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Anaerobic induction in conifers: Expression of endogenous and chimeric anaerobically-induced genes
Authors:A G Good  F Bekkaoui  G Pilate  D I Dunstan  W L Crosby
Institution:Plant Biotechnology Inst., National Research Council of Canada, 110 Gymnasium Road, Saskatoon, Saskatchewan, Canada S7N OW9
Abstract:Alcohol dehydrogenase (ADH; EC 1.1.1.1) activity was measured in Picea glauca (Moench) Voss cell suspensions under differing conditions of hypoxia. ADH activity increased 4.5 fold after 48 h of induction. When cells were induced under different levels of hypoxia (2, 5 and 20% O2) changes in ADH activity were found to increase with lower levels of oxygen. Alanine aminotransferase (AlaAT; EC 2.6.1.2) activity increased under hypoxia in a pattern similar to ADH, however lactate dehydrogenase (LDH; EC 1.1.1.27) activity did not increase under hypoxic conditions. The ability of white spruce cells to accurately regulate heterologous anaerobic promotors was tested by electroporating chimeric ADH reporter genes into protoplasts. While protoplasts were capable of anaerobically regulating a maize ADH reporter construct, constructs with dicotyledonous promoters (pea and Arabidopsis ) were not expressed.
Keywords:Alcohol dehydrogenase  anaerobiosis  conifers  electroporation  hypoxia  protoplasts  transient expression
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