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Over-expression of a cacao class I chitinase gene in Theobroma cacao L. enhances resistance against the pathogen, Colletotrichum gloeosporioides

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Abstract

Theobroma cacao L. plants over-expressing a cacao class I chitinase gene (TcChi1) under the control of a modified CaMV-35S promoter were obtained by Agrobacterium-mediated transformation of somatic embryo cotyledons. Southern blot analysis confirmed insertion of the transgene in eight independent lines. High levels of TcChi1 transgene expression in the transgenic lines were confirmed by northern blot analysis. Chitinase activity levels were measured using an in vitro fluorometric assay. The transgene was expressed at varying levels in the different transgenic lines with up to a sixfold increase of endochitinase activity compared to non-transgenic and transgenic control plants. The in vivo antifungal activity of the transgene against the foliar pathogen Colletotrichum gloeosporioides was evaluated using a cacao leaf disk bioassay. The assay demonstrated that the TcChi1 transgenic cacao leaves significantly inhibited the growth of the fungus and the development of leaf necrosis compared to controls when leaves were wound inoculated with 5,000 spores. These results demonstrate for the first time the utility of the cacao transformation system as a tool for gene functional analysis and the potential utility of the cacao chitinase gene for increasing fungal pathogen resistance in cacao.

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Acknowledgements

We would like to thank Gabriela Antunez de Mayolo Wilmking for her contribution to the construction of vector pGAM00.0511 and Sara Milillo and Amanda Thompson for their technical assistance with performing and data analysis of the chitinase protein activity assay.

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Authors and Affiliations

  1. The Department of Horticulture, The Pennsylvania State University, University Park, Chester, PA, 16802, USA

    Siela N. Maximova, Ann Young, Sharon Pishak, Joseph A. Verica & Mark J. Guiltinan

  2. The Department of Plant Pathology, The Pennsylvania State University, Chester, PA, 16802, USA

    Jean-Philippe Marelli

Authors
  1. Siela N. Maximova
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  2. Jean-Philippe Marelli
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  3. Ann Young
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  4. Sharon Pishak
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  6. Mark J. Guiltinan
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Correspondence to Mark J. Guiltinan.

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Maximova, S.N., Marelli, JP., Young, A. et al. Over-expression of a cacao class I chitinase gene in Theobroma cacao L. enhances resistance against the pathogen, Colletotrichum gloeosporioides . Planta 224, 740–749 (2006). https://doi.org/10.1007/s00425-005-0188-6

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  • DOI: https://doi.org/10.1007/s00425-005-0188-6

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