Location: Crop Improvement and Protection ResearchTitle: Molecular quantification of Fusarium oxysporum f. sp. fragariae in soil
|Matson, Michael - Mike|
|CROUCH, UMA - Former ARS Employee|
|MARTIN, LEAH - Former ARS Employee|
|JACQUEZ, MARC - Former ARS Employee|
Submitted to: American Phytopathological Society Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 4/15/2021
Publication Date: 8/2/2021
Citation: Matson, M.E., Crouch, U.T., Martin, L.R., Jacquez, M.A., Zepeda, S., Dorn, J.R., Goldman, P.H., Henry, P.M., Martin, F.N. 2021. Molecular quantification of Fusarium oxysporum f. sp. fragariae in soil. American Phytopathological Society Annual Meeting, August 2-6, 2021 (virtual).
Technical Abstract: Informed management decisions with early pathogen detection is a main line of defense against soilborne pathogens and their respective diseases. Early detection is especially important for soilborne pathogens that have few control measures following disease incidence. While some fungal pathogens such as Verticillium dahliae (Vd) and Macrophomina phaseolina (Mps) can already be detected from soil DNA at inoculum levels approaching their limit of pathogenicity, currently available methods do not achieve this for Fusarium oxysporum f.sp. fragariae (Fof). Difficulties of molecular-based detection and quantification of Fof include a uninucleate long-term resting chlamydospore, a qPCR TaqMan assay targeting'a single copy gene, and the lack of effective DNA extraction methods'that overcomes PCR inhibitors commonly found in soil/environmental samples.''Here, we describe a new technique to extract DNA from larger masses of soils, compared to commercial kits, and'a cleanup process that is effective and automated.' Fof'could be detected through qPCR at counts below 10 CFU/g of soil. In some instances, soil samples were positive using molecular detection and negative for Fof in plating assays. Compared to the MP'FastPrep'Soil kit'that extracts DNA from 0.5 g of soil, our method yields an average of 2.5x more DNA'from 15.0 g of soil.' The application of'our method is extended to'Vd'and'Mps and'increases the sensitivity of qPCR assays for those pathogens, thereby opening the possibility of pooling multiple soil samples and reducing costs for pathogen detection.