|URNER, MICHAEL - California State University|
|HUTMACHER, ROBERT - University Of California, Davis|
|ELLIS, MARGARET - California State University|
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 8/6/2022
Publication Date: 8/10/2022
Citation: Urner, M., Hutmacher, R., Ulloa, M., Ellis, M. 2022. Improving Fusarium oxysporum f. sp. vasinfectum race 4 detection with the development of a new quantitative PCR assay. Plant Health 2022, Pittsburgh, Pennsylvania, August 6-10, 2022. ID: 22179.
Technical Abstract: Fusarium oxysporum f. sp. vasinfectum (FOV) is a pathogenic soilborne fungus responsible for Fusarium wilt in cotton (Gossypium spp.). In California, FOV race 4 is of economic importance, and can currently be detected using conventional PCR and race specific primers. However, the current protocol is time consuming, requiring isolation of the fungus from soil or plant tissue, followed by DNA extraction, before confirmation with PCR. Therefore, our goal was to improve the speed and accuracy of detecting FOV race 4 by developing a quantitative PCR (qPCR) assay. FOV race 4 specific primers were developed based on the unique Tfo1 insertion event at the N-acetyl transferase gene (NAT) locus. The primers were able to amplify all three unique genotypes of FOV race 4. FOV race 3 and Fusarium solani were used as controls to determine specificity of the primers to preclude false positives. Once the specificity of the FOV race 4 primers was determined a qPCR assay was developed using SYBR Green technology. Additional work is currently being conducted to determine the utility of the qPCR assay in detecting and quantifying FOV race 4 from plant tissue and soil samples from commercial fields in CA.