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Superoxide dismutase and glutathione reductase overexpression in wheat protoplast: photooxidative stress tolerance and changes in cellular redox state

Abstract

In previous works, we have established a correlation between antioxidant system response and tolerance to drought, osmotic stress and photooxidative stress of different wheat cultivars with contrasting drought tolerance. In the present work, a protocol to obtain and transform wheat protoplasts was established. Transgenic protoplasts with Manganese Superoxide Dismutase (Mn-SOD) (E.C.: 1.15.1.1) and Glutathione Reductase (GR) (E.C.: 1.6.4.2) overexpression in chloroplasts were obtained, and their responses to photooxidative stress were characterized. Protoplasts with Mn-SOD or GR overexpression, showed different responses and tolerance to photooxidative stress. Protoplasts with Mn-SOD overexpression showed lower levels of oxidative damage, higher level of endogenous hydrogen peroxide and a great induction of total SOD and GR activities during photooxidative treatments. In protoplasts with GR overexpression the oxidative damage provoked by the photooxidative treatment was similar to control protoplasts, the GSH content and GSH/GSH + GSSG ratio were higher than control and Mn-SOD transformed protoplast, and total SOD and GR activities were not induced. Our results suggest that the differential responses and tolerance to photooxidative stress given by Mn-SOD or GR overexpression, also depend on the effects of these enzyme activities over the cellular redox state balance, which modulate the responses to photooxidative stress.

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Abbreviations

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

GR:

Glutathione reductase

GSH:

Reduced glutathione

GSSG:

Glutathione disulfide

MDA:

Malondialdehyde

H2O2 :

Hydrogen peroxide

O2 :

Superoxide radical

PEG:

Polyethylene glycol

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Acknowledgements

We thank Prof. Alejandra Alamo for the English corrections. This work was supported by grants from CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas-Argentina-PID 8170 and 0370). MM, RL, VST, are researchers from CONICET GR are INTA fellowshiper.

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Correspondence to H. Ramiro Lascano.

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Melchiorre, M., Robert, G., Trippi, V. et al. Superoxide dismutase and glutathione reductase overexpression in wheat protoplast: photooxidative stress tolerance and changes in cellular redox state. Plant Growth Regul 57, 57 (2009). https://doi.org/10.1007/s10725-008-9322-3

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  • DOI: https://doi.org/10.1007/s10725-008-9322-3

Keywords

  • Glutathione reductase
  • Photooxidative stress
  • Reactive oxygen species
  • Superoxide dismutase
  • Wheat protoplasts