Ezzeddine El Ferjani

Nickel-induced oxidative damage and antioxidant responses in Zea mays shoots

Abstract Zea mays (cv. LG 23/01) seedlings precultured in hydroponic culture for seven days were treated with 250 μM NiCl 2 for five days. Several metabolic parameters representative of oxidative damage and antioxidant activity in shoots were regularly studied after the metal treatment. Two days after Ni-application, membrane lipid peroxidation intensified. A stimulation of guaiacol peroxidase (EC 1.11.1.7) activity was recorded. Analysis of the profile of anionic isoenzymes of guaiacol peroxidase revealed quantitative changes occurring during Ni-exposure. Superoxide dismutase (EC 1.15.1.1) activity was stimulated early and transiently. Later, the activities of ascorbate peroxidase (EC 1.11.1.11), catalase (EC 1.11.1.6) and glutathione reductase (EC 1.6.4.2) increased. The results are discussed with regard to nickel-induced oxidative stress and potential antioxidant reactions.

NICKEL TOXICITY INDUCES OXIDATIVE DAMAGE IN ZEA MAYS ROOTS

Seven day-old maize (Zea mays cv. LG 23/01) plants were treated for five days with 250 μM NiCl2. The relationship between nickel (Ni) toxicity and oxidative reactions were studied in roots during metal accumulation. Membrane lipid peroxidation enhanced only 6 h after metal treatment before roots revealed a decrease in growth. The activities of superoxide dismutase (SOD), EC 1.15.1.1, guaiacol peroxidase (GPX), EC 1.11.1.7, were unaffected by Ni stress. However, catalase (CAT), EC 1.11.1.6, activity was increased from 24 h after metal treatment. In the ascorbate gluthatione cycle, only two enzymes, the ascorbate peroxidase (APX), EC 1.11.1.11, and monodehydroascorbate reductase (MDHAR), EC 1.6.5.4, were stimulated, while the activities of dehydroascorbate reductase (DHAR), EC 1.8.5.1, and glutathione reductase (GR), EC 1.6.4.2, remained around the control values. The results suggest that oxidative disorder is part of the overall expression of Ni toxicity in roots of Zea mays and that enhanced lipid peroxidation could be a consequence of primary effects of Ni stress.

Plant peroxidases: biomarkers of metallic stress

The term “peroxidase” designs a group of hemoproteins with a wide structural variability. These enzymes catalyze the redox reaction between hydrogen peroxide and some reductors. They can be found in animals, plants and microorganisms. In plants, peroxidases are involved in numerous cellular processes such as development and stress responses. In fact, they are involved in growth regulation by controlling hormonal and cell wall metabolism and antioxidant defense. On the other hand, these enzymes are considered as a biomarker indicating biotic and abiotic stresses. Under metallic stress conditions, the quantitative and qualitative profiles of peroxidases are generally modified. Such modulations could prove the major role played by these enzymes in the defense mechanism. In this paper, we discussed the variation of isoperoxidases behavior under metallic stress conditions.

Membrane damage and solute leakage from germinating pea seed under cadmium stress.

Seed germination represents a limiting stage of plant life cycle under heavy metal stress situation. Delay in germination can be associated with disorders in the event chain of germinative metabolism which is a highly complex multistage process, but one of underlying metabolic activities following imbibition of seed is the storage mobilization. The influence of cadmium on carbohydrates and aminoacids export from cotyledon to embryonic axis during germination of pea seed was investigated. Compared to the control, Cd caused a restriction in reserve mobilization as evidenced by the pronounced increase in cotyledon/embryo ratios of total soluble sugars, glucose, fructose and aminoacids. Moreover, the nutrient concentrations, as well as the electrical conductivity of germination medium were determined to quantify the extent of solute leakage. Such nutrients were lost into the imbibition medium at the expense of suitable mobilization to the growing embryonic axis. This was concomitant with an over-accumulation of lipid peroxidation products in Cd-poisoned embryonic tissues. However, the impairment of membrane integrity cannot be due to a stimulation in lipoxygenase activity, since the later was markedly inhibited after Cd exposure.

Effect of copper excess on superoxide dismutase, catalase, and peroxidase activities in sunflower seedlings (Helianthus annuus L.)

The changes in lipid peroxidation, antioxidative and lignifying enzyme activities were studied in leaves and stems of Cu-stressed sunflower seedlings. In both organs, membrane lipid peroxidation was enhanced by copper treatment. Additionally, catalase (EC 1.11.1.6) and superoxide dismutase (EC 1.15.1.1) activities were modulated: The activity of superoxide dismutase was enhanced in both plant organs. Differently, catalase activity was not affected in leaves but significantly reduced in stems. Peroxidase (EC 1.11.1.7) activities were also changed. Guaiacol peroxidase activity was increased in leaves and stems. In the same way, electrophoretic analysis of the anionic isoperoxidases involved in lignification (syringaldazine peroxidase) revealed qualitative and quantitative changes on the isoenzyme patterns. These modifications were accompanied by the increase of the NADH-oxidase activity in ionically cell wall bound fraction. It appeared that the growth delay caused by copper excess could be related to the activation of lignifying peroxidases.

Oxidative injury and antioxidant genes regulation in cadmium-exposed radicles of six contrasted Medicago truncatula genotypes.

