Biochemical characterization of transgenic tobacco plants expressing a human dehydroascorbate reductase gene

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Title
Biochemical characterization of transgenic tobacco plants expressing a human dehydroascorbate reductase gene
Author(s)
Suk Yoon Kwon; Young Ock Ahn; Heang Soon Lee; Sang Soo Kwak
Bibliographic Citation
BMB Reports, vol. 34, no. 4, pp. 316-321
Publication Year
2001
Abstract
Dehydroascorbate (DHA) reductase (DHAR, EC 1.8.5.1) catalyzes the reduction of DHA to reduced ascorbate (AsA) using glutathione (GSH) as the electron donor in order to maintain an appropriate level of ascorbate in plant cells. To analyze the physiological role of DHAR in environmental stress adaptation, we developed transgenic tobacco (Nicotiana tabacum cv. Xanthi) plants that express a human DHAR gene isolated from the human fetal liver cDNA library in the chloroplasts. We also investigated the DHAR activity, levels of ascorbate, and GSH. Two transgenic plants were successfully developed by Agrobacterium-mediated transformation and were confirmed by PCR and Southern blot analysis. DHAR activity and AsA content in mature leaves of transgenic plants were approximately 1.41 and 1.95 times higher than in the non-transgenic (NT) plants, respectively. In addition, the content of oxidized glutathione (GSSG) in transgenic plants was approximately 2.95 times higher than in the NT plants. The ratios of AsA to DHA and GSSG to GSH were changed by overexpression of DHAR, as expected, even though the total content of ascorbate and glutathione was not significantly changed. When tobacco leaf discs were subjected to methyl viologen at 5 μM, T0 transgenic plants showed about a 50% reduction in membrane damage compared to the NT plants.
Keyword
ascorbic aciddehydroascorbate reductaseglutathioneoxidative stresstransgenic tobacco plant
ISSN
1225-8687
Publisher
Korea Soc-Assoc-Inst
Type
Article
Appears in Collections:
Division of Research on National Challenges > Plant Systems Engineering Research > 1. Journal Articles
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