Planta

, Volume 235, Issue 2, pp 253–266

Cloning and characterization of a maize bZIP transcription factor, ZmbZIP72, confers drought and salt tolerance in transgenic Arabidopsis

Original Article

Abstract

In plants, the bZIP (basic leucine zipper) transcription factors regulate diverse functions, including processes such as plant development and stress response. However, few have been functionally characterized in maize (Zea mays). In this study, we cloned ZmbZIP72, a bZIP transcription factor gene from maize, which had only one copy in the maize genome and harbored three introns. Analysis of the amino acid sequence of ZmbZIP72 revealed a highly conserved bZIP DNA-binding domain in its C-terminal region, and four conserved sequences distributed in N- or C-terminal region. The ZmbZIP72 gene expressed differentially in various organs of maize plants and was induced by abscisic acid, high salinity, and drought treatment in seedlings. Subcellular localization analysis in onion epidermal cells indicated that ZmbZIP72 was a nuclear protein. Transactivation assay in yeast demonstrated that ZmbZIP72 functioned as a transcriptional activator and its N terminus (amino acids 23–63) was necessary for the transactivation activity. Heterologous overexpression of ZmbZIP72 improved drought and partial salt tolerance of transgenic Arabidopsis plants, as determined by physiological analyses of leaf water loss, electrolyte leakage, proline content, and survival rate under stress. In addition, the seeds of ZmbZIP72-overexpressing transgenic plants were hypersensitive to ABA and osmotic stress. Moreover, overexpression of ZmbZIP72 enhanced the expression of ABA-inducible genes such as RD29B, RAB18, and HIS1-3. These results suggest that the ZmbZIP72 protein functions as an ABA-dependent transcription factor in positive modulation of abiotic stress tolerance and may be a candidate gene with potential application in molecular breeding to enhance stress tolerance in crops.

Keywords

Abiotic stress bZIP-transcription factor Maize Stress tolerance 

Abbreviations

ABA

Abscisic acid

ABRE

ABA responsive element

Arabidopsis

Arabidopsis thaliana

bZIP

Basic leucine zipper

CaMV 35S

Cauliflower mosaic virus 35S promoter

GFP

Green fluorescent protein

PCR

Polymerase chain reaction

PPT

Phosphinothricin

qRT-PCR

Quantitative real-time PCR

RACE

Rapid amplification of cDNA ends

ROS

Reactive oxygen species

SnRK

Sucrose non-fermenting-1 related protein kinase

UTR

Untranslated region

WT

Wild-type

Supplementary material

425_2011_1496_MOESM1_ESM.doc (24 kb)
Expression analysis of ZmbZIP72 in transgenic lines. qRT-PCR analysis of ZmbZIP72 expression in WT and transgenic lines TL5-2, TL6-1, TL11-3 and TL12-3. The transcript level of ZmbZIP72 in TL11-3 was used as the calibrator. The Arabidopsis actin gene (NM_112764) was used as the internal control for normalization. Error bars indicate SD (n = 4) (DOC 23 kb)
425_2011_1496_MOESM2_ESM.doc (138 kb)
Phenotype of ZmbZIP72-overexpressed transgenic Arabidopsis plants under normal growth conditions. a Morphological comparisons of 2-week-old plants of WT and ZmbZIP72-overexpressed transgenic plants under normal growth conditions. b The plant height of WT and ZmbZIP72-overexpressed transgenic plants as described in a. Error bars indicate SD (n = 8) (DOC 137 kb)
425_2011_1496_MOESM3_ESM.doc (49 kb)
Free proline content in plants. Relative content of proline in 2-week-old transgenic seedlings. Error bars indicate SD (n = 6). * indicates significant difference from WT at P < 0.05 by student’s t-test (DOC 49 kb)
425_2011_1496_MOESM4_ESM.doc (130 kb)
Salt-stress tolerance of WT and ZmbZIP72-overexpressed Arabidopsis seedlings. Two-week-old seedlings grown in soil were treated with 300 mM NaCl solution for 2 weeks, and then recovered for 7 days (DOC 130 kb)
425_2011_1496_MOESM5_ESM.doc (58 kb)
Supplementary material 5 (DOC 57 kb)
425_2011_1496_MOESM6_ESM.doc (30 kb)
Supplementary material 6 (DOC 29 kb)

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Institute of Crop ScienceChinese Academy of Agricultural SciencesBeijingChina
  2. 2.College of Biological SciencesChina Agricultural UniversityBeijingChina

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