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Evaluation of Transgenic ‘Chardonnay’ (Vitis vinifera) Containing Magainin Genes for Resistance to Crown Gall and Powdery Mildew

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Abstract

Magainins, short peptides with broad-spectrum antimicrobial activity in vitro, were assayed for their ability to confer resistance to pathogens in transgenic grapevines. Embryogenic cell suspensions of ‘Chardonnay’ (Vitis vinifera L.) were co-transformed by microprojectile bombardment with a plasmid carrying the npt-II gene and a second plasmid harboring either a natural magainin-2 (mag2) or a synthetic derivative (MSI99) gene. Magainin genes and the marker gene were driven by Arabidopsis ubiquitin-3 and ubiquitin-11 promoters, respectively. A total of 10 mag2 and 9 MSI99 regenerated lines were studied by Southern blot hybridization, which showed 1–6 transgene integration events into the plant genome. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) revealed a variable range in transcription levels among mag2 and MSI99 lines. A positive correlation between number of integration events and transcription level was observed (p<0.05). Plants were acclimated and challenged in the greenhouse with either Agrobacterium vitis strains (bacterial crown gall pathogen) at 108 cfu/ml or Uncinula necator (fungal powdery mildew pathogen) at 105 conidia/ml for evaluation of disease resistance. A total of 6 mag2 and 5 MSI99 lines expressing the antimicrobial genes exhibited significant reductions of crown gall symptoms as compared to non-transformed controls. However, only two mag2 lines showed measurable symptom reductions in response to U. necator, but not strong resistance. Our results suggest that the expression of magainin-type genes in grapevines may be more effective against bacteria than fungi. Additional strategies to enhance transgene expression and the spectrum of resistance to grape diseases are suggested.

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Abbreviations

AMP:

antimicrobial peptide

mag2 :

natural magainin-2 gene

MSI99 :

synthetic analog of magainin-2 gene

RT-PCR:

reverse transcription-polymerase chain reaction

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Authors and Affiliations

  1. Department of Horticultural Sciences, New York State Agricultural Experiment Station, Cornell University, Geneva, NY, 14456, USA

    José R. Vidal, Julie R. Kikkert, Patricia G. Wallace & Bruce I. Reisch

  2. Department of Plant Pathology, New York State Agricultural Experiment Station, Cornell University, Geneva, NY, 14456, USA

    Mickael A. Malnoy

  3. Computer Center, New York State Agricultural Experiment Station, Cornell University, Geneva, NY, 14456, USA

    John Barnard

  4. Departamento de Biotecnología, ETS Ingenieros Agrónomos, Universidad Politécnica de Madrid, 28040, Madrid, Spain

    José R. Vidal

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  1. José R. Vidal
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  2. Julie R. Kikkert
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  3. Mickael A. Malnoy
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  4. Patricia G. Wallace
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  5. John Barnard
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  6. Bruce I. Reisch
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Correspondence to José R. Vidal.

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Vidal, J.R., Kikkert, J.R., Malnoy, M.A. et al. Evaluation of Transgenic ‘Chardonnay’ (Vitis vinifera) Containing Magainin Genes for Resistance to Crown Gall and Powdery Mildew. Transgenic Res 15, 69–82 (2006). https://doi.org/10.1007/s11248-005-4423-5

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  • DOI: https://doi.org/10.1007/s11248-005-4423-5

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