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The grapevine polygalacturonase-inhibiting protein (VvPGIP1) reduces Botrytis cinerea susceptibility in transgenic tobacco and differentially inhibits fungal polygalacturonases

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Abstract

Polygalacturonase-inhibiting proteins (PGIPs) selectively inhibit polygalacturonases (PGs) secreted by invading plant pathogenic fungi. PGIPs display differential inhibition towards PGs from different fungi, also towards different isoforms of PGs originating from a specific pathogen. Recently, a PGIP-encoding gene from Vitis vinifera (Vvpgip1) was isolated and characterised. PGIP purified from grapevine was shown to inhibit crude polygalacturonase extracts from Botrytis cinerea, but this inhibitory activity has not yet been linked conclusively to the activity of the Vvpgip1 gene product. Here we use a transgenic over-expression approach to show that the PGIP encoded by the Vvpgip1 gene is active against PGs of B. cinerea and that over-expression of this gene in transgenic tobacco confers a reduced susceptibility to infection by this pathogen. A calculated reduction in disease susceptibility of 47–69% was observed for a homogeneous group of transgenic lines that was statistically clearly separated from untransformed control plants following infection with Botrytis over a 15-day-period. VvPGIP1 was subsequently purified from transgenic tobacco and used to study the specific inhibition profile of individual PGs from Botrytis and Aspergillus. The heterologously expressed and purified VvPGIP1 selectively inhibited PGs from both A. niger and B.␣cinerea, including BcPG1, a PG from B. cinerea that has previously been shown to be essential for virulence and symptom development. Altogether our data confirm the antifungal nature of the VvPGIP1, and the in vitro inhibition data suggest at least in part, that the VvPGIP1 contributed to the observed reduction in disease symptoms by inhibiting the macerating action of certain Botrytis PGs in planta. The ability to correlate inhibition profiles to individual PGs provides a more comprehensive analysis of PGIPs as antifungal genes with biotechnological potential, and adds to our understanding of the importance of PGIP:PG interactions during disease and symptom development in plants.

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Acknowledgements

This work was supported by grants from the South African Wine Industry (Winetech), the National research foundation of South Africa (NRF), the US Department of Energy (DOE) (DE-FG02-96ER20221) and the DOE-funded Center for Plant and Microbial Complex Carbohydrates (DE-FG02-93ER20097).

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Authors and Affiliations

  1. Department of Viticulture & Oenology, Institute for Wine Biotechnology, Stellenbosch University, Stellenbosch, 7600, South Africa

    Dirk A. Joubert, John V. W. Becker & Melané A. Vivier

  2. Department of Biochemistry, Institute of Botany, University of Neuchatel, Rue Emile-Argand 11, CH-2007, Neuchatel, Switzerland

    Ana R. Slaughter

  3. Department of Plant Sciences, University of the Free State, Bloemfontein, 9300, South Africa

    Gabré Kemp

  4. Complex Carbohydrate Research Centre, University of Georgia, Athens, Georgia, 30602-4712, USA

    Carl Bergmann

  5. Laboratory of Microbiology, Wageningen University, Wageningen, 6703 HA, The Netherlands

    Geja H. Krooshof & Jacques Benen

  6. The Australian Wine Research Institute, Waite Road, Urrbrae, Adelaide, SA, 5064, Australia

    Isak S. Pretorius

Authors
  1. Dirk A. Joubert
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  2. Ana R. Slaughter
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  3. Gabré Kemp
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  4. John V. W. Becker
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  5. Geja H. Krooshof
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  6. Carl Bergmann
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  7. Jacques Benen
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  8. Isak S. Pretorius
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  9. Melané A. Vivier
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Correspondence to Melané A. Vivier.

Additional information

Dirk A. Joubert and Ana R. Slaughter contributed equally to this work.

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Joubert, D.A., Slaughter, A.R., Kemp, G. et al. The grapevine polygalacturonase-inhibiting protein (VvPGIP1) reduces Botrytis cinerea susceptibility in transgenic tobacco and differentially inhibits fungal polygalacturonases. Transgenic Res 15, 687–702 (2006). https://doi.org/10.1007/s11248-006-9019-1

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  • DOI: https://doi.org/10.1007/s11248-006-9019-1

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