The Exocyst Regulates Hydrolytic Enzyme Secretion At Hyphal Tips And Septa In The Banana Fusarium Wilt Fungus Fusarium Odoratissimum

Hyphal polarized growth in filamentous fungi requires tip-directed secretion, while additional evidence suggests that fungal exocytosis for the hydrolytic enzyme secretion can occur at other sites in hyphae, including the septum. In this study, we analyzed the role of the exocyst complex involved in the secretion in the banana wilt fungal pathogen Fusarium odoratissimum. All eight exocyst components in F. odoratissimum not only localized to the tips ahead of the Spitzenkorper in growing hyphae but also localized to the outer edges of septa in mature hyphae. To further analyze the exocyst in F. odoratissimum, we attempted single gene deletion for all the genes encoding the eight exocyst components and only succeeded in constructing the gene deletion mutants for exo70 and sec5; we suspect that the other 6 exocyst components are encoded by essential genes. Deletion of exo70 or sec5 led to defects in vegetative growth, conidiation, and pathogenicity in F. odoratissimum. Notably, the deletion of exo70 resulted in decreased activities for endoglucosidase, filter paper enzymes, and amylase, while the loss of sec5 only led to a slight reduction in amylase activity. Septum-localized alpha-amylase (AmyB) was identified as the marker for septum-directed secretion, and we found that Exo70 is essential for the localization of AmyB to septa. Meanwhile the loss of Sec5 did not affect AmyB localization to septa but led to a higher accumulation of AmyB in cytoplasm. This suggested that while Exo70 and Sec5 both take part in the septum-directed secretion, the two play different roles in this process.IMPORTANCE The exocyst complex is a multisubunit tethering complex (MTC) for secretory vesicles at the plasma membrane and contains eight subunits, Sec3, Sec5, Sec6, Sec8, Sec10, Sec15, Exo70, and Exo84. While the exocyst complex is well defined in eukaryotes from yeast (Saccharomyces cerevisiae) to humans, the exocyst components in filamentous fungi show different localization patterns in the apical tips of hyphae, which suggests that filamentous fungi have evolved divergent strategies to regulate endo-membrane trafficking. In this study, we demonstrated that the exocyst components in Fusarium odoratissimum are localized not only to the tips of growing hyphae but also to the outer edge of the septa in mature hyphae, suggesting that the exocyst complex plays a role in the regulation of septum-directed protein secretion in F. odoratissimum. We further found that Exo70 and Sec5 are required for the septum-directed secretion of alpha-amylase in F. odoratissimum but with different influences.