Phytophthora Ramorum and Grosmannia Clavigera 3
bioRxiv preprint doi: https://doi.org/10.1101/736637; this version posted August 15, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 1 Molecular assays to detect the presence and viability of 2 Phytophthora ramorum and Grosmannia clavigera 3 4 Barbara Wonga,b*, Isabel Lealc*, Nicolas Feaub, Angela Daleb, Adnan Uzunovicd, 5 Richard C. Hamelina,b 6 aFaculté de foresterie et géomatique, Institut de Biologie Intégrative et des Systèmes (IBIS), 7 Université Laval, Québec, QC, Canada 8 bDepartment of Forest and Conservation Sciences, University of British Columbia, Vancouver, BC, 9 Canada 10 cPacific Forestry Centre, Natural Resources Canada, Victoria, BC, Canada 11 dFPInnovations, Vancouver, BC, Canada 12 *These authors contributed equally to this work 13 Abstract/Keywords 14 To determine if living microorganisms of phytosanitary concern are present in wood after 15 eradication treatment and to evaluate the efficacy of such treatments, the method of choice is to grow 16 microbes in petri dishes for subsequent identification. However, some plant pathogens are difficult or 17 impossible to grow in axenic cultures. A molecular methodology capable of detecting living fungi and 18 fungus-like organisms in situ can provide a solution. RNA represents the transcription of genes and can 19 therefore only be produced by living organisms, providing a proxy for viability. We designed and used 20 RNA-based molecular diagnostic assays targeting genes essential to vital processes and assessed their 21 presence in wood colonized by fungi and oomycetes through reverse transcription and real-time 22 polymerase chain reaction (PCR).
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