Quantification of disease progression of several microbial pathogens on Arabidopsis thaliana using real-time fluorescence PCR

Authors

  • Margreet Brouwer,

    1. Centre of Microbial and Plant Genetics (CMPG), Katholieke Universiteit Leuven, Kasteelpark Arenberg 20, B-3001 Heverlee-Leuven, Belgium
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  • Bart Lievens,

    1. Centre of Microbial and Plant Genetics (CMPG), Katholieke Universiteit Leuven, Kasteelpark Arenberg 20, B-3001 Heverlee-Leuven, Belgium
    2. Scientia Terrae Research Institute, Fortsesteenweg 30A, B-2860 Sint-Katelijne-Waver, Belgium
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  • Wendy Van Hemelrijck,

    1. Centre of Microbial and Plant Genetics (CMPG), Katholieke Universiteit Leuven, Kasteelpark Arenberg 20, B-3001 Heverlee-Leuven, Belgium
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  • Guido Van den Ackerveken,

    1. Department of Molecular and Cellular Biology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands
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  • Bruno P.A Cammue,

    1. Centre of Microbial and Plant Genetics (CMPG), Katholieke Universiteit Leuven, Kasteelpark Arenberg 20, B-3001 Heverlee-Leuven, Belgium
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  • Bart P.H.J Thomma

    Corresponding author
    1. Centre of Microbial and Plant Genetics (CMPG), Katholieke Universiteit Leuven, Kasteelpark Arenberg 20, B-3001 Heverlee-Leuven, Belgium
    2. Laboratory of Phytopathology, Wageningen University, Binnenhaven 5, 6709 PD Wageningen, The Netherlands
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*Corresponding author. Tel.: +31 (317) 484536; Fax: +31 (317) 483412, E-mail address: bart.thomma@wur.nl

Abstract

An accurate monitoring of disease progression is important to evaluate disease susceptibility phenotypes. Over the years, Arabidopsis thaliana has become the model species to serve as a host in plant–pathogen interactions. Despite the efforts to study genetic mechanisms of host defense, little efforts are made for a thorough pathogen assessment, often still depending on symptomology. This manuscript describes the use of real-time polymerase chain reaction (PCR) to assess pathogen growth in the host Arabidopsis for a number of frequently studied pathogens. A wide range of correlations between pathogen biomass and fluorescence is demonstrated, demonstrating the theoretical sensitivity of the technique. It is also demonstrated that host DNA does not interfere with the quantification of pathogen DNA over a wide range. Finally, quantification of pathogen biomass in different plant genotypes with a varying degree of resistance shows the capability of this technique to be used for assessment of pathogen development in disease progression.

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