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Over-expression of a glutamate dehydrogenase gene, MgGDH, from Magnaporthe grisea confers tolerance to dehydration stress in transgenic rice

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Abstract

Main conclusion

Heterologous expression of a fungal NADP(H)-GDH gene ( MgGDH ) from Magnaporthe grisea can improve dehydration stress tolerance in rice by preventing toxic accumulation of ammonium.

Glutamate dehydrogenase (GDH; EC 1.4.1.2 and EC 1.4.1.4) may act as a stress-responsive enzyme in detoxification of high intracellular ammonia and production of glutamate for proline synthesis under stress conditions. In present study, a fungal NADP(H)-GDH gene (MgGDH) from Magnaporthe grisea was over-expressed in rice (Oryza sativa L. cv. ‘kitaake’), and the transgenic plants showed the improvement of tolerance to dehydration stress. The kinetic analysis showed that His-TF-MgGDH preferentially utilizes ammonium to produce l-glutamate. Moreover, the affinity of His-TF-MgGDH for ammonium was dramatically higher than that of His-TF-OsGDH for ammonium. Over-expressing MgGDH transgenic rice plants showed lower water-loss rate and higher completely close stomata than the wild-type plants under dehydration stress conditions. In transgenic plants, the NADP(H)-GDH activities were markedly higher than those in wild-type plants and the amination activity was significantly higher than the deamination activity. Compared with wild-type plants, the transgenic plants accumulated much less NH4 + but higher amounts of glutamate, proline and soluble sugar under dehydration stress conditions. These results indicate that heterologous expression of MgGDH can prevent toxic accumulation of ammonium and in return improve dehydration stress tolerance in rice.

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Abbreviations

Mg:

Magnaporthe grisea

GDH:

Glutamate dehydrogenase

GOGAT:

Glutamate synthase

GS:

Glutamine synthetase

2-OG:

2-oxoglutarate

TF:

Trigger factor

P5CS:

△1-Pyrroline-5-carboxylate synthetase

P5CR:

△1-Pyrroline-5-carboxylic acid reductase

GSA:

Glutamic γ-semialdehyde

PCR:

Polymerase chain reaction

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Acknowledgments

The authors thank Dr. Hong Liu for providing the fungi Magnaporthe grisea and assistance of cloning MgGDH. This research was supported by Important National Science and Technology Specific Projects (No. 2009ZX08001-030B), National Science Foundation of China (No. 31170172), Hunan Provincial Natural Science Foundation of China (No.12JJ3024), Cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province (No. 20134486), Planned Science and Technology Project of Hunan Province (No. 2014WK2005 and No. 2014NK3001).

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  1. Hunan Province Key Laboratory of Plant Functional Genomics and Developmental Regulation, Hunan University, Changsha, 410082, China

    Yanbiao Zhou, Caisheng Zhang, Jianzhong Lin, Yuchong Peng, Dongying Tang, Xiaoying Zhao, Yonghua Zhu & Xuanming Liu

  2. State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Biology, Hunan University, Changsha, 410082, China

    Yanbiao Zhou, Caisheng Zhang, Jianzhong Lin, Yuchong Peng, Dongying Tang, Xiaoying Zhao, Yonghua Zhu & Xuanming Liu

  3. Academy of Seed Industry of Hunan Yahua, Changsha, 410001, Hunan, China

    Yuanzhu Yang

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  1. Yanbiao Zhou
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Correspondence to Xuanming Liu.

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Y. Zhou, C. Zhang and J. Lin contributed equally to this work.

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Zhou, Y., Zhang, C., Lin, J. et al. Over-expression of a glutamate dehydrogenase gene, MgGDH, from Magnaporthe grisea confers tolerance to dehydration stress in transgenic rice. Planta 241, 727–740 (2015). https://doi.org/10.1007/s00425-014-2214-z

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