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Plant Molecular Biology Reporter

, Volume 34, Issue 1, pp 62–75 | Cite as

A Phytocystatin Gene from Malus prunifolia (Willd.) Borkh., MpCYS5, Confers Salt Stress Tolerance and Functions in Endoplasmic Reticulum Stress Response in Arabidopsis

  • Yanxiao Tan
  • Xiaoyu Wei
  • Ping Wang
  • Xun Sun
  • Mingjun Li
  • Fengwang MaEmail author
Original Paper

Abstract

Cystatins, or phytocystatins (PhyCys), comprise a family of plant-specific inhibitors of cysteine proteinases. They are thought to help regulate endogenous processes and protect plants against biotic or abiotic stresses, such as heat, salinity, cold, water deficit, chilling, and abscisic acid (ABA) treatment. We isolated and identified a novel cystatin gene from Malus prunifolia, MpCYS5. Its expression was typically induced by salt stress treatment; ectopic expression in Arabidopsis enhanced salt tolerance. Physiological parameters confirmed this phenotype, with the transgenics having remarkably lower electrolyte leakage (EL) values, higher chlorophyll concentrations, and lower levels of malondialdehyde (MDA) upon salt treatment. In addition, the accumulation of reactive oxygen species was markedly regulated by MpCYS5 under stress conditions, as shown by fluctuations in the concentrations of hydrogen peroxide and superoxide radicals and the activities of antioxidant enzymes. We also noted that this gene modulated tunicamycin (TM)-induced endoplasmic reticulum (ER) stress tolerance and functioned in the unfolded protein response (UPR)-signaling pathway in Arabidopsis. This was confirmed by the expression of eight ER stress-responsive genes. All marker genes examined were strongly induced in the wild type, while most of them maintained relatively stable over time in the transgenics. These results demonstrated that ectopic expression of a cystatin gene is associated with salt-tolerant and TM-tolerant phenotypes. Therefore, the discovery of MpCYS5 from M. prunifolia might establish a molecular link between the ER stress response and salt tolerance in plants.

Keywords

Endoplasmic reticulum stress Malus prunifolia Plant cystatin Reactive oxygen species Salt stress Unfolded protein response 

Abbreviations

ABA

Abscisic acid

APX

Ascorbate peroxidase

CAT

Catalase

DAB

Diaminobenzidine

EL

Electrolyte leakage

ER

Endoplasmic reticulum

Km

Kanamycin

MDA

Malondialdehyde

MS

Murashige and Skoog

MV

Methyl viologen

NBT

Nitro blue tetrazolium

ORF

Open reading frame

PhyCys

Phytocystatins

POD

Peroxidase

qRT-PCR

Quantitative real-time PCR

SOD

Superoxide dismutase

TM

Tunicamycin

UPR

Unfolded protein response

Notes

Acknowledgments

This work was supported by the National High Technology Research and Development Program of China (863 Program) (2011AA100201), and by the earmarked fund for China Agriculture Research System (CARS-28). The authors are grateful to Priscilla Licht for help in revising our English composition and to Mr. Zhengwei Ma for management of the potted apple plants.

Supplementary material

11105_2015_900_MOESM1_ESM.doc (30 kb)
Supplementary Table S1 Primers used in this study (DOC 30 kb)
11105_2015_900_MOESM2_ESM.doc (36 kb)
Supplementary Table S2 GenBank Accession Numbers or Locus IDs for genes from Arabidopsis thaliana (At)a, Oryza sativa (Os)b, Hordeum vulgare (Hv)c, and Malus prunifolia c (DOC 36 kb)

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Yanxiao Tan
    • 1
  • Xiaoyu Wei
    • 1
  • Ping Wang
    • 1
  • Xun Sun
    • 1
  • Mingjun Li
    • 1
  • Fengwang Ma
    • 1
    Email author
  1. 1.State Key Laboratory of Crop Stress Biology for Arid Areas, College of HorticultureNorthwest A&F UniversityYanglingPeople’s Republic of China

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