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Induced ectopic expression of At-CBF1 in marker-free transgenic tomatoes confers enhanced chilling tolerance

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Abstract

In an attempt to improve chilling stress tolerance, an Arabidopsis C-repeat binding factor 1 (At-CBF1) gene driven by the inducible promoter RD29A was co-transferred into tomato var. Shalimar. Marker (NPTII)-free transgenic were obtained in T1 generation because of unlinked integration of CBF1 and NPTII genes. Reverse transcription-polymerase chain reaction confirmed the expression of CBF1 in T1 transgenic lines. Study of expression pattern in T1 transgenic line showed a gradual increase with increasing chilling stress period and also confirmed the reversibility of expression on removal of stress. The transgenic plants exhibited no morphological and agronomical differences as compared to non-transformed plants. When young transgenic plants were exposed to chilling stress (4°C) for 3 days, increased survival (50%) was observed in transgenic lines than non-transformed plants (10%). Transgenic plants subjected to the chilling stress showed a significant decrease in membrane injury index and lipid peroxidation and also increased significantly free proline content in the leaf tissues as compared to non-transformed plants. Thus, these findings indicate that marker-free transgenic tomato plants expressing Arabidopsis CBF1 gene provided protection and conferred cold tolerance to transgenic tomato without any phenotypic variation.

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Abbreviations

BAP:

6-Benzylaminopurine

CBF1 :

C-repeat binding factor-1

CTAB:

Cetyl tri-methyl ammonium bromide

IAA:

Indole acetic acid

MII:

Membrane injury index

MS:

Murashige and Skoog

NPTII :

Neomycin phosphotransferase-II

RT-PCR:

Reverse transcription-polymerase chain reaction

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Acknowledgments

Authors are grateful to DRDO HQ, Ministry of Defence, Govt. of India for the financial help. The generous gift of the co-transformation constructs by Dr. A. K. Sharma (Centre for Plant Molecular Biology, University of Delhi, South Campus, India) is gratefully acknowledged.

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Authors and Affiliations

  1. Molecular Biology and Genetic Engineering Division, Defence Institute of Bio-Energy Research, Haldwani, 263 139, Uttarakhand, India

    Shweta Singh, Meenal Rathore, Danswrang Goyary, Rupesh Kumar Singh, Sivalingam Anandhan & Zakwan Ahmed

  2. Department of Zoology, HNB Garhwal University, SRT Campus, Badshahithal, Tehri, Uttarakhand, India

    Dinesh K. Sharma

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  1. Shweta Singh
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  2. Meenal Rathore
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  3. Danswrang Goyary
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  6. Dinesh K. Sharma
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  7. Zakwan Ahmed
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Correspondence to Danswrang Goyary.

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Communicated by A. Feher.

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Singh, S., Rathore, M., Goyary, D. et al. Induced ectopic expression of At-CBF1 in marker-free transgenic tomatoes confers enhanced chilling tolerance. Plant Cell Rep 30, 1019–1028 (2011). https://doi.org/10.1007/s00299-011-1007-0

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  • DOI: https://doi.org/10.1007/s00299-011-1007-0

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