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Risultati sfruttabili
Many environmental stress conditions such as high or low temperatures, drought and pollutants lead to the excess production of active oxygen species (AOS) such as superoxide radicals (O2-.), hydrogenperoxide (H2O2) and hydroxyl radicals. These so-called AOS react with lipids, proteins and DNA and are thought to be the major cause irreversible damage caused by environmental stress to plants. The last year, our knowledge on the mechanisms which limit the production of AOS has been greatly improved. Extensive knowledge is available on the structure, complexity and expression levels of various antioxidant genes, such as those encoding superoxide dismutases, catalyses, ascorbate peroxidase, glutathion peroxidase,...However, little information is available on how plants sense oxidative stress and on how this signal is transmitted to turn on the appropriate defense genes. In addition, methods needed to be developed to address the antioxidant capacity of individual cells. The current INTAS project has successfully addressed these questions.
Several Arabidopsis and tobacco mutants were identified with enhanced tolerance to experimental conditions which cause oxidative damage. These mutants are of great scientific interest and might, upon further characterization, provide novel insights in the mechanisms which control the general defense reactions of plants to stress.
Glutathione levels and the pathway of glutathione-mediated detoxification in plants were followed in vivo using monochlorobimane (MCB) as a substrate for GSH conjugation. The glutathione-bimane (GS-B) conjugate was fluorescent and was imaged in suspension cultured cells and intact roots of Arabidopsis using confocal scanning laser microscopy with excitation at 442 nm. The average glutathione concentration in suspension cultured cells and different cell types along the Arabidopsis root was determined after calibration against GS-B standards and correction for tissue and depth dependent attenuation. However, the values reported represent averages over the whole cell rather than just the GSH concentration in the cytoplasm. Direct measurement of cytoplasmic and vacuolar volumes was not straightforward. As an alternative strategy we have imaged the protein distribution in the same root and normalized the GSH levels to average cellular protein content. There was considerable variation in the levels of GSH on a protein basis. Most notably the cells in the quiescent center had lower levels that the neighbouring initial cells. The difference in GSH levels appears to be a critical part of a redox control system affecting root development.
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