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Constitutive overexpression of small HSP24.4 gene in transgenic tomato conferring tolerance to high-temperature stress

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Abstract

Acquired thermotolerance in plants refers to the ability to cope with lethal high temperatures and it reflects an actual tolerance mechanism that occurs naturally in plants. Tomato (Solanum lycopersicum syn. Lycopersicon esculentum L.) is sensitive to high temperature at all stages of its growth and development. Considering the important role of the heat shock protein gene (sHSP24.4 gene) in imparting tolerance to high temperature stress in the cells and tissues, we isolated small HSP24.4 (MasHSP24.4) cDNA from wild banana (Musa accuminata) and introduced it into the cultivated tomato cv. PKM1 by using Agrobacterium tumefaciens-mediated genetic transformation. Stable integration and expression of the transgene in the tomato genome was demonstrated by Southern, Northern and Western blot analyses. There was no adverse effect of transgene expression on overall growth and development of the transgenic plants. The genetic analysis of the transgenic T2 lines showed that the transgene segregated in a Mendelian ratio. We compared the survival of T2 transgenic lines compared to the control plants after exposure to different levels of high temperature. The gene MasHSP24.4 was expressed in root, shoot and stem tissues under 45 °C treatment and conferred tolerance to high-temperature stress as shown by increased seed germination, healthy vegetative growth and normal fruit and seed setting. The transgenic tomato plants showed significantly better growth performance in the recovery phase following the stress. This thermotolerance appeared to be solely due to overexpression of the sHSP24.4 gene. Thus, the transgenic tomato plants developed during the present investigations can be grown at high temperatures.

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Acknowledgments

Upender Mahesh is grateful to Andhra Pradesh Netherlands Biotechnology Research Programme sanctioned by Ministry of External Affairs, The Netherlands (BTU/PhD/Fellowship/2004-05/1372) for providing Junior and Senior Research Fellowships, and Department of Science and Technology, New Delhi, India for Young Scientist Fellowship and UGC for awarding Dr. S. Kothari Post-Doctoral Fellowship, India.

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  1. Upender Mahesh

    Present address: Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad, 500 046, India

Authors and Affiliations

  1. Department of Biotechnology, Plant Biotechnology Research Group, Kakatiya University, Warangal, 506 009, Andhra Pradesh, India

    Upender Mahesh, Praveen Mamidala & Rama Swamy Nanna

  2. National Centre for Genetic Resources and Biotechnology, Embrapa-Cenargen, Brasilia, Brazil

    Francisco J. L. Aragao & M. T. Souza

  3. Biotechnology Research Laboratory, Regional Agricultural Research Station, Acharya NG Ranga Agricultural University, Warangal, 506 007, Andhra Pradesh, India

    Upender Mahesh, Savitha Rapolu & P. J. M. Rao

  4. Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad, 500 046, India

    P. B. Kirti

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  1. Upender Mahesh
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Correspondence to Rama Swamy Nanna.

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Mahesh, U., Mamidala, P., Rapolu, S. et al. Constitutive overexpression of small HSP24.4 gene in transgenic tomato conferring tolerance to high-temperature stress. Mol Breeding 32, 687–697 (2013). https://doi.org/10.1007/s11032-013-9901-5

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