Developing molecular tools to simultaneously monitor deoxynivalenol and zearalenone production by Fusarium fungi
Deoxynivalenol (DON) and zearalenone (ZON) are mycotoxins frequently detected in Fusarium -infected cereal grain. Both toxins are produced by Fusarium culmorum and F. graminearum during the development of Fusarium Head Blight (FHB) disease. We are developing tools to allow us to simultaneously detect DON and ZON production by Fusarium species. We generated a transgenic F. culmorum strain that expresses fluorescent proteins under the control of promoters of genes that are essential for the biosynthesis of DON (trichodiene synthase; Tri5) and ZON (a promoter that drives two polyketide synthetases, Zea1 and Zea2). We developed a duplex real time PCR assays for the concurrent analysis of Zea1 and Tri4 (another gene essential for DON production). We are currently generating a multiplex real time PCR version of this assay to detect Zea1, Tri4 and a plant actin (Act1) gene. This assay can be used to detect DON and ZON producers in grain; Act1 serves as a positive control plant gene in the reaction. We are also developing a multiplex real time RT-PCR assay to detect Zea1, Tri4 and a F. culmorum/F. graminearum beta-tubulin (Btub) gene. Incorporation of the Btub enables normalization of Tri4 and Zea1 transcript expression, relative to fungal abundance.
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