Abstract
Fusarium graminearum, a member of the F. graminearum
species complex, is a filamentous ascomycetous group that
causes serious diseases in cereal crops. A screen of insertional
mutants of F. graminearum, generated using a restriction
enzyme-mediated integration method, identified a mutant
designated R7048 showing pleiotropic phenotypes in several
mycological traits. The vector insertion site in the R7048 genome
was identified as the KpnI site within an ORF annotated
as FGSG_06346 (designated FgVPS74), which showed
similarity to vacuolar protein sorting-associated protein 74
in the baker yeast. Both targeted gene deletion and complementation
analyses confirmed that FgVPS74 was involved
in hyphal growth, conidiation, sexual development, mycotoxin
production, and virulence towards host plants in F.
graminearum. Electron microscopy analysis revealed no significant
changes in morphology of the vacuole or other organelles,
but a greater number of mitochondria were produced
in the ΔFgVPS74 strain compared to the wild-type
progenitor. Expression of a GFP-tagged FgVPS74 construct
under its native promoter in the ΔFgVPS74 strain exhibited
localization of GFP signal to putative vesicle structures, but
not to the vacuolar membrane. Taken together, these findings
demonstrated that a functional vacuolar protein-sorting
pathway mediated by FgVPS74 is crucial for fungal growth
and development in F. graminearum.
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