Development of a quick and reliable molecular detection system for Sclerotinia stem rot of canola in western Canada
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Sclerotinia stem rot is an important, yield-limiting disease of canola caused by the necrotrophic fungus Sclerotinia sclerotiorum. The development of stem rot is highly dependent on environmental conditions and inoculum levels, and as a result disease occurrence and severity are sporadic in nature. In western Canada, stem rot is primarily controlled through the routine application of fungicides, which are often applied with no indication of disease risk. In order to reduce non-economical application of fungicides, a reliable forecasting system is needed. Since the infection of canola petals in an important step in the stem rot disease cycle, a quantitative (q)PCR-based system that can give an estimate of petal infestation rates could form the basis of a reliable forecasting system to help guide fungicide spray decisions. A primer and TaqMan probe have been developed based on the sequence of a novel protein that is specific to S. sclerotiorum. This primer-probe system, when used in a TaqMan qPCR assay, can quantify S. sclerotiorum DNA with a high level of sensitivity and specificity. The development of an internal amplification control will further increase the reliability of the assay by accounting for false negatives. This assay shows great potential for use as the basis of a stem rot forecasting system for canola in western Canada.
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