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

  • Rongrong Guo
  • Jiao Zhao
  • Xianhang Wang
  • Chunlei Guo
  • Zhi Li
  • Yuejin Wang
  • Xiping WangEmail author
Original Paper

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.

Keywords

ABA biosynthetic AP17 Aspartic protease Drought Grape Salt 

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

Notes

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.

Supplementary material

11240_2014_699_MOESM1_ESM.pdf (458 kb)
Supplementary material 1 (PDF 458 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Rongrong Guo
    • 1
    • 2
  • Jiao Zhao
    • 1
    • 2
  • Xianhang Wang
    • 1
    • 2
  • Chunlei Guo
    • 1
    • 2
  • Zhi Li
    • 1
    • 2
  • Yuejin Wang
    • 1
    • 2
  • Xiping Wang
    • 1
    • 2
    Email author
  1. 1.State Key Laboratory of Crop Stress Biology in Arid Areas, College of HorticultureNorthwest A&F UniversityYanglingChina
  2. 2.Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of AgricultureNorthwest A&F UniversityYanglingChina

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