Multiple Clonostachys rosea UDP-Glycosyltransferases Contribute to the Production of 15-Acetyl-Deoxynivalenol-3-O-Glycoside When Confronted with Fusarium graminearum

Mycotoxins, derived from toxigenic fungi such as Fusarium, Aspergillus, and Penicillium species have impacted the human food chain for thousands of years. Deoxynivalenol (DON), is a tetracyclic sesquiterpenoid type B trichothecene mycotoxin predominantly produced by F. culmorum graminearum during infection corn, wheat, oats, barley, rice. Glycosylation DON protective detoxification mechanism employed plants. More recently, glycosylating activity has also been detected in fungal microparasitic (biocontrol) organisms. Here we follow up on reported conversion 15-acetyl-DON (15-ADON) into 15-ADON-3-O-glycoside (15-ADON-3G) Clonostachys rosea. Based hypothesis that reaction likely being carried out uridine diphosphate glycosyl transferase (UDP-GTase), applied protein structural comparison strategy, leveraging availability crystal structure rice Os70 to identify subset potential C. rosea UDP-GTases might against 15-ADON. Using CRISPR/Cas9 technology, knocked several selected strain ACM941. Evaluation impact knockouts production 15-ADON-3G confrontation assays with revealed multiple UDP-GTase enzymes, each contributing partial activities. The relationship between these positive hits other plant discussed.