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Constitutive expression of a grape aspartic protease gene in transgenic Arabidopsis confers osmotic stress tolerance

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Abstract

Aspartic proteases are involved in various processes of plant senescence, programmed cell death, reproduction and stress responses. We previously identified a salt and drought stress induced gene, VlAP17, encoding a Group C aspartic protease. Here, we report its functional analysis through the characterization of transgenic Arabidopsis thaliana plants overexpressing VlAP17 under the control of constitutive promoter. The transgenic plants showed enhance salt and drought stress tolerance during seed germination as well as the seedling and mature plant stage. In additional, various stress responses indicators were analyzed and results suggested that osmotic stress caused less damage to the plasma membrane of transgenic seedlings than to that of wild type plants. VlAP17 overexpression also resulted in increased ABA levels, a reduction in average stomatal aperture size, and elevated expression levels of stress-response genes involved in the ABA-dependent pathway, as well as higher activities of several antioxidases: superoxide dismutase, catalase and peroxidase. Taken together, these findings suggest that VlAP17 plays a role in protecting the integrity of plasma membrane and it may also be involved in, or have a function upstream of, the ABA biosynthetic pathway.

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Abbreviations

AP:

Aspartic protease

WT:

Wild type

CaMV:

Cauliflower mosaic virus

ORF:

Open reading frame

RT-PCR:

Reverse transcription-polymerase chain reaction

MS:

Murashige and Skoog

MDA:

Malondialdehyde

TCA:

Trichloroacetic acid

TBA:

Thiobarbituric acid

ELISA:

Enzyme-linked immunosorbent assay

NBT:

Nitro blue tetrazolium

DAB:

Diaminobenzidine

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

CAT:

Catalase

POD:

Peroxidase

H2O2 :

Hydrogen peroxide

O2 − :

Superoxide anion

NCED3:

9-cis-epoxycarotenoid dioxygenase

ADH:

Alcohol dehydrogenase gene

FRY1:

Fiery 1

ERD:

Early responsive to dehydration

SOS:

Salt overly sensitive

DREB:

Dehydration responsive element binding factor

RD:

Responsive to dehydration

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (31272136), the 948 Project from the Ministry of Agriculture of China (2012-S12), as well as the Program for Innovative Research Team of Grape Germplasm Resources and Breeding (2013KCT-25). We thank PlantScribe (www.plantscribe.com) for editing this manuscript.

Conflict of interest

There are no competing interests in this paper, and the authors do not have any possible conflicts of interest.

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Authors and Affiliations

  1. State Key Laboratory of Crop Stress Biology in Arid Areas, College of Horticulture, Northwest A&F University, Yangling, 712100, Shaanxi, China

    Rongrong Guo, Jiao Zhao, Xianhang Wang, Chunlei Guo, Zhi Li, Yuejin Wang & Xiping Wang

  2. Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, Northwest A&F University, Yangling, 712100, Shaanxi, China

    Rongrong Guo, Jiao Zhao, Xianhang Wang, Chunlei Guo, Zhi Li, Yuejin Wang & Xiping Wang

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  1. Rongrong Guo
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  2. Jiao Zhao
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  3. Xianhang Wang
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  4. Chunlei Guo
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  5. Zhi Li
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  7. Xiping Wang
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Corresponding author

Correspondence to Xiping Wang.

Additional information

The nucleotide sequence reported in this paper has been submitted to GeneBank with accession numbers KM210287.

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Guo, R., Zhao, J., Wang, X. et al. Constitutive expression of a grape aspartic protease gene in transgenic Arabidopsis confers osmotic stress tolerance. Plant Cell Tiss Organ Cult 121, 275–287 (2015). https://doi.org/10.1007/s11240-014-0699-6

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