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Transgenic tomato plants expressing strawberry d-galacturonic acid reductase gene display enhanced tolerance to abiotic stresses

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Abstract

After analyzing tomato plants transformed with GalUR gene for their ascorbic acid contents, it was found that some transgenic lines contained higher levels of ascorbic acid compared to control plants. In the present study, callus induction rate was 50.2 % in the explant and shoot regeneration rate was 51.5 % from the callus with transformation efficiency of 3.0 %. Based on PCR and Southern blot analysis, three independent transformants containing the insert gene were selected. Phenotypic traits of these transgenic progeny were similar to those of control tomatoes. Tomatoes (H15) with high fruit ascorbic acid contents were selected for next generation (GalUR T3) analysis. Transgenic tomatoes with increased ascorbic acid contents were found to be more tolerant to abiotic stresses induced by viologen, NaCl, or mannitol than non-transformed plants. In leaf disc senescence assay, the tolerance of these transgenic plants was better than control plants because they could retain higher chlorophyll contents. Under salt stress of less than 200 mM NaCl, these transgenic plants survived. However, control plants were unable to survive such high salt stress. Ascorbic acid contents in the transgenic plants were inversely correlated with MDA contents, especially under salt stress conditions. The GalUR gene was expressed in H15 tomatoes, but not in control plants. Higher expression levels of antioxidant genes (APX and CAT) were also found in these transgenic plants compared to that in the control plants. However, no detectable difference in SOD expression was found between transgenic plants and control plants. Results from this study suggest that the increase in ascorbic acid contents in plants could up-regulate the antioxidant system to enhance the tolerance of transgenic tomato plants to various abiotic stresses.

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Abbreviations

APX:

Ascorbate peroxidase

AsA:

Ascorbic acid

CAT:

Catalase

EtBr:

Ethidium bromide

GalUR :

d-Galacturonic acid reductase

GLOase :

l-Gulono-γ-lactone oxidase

GSG:

Glutathione

IAA:

Indole acetic acid

MDA:

Malondialdehyde

MS:

Murashige and Skoog medium

MV:

Methyl viologen

PCR:

Polymerase chain reaction

POD:

Peroxidase

ROS:

Reactive oxygen species

RT-PCR:

Reverse transcriptase-polymerase chain reaction

SOD:

Superoxide dismutase

TBA:

Thiobarbituric acid

TCA:

Trichloroacetic acid

YEP:

Yeast extract peptone

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Acknowledgments

This research was supported by Crop Functional Genomics Center of the 21st century Frontier Research Program funded by the Ministry of Education, Science and Technology, Republic of Korea. It was also partially supported by BioGreen 21 Research Program, Rural Development of Administration, Republic of Korea.

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Correspondence to Byoung Ryong Jeong or Chee Hark Harn.

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Lim, M.Y., Jeong, B.R., Jung, M. et al. Transgenic tomato plants expressing strawberry d-galacturonic acid reductase gene display enhanced tolerance to abiotic stresses. Plant Biotechnol Rep 10, 105–116 (2016). https://doi.org/10.1007/s11816-016-0392-9

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  • DOI: https://doi.org/10.1007/s11816-016-0392-9

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