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Stable expression of a defense-related gene in wheat epidermis under transcriptional control of a novel promoter confers pathogen resistance

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Abstract

Tissue-specific or regulated expression of transgenes is desirable in order to prevent pleiotropic side effects of putatively harmful transgene products as well as loss of energy resources due to unnecessary accumulation of transgene products. Epidermis-specific expression would be useful for many defense-related genes directed against attack by fungal pathogens that enter the plant body by direct penetration through the epidermis. In an approach to enhance resistance of wheat to the powdery mildew fungus Blumeria graminis f.sp. tritici, a novel epidermis-specific promoter was developed and used for expression of two defense-related genes. A 2.3 kb fragment of the wheat GstA1 promoter in combination with an intron-containing part of the wheat WIR1a gene was found to drive strong and constitutive transient expression in wheat epidermis. This promoter–intron combination was used for overexpression of oxalate oxidase9f-2.8 and TaPERO peroxidase, two defense-related wheat genes expressed in inner leaf tissues. Expression studies of several transgenic lines by in situ oxalate-oxidase staining, RNA and protein blot analyses, as well as real-time PCR, demonstrated strong and constitutive transgene expression in the shoot epidermis. Transient as well as stable over-expression of the TaPERO peroxidase gene in wheat epidermis under the control of the GstA1i promoter resulted in enhanced resistance against Blumeria graminis f.sp. tritici, whereas oxalate-oxidase overexpression had no effect in either system. The data suggest that the wheat GstA1 promoter in combination with the WIR1a intron is useful for transgenic approaches to fungal disease resistance in cereals.

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Authors and Affiliations

  1. Agronomy Department, PMCB, Laboratory of Molecular Plant Physiology, University of Florida - IFAS, 2191 McCarty Hall, P.O. Box 110300, Gainesville, FL, 32611-0300, USA

    Fredy Altpeter

  2. Institute of Plant Genetics and Crop Plant Research, Corrensstrasse 3, D-06466, Gatersleben, Germany

    Alok Varshney, Dimitar Douchkov, Jochen Kumlehn & Patrick Schweizer

  3. Institute of Plant Biology, University of Zürich, Zollikerstrasse 107, CH-8000, Zürich, Switzerland

    Olaf Abderhalden & Robert Dudler

  4. ETH Zürich, Institut für Pflanzenwissenschaften, Universitätstr. 2, CH-8092, Zürich, Switzerland

    Christof Sautter

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  1. Fredy Altpeter
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  2. Alok Varshney
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  3. Olaf Abderhalden
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  4. Dimitar Douchkov
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  7. Robert Dudler
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  8. Patrick Schweizer
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Correspondence to Patrick Schweizer.

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Altpeter, F., Varshney, A., Abderhalden, O. et al. Stable expression of a defense-related gene in wheat epidermis under transcriptional control of a novel promoter confers pathogen resistance. Plant Mol Biol 57, 271–283 (2005). https://doi.org/10.1007/s11103-004-7564-7

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  • DOI: https://doi.org/10.1007/s11103-004-7564-7

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