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  • 1
    ISSN: 1432-2048
    Keywords: Chitinase ; β-1,3-Glucanase-Epidermus(enzyme induction) ; Ethylene (enzyme induction) ; Vacuole (enzyme accumulation) ; Phaseolus (ethylene and enzyme induction)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have studied the spatial pattern of accumulation of chitinase (EC 3.2.1.14) and β-1,3-glucanase (EC 3.2.1.39) in ethylene-treated leaves of bean (Phaseolus vulgaris L.). Electron-microscopical examination of chemically fixed tissue demonstrated the presence of large electron-dense aggregates in the vacuoles of ethylene-treated leaf cells. No such vacuolar structures were observed in untreated control cells. Immunogold labelling with antisera directed against the basic forms of chitinase and β-1,3-glucanase indicated that the vacuolar aggregates were the major site of accumulation of chitinase and β-1,3-glucanase. The chitinase- and β-1,3-glucanase-containing vacuolar aggregates were not randomly distributed within the leaf tissue but were restricted to the lower epidermal cells and to parenchyma cells adjacent to vascular strands. In addition, heavy β-1,3-glucanase labelling was observed over spongy plugs of expanded middle-lamella material that appear to occlude the transition regions between the airspaces underlying the stomata and those throughout the rest of the leaf. Some labelling was also seen to extend along the surface layer of the cell walls lining all of the airspaces. Protein analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting as well as enzyme-activity measurements showed that the peeled lower epidermis of the ethylene-treated leaves contained on a protein and on a per-weight basis several times more chitinase and β-1,3-glucanase than the remainder of the leaf.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2048
    Keywords: Chitinase ; Defense (against bacteria, fungi) ; Enzyme induction ; Ethylene ; Lysozyme ; Phaseolus (chitinase)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ethylene induced an endochitinase in primary leaves of Phaseolus vulgaris L. The enzyme formed chitobiose and higher chitin oligosaccharides from insoluble, colloidal or regenerated chitin. Less than 5% of the total chitinolytic activity was detected in an exochitinase assay proposed by Abeles et al. (1970, Plant Physiol. 47, 129–134) for ethylene-induced chitinase. In ethylene-treated plants, chitinase activity started to increase after a lag of 6 h and was induced 30 fold within 24 h. Exogenously supplied ethylene at 1 nl ml−1 was sufficient for half-maximal induction, and enhancement of the endogenous ethylene formation also enhanced chitinase activity. Cycloheximide prevented the induction. Among various hydrolases tested, only chitinase and, to a lesser extent, β-1,3-glucanase were induced by ethylene. Induction of chitinase by ethylene occurred in many different plant species. Ethylene-induced chitinase was purified by affinity chromatography on a column of regenerated chitin. Its apparent molecular weight obtained by sodium dodecyl sulfate-gel electrophoresis was 30,000; the molecular weight determined from filtration through Sephadex G-75 was 22,000. The purified enzyme attacked chitin in isolated cell walls of Fusarium solani. It also acted as a lysozyme when incubated with Micrococcus lysodeikticus. It is concluded that ethylene-induced chitinase functions as a defense enzyme against fungal and bacterial invaders.
    Type of Medium: Electronic Resource
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