Abstract
Fusarium graminearum is a fungal pathogen that causes Fusarium head blight (FHB) in wheat and barley. Autophagy is a highly conserved vacuolar degradation pathway essential for cellular homeostasis in which Atg9 serves as a multispanning membrane protein important for generating membranes for the formation of phagophore assembly site. However, the mechanism of autophagy or autophagosome formation in phytopathogens awaits further clarifications. In this study, we identified and characterized the Atg9 homolog (FgAtg9) in F. graminearum by live cell imaging, biochemical and genetic analyses. We find that GFP-FgAtg9 localizes to late endosomes and trans-Golgi network under both nutrient-rich and nitrogen starvation conditions and also show its dynamic actin-dependent trafficking in the cell. Further targeted gene deletion of FgATG9 demonstrates that it is important for growth, aerial hyphae development, and pathogenicity in F. graminearum. Furthermore, the deletion mutant (ΔFgatg9) shows severe defects in autophagy and lipid metabolism in response to carbon starvation. Interestingly, small GTPase FgRab7 is found to be required for the dynamic trafficking of FgAtg9, and co-immunoprecipitation (Co-IP) assays show that FgAtg9 associates with FgRab7 in vivo. Finally, heterologous complementation assay shows that Atg9 is functionally conserved in F. graminearum and Magnaporthe oryzae. Taken together, we conclude that FgAtg9 is essential for autophagy-dependent development and pathogenicity of F. graminearum, which may be regulated by the small GTPase FgRab7.
Highlights
Pathogenic fungi are great threats to both plants and animals, jeopardizing food security [1]
Autophagy is an intracellular degradation pathway conserved in eukaryotes, but the mechanism of autophagy or autophagosome formation in the wheat head blight fungus Fusarium graminearum remains unclear
We observed the localization pattern of FgAtg9 in F. graminearum by live cell imaging and demonstrated that it is essential for autophagy, development and pathogenicity in F. graminearum
Summary
Pathogenic fungi are great threats to both plants and animals, jeopardizing food security [1]. During Fusarium head blight infection, the fungus forms lobate appressoria and infection cushions which help it gain entry into the host cell, or may enter the cell through vulnerable openings and the stomata, and colonizes the host cells through hyphal elongation [5,6]. It produces mycotoxins such as deoxynivalenol (DON) and zearalenone in cereal grains and animal feeds making them unfit for consumption [7,8]. Atg is a core component of the ubiquitin-like protein conjugation systems that are essential for autophagosome formation [23]
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