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Permeabilization of plant cells for release of intracellularly stored products: viability studies
Authors:Peter Brodelius
Affiliation:(1) Institute of Biotechnology, Swiss Federal Institute of Technology, ETH-Hönggerberg, CH-8093 Zürich, Switzerland
Abstract:Summary The effects of various chemical substances on the permeability of plasma membranes and tonoplasts of three suspension cultures (Catharanthus roseus, Thalictrum rugosum and Chenopodium rubrum) have been studied. The permeability of the plasma membrane is monitored by measuring the activity of the cytosolic enzyme isocitrate dehydrogenase and the permeability of the tonoplast is measured by determining the release of substances stored in the vacuoles (inorganic phosphate, berberine and betanin for the three cell lines, respectively). The minimum concentration required for quantitative release of vacuolar products have been established for five different permeabilization agents. Cell viability is lost upon permeabilization except for treatment of Catharanthus roseus with DMSO and Triton X-100.Abbreviations DMSO dimethylsulfoxide - PEA phenethylalcohol - HDTMAB hexadecyltrimethylammonium bromide - ICDH isocitrate dehydrogenase
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