Sequential effects of cadmium on genotoxicity and lipoperoxidation in Vicia faba roots.

Kinetics of stress responses to Cd exposure (50, 100 and 200 µM) expanding from 12 to 48 h were studied in roots of hydroponically cultivated-Vicia faba seedlings. The heavy metal induced toxicity symptoms and growth arrest of Vicia roots gradually to the Cd concentration and duration of the treatment. The intracellular oxidative stress was evaluated with the H(2)O(2) production. The H(2)O(2) content increased gradually with the sequestered Cd and root growth inhibition. Lipid peroxidation-evidenced by malondialdehyde (MDA) content and Evans blue uptake-and genotoxicity-evidenced by mitotic index (MI) and micronuclei (MCN) values-were concomitantly investigated in root tips. By 12 h, root meristematic cells lost 15% of their mitotic activity under 50 or 100 µM Cd treatment and 50% under 200 µM Cd treatment and led cells with MCN, while the MDA content and Evans blue absorption were not affected. The loss of membrane integrity occurred subsequently by 24 h. The increase in MDA content in root cells treated with 50, 100 and 200 µM Cd was significantly higher than the control. By 48 h, the MDA content increased 134, 178 or 208% in root cells treated with 50, 100 and 200 µM Cd, respectively. The Evans blue absorption was also affected by 24 h in roots when treated with 200 µM Cd and gradually increase by 48 h with the Cd concentration of the treatment. The decrease of mitotic activity triggered by 12 h was even higher by 24 h and the MI reduced to 44, 56 or 80% compared to the control in the three different Cd concentrations tested. The different kinetics of early in vivo physiological and cytogenetic responses to Cd might be relevant to the characterization of its toxicity mechanisms in disrupting primarily the mitosis process.

A comparative analysis of fatty acid composition of root and shoot lipids in Zea mays under copper and cadmium stress.

A comparative analysis of fatty acid composition was conducted in maize (Zea mays L.) under copper and cadmium stress. The unsaturation level (double-bond index) of phosphatidylethanolamine (PE) and digalactosyldiacylglycerol (DGDG) was increased in response to both metal treatments, whereas the phosphatidylinositol (PI), the phosphatidylcholine (PC) showed no significant changes. The Cu-treated roots showed a marked increase (about 2-fold) in the phospholipid (PL) content, while the Cd-treated roots showed a slight but insignificant increase. The steryl lipid SL/PL ratio was markedly decreased in response to Cu stress, and therefore, may indicate an activated phospholipid biosynthesis and turnover, in response to damage caused by Cu stress. The double bond indices of chloroplastic lipids: phosphatidylglycerol (PG), monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG) and sulfoquinovosyldiacylglycerol (SQDG) revealed a similar but not identical pattern of change. The PG and MGDG contents in shoots were markedly decreased under Cu (by 53 and 48%) and Cd (by 78 and 65%) stress. The increase in the malondialdehyde (MDA) content in roots induced by both metals indicates lipid peroxidation. Generally, in the presence of Cu fatty acid composition was markedly modulated but to lesser extent under Cd stress. These results suggest that changes in the fatty acid composition under Cu and Cd stress conditions are metal-specific and may therefore result in differential metal tolerance.

Effect of copper excess on H2O2 accumulation and peroxidase activities in bean roots.

We studied oxidative stress and peroxidase activity resulting from application of excess copper in the nutrient medium on the roots of young bean seedlings. The change in H2O2 content, lipid peroxidation and antioxidant enzymes activities were quantified and located. Excess of copper caused a loss of membrane integrity and the formation of hydrogen peroxide (H2O2) as visualized in the transmission electron microscopy and measured using spectrophotometry. H2O2 accumulated in the intercellular spaces and in the cell wall. The production of H2O2 was accompanied by an increase in the activity of soluble and ionic GPX (guaiacol peroxidase, EC 1.11.17), CAPX (coniferyl alcohol peroxidase) and NADH oxidase.

Copper-induced oxidative stress in maize shoots (Zea mays L.): H2O2 accumulation and peroxidases modulation.

