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ARS Home » Plains Area » Lubbock, Texas » Cropping Systems Research Laboratory » Plant Stress and Germplasm Development Research » Research » Publications at this Location » Publication #403483
Research Project: Development of Economically Important Row Crops that Improve the Resilience of U.S. Agricultural Production to Present and Future Production Challenges

Location: Plant Stress and Germplasm Development Research

Title: A new quantitative PCR assay for the detection of Fusarium oxysporum f. sp. vasinfectum race 4 from cotton

Author
item URNER, MICHAEL - Fresno State University
item Ulloa, Mauricio
item HUTMACHER, ROBERT - University Of California, Davis
item ELLIS, MARGARET - Fresno State University

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 8/12/2023
Publication Date: 8/16/2023
Citation: Urner, M., Ulloa, M., Hutmacher, R.B., Ellis, M.L. 2023. A new quantitative PCR assay for the detection of Fusarium oxysporum f. sp. vasinfectum race 4 from cotton. Meeting Abstract.

Interpretive Summary:

Technical Abstract: Fusarium oxysporum f. sp. vasinfectum (FOV) is a pathogenic soil borne fungus responsible for Fusarium wilt in cotton (Gossypium spp.). FOV race 4 is economically damaging race that threatens California cotton production. Today, there is not a quantitative PCR method to precisely identify race 4. Currently a PCR protocol utilizes race-specific primers that can be used to identify FOV race 4 from other FOV races. However, this PCR method is time-consuming and requires the fungal isolation from plants or soil prior to DNA isolation and PCR. Therefore, the goal of this research was to improve the speed and accuracy of detecting FOV race 4 by developing a quantitative PCR (qPCR) assay. FOV race 4 specific primers for the qPCR assay 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. The qPCR assay was also able to detect FOV race 4 directly from plant tissue without the need for fungal isolation. Further studies will be done to determine if the qPCR assay can be utilized to quantify the inoculum load in infested plant tissues.