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Homologous expression of cytosolic dehydroascorbate reductase increases grain yield and biomass under paddy field conditions in transgenic rice (Oryza sativa L. japonica)

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Abstract

Dehydroascorbate reductase (DHAR, EC 1.8.5.1) maintains redox pools of ascorbate (AsA) by recycling oxidized AsA to reduced AsA. To investigate whether DHAR affects rice yield under normal environmental conditions, cDNA-encoding DHAR (OsDHAR1) was isolated from rice and used to develop OsDHAR1-overexpressing transgenic rice plants, under the regulation of a maize ubiquitin promoter. Incorporation and expression of the transgene in transgenic rice plants was confirmed by genomic polymerase chain reaction (PCR), semi-quantitative reverse transcription PCR (RT-PCR), western blot, and enzyme activity. The expression levels were at least twofold higher in transgenic (TG) rice plants than in control wild-type (WT) rice plants. In addition, OsDHAR1-overexpression in seven-independent homologous transgenic plants, as compared to WT plants, increased photosynthetic capacity and antioxidant enzyme activities under paddy field conditions, which led to an improved AsA pool and redox homeostasis. Furthermore, OsDHAR1 overexpression significantly improved grain yield and biomass due to the increase of culm and root weights and to enhance panicle and spikelet numbers in the same seven independent TG rice plants during the farming season (2010 and 2011) in South Korea. The OsDHAR protein contained the redox-active site (Cys20), as well as the conserved GSH-binding region, GSH-binding motif, glutathione-S-transferase (GST) N-terminal domain, C-terminal domain interface, and GST C-terminal domain. Therefore, our results indicate that OsDHAR1 overexpression, capable of functioning in AsA recycling, and protein folding increases environmental adaptation to paddy field conditions by the improving AsA pool and redox homeostasis, which enhances rice grain yield and biomass.

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Abbreviations

APX:

Ascorbate peroxidase

AsA:

Ascorbate

AsA/DHA:

Ascorbate to dehydroascorbate ratio

DHAR:

Dehydroascorbate reductase

Fv/Fm :

Maximum quantum yield of PSII

GR:

Glutathione reductase

GSH:

Reduced glutathione

MDHAR:

Monodehydroascorbate reductase

Os:

Oryza sativa

OX:

Overexpression

ROS:

Reactive oxygen species

TG:

Transgenic

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Acknowledgments

This work was supported by a grant from the Next-Generation BioGreen 21 Program (No. PJ008115012013), Rural Development Administration, and funded by the Priority Research Centers Program (2011-0030748) through the National Research Foundation (NRF) of the Ministry of Education, Science and Technology, Korea.

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Authors and Affiliations

  1. Department of Biology, College of Natural Sciences, Kyungpook National University, Daegu, 702-701, South Korea

    Young-Saeng Kim, Il-Sup Kim, Mi-Jung Bae, Yong-Hoe Choe & Ho-Sung Yoon

  2. National Institute of Crop Science, Rural Development Administration, Suwon, 441-857, South Korea

    Yul-Ho Kim & Hyang-Mi Park

  3. Subtropical Horticulture Research Institute, Jeju National University, Jeju, 690-756, South Korea

    Hong-Gyu Kang

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  1. Young-Saeng Kim
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  2. Il-Sup Kim
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Correspondence to Hong-Gyu Kang or Ho-Sung Yoon.

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Y.-S. Kim and I. S. Kim contributed equally to this work.

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Kim, YS., Kim, IS., Bae, MJ. et al. Homologous expression of cytosolic dehydroascorbate reductase increases grain yield and biomass under paddy field conditions in transgenic rice (Oryza sativa L. japonica). Planta 237, 1613–1625 (2013). https://doi.org/10.1007/s00425-013-1862-8

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