The effect of copper excess on growth, H2O2 level and peroxidase activities were studied in maize shoots. Ten-day-old seedlings were cultured in nutrient solution that contained Cu2+ ions at various concentrations (50 and 100 microM) for seven days. High concentrations of Cu2+ ions caused significant decrease both in matter production and elongation of maize shoots. In addition, treatment with CuSO4 increased levels of H2O2 and induced changes in several peroxidase activities. Moreover, the disturbance of the physiological parameters was accompanied by the modulation of the peroxidase activities: GPX (Guaiacol peroxidase, EC 1.11.1.7), CAPX (Coniferyl alcohol peroxidase, EC 1.11.1.4) and APX (Ascorbate peroxidase, EC. 1.11.1.11). Furthermore, this modulation becomes highly significant, especially, in the presence of 100 microM of CuSO4.

Interaction between heavy metals and thiol-linked redox reactions in germination.

Thioredoxin (TRX) proteins perform important biological functions in cells by changing the redox state of proteins via dithiol disulfide exchange. Several systems are able to control the activity, stability, and correct folding of enzymes through dithiol/disulfide isomerization reactions including the enzyme protein disulfide-isomerase, the glutathione-dependent glutaredoxin system, and the thioredoxin systems. Plants have devised sophisticated mechanisms to cope with biotic and abiotic stresses imposed by their environment. Among these mechanisms, those collectively referred to as redox reactions induced by endogenous systems. This is of agronomical importance since a better knowledge of the involved mechanisms can offer novel means for crop protection. In the plant life cycle, the seed and seedling stages are key developmental stages conditioning the final yield of crops. Both are very sensitive to heavy metal stress. Plant redox reactions are principally studied on adult plant organs and there is only very scarce informations about the onset of redox regulation at the level of seed germination. In the here presented study, we discussed the importance of redox proteins in plant cell metabolism and defence. Special focus is given to TRX, which are involved in detoxification of ROS and also to their targets.

Exposure of Vicia faba and Pisum sativum to copper-induced genotoxicity.

The potential genotoxicity of Cu(2+) was investigated in Vicia faba and Pisum sativum seedlings in hydroponic culture conditions. Cu(2+) caused a dose-dependent increase in micronuclei frequencies in both plant models. Cytological analysis of root tips cells showed clastogenic and aneugenic effects of this heavy metal on V. faba root meristems. Cu(2+) induced chromosomal alterations at the lowest concentration used (2.5 mM) when incubated for 42 h, indicating the potent mutagenic effect of this ion. A spectrum of chromosomal abnormalities was observed in V. faba root meristems, illustrating the genotoxic events leading to micronuclei formation.

[Zinc phytotoxicity and induction of stress proteins in bean (Phaseolus vulgaris L.)]. / Phytotoxicité du zinc et induction de protéine(s) de stress chez le haricot (Phaseolus vulgaris L.).

Bean seedlings (Phaseolus vulgaris L., cv. fin de Bagnol) were grown for ten days in nutrient solutions at 0, 50, 100, 500 and 1000 microM ZnS0(4). The results obtained show that Zn treatment is followed by a pathological behaviour, the precocity and intensity of which are dependent on the applied dose. In addition to the depressive effects on the biomass yield and the tissue water content, a decrease in the proteins contents was also observed. Zinc toxicity seems to induce synthesis of low molecular weight proteins in Zn-treated bean stems, but not in the roots and leaves. These peptides, in stems, may protect, hypothetically, the susceptibles metabolic sites in foliar parenchyma against the zinc deleterious effects.

Structural changes induced by NaCl in companion and transfer cells of Medicago sativa blades.

Medicago sativa var. Gabes is a perennial glycophyte that develops new shoots even in high salinity (150 mM NaCl). In the upper exporting leaves, K(+) is high and Na(+) is low by comparison with the lower leaves, where Na(+) accumulation induces chlorosis after 4 weeks of NaCl treatment. By secondary ion mass spectroscopy, a low Na(+)/K(+) ratio was detected in the phloem complex of blade veins in these lower leaves. By transmission electron microscopy, the ultrastructural features were observed in the phloem complex. In the upper leaves of both control and NaCl-treated plants, companion cells in minor veins were found to be transfer cells. These cells may well be involved in the intravenous recycling of ions and in Na(+) flowing out of exporting leaves. Under the effect of NaCl, companion cells in the main veins develop transfer cell features, which may favor the rate of assimilate transport from exporting leaves toward meristems, allowing the positive balance necessary for the survival in salt conditions. These features no longer assist the lower leaves when transfer cells are necrotized in both minor and main veins of NaCl-treated plants. As transfer cells are the only degenerating phloem constituent, our observations emphasize their role in controlling nutrient (in particular, Na(+)) fluxes associated with the stress response.