Plant Molecular Biology

, Volume 68, Issue 1, pp 131–143

Two cysteine proteinase inhibitors from Arabidopsis thaliana, AtCYSa and AtCYSb, increasing the salt, drought, oxidation and cold tolerance

Article

DOI: 10.1007/s11103-008-9357-x

Cite this article as:
Zhang, X., Liu, S. & Takano, T. Plant Mol Biol (2008) 68: 131. doi:10.1007/s11103-008-9357-x

Abstract

Two cysteine proteinase inhibitors (cystatins) from Arabidopsis thaliana, designated AtCYSa and AtCYSb, were characterized. Recombinant GST-AtCYSa and GST-AtCYSb were expressed in Escherichia coli and purified. They inhibit the catalytic activity of papain, which is generally taken as evidence for cysteine proteinase inhibitor function. Northern blot analyses showed that the expressions of AtCYSa and AtCYSb gene in Arabidopsis cells and seedlings were strongly induced by multiple abiotic stresses from high salt, drought, oxidant, and cold. Interestingly, the promoter region of AtCYSa gene contains a dehydration-responsive element (DRE) and an abscisic acid (ABA)-responsive element (ABRE), which identifies it as a DREB1A and AREB target gene. Under normal conditions, AtCYSa was expressed in 35S: DREB1A and 35S:AREB1 plants at a higher level than in WT plants, while AtCYSa gene was expressed in 35S:DREB2A plants at the same level as in WT plants. Under stress conditions (salt, drought and cold), AtCYSa was expressed more in all three transgenic plants than in WT plants. Over-expression of AtCYSa and AtCYSb in transgenic yeast and Arabidopsis plants increased the resistance to high salt, drought, oxidative, and cold stresses. Taken together, these data raise the possibility of using AtCYSa and AtCYSb to genetically improve environmental stresses tolerance in plants.

Keywords

Cysteine proteinase inhibitorsGene expressionInhibitory activityStress toleranceTransgenic Arabidopsis

Abbreviations

ABA

Abscisic acid

ABRE

ABA-responsive element

AREB

ABRE binding protein

Cystatins

Cysteine proteinase inhibitors

DRE

Dehydration-responsive element

DREB

DRE binding protein

GST

Glutathione S-transferase

PCD

Programmed cell death

Pis

Proteinase inhibitors

PMSF

Phenylmethylsulfonyl fluoride

TCA

Trichloroacetic acid

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  1. 1.Asian Natural Environment Science Center (ANESC)The University of TokyoNishitokyo CityJapan
  2. 2.Alkali Soil Natural Environmental Science Center (ASNESC), Stress Molecular Biology LaboratoryNortheast Forestry UniversityHarbinPeoples’ Republic of China