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Enhanced sensitivity to oxidative stress in transgenic tobacco plants with decreased glutathione reductase activity leads to a decrease in ascorbate pool and ascorbate redox state

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Abstract

To investigate the possible mechanisms of glutathione reductase (GR) in protecting against oxidative stress, we obtained transgenic tobacco (Nicotiana tabacum) plants with 30–70% decreased GR activity by using a gene encoding tobacco chloroplastic GR for the RNAi construct. We investigated the responses of wild type and transgenic plants to oxidative stress induced by application of methyl viologen in vivo. Analyses of CO2 assimilation, maximal efficiency of photosystem II photochemistry, leaf bleaching, and oxidative damage to lipids demonstrated that transgenic plants exhibited enhanced sensitivity to oxidative stress. Under oxidative stress, there was a greater decrease in reduced to oxidized glutathione ratio but a greater increase in reduced glutathione in transgenic plants than in wild type plants. In addition, transgenic plants showed a greater decrease in reduced ascorbate and reduced to oxidized ascorbate ratio than wild type plants. However, there were neither differences in the levels of NADP and NADPH and in the total foliar activities of monodehydroascorbate reductase and dehydroascorbate reductase between wild type and transgenic plant. MV treatment induced an increase in the activities of GR, ascorbate peroxidase, superoxide dismutase, and catalase. Furthermore, accumulation of H2O2 in chloroplasts was observed in transgenic plants but not in wild type plants. Our results suggest that capacity for regeneration of glutathione by GR plays an important role in protecting against oxidative stress by maintaining ascorbate pool and ascorbate redox state.

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Acknowledgments

We thank CSIRO Plant Industry for providing the RNAi vectors and Prof. Jinxin Lin for help with cytochemical analysis. This work was supported by the Frontier Project of the Knowledge Innovation Engineering of Chinese Academy of Sciences (KSCX2-YW-N-042), the National Natural Science Foundation of China (30725024), and the Major Key Basic Research and Development Plan of China (2009CB118500).

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  1. Photosynthesis Research Center, Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, 100093, Beijing, China

    Shunhua Ding, Qingtao Lu, Yan Zhang, Zhipan Yang, Xiaogang Wen, Lixin Zhang & Congming Lu

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  1. Shunhua Ding
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  2. Qingtao Lu
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Correspondence to Congming Lu.

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Ding, S., Lu, Q., Zhang, Y. et al. Enhanced sensitivity to oxidative stress in transgenic tobacco plants with decreased glutathione reductase activity leads to a decrease in ascorbate pool and ascorbate redox state. Plant Mol Biol 69, 577–592 (2009). https://doi.org/10.1007/s11103-008-9440-3

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