Project Number: 3091-22000-031-29-R
Project Type: Reimbursable Cooperative Agreement
Start Date: Jan 1, 2014
End Date: Dec 31, 2014
Recent work at the USDA-ARS Cotton Pathology Laboratory suggests that fusaric acid (FA) production is a common feature of highly virulent Fusarium oxysporum fsp. vasinfectum (FOV) strains. Therefore, the objective of this project is to determine the role of FA in the pathogenicity of FOV strains and identify the genes involved in FA biosynthesis by FOVs.
FOV strains will be grown in the lab under conditions that induce FA synthesis. mRNA will be extracted from cultures at times of rapidly increasing FA synthesis. Degenerate primers will be prepared to correspond with the conserved region of the ketoacyl synthases (KS) domain of the polyketide synthase (PKS) genes. Gene fragments of expected lengths will be excised, cloned, and transferred into Escherichia coli cells using appropriate vectors. Plasmids will be isolated from the transformed cells and the insert sequenced. The cloned PKS gene fragments will be identified using BLAST searches. Once PKS genes are identified by sequence homology, primers will be synthesized, and mRNA from cultures will be used as a template for polymerase chain reaction (PCR) analysis of the expression profile.