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The responses of the enzymes related with ascorbate–glutathione cycle during drought stress in apple leaves

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Abstract

To explore the significance of the ascorbate–glutathione cycle under drought stress, the leaves of 2-year-old potted apple (Malus domestica Borkh.) plants were used to investigate the changes of each component of the ascorbate–glutathione cycle as well as the gene expression of dehydroascorbate reductase (DHAR, EC 1.8.5.1), ascorbate peroxidase (APX, EC 1.11.1.11) and glutathione reductase (GR, EC 1.6.4.2) under drought stress. The results showed that the malondialdehyde (MDA) and H2O2 concentrations in apple leaves increased during drought stress and began to decrease after re-watering. The contents of total ascorbate, reduced ascorbic acid (AsA), total glutathione and glutathione (GSH) were obviously upregulated in apple leaves when the soil water content was 40–45%. With further increase of the drought level, the contents of the antioxidants and especially redox state of AsA and GSH declined. However, levels of them increased again after re-watering. Moreover, drought stress induced significant increase of the activities of enzymes such as APX, scavenging H2O2, and also of monodehydroascorbate reductase (MDHAR, EC 1.6.5.4), DHAR and GR used to regenerate AsA and GSH, especially when the soil water content was above 40–45%. During severe drought stress, activities of the enzymes were decreased and after re-watering increased again. Gene expression of cytoplasmic DHAR, cytoplasmic APX and cytoplasmic GR showed similar changes as the enzyme activities, respectively. The results suggest that the ascorbate–glutathione cycle is up-regulated in response to drought stress, but cannot be regulated at severe drought stress conditions.

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Abbreviations

APX:

Ascorbate peroxidase

AsA:

Reduced ascorbic acid

BSA:

Bovine serum albumin

CP:

Control plants

DHA:

Dehydroascorbate

DHAR:

Dehydroascorbate reductase

DP:

Drought stressed plants

DTT:

Dithiothreitol

GR:

Glutathione reductase

GSH:

Glutathione

GSSG:

Oxidized glutathione

H2O2 :

Hydrogen peroxide

KOH:

Potassium hydroxide

MDHAR:

Monodehydroascorbate reductase

MDA:

Malondialdehyde

NADH:

β-Nicotinamide adenine dinucleotide

NADPH:

β-Nicotinamide adenine dinucleotide 2′-phosphate

NEM:

N-Ethylmaleimide

PCR:

Polymerase chain reaction

ROS:

Reactive oxygen species

SD:

Standard deviation

SDS:

Sodium dodecyl sulfate

TBA:

2-Thiobarbituric acid

TCA:

Trichloroacetic acid

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Acknowledgments

This work were supported by the Natural Science Fund of Shannxi province, Talent Support Program of Northwest A&F University and Special Fund of Guizhou province Academy of Agricultural Sciences([2009]024). The authors are grateful to Prof. Karin Krupinska for help in revising our English composition and to Mr Xuanchang Fu for management on the plant materials.

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Authors and Affiliations

  1. College of Horticulture, Northwest A&F University, Yangling, 712100, Shaanxi, China

    Yu-Hua Ma, Feng-Wang Ma & Jun-Ke Zhang

  2. Guizhou Horticulture Institute, Guiyang, 550006, Guizhou, China

    Yu-Hua Ma

  3. Guizhou Institute of Environmental Science and Desiging, Guiyang, 550002, Guizhou, China

    Yong-Hong Wang

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  1. Yu-Hua Ma
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  2. Feng-Wang Ma
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Correspondence to Feng-Wang Ma.

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Communicated by K. Krupinska.

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Ma, YH., Ma, FW., Wang, YH. et al. The responses of the enzymes related with ascorbate–glutathione cycle during drought stress in apple leaves. Acta Physiol Plant 33, 173–180 (2011). https://doi.org/10.1007/s11738-010-0535-5

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  • DOI: https://doi.org/10.1007/s11738-010-0535-5

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