epon-post-embedding-correlative-light-and-electron-microscopy

Epon Post Embedding Correlative Light and Electron Microscopy

Correlative light and electron microscopy (CLEM) is a comprehensive microscopy that combines the localization information provided by fluorescence ...

Fujian Medical University

The AP1 complex is a highly conserved clathrin adaptor that plays important roles in regulating cargo protein sorting and intracellular vesicle trafficking in eukaryotes. However, the functions of the AP1 complex in the plant pathogenic fungi including the devastating wheat pathogen  are still unclear. In this study, we investigated the biological functions of FgAP1, a subunit of the AP1 complex in . Disruption of FgAP1 causes seriously impaired fungal vegetative growth, conidiogenesis, sexual development, pathogenesis, and deoxynivalenol (DON) production. The Δ mutants were found to be less sensitive to KCl- and sorbitol-induced osmotic stresses but more sensitive to SDS-induced stress than the wild-type PH-1. Although the growth inhibition rate of the Δ mutants was not significantly changed under calcofluor white (CFW) and Congo red (CR) stresses, the protoplasts released from Δ hyphae were decreased compared with the wild-type PH-1, suggesting that FgAP1 is necessary for cell wall integrity and osmotic stresses in . Subcellular localization assays showed that FgAP1 was predominantly localized to endosomes and the Golgi apparatus. In addition, FgAP1-GFP, FgAP1-GFP, and FgAP1-GFP also localize to the Golgi apparatus. FgAP1 interacts with FgAP1, FgAP1, and FgAP1, while FgAP1 regulates the expression of , , and  in . Furthermore, the loss of  blocks the transportation of the v-SNARE protein FgSnc1 from the Golgi to the plasma membrane and delays the internalization of FM4-64 dye into the vacuole. Taken together, our results demonstrate that FgAP1 plays vital roles in vegetative growth, conidiogenesis, sexual reproduction, DON production, pathogenicity, cell wall integrity, osmotic stress, exocytosis, and endocytosis in . These findings unveil the functions of the AP1 complex in filamentous fungi, most notably in , and lay solid foundations for effective prevention and control of Fusarium head blight (FHB).

FgAP1 Is Critical for Vegetative Growth, Conidiation, Virulence, and DON Biosynthesis in .

Secretion is a fundamental process that plant pathogens utilize to deliver effectors into the host to downregulate immunity and promote infection. Here, we uncover a fascinating membrane trafficking and delivery route that originates from vacuolar membranes in Magnaporthe oryzae and conduits to the host interface and plasma membrane. To perform such secretory/trafficking function, MoRab7 first recruits the retromer complex to the vacuolar membrane, enabling recognition of a family of SNARE proteins, including MoSnc1. Live-cell imaging confirmed a highly dynamic vesicular trafficking of the retromer complex component(s) and MoSnc1 toward and across the host interface or plasma membrane, and subsequent fusion with target membranes. Interestingly, disruption of the MoRab7/Retromer/MoSnc1-based endolysosomal cascade affects effector secretion and fungal pathogenicity. Taken together, we discovered an unconventional protein and membrane trafficking route starting from the fungal endolysosomes to the M. oryzae-rice interaction interface and dissect the role of MoRab7/Retromer/MoSnc1 sorting machinery in effector secretion during biotrophy and invasive growth in rice blast fungus.

Congxian Wu

Public Technology Service Center

Fujian Medical University

Epon Post Embedding Correlative Light and Electron Microscopy

Congxian Wu

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Fujian Medical University

Epon Post Embedding Correlative Light and Electron Microscopy

Public Technology Service Center