation state of the xanthophyll cycle. In parallel, stomatal conductance for water vapour, net photosynthesis, intercellular CO₂ concentration, chlorophyll fluorescence parameters, as well as antioxidant levels were hardly affected. It is concluded that presently prevailing O₃ levels do not impose severe stress on P. canariensis seedlings.     

Belowground effects of enhanced tropospheric ozone and drought in a beech/spruce forest (Fagus sylvatica L./Picea abies [L.] Karst)

The effects of experimentally elevated O₃ on soil respiration rates, standing fine-root biomass, fine-root production and δ¹³C signature of newly produced fine roots were investigated in an adult European beech/Norway spruce forest in Germany during two subsequent years with contrasting rainfall patterns. During humid 2002, soil respiration rate was enhanced under elevated O₃ under beech and spruce, and was related to O₃-stimulated fine-root production only in beech. During dry 2003, the stimulating effect of O₃ on soil respiration rate vanished under spruce, which was correlated with decreased fine-root production in spruce under drought, irrespective of the O₃ regime. δ¹³C signature of newly formed fine-roots was consistent with the differing g_s of beech and spruce, and indicated stomatal limitation by O₃ in beech and by drought in spruce. Our study showed that drought can override the stimulating O₃ effects on fine-root dynamics and soil respiration in mature beech and spruce forests.     

Stomatal characteristics and infection biology of Pyrenopeziza betulicola in Betula pendula trees grown under elevated CO2 and O3

Two silver birch clones were exposed to ambient and elevated concentrations of CO₂ and O₃, and their combination for 3 years, using open-top chambers. We evaluated the effects of elevated CO₂ and O₃ on stomatal conductance (g_s), density (SD) and index (SI), length of the guard cells, and epidermal cell size and number, with respect to crown position and leaf type. The relationship between the infection biology of the fungus (Pyrenopeziza betulicola) causing leaf spot disease and stomatal characteristics was also studied. Leaf type was an important determinant of O₃ response in silver birch, while crown position and clone played only a minor role. Elevated CO₂ reduced the g_s, but had otherwise no significant effect on the parameters studied. No significant interactions between elevated CO₂ and O₃ were found. The infection biology of P. betulicola was not correlated with SD or g_s, but it did occasionally correlate positively with the length of the guard cells.     

Detoxification and repair process of ozone injury: from O3 uptake to gene expression adjustment

Plants react to O₃ threat by setting up a variety of defensive strategies involving the co-ordinated modulation of stress perception, signalling and metabolic responses. Although stomata largely controls O₃ uptake, differences in O₃ tolerance cannot always be ascribed to changes in stomatal conductance but cell protective and repair processes should be taken into account. O₃-driven ROS production in the apoplast induces a secondary, active, self-propagating generation of ROS, whose levels must be finely tuned, by many enzymatic and non-enzymatic antioxidant systems, to induce gene activation without determining uncontrolled cell death. Additional signalling molecules, as ethylene, jasmonic and salicylic acid are also crucial to determine the spreading and the containment of leaf lesions. The main recent results obtained on O₃ sensing, signal transduction, ROS formation and detoxification mechanisms are here discussed.     

A study of the ozone formation by ensemble back trajectory-process analysis using the Eta–CMAQ forecast model over the northeastern U.S. during the 2004 ICARTT period

The integrated process rates (IPRs) estimated by the Eta–CMAQ model at grid cells along the trajectory of the air mass transport path were analyzed to quantitatively investigate the relative importance of physical and chemical processes for O₃ formation and evolution over the northeastern U.S. during the 2004 International Consortium for Atmospheric Research on Transport and Transformation (ICARTT) period. The Hybrid Single Particle Lagrangian Integrated Trajectory (HYSPLIT) model is used to determine the back trajectory of air masses reaching the northeast by linking a downwind receptor to upwind source areas. The process analysis is applied to a high O₃ episode occurring on July 22, 2004 at three selected sites in the northeastern U.S. The process analysis at the location of the site shows that during the daytime, the O₃ concentrations in the surface layer are mainly enhanced by the vertical diffusion of O₃-rich air from aloft, followed by horizontal advection (HADV) and chemical production (CHEM), whereas dry deposition (DDEP) and vertical advection (ZADV) mainly deplete O₃ concentrations at the sites of Valley Central (VC), NY and Castle Spring (CS), NH. By integrating the effects of each process over the depth of the daytime planetary boundary layer (PBL), it was found that at the VC site, CHEM and HADV contributed about 53% and 41%, respectively, to O₃ levels within the PBL. This confirms the significance of regional transport of O₃ from the industrialized areas into the Northeast. On the other hand, the process analysis results for O₃ formation in moving air masses indicate that on July 22, large chemical production of O₃ along the transport path over the polluted urban regions leads to significant increase in O₃ in the air mass reaching the VC site, whereas the low chemical production of O₃ along the transport path over the low emission regions leads to the low O₃ concentration at the site of Belleayre Mountain (BM), NY. The dramatic buildup of O₃ concentration from 50 ppb to 102 ppb in the air masses before reaching the VC site after 12:00 EST on 7/22 indicates the significant impact of pollution from the northeastern urban corridor at this site. On the basis of the results at the CS site, it was found that high NO_x emissions along the transport path led to large chemical production of O₃ in the air mass reaching the CS site on July 22. In contrast, the low chemical production of O₃ associated with low emission (relatively clean conditions) along the transport path over the northern portions of the domain is responsible for the low O₃ concentration at the CS site on July 26.     

Cornmeal-induced resistance to ciprofloxacin and erythromycin in enterococci

Triclosan is used as an antibacterial agent in household items and personal care products. Since this compound is found in maternal milk of humans and bodies of wild animals, there is growing concern among some consumer groups and scientific community that triclosan is adverse for humans and wild animals. In order to estimate adverse actions of triclosan, the effects of triclosan on intracellular Zn²⁺ concentration and cellular thiol content were studied in rat thymocytes by the use of flow cytometer with appropriate fluorescent probes. Triclosan at 1-3 μM (sublethal concentrations) increased the intensity of FluoZin-3 fluorescence (intracellular Zn²⁺ concentration) and decreased the intensity of 5-chloromethylfluorescein (5-CMF) fluorescence (cellular thiol content). Negative correlation (r = −0.985) between triclosan-induced changes in FluoZin-3 and 5-CMF fluorescences was found. Removal of external Zn²⁺ did not significantly affect the triclosan-induced augmentation of FluoZin-3 fluorescence, suggesting an intracellular Zn²⁺ release by triclosan. These actions of triclosan were similar to those of H₂O₂ and triclosan significantly potentiated the cytotoxicity of H₂O₂. Therefore, the results may suggest that triclosan at sublethal concentrations induces oxidative stress that decreases cellular thiol content, resulting in an increase in intracellular Zn²⁺ concentration by Zn²⁺ release from intracellular store(s). Since recent studies show many physiological roles of intracellular Zn²⁺ in cellular functions, the triclosan-induced disturbance of cellular Zn²⁺ homeostasis may induce adverse actions on the cells.