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A bifunctional TPS–TPP enzyme from yeast confers tolerance to multiple and extreme abiotic-stress conditions in transgenic Arabidopsis

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Abstract

Improving stress tolerance is a major goal for agriculture. Trehalose is a key molecule involved in drought tolerance in anhydrobiotic organisms. Here we describe the construction of a chimeric translational fusion of yeast trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase. This construct was overexpressed in yeast cells displaying both TPS and TPP enzyme activities and trehalose biosynthesis capacity. In Arabidopsis thaliana, the gene fusion was overexpressed using either the 35S promoter or the stress-regulated rd29A promoter. Transgene insertion in the genome was checked by PCR and transcript expression by RT-PCR. Several independent homozygous lines were selected in the presence of kanamycin and further analyzed. Trehalose was accumulated in all these lines at low levels. No morphological or growth alterations were observed in lines overexpressing the TPS1–TPS2 construct, whereas plants overexpressing the TPS1 alone under the control of the 35S promoter had aberrant growth, color and shape. TPS1–TPS2 overexpressor lines were glucose insensitive, consistent with a suggested role of trehalose/T6P in modulating sugar sensing and carbohydrate metabolism. Moreover, TPS1–TPS2 lines displayed a significant increase in drought, freezing, salt and heat tolerance. This is the first time that trehalose accumulation in plants is shown to protect against freezing and heat stress. Therefore, these results demonstrate that engineering trehalose metabolism with a yeast TPS–TPP bifunctional enzyme confers multiple stress protection in plants, comprising a potential tool to improve stress-tolerance in crops.

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Abbreviations

T6P:

Trehalose-6-phosphate

TPS:

Trehalose-6-phosphate synthase

TPS1 :

Yeast gene encoding TPS

TPP:

Trehalose-6-phosphate phosphatase

TPS2 :

Yeast gene encoding TPP

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Acknowledgments

We are indebted to Dr. Patricia León (Instituto de Biotecnología-UNAM, Cuernavaca, Mexico) for her kind gift of Col-0 Arabidopsis thaliana seeds and the abi4–1 mutant. We also thank Paul Gaytán and Eugenio López (Instituto de Biotecnología-UNAM, Cuernavaca, Mexico) for oligonucleotide synthesis. G.I. received funding from CONACYT (No. 2004-CO1-46078) and R.S. from PROMEP (No. PTC-137).

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Authors and Affiliations

  1. Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Av. Universidad 1001, Col. Chamilpa, Cuernavaca Mor, 62209, Mexico

    José A. Miranda, Nelson Avonce, Ramón Suárez & Gabriel Iturriaga

  2. VIB Department of Molecular Microbiology, K.U. Leuven, Kasteelpark Arenberg 31, 3001, Leuven-Heverlee, Flanders, Belgium

    Nelson Avonce, Johan M. Thevelein & Patrick Van Dijck

  3. Laboratory of Molecular Cell Biology, K.U. Leuven, Kasteelpark Arenberg 31, 3001, Leuven-Heverlee, Flanders, Belgium

    Nelson Avonce, Johan M. Thevelein & Patrick Van Dijck

Authors
  1. José A. Miranda
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  2. Nelson Avonce
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  3. Ramón Suárez
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  4. Johan M. Thevelein
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  5. Patrick Van Dijck
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  6. Gabriel Iturriaga
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Corresponding author

Correspondence to Gabriel Iturriaga.

Additional information

J.A. Miranda, and N. Avonce contributed equally to the present work.

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Miranda, J.A., Avonce, N., Suárez, R. et al. A bifunctional TPS–TPP enzyme from yeast confers tolerance to multiple and extreme abiotic-stress conditions in transgenic Arabidopsis . Planta 226, 1411–1421 (2007). https://doi.org/10.1007/s00425-007-0579-y

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