Functional characterization of FgERG3 and FgERG5 associated with ergosterol biosynthesis, vegetative differentiation and virulence of Fusarium graminearum.

The ergosterol biosynthesis pathway is well characterized in Saccharomyces cerevisiae, while little is known about the pathway in filamentous fungi. In this study, we isolated and genetically documented biological functions of FgErg3 and FgErg5, which are located upstream of FgErg4, the enzyme catalyzing the final step of ergosterol synthesis in Fusarium graminearum. Our results demonstrated that F. graminearum contains two paralogous FgERG3 and two FgERG5 genes. FgErg3, but not FgErg5, is involved in ergosterol biosynthesis. Double deletion mutants of FgERG3 alleles or the double deletion mutants of FgERG5 alleles showed decreased conidiation and produced abnormal conidia. Fungicide susceptibility tests revealed that FgERG3 and FgERG5 mutants have increased resistance towards triadimefon. However, FgERG3 mutants exhibited increased susceptibility to tebuconazole as well as increased susceptibility to oxidative stress, paraquat and to Mg(2+). Pathogenicity tests showed that the FgERG3 and FgERG5 double deletion mutant displayed dramatically attenuated virulence although they were able to successfully colonize flowering wheat head. In addition, complementation of FgERG3 and FgERG5 genes into S. cerevisiae partially rescued the susceptibility of S. cerevisiae ERG3 and ERG5 deletion mutants towards hydroxyurea and caffeine. Taken together, our results indicate that FgERG3 and FgERG5 play a crucial role in vegetative differentiation, resistance to fungicides and virulence in F. graminearum. FgErg3 alleles, but not FgErg5 alleles, are required for ergosterol biosynthesis in the filamentous fungus F. graminearum.