The production of an inducible antisense alternative oxidase (Aox1a) plant |
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Authors: | Felicity Johnson Potter Joseph T Wiskich Ian B Dry |
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Institution: | (1) Colorado State University, Department of Horticulture and Landscape Architecture, Fort Collins, CO 80525, USA, US;(2) The University of Adelaide, Department of Environmental Biology, Adelaide 5005, South Australia, AU;(3) CSIRO Plant Industry, Horticulture Unit, Hartley Grove, Urrbrae 5064, South Australia, AU |
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Abstract: | Plant mitochondria contain an alternative oxidase (AOX) acting as a terminal electron acceptor of the alternative pathway
in the electron transport chain. Here we describe the production of inducible antisense Aox1a plants of Arabidopsis thaliana (L.) Heynh. and the procedures used to determine the resulting alternative pathway activity. The Arabidopsis Aox1a cDNA sequence was cloned behind a copper-inducible promoter system in the antisense orientation. Arabidopsis thaliana (Columbia) plants were transformed by in-planta vacuum infiltration with Agrobacterium containing the antisense construct. Whole-leaf ethanol production was used as a measure to investigate alternative pathway
activity in the presence of antimycin A. After 24 h, leaves from the copper-induced, antisense line F1.1 produced up to 8.8
times more ethanol (via aerobic fermentation) than the non-induced and wild-type leaves, indicating effective cytochrome pathway
inhibition by antimycin A and a decreased alternative pathway activity in induced F1.1 leaves. Transgene expression studies
also revealed no expression in non-induced leaves and up until 24 h post-induction. Copper-induced transgenic leaves were
less susceptible to alternative pathway inhibition than non-induced transgenic leaves, as seen via tissue-slice respiratory
studies, and mitochondrial respiration, using F1.1 cell cultures, also supported this. These results demonstrate the successful
production of a transgenic line of Arabidopsis in which the alternative pathway activity can be genetically manipulated with an inducible antisense system.
Received: 31 March 2000 / Accepted: 10 May 2000 |
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Keywords: | : Alternative oxidase Antisense Aox1a Arabidopsis (alternative oxidase) Copper-inducible gene Electron transport Mitochondrial respiration |
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