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Nitrate reductase-mediated nitric oxide production is involved in copper tolerance in shoots of hulless barley

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Abstract

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An NR-mediated early NO production in the shoots of hulless barley plays an important role in protecting hulless barley from Cu toxicity through enhanced antioxidant enzyme activities and antioxidant pools.

Abstract

Nitric oxide (NO) has been identified as an important signaling molecule that is involved in multiple plant physiological responses, especially under some abiotic stress. Here, we investigated NO production and its effects on copper (Cu) excess in hulless barley shoots. An early NO burst at 24 h was observed in shoots of hulless barley, and the synthesis of early NO was mediated through nitrate reductase (NR), but not through nitric oxide synthase (NOS). Application of the NO donor sodium nitroprusside (SNP) efficiently alleviated Cu-induced shoot inhibition and decrease in chlorophyll content, as well as oxidative damage and reactive oxygen species (ROS) accumulation, while inhibiting NO accumulation by a specific NO scavenger or a NR inhibitor aggravated shoot inhibition as well as the increase of hydrogen peroxide (H2O2) content, supporting the role of an NR-mediated early NO production in hulless barley responses to Cu toxicity. Furthermore, elevated antioxidant enzyme activities were induced by Cu stress in the shoots of hulless barley and further significantly enhanced by NO donor, whereas suppressed by NO scavenger or NR inhibitor. On the other hand, the application of NO scavenger significantly reduced Cu-induced accumulation of glutathione (GSH) and ascorbate (Asc) in the shoots of hulless barley. Taken together, our results indicate that NO may induce hulless barley seedling tolerance to Cu toxicity through modulating antioxidant enzyme activity and antioxidants accumulation.

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Abbreviations

Asc:

Ascorbate

APX:

Ascorbate peroxidase

CAT:

Catalase

cPTIO:

2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide

DTT:

Dithiothreitol

FAD:

Flavin adenine dinucleotide

GPX:

Glutathione peroxidase

GR:

Glutathione reductase

GSH:

Glutathione

HbO2:

Oxyhemoglobin

H2O2 :

Hydrogen peroxide

MetHb:

Methemoglobin

L-NAME:

Nω-nitro-l-arginine methyl ester hydrochloride

MDA:

Malondialdehyde

NR:

Nitrate reductase

NOS:

Nitric oxide synthase

NO:

Nitric oxide

O2 − :

Superoxide anion radical

POD:

Peroxidase

PVP:

Polyvinylpyrrolidone

ROS:

Reactive oxygen species

SNP:

Sodium nitroprusside

SOD:

Superoxide dismutase

TCA:

Trichloroacetic acid

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Acknowledgments

This work was supported by Open Funds for Key Laboratory of Mollisols Agroecology of Chinese Academy of Sciences and the National Nature Science Foundation of China (No.31301252).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Key Laboratory of Mollisols Agroecology, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, Harbin, 150000, China

    Yanfeng Hu

  2. School of Life Science, Northwest Normal University, Lanzhou, 730070, China

    Jia You

  3. Key Laboratory of Cell Activities and Stress Adaptations, Ministry of Education, School of Life Sciences, Lanzhou University, Lanzhou, 730000, Gansu, China

    Xiaolei Liang

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  1. Yanfeng Hu
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  2. Jia You
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  3. Xiaolei Liang
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Correspondence to Yanfeng Hu.

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Communicated by Kang Chong.

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Hu, Y., You, J. & Liang, X. Nitrate reductase-mediated nitric oxide production is involved in copper tolerance in shoots of hulless barley. Plant Cell Rep 34, 367–379 (2015). https://doi.org/10.1007/s00299-014-1715-3

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  • DOI: https://doi.org/10.1007/s00299-014-1715-3

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