| Detection of Puccinia horiana, the causal agent of Chrysanthemum white rust, with PCR and chlorophyll fluorescence image analysis. S., , E., , P., , M., , M., & , K. 55th International Symposium on Crop Protection; May 6, 2003, Gent, Belgium (2005) | |||||||||
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| Chrysanthemum white rust is the most important foliar disease of pot chrysanthemum and is often controlled through intensive preventive fungicide applications. The objective of this research was to develop methods for early and sensitive detection of this pathogen in plant and air samples. These methods may be applied in more sustainable disease management and will serve as research tools for the study of the biology of the pathogen. Two methods were evaluated. The first method was chlorophyll fluorescence image analysis. With this method, the pathogen could be detected in infected leaves before visual symptoms were apparent. However, the amount of time gained was limited to 2 days maximum. The second method was PCR detection based on molecular analysis of the internal transcribed spacer (ITS) region of the rDNA. DNA was isolated from the basidiospores from several strains and the ITS region was cloned and sequenced. Several PCR and real-time PCR primers were designed based on the ITS1 and ITS2 sequences. As little as 10 fg of genomic DNA could be detected. First tests show that these primers may be useful in the sensitive detection of P. horiana in DNA extracted from infected plants. Specificity of detection was cross-checked against a variety of other fungi (saprophytes and other rusts) that may occur in the same environment. | |||||||||
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