Characterization of the fusaric acid gene cluster in Fusarium fujikuroi

Abstract

The "bakanae" fungus Fusarium fujikuroi is a common pathogen of rice and produces a variety of mycotoxins, pigments, and phytohormones. Fusaric acid is one of the oldest known secondary metabolites produced by F. fujikuroi and some other Fusarium species. Investigation of its biosynthesis and regulation is of great interest due to its occurrence in cereal-based food and feed. This study describes the identification and characterization of the fusaric acid gene cluster in F. fujikuroi consisting of the PKS-encoding core gene and four co-regulated genes, FUB1-FUB5. Besides fusaric acid, F. fujikuroi produces two fusaric acid-like derivatives: fusarinolic acid and 9,10-dehydrofusaric acid. We provide evidence that these derivatives are not intermediates of the fusaric acid biosynthetic pathway, and that their formation is catalyzed by genes outside of the fusaric acid gene cluster. Target gene deletions of all five cluster genes revealed that not all of them are involved in fusaric acid biosynthesis. We suggest that only two genes, FUB1 and FUB4, are necessary for the biosynthesis. Expression of the FUB genes and production of fusaric acid and the two derivatives are favored under high nitrogen. We show that nitrogen-dependent expression of fusaric acid genes is positively regulated by the nitrogen-responsive GATA transcription factor AreB, and that pH-dependent regulation is mediated by the transcription factor PacC. In addition, fusaric acid production is regulated by two members of the fungal-specific velvet complex: Vel1 and Lae1. In planta expression studies show a higher expression in the favorite host plant rice compared to maize.