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Constitutive expression of a fungal glucose oxidase gene in transgenic tobacco confers chilling tolerance through the activation of antioxidative defence system

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Abstract

Scientific evidences in the literature have shown that plants treated exogenously with micromole concentration of hydrogen peroxide (H2O2) acquire abiotic stress tolerance potential, without substantial disturbances in the endogenous H2O2 pool. In this study, we enhanced the endogenous H2O2 content of tobacco (Nicotiana tabaccum L. cv. SR1) plants by the constitutive expression of a glucose oxidase (GO; EC 1.1.3.4) gene of Aspergillus niger and studied their cold tolerance level. Stable integration and expression of GO gene in the transgenic (T0–T2) tobacco lines were ascertained by molecular and biochemical tests. Production of functionally competent GO in transgenic plants was confirmed by the elevated levels of H2O2 in the transformed tissues. When three homozygous transgenic lines were exposed to different chilling temperatures for 12 h, the electrolyte conductivity was significantly lower in GO-expressing tobacco plants than the control plants; in particular, chilling protection was more prominent at −1°C. In addition, most transgenic lines recovered within a week when returned to normal culture conditions after −1°C–12 h cold stress. However, control plants displayed symptoms of chilling injuries such as necrosis of shoot tip, shoots and leaves, consequently plant death. The protective effect realized in the transgenic plants was comparable to cold-acclimatized wild tobacco. The chilling tolerance of transgenic lines was found associated, at least in part, with elevated levels of total antioxidant content, CAT and APX activities. Based on our findings, we predict that the transgenic expression of GO may be deployed to improve cold tolerance potential of higher plants.

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Abbreviations

APX:

Ascorbate peroxidase

CAT:

Catalase

FRAP:

Ferric reducing antioxidant power

GO:

Glucose oxidase

GR:

Glutathione reductase

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

TPTZ:

2,4,6-Tri(2-pyridyl)-s-triazine

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Acknowledgments

This work was supported by the Taiwan-Philippine collaborative research grant (NSC99-2923-B-005-001) to CHL from the National Science Council, Taiwan, Republic of China.

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Authors and Affiliations

  1. Department of Life Sciences, National Chung Hsing University, Taichung, 402, Taiwan, ROC

    Subbiyan Maruthasalam, Yi Lun Liu, Ching Mei Sun, Pei Ying Chen & Chin Ho Lin

  2. Department of Molecular Biotechnology, Da Yeh University, Changhua, 515, Taiwan, ROC

    Chih Wen Yu

  3. Department of Biotechnology, Fooyin University, Kaohsiung, 831, Taiwan, ROC

    Pei Fang Lee

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  1. Subbiyan Maruthasalam
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  2. Yi Lun Liu
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  3. Ching Mei Sun
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  6. Pei Fang Lee
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  7. Chin Ho Lin
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Correspondence to Chin Ho Lin.

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Communicated by P. Lakshmanan.

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Maruthasalam, S., Liu, Y.L., Sun, C.M. et al. Constitutive expression of a fungal glucose oxidase gene in transgenic tobacco confers chilling tolerance through the activation of antioxidative defence system. Plant Cell Rep 29, 1035–1048 (2010). https://doi.org/10.1007/s00299-010-0889-6

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