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

被引:116
作者
Brouwer, M
Lievens, B
Van Hemelrijck, W
Van den Ackerveken, G
Cammue, BPA
Thomma, BPHJ
机构
[1] Katholieke Univ Leuven, Ctr Microbial & Plant Genet, B-3001 Heverlee, Belgium
[2] Sci Terrae Res Inst, B-2860 St Katelijne Waver, Belgium
[3] Univ Utrecht, Dept Mol & Cellular Biol, NL-3584 CH Utrecht, Netherlands
[4] Wageningen Univ, Lab Phytopathol, NL-6709 PD Wageningen, Netherlands
关键词
LightCycler; SYBR green I; Alternaria brassicicola; Arabidopsis thaliana; Botrytis cinerea; Erwinia carotovora; Peronospora parasitica; Pseudomonas syringae;
D O I
10.1016/S0378-1097(03)00759-6
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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. (C) 2003 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.
引用
收藏
页码:241 / 248
页数:8
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