Volume 19 Part 1 Article 20
Title: Multiplexed Detection and Relative Quantitation of Bacterial And Fungal Pathogens of Mushrooms
Author: Lee Smith and Keith Stanley
PCR-based methods for pathogen detection are fast, sensitive and specific. However, given the diversity of pathogens capable of infecting mushroom crops and affecting crop value, the capacity to combine or multiplex multiple PCR tests is essential to allow the implementation of these methods within the mushroom cultivation industry. Using the Multiplex-Tandem PCR technology, we have developed systems for the rapid detection of up to 8 different pathogens (including pathogenic Trichoderma, Lecanicilium, Cladobotryum, and Pseudomonas tolaasii) with concurrent differentiation of Trichoderma aggressivum (v. aggressivum and v. europaeum) and Lecanicillium fungicola (v. aleophilum and v. fungicola) variants. These assays enable relative quantitation of each pathogen and can be applied to compost and casing in addition to environmental swabs. Each assay within the multiplex has been validated by nucleic acid sequencing and trials on mushroom swabs. The assay does not necessarily require extraction, and the results on diseased mushrooms produce the expected result. The utility of this multiplex assay to mushroom farms will be discussed.