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Biochemical analysis of enhanced tolerance in transgenic potato plants overexpressing d-galacturonic acid reductase gene in response to various abiotic stresses

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Abstract

Upregulation of the antioxidant enzyme system in plants provides protection against various abiotic stresses. Transgenic potato plants overexpressing the strawberry d-galacturonic acid reductase (GalUR) gene with enhanced accumulation of ascorbate (AsA) were used to study the antioxidant system involving the ascorbate–glutathione cycle in order to understand the tolerance mechanism in plants in response to various abiotic stresses under in vitro conditions. Transgenic potato tubers subjected to various abiotic stresses induced by methyl viologen, sodium chloride and zinc chloride showed enhanced activities of superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.1.1.1.6) and enzymes of the ascorbate–glutathione cycle such as ascorbate peroxidase (APX, EC 1.11.1.11), dehydroascorbate reductase (DHAR, EC 1.8.5.1) and glutathione reductase (GR, EC 1.8.1.7), as well as increased levels of ascorbate, glutathione (GSH) and proline when compared to untransformed tubers. The increased enzyme activities correlated with the mRNA transcript levels in the stressed transgenic tubers. Significant differences in redox status of AsA and GSH were also observed in stressed transgenic potato tubers that showed increased tolerance to abiotic stresses compared to untransformed tubers. This study suggests that the increased accumulation of AsA could upregulate the antioxidant system which imparts improved tolerance against various abiotic stresses in transgenic tubers compared to untransformed tubers.

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Abbreviations

APX:

Ascorbate peroxidase

CAT:

Catalase

DHAR:

Dehydroascorbate reductase

GalUR:

d-galacturonic acid reductase

GR:

Glutathione reductase

GSH:

Reduced glutathione

GSSG:

Oxidized glutathione

MDA:

Malondialdehyde

MDHAR:

Monodehydroascorbate reductase

MV:

Methyl viologen

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

ZnCl:

Zinc chloride

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Acknowledgments

This research was supported by Konkuk University research fund. The research fellowship from Konkuk University to Hemavathi as research fellow is gratefully acknowledged. We thank Mayank A. Gururani and Shashank K. Pandey for experimental assistance and proof-reading of the manuscript.

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Correspondence to Chandrama Prakash Upadhyaya or Se Won Park.

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Hemavathi and Chandrama Prakash Upadhyaya contributed equally to this work.

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11032_2010_9465_MOESM1_ESM.tif

Fig. A. Antioxidant defense system in plants (Halliwell-Asada pathway; modified from Bowler et al. 1992). The dismutation of superoxide radicals (O2 ) occurs in this pathway. APX-ascorbate peroxidase; DHAR-dehydroascorbate reductase; GR-glutathione reductase; GSH-glutathione (red.); GSSG-glutathione (ox.); SOD-superoxide dismutase (TIFF 176 kb)

11032_2010_9465_MOESM2_ESM.tif

Fig. B. Quantitative real time PCR analysis of antioxidant protein gene expression profiling during various stress treatments. The cDNA was normalized in dependence of the transcript level of actin mRNA. Antioxidant enzyme gene expression in transformed and untransformed potato tubers under control conditions (A), Methyl viologen i.e. Oxidative stress (B); NaCl i.e. salt stress (C) and ZnCl i.e. heavy metal stress (D). The values are presented as the mean ± SEM of three replicates (TIFF 201 kb)

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Hemavathi, Upadhyaya, C.P., Akula, N. et al. Biochemical analysis of enhanced tolerance in transgenic potato plants overexpressing d-galacturonic acid reductase gene in response to various abiotic stresses. Mol Breeding 28, 105–115 (2011). https://doi.org/10.1007/s11032-010-9465-6

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