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Yellow lupin (Lupinus luteus) cv. Juno seedlings exposed to increasing concentrations of Pb²⁺ (50-350 mg l⁻¹) were analysed in respect to its effect on the degradation of lipids, the content of antioxidants (ascorbate, α-tocopherol) and the activity of the ascorbate glutathione cycle enzymes (dehydroascorbate reductase DAR; EC 1.8.5.1 and glutathione reductase GR; EC 1.6.4.2). Lipid peroxidation, expressed as the content of TBArm (thiobarbituric acid reactive metabolites), increased only at 50 and 100 mgl⁻¹ Pb²⁺, whereas at higher lead concentrations it decreased as compared with the control. The level of free fatty acids was not significantly affected as a result of Pb²⁺ exposure, except for 300 mgl⁻¹ Pb²⁺. The content of α-tocopherol increased significantly at the range of concentrations between 150 and 300 mgl⁻¹ and at these concentrations lipid peroxidation was inhibited. Ascorbic acid (AA) and dehydroascorbic acid (DHA) content increased considerably in roots treated with 100 and 150 mgl⁻¹ Pb²⁺. In general the activity of DHAR and GR was stimulated by lead, however at higher Pb²⁺ concentrations (300 and 350 mgl⁻¹) GR revealed lower enzymatic activtty. Our results suggest that in lead-treated roots lipids might be protected against reactive oxygen species (ROS) by lipid-soluble molecules of α-tocopherol and this direct defense seems to be supported by AA as well as the enzymes: DHAR and GR.
In an earlier work using tissue printing method, we found that the PR-10 stress protein was observed in leaf petiole of lupin seedling where lead was not detected (Przymusiński et al. 2001). These results suggested the presence of substance(s) mediating a signal transduction from directly affected cells to distant organs. As the hydrogen peroxide was found to be involved in signal transduction pathway, in the present paper, we analysed the level of H₂O₂ in the organ of lupin seedlings exposed to Pb²⁺ with spectrophotometric method and tissue printing technique. It was unequivocally demonstrated that the level of H₂O₂ and the activity of peroxidase increased in every tested organ of lead-treated lupin seedling. Both the level of H₂O₂ and the activity of POX were correlated with amount of Pb²⁺ ions in the cells (Przymusiński et al. 2001) and decreased in tissues more and more distant from the site of metal application. On the other hand, there was no correlation between the histological localization of H₂O₂ and peroxidase. Our results seem to confirm the hypothesis that H₂O₂ may act as a signalling substance involved in the induction of PR protein synthesis. It was indicated that there is high degree of correlation between the localization of H₂O₂ and the histological localization of PR-10 proteins (Przymusiński et al. 2001) in every tested organ of lupin seedling. The presented hypothesis is also supported by the fact that H₂O₂ and PR-10 proteins are detected in organs and tissues where Pb²⁺ was not found at all.
Embryonic root is the primary site of salinity perception in germinating seeds. To understand better the NaCl stress response of lupine embryo axes, ultrastructural approach combined with analysis of DNA degradation was used. In this study lupine embryo axes were cultured in vitro on the medium supplemented with two salt concentrations 250 and 500 mM to differ the reaction. To assess the rate of DNA damage, alkaline electrophoresis of isolated nuclei and DNA fragmentation analysis were performed. Results of these studies suggest programmed cell death induction under salinity stress. Moreover, ultrastructure observations revealed other characteristic features of programmed cell death like endoplasmic reticulum reorganization, increased level of vacuolization, chromatin condensation and starch grains degradation. Our comparative analysis of ultrastructure changes and DNA fragmentation speak in favour of programmed cell death in lupine (Lupinus luteus L. ‘Mister’) embryo axes treated for 12 h with 250 and 500 mM NaCl.
The present study investigated the potential role of selected antioxidant enzymes: superoxide dismutase (SOD) and peroxidase (POX) as biochemical markers of the impact of tropospheric ozone. Experiments were carried out in ambient air conditions with two tobacco cultivars: Bel W3 and Bel B, which are sensitive and resistant to ozone, respectively. In this study, the degree of leaf injury of the sensitive cultivar was used as an indicator of the ozone level in correlation to the enzyme activity of both tobacco cultivars. In spite of low levels of tropospheric ozone during experimental season, the increase of antioxidant enzyme (SOD and POX) activity concomitant with the increase of ozone concentration was noticed in the sensitive cultivar as well as in the resistant one. This observation is especially important for the resistant tobacco, which does not exhibit any visual effects of ozone influence. Our results could be extrapolated to other plant species (i.e. Poaceae, Fabaceae, Solanaceae, Betulaceae, Salicaceae, Pinaceae), which do not reveal visible lesions in response to ozone stress.
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