Oxidative disorders were triggered in the presence of Cd toxicity in early seedling growth of six Medicago truncatula genotypes. Interactions between root growth inhibition, cadmium uptake, as well as the occurrence of oxidative injury suggest differential responses of the genotypes, with susceptible or tolerant accessions. ROS enhancement was observed in situ and H2O2 content was measured, that did not seem related to tolerance or susceptibility. Oxidative burst impact on cell membrane integrity was analyzed in agreement with MDA content and glucose exudation, which suggest an active role of this burst in susceptible lines. Transcriptional changes in response to cadmium treatment were analyzed on target genes involved in (1) ROS-scavenging enzymes (superoxide dismutase (SOD; EC1.15.1.1) and peroxidase (PRX; EC 1.11.1.7)), (2) reduced glutathione (γ-Glu-Cys-Gly, GSH) metabolism (glutathione-S-transferase (GST; EC: 2.5.1.18) and glutathione reductase (GR; EC 1.8.1.7)), and (3) metal-chelating metabolism (PCS). The susceptible line shows no response or non-timely gene expression patterns. This research work gave an overview of the deleterious effects and oxidative injury of cadmium stress in Medicago truncatula. Oxidative defense efficiency and gene upregulation should explain relative tolerance in tested genotypes.

Unsuitable Availability of Nutrients in Germinating Bean Embryos Exposed to Copper Excess

The influence of copper excess on germination rate, growth, minerals, carbohydrates, and amino acids supply in embryonic axis of bean seed was investigated. Compared to the control, Cu treatment caused a reduction in germination percent, embryo length, and accumulation of Ca, Fe, K, Mn, Zn, total soluble sugars, glucose, fructose, sucrose, and amino acids. Moreover, the nutrient concentrations, as well as the electrical conductivity were determined in the germination medium to quantify the extent of solute leakage. Such nutrients were lost in the imbibition medium at the expense of suitable mobilization to the growing embryonic axis. This was associated with an enhancement in accumulation of malondialdehyde, major product of lipoperoxidation process which can be due to the stimulation in lipoxygenase activity in Cu-poisoned tissues.

Partial purification and characterization of a copper-induced anionic peroxidase of sunflower roots.

Treatment of 14-day-old sunflower seedlings with a toxic amount of copper (50 microM of CuSO(4)) during 5days caused significant increase in peroxidase activity in roots. Qualitative analysis of soluble proteins using native anionic PAGE followed by detection of peroxidase activity with guaïacol as electron donor in the presence of H(2)O(2) revealed five stimulated peroxidases, named A1, A2, A3, A4, and A5. These peroxidases had differential behavior during the period of treatment. A1, A2, A3 and A4 were stimulated in the first period of stress, but rapidly suppressed at 72h. A5 showed a progressive stimulation which was even increased at 120h. A1 was partially purified, identified using liquid chromatography coupled to mass spectrometry (LC-MS/MS), and characterized. Effects of pH and temperature on its activity were determined with guaïacol as electron donor. Optima were obtained at pH 8 and at 40 degrees C. Analysis of substrate specificity showed that A1 was active on coniferyl alcohol but not on IAA. Enzymatic activity was inhibited by a high concentration of H(2)O(2).

Toxicity of sulcotrione and grape marc on Vicia faba cells.

The cell toxicity of sulcotrione, a selective triketone herbicide, was evaluated on Vicia faba. Sulcotrione, trademark Mikado, grape marc, and mixtures of sulcotrione or Mikado with grape marc induced cell death. Addition of grape marc to either sulcotrione or Mikado enhanced cell death, especially with Mikado. Addition of grape marc to herbicides, sulcotrione, or Mikado resulted in different expression of genes usually associated with cell stress. Mixtures of grape marc and herbicides enhanced transcript accumulation for ubiquitin, hsp 70, and cytosolic superoxide dismutase, but did not change ascorbate peroxidase transcript accumulation. The results thus provide evidence that sulcotrione, Mikado, and mixtures with grape marc can trigger cell death and specific gene expressions. Cocktails of products with sulcotrione, such as commercial additives and grape marc, can modify biological features of pesticide. Moreover, grape marc differently enhanced cell toxicity of sulcotrione and Mikado, suggesting a synergy between pesticide products and grape marc.

Redox biology response in germinating Phaseolus vulgaris seeds exposed to copper: evidence for differential redox buffering in seedlings and cotyledon

In agriculture, heavy metal contamination of soil interferes with processes associated with plant growth, development and productivity. Here, we describe oxidative and redox changes, and deleterious injury within cotyledons and seedlings caused by exposure of germinating (Phaseolus vulgaris L. var. soisson nain hâtif) seeds to copper (Cu). Cu induced a marked delay in seedling growth, and was associated with biochemical disturbances in terms of intracellular oxidative status, redox regulation and energy metabolism. In response to these alterations, modulation of activities of antioxidant proteins (thioredoxin and glutathione reductase, peroxiredoxin) occurred, thus preventing oxidative damage. In addition, oxidative modification of proteins was detected in both cotyledons and seedlings by one- and two-dimensional electrophoresis. These modified proteins may play roles in redox buffering. The changes in activities of redox proteins underline their fundamental roles in controlling redox homeostasis. However, observed differential redox responses in cotyledon and seedling tissues showed a major capacity of the seedlings’ redox systems to protect the reduced status of protein thiols, thus suggesting quantitatively greater antioxidant protection of proteins in seedlings compared to cotyledon. To our knowledge, this is the first comprehensive redox biology investigation of the effect of Cu on seed germination.