Abstract
A polymerase chain reaction (PCR) assay was developed that amplified a 170-bp fragment of the intergenic spacer region of Sclerotinia sclerotiorum, the cause of white mould. Sensitivity was 10 S. sclerotiorum ascospores per DNA extraction (0.2 ascospores per PCR reaction). The presence of soil did not affect sensitivity at 50, 100 and 500 ascospores/DNA extraction, but reduced sensitivity at 25 and 10 ascospores/DNA extraction by 10% and 30%, respectively. The assay did not amplify DNA of Botrytis cinerea but detected S. minor and S. trifoliorum. Utility of the test for detection of S. sclerotiorum ascospores in bean fields was demonstrated using rotating impaction samplers over two seasons. The use of the test in combination with an impaction sampler may provide benefits in time, sensitivity and specificity compared with visual identification and enumeration of spores from traps only. This system may provide an opportunity to schedule fungicides during periods of inoculum presence for disease management.
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