Quantification of disease progression of several microbial pathogens on Arabidopsis thaliana using real-time fluorescence PCR.
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.
Brouwer
Margreet
M
Centre of Microbial and Plant Genetics, Katholieke Universiteit Leuven, Kasteelpark Arenberg 20, B-3001 Heverlee-Leuven, Belgium.
Lievens
Bart
B
Van Hemelrijck
Wendy
W
Van den Ackerveken
Guido
G
Cammue
Bruno P A
BP
Thomma
Bart P H J
BP
D000528:Alternaria / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D017360:Arabidopsis / Q000382:microbiology*; D001419:Bacteria / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification; D018533:Biomass; D020171:Botrytis / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D004247:DNA / Q000032:analysis / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D004271:DNA, Fungal / Q000032:analysis / Q000302:isolation & purification; D005453:Fluorescence; D005658:Fungi / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification; D016973:Pectobacterium carotovorum / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D044742:Peronospora / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification; D010935:Plant Diseases / Q000382:microbiology*; D018515:Plant Leaves / Q000382:microbiology; D016133:Polymerase Chain Reaction; D044224:Pseudomonas syringae / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification
D004269:DNA, Bacterial; D004271:DNA, Fungal; D004247:DNA