Journal
EUROPEAN JOURNAL OF PLANT PATHOLOGY
Volume 126, Issue 1, Pages 129-133Publisher
SPRINGER
DOI: 10.1007/s10658-009-9522-3
Keywords
Detection; Identification; Potato wart disease; TaqMan PCR
Categories
Funding
- Dutch Ministry of Agriculture, Nature and Food Quality
- EU [SSPE-CT-2004-502348]
Ask authors/readers for more resources
Real-time PCR was used for quantitative detection of the potato pathogen, Synchytrium endobioticum, in different substrates: zonal centrifuge extracts, warts and different plant parts of potato. Specific primers and a TaqMan probe, designed from the internal transcribed spacer region of the multi-copy rDNA gene were tested in extracts from artificially and naturally infested soil. Co-amplification of target DNA along with an internal competitor DNA fragment made the diagnostic assay more reliable by guarding against false negative results. A calibrations curve was created by spiking zonal centrifuge fractions of clean soil samples with a dilution series of winter spores. The Taqman assay was also performed on infected potato plant material (stolons) along with the detection of the cytochrome oxidase gene as a potato endogenous control. Sensitivity of the TaqMan assay was improved at least 100-fold and proved to be reliable for accurate diagnosis of the disease.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available