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Transgenic Research

, Volume 22, Issue 2, pp 327–341 | Cite as

Co-expression of AtbHLH17 and AtWRKY28 confers resistance to abiotic stress in Arabidopsis

  • K. C. Babitha
  • S. V. Ramu
  • V. Pruthvi
  • Patil Mahesh
  • Karaba N. Nataraja
  • M. UdayakumarEmail author
Original Paper

Abstract

Stress adaptation in plants involves altered expression of many genes through complex signaling pathways. To achieve the optimum expression of downstream functional genes, we expressed AtbHLH17 (AtAIB) and AtWRKY28 TFs which are known to be upregulated under drought and oxidative stress, respectively in Arabidopsis. Multigene expression cassette with these two TFs and reporter gene GUS was developed using modified gateway cloning strategy. The GUS assay and expression analysis of transgenes in transgenic plants confirmed the integration of multigene cassette. The transgenic lines exhibited enhanced tolerance to NaCl, Mannitol and oxidative stress. Under mannitol stress condition significantly higher root growth was observed in transgenics. Growth under stress and recovery growth was substantially superior in transgenics exposed to gradual long term desiccation stress conditions. We demonstrate the expression of several downstream target genes under various stress conditions. A few genes having either WRKY or bHLH cis elements in their promoter regions showed higher transcript levels than wild type. However, the genes which did not have either of the motifs did not differ in their expression levels in stress conditions compared to wild type plants. Hence co-expressing two or more TFs may result in upregulation of many downstream target genes and substantially improve the stress tolerance of the plants.

Keywords

Drought Methyl viologen Multigene-construct Salinity Transcription factors 

Abbreviations

bHLH

Basic helix loop helix

FC

Field capacity

GUS

β-glucuronidase

MV

Methyl viologen

NBT

Nitroblue tetrazolium

TF

Transcription factor

H2O2

Hydrogen peroxide

Notes

Acknowledgments

Authors acknowledge the financial support from Department of Biotechnology, Centre of Excellence programme support (BT/01/COE/05/03), Indian Council of Agricultural Research—Niche area of Excellence programme (F.No. 10-(6)/2005 EP&D) and Department of Science and Technology—Fund for Improvement of Science and Technology (SR/FST/LSI-051/2002).

Supplementary material

11248_2012_9645_MOESM1_ESM.pdf (358 kb)
Supplementary material 1 (PDF 357 kb)

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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • K. C. Babitha
    • 1
  • S. V. Ramu
    • 1
  • V. Pruthvi
    • 1
  • Patil Mahesh
    • 1
  • Karaba N. Nataraja
    • 1
  • M. Udayakumar
    • 1
    Email author
  1. 1.Department of Crop PhysiologyUniversity of Agricultural Sciences, GKVKBangaloreIndia

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