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The expression of BrMDHAR gene in chloroplasts and mitochondria enhances tolerance to freezing stress in Arabidopsis thaliana

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Biologia Plantarum

Abstract

In chloroplasts and mitochondria, antioxidant mechanisms include the ascorbate-glutathione cycle, and monodehydroascorbate reductase (MDHAR) is important for regeneration of ascorbate (AsA) from monodehydroascorbate (MDHA). To improve detoxification of reactive oxygen species (ROS), we established a construct of the MDHAR gene from Brassica rapa fused to the targeting signal peptides of Pisum sativum glutathione reductase (GR), which was controlled by a stress-inducible SWPA2 promoter, and introduced this expression system into Arabidopsis thaliana. Transgenic (TG) plants overexpressing BrMDHAR targeted to chloroplasts and mitochondria through signal peptides showed an elevated MDHAR activity and an increased ratio of AsA to dehydroascorbate (DHA) when compared to wild-type (WT) plants under a freezing stress. These led to increased photosynthetic parameters, redox homeostasis, and biomass in TG plants when compared to the WT plants. Our results suggest that the overexpression of the BrMDHAR gene targeted to chloroplasts and mitochondria conferred an enhanced tolerance against the freezing stress, and a stress adaptation of dual-targeted BrMDHAR was better than that of single BrMDHAR.

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Abbreviations

APX:

ascorbate peroxidase

AsA:

ascorbate

BrMHAR :

Brassica rapa monodehydroascorbate reductase gene

DCFH-DA:

6-carboxy-2′,7′-dichlorofluorescin diacetate

DHA:

dehydroascorbate

DHAR:

dehydroascorbate reductase

GPX:

glutathione peroxidase

GR:

glutathione reductase

GSH:

glutathione

MDA:

malondialdehyde

MDHA:

monodehydroascorbate

MDHAR:

monodehydroascorbate reductase

Prx Q:

peroxiredoxin Q

ROS:

reactive oxygen species

SOD:

superoxide dismutase

SWPA2 promoter:

stress-inducible promoter from sweet potato ascorbate peroxidase

TBA:

thiobarbituric acid

TCA:

trichloroacetic acid

TG:

transgenic

WT:

wild type

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Authors and Affiliations

  1. Advanced-Bioresource Research Center, Kyungpook National University, Daegu, 702-701, Republic of Korea

    S. Y. Shin, Y. S. Kim, I. S. Kim & H. S. Yoon

  2. Department of Biology, Kyungpook National University, Daegu, 702-701, Republic of Korea

    H. S. Yoon

  3. National Institute of Crop Science, Rural Development Administration, Suwon, 441-857, Republic of Korea

    Y. H. Kim & H. M. Park

  4. National of Horicultural & Herbal Science, Rural Development Administration, Suwon, 440-706, Republic of Korea

    S. Y. Shin

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  1. S. Y. Shin
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  3. I. S. Kim
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Correspondence to H. S. Yoon.

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Acknowledgements: This work was supported by a grant from the Next-Generation BioGreen 21 Program (No. PJ008060), Rural Development Administration, South Korea. I.S. Kim and H.S. Yoon equally contributed to this study.

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Shin, S.Y., Kim, Y.S., Kim, I.S. et al. The expression of BrMDHAR gene in chloroplasts and mitochondria enhances tolerance to freezing stress in Arabidopsis thaliana . Biol Plant 58, 456–468 (2014). https://doi.org/10.1007/s10535-014-0416-7

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