Autophagy is a conserved and unique degradation system in eukaryotic cells, which plays crucial roles in the growth, development and pathogenesis of Fungi. Despite that, it is poorly understood in Fusarium graminearum currently. Here, we identified an autophagy gene FgAtg27 from F. graminearum, and investigated its possible roles in regulating morphogenesis and pathogenicity. Results showed that FgAtg27 is homologous to Saccharomyces cerevisiae Atg27 and with an active signal peptide at N-terminal. Then, the ΔFgAtg27 mutant was generated and gene deletion did not change growth and sporulation, whereas significantly decreased pathogenicity. FgAtg27 showed subcellular localization at pre-autophagosomal structure (PAS). After starvation induction, amount of autophagosomes in ΔFgAtg27 was significantly less than wild type and complemented strain, indicating that FgAtg27 deletion affects the autophagosome formation in F. graminearum. Meanwhile, under high Ca