Genomic Identification of the TOR Signaling Pathway As a Target of the Plant Alkaloid Antofine in the Phytopathogen Fusarium Graminearum

Antofine, a phenanthroindolizidine alkaloid, is a bioactive natural product isolated from milkweeds that exhibits numerous biological activities, including anticancer, antimicrobial, antiviral, and anti-inflammatory properties. However, the direct targets and mode of action of antofine have not been determined. In this report, we show that antofine displays antifungal properties against the phytopathogen , the cause of head blight disease (FHB). FHB does devastating damage to agriculture, causing billions of dollars in economic losses annually. We therefore sought to understand the mode of action of antofine in using insights from yeast chemical genomic screens. We used haploinsufficiency profiling (HIP) to identify putative targets of antofine in yeast and identified three candidate targets, two of which had homologs in The homologues of two targets, glutamate dehydrogenase (GDH) and resistance to rapamycin deletion 2 (RRD2), can bind antofine. Of the two genes, only the knockout displayed a loss of virulence in wheat, indicating that RRD2 is an antivirulence target of antofine in Mechanistically, we demonstrate that antofine disrupts the interaction between RRD2 and Tap42, which is part of the Tap42-phosphatase complex in the target of rapamycin (TOR) signaling pathway, a central regulator of cell growth in eukaryotes and a pathway of extensive study for controlling numerous pathologies. head blight caused by the fungal pathogen is a devastating disease of cereal crops worldwide, with limited effective chemical treatments available. Here we show that the natural alkaloid compound antofine can inhibit fusarium head blight in wheat. Using yeast genomic screening, we identified the TOR pathway component RRD2 as a target of antofine that is also required for pathogenicity.