Involvement of the autophagy-related gene BdATG8 in development and pathogenicity in Botryosphaeria dothidea

Botryosphaeria dothidea is a destructive fungal pathogen that causes Botryosphaeria canker and fruit ring rot on apple worldwide. Autophagy is a process of self-degradation that maintains intracellular homeostasis via lysosomal pathway. To date, the biological role of autophagy in B. dothidea remains unknown. In this study, we identified and characterized the autophagy-related gene BdATG8 in B. dothidea. BdATG8 was able to restore the defect in nitrogen starvation tolerance of Saccharomyces cerevisiae ATG8 deletion mutant. GFP-BdAtg8 was shown to be a useful marker for monitoring autophagy in B. dothidea. Target deletion of BdATG8 (ΔBdAtg8) blocked autophagy and significantly impaired mycelial growth, conidiation and perithecium formation. In addition, ΔBdAtg8 showed significantly increased sensitivity to phytoalexin and oxidative stress, suggesting that BdATG8 plays critical roles in overcoming phytoalexin and reactive oxygen species (ROS)-mediated plant immunity. Pathogenicity assays revealed that ΔBdAtg8 almost lost ability to infect hosts. Overall, our results indicate that BdATG8 plays an important role in fungal development, stress responses and pathogenesis in B. dothidea.