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Title 

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

Authors 

Suk Yoon KwonYoung Ock AhnHeang Soon LeeSang Soo Kwak

Publisher 

Springer Verlag (Germany)

Issue Date 

2001

Citation 

Journal of Biochemistry and Molecular Biology, vol. 34, no. 4, pp. 316-321

Keywords 

ascorbic aciddehydroascorbate reductaseglutathioneoxidative stresstransgenic tobacco plant

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.

ISSN 

1225-8687

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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