Yeast Acetyltransferase for Engineering Deoxynivalenol Metabolism in Tobacco

Trichothecene mycotoxins such as Deoxynivalenol or DON are potent protein synthesis inhibitors for eukaryotic organisms and Tri101 coded a 3-O-acetyl transferase works as a resistance mechanism for Fusarium species that produce t-type trichothecenes. AYT1 is a homologous gene to Tri101 capable of trichothecene 3-O-acetylation. Previous studies have indicated that over-expression of yeast acetyl transferase gene (AYT1) encoding a 3-OH trichothecene acetyl transferase that converts DON to a less toxic acetylated form, leads to suppression of the DON sensitivity. The aim of this study was to evaluate transgenic tobacco model plants to deoxynivalenol (DON). In order to detect expression of the transgene, we added cMyc tag to AYT1 and introduced it into the model tobacco plants through Agrobacterium-mediated transformation in an attempt to detoxify DON. After confirmation of integrations of AYT1-cMyc into the tobacco genome by molecular analyses, Immuno-blotting and serological protein studies and trichothecene acetyltransferase activity analyses confirmed the expression of AYT1 in five independent transgenic lines.

Keywords:

Fusarium head blight , DON , ScAYT1 , PCR-tagging , indirect-ELISA , TLC