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Applied and Environmental Microbiology, March 2005, p. 1546-1552, Vol. 71, No. 3
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.3.1546-1552.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

In Vivo Monitoring of Obligate Biotrophic Pathogen Growth by Kinetic PCR

Brian Boyle, Richard C. Hamelin, and Armand Séguin*

Natural Resources Canada, Canadian Forest Service, Laurentian Forestry Centre, Sainte-Foy, Quebec, Canada

Received 15 June 2004/ Accepted 5 October 2004

The plant kingdom is constantly challenged by a battery of evolving pathogens. New species or races of pathogens are discovered on crops that were initially bred for disease resistance, and globalization is facilitating the movement of exotic pests. Among these pests, obligate biotrophic parasites make up some of the most damaging groups and have been particularly challenging to study. Here we demonstrate the utility of kinetic PCR (kPCR) (real-time PCR, quantitative PCR) to assess the growth of poplar rust, caused by Melampsora species, by quantification of pathogen DNA. kPCR allowed the construction of reliable growth curves from inoculation through the final stages of uredinial maturation, as well as pathogen monitoring before symptoms become visible. Growth parameters, such as latency period, generation time in logarithmic growth, and the increase in DNA mass at saturation, were compared in compatible, incompatible, and nonhost interactions. Pathogen growth was monitored in different applications dealing with plant pathology, such as host and pathogen diversity and transgenic crop improvement. Finally, the capacity of kPCR to differentiate pathogens in the same sample has broad molecular ecology applications for dynamically monitoring the growth of fungi in their environments or in mixed populations or to measure the efficacy of pest control strategies.

* Corresponding author. Mailing address: Natural Resources Canada, Canadian Forest Service, Laurentian Forestry Centre, 1055 du P.E.P.S., P.O. Box 3800, Sainte-Foy, Quebec, Canada G1V 4C7. Phone: (418) 648-5832. Fax: (418) 648-5849. E-mail: Armand.Seguin{at}NRCan.gc.ca.

Applied and Environmental Microbiology, March 2005, p. 1546-1552, Vol. 71, No. 3
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.3.1546-1552.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Rinaldi, C., Kohler, A., Frey, P., Duchaussoy, F., Ningre, N., Couloux, A., Wincker, P., Le Thiec, D., Fluch, S., Martin, F., Duplessis, S. (2007). Transcript Profiling of Poplar Leaves upon Infection with Compatible and Incompatible Strains of the Foliar Rust Melampsora larici-populina. Plant Physiol. 144: 347-366 [Abstract] [Full Text]  


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