title: Autophagy contributes to regulation of nuclear dynamics during vegetative growth and hyphal fusion in Fusarium oxysporum.
abstract: In the fungal pathogen Fusarium oxysporum, vegetative hyphal fusion triggers nuclear mitotic division in the invading hypha followed by migration of a nucleus into the receptor hypha and degradation of the resident nucleus. Here we examined the role of autophagy in fusion-induced nuclear degradation. A search of the F. oxysporum genome database for autophagy pathway components identified putative orthologs of 16 core autophagy-related (ATG) genes in yeast, including the ubiquitin-like protein Atg8, which is required for the formation of autophagosomal membranes. F. oxysporum Foatg8Δ mutants were generated in a strain harboring H1-cherry fluorescent protein (ChFP)-labeled nuclei to facilitate analysis of nuclear dynamics. The Foatg8Δ mutants did not show MDC-positive staining in contrast to the wild type and the FoATG8-complemented (cFoATG8) strain, suggesting that FoAtg8 is required for autophagy in F. oxysporum. The Foatg8Δ strains displayed reduced rates of hyphal growth, conidiation, and fusion, and were significantly attenuated in virulence on tomato plants and in the nonvertebrate animal host Galleria mellonella. In contrast to wild-type hyphae, which are almost exclusively composed of uninucleated hyphal compartments, the hyphae of the Foatg8Δ mutants contained a significant fraction of hyphal compartments with 2 or more nuclei. The increase in the number of nuclei per hyphal compartment was particularly evident after hyphal fusion events. Time-lapse microscopy analyses revealed abnormal mitotic patterns during vegetative growth in the Foatg8Δ mutants. Our results suggest that autophagy mediates nuclear degradation after hyphal fusion and has a general function in the control of nuclear distribution in F. oxysporum.
authors.affiliation: a Departamento de Genética; Universidad de Córdoba; Campus de Excelencia Agroalimentario ; Córdoba , Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Corral-Ramos
authors.forename: M Gabriela
authors.initials: MG
authors.lastname: Roca
authors.forename: Antonio
authors.initials: A
authors.lastname: Di Pietro
authors.forename: M Isabel G
authors.initials: MI
authors.lastname: Roncero
authors.forename: Carmen
authors.initials: C
authors.lastname: Ruiz-Roldán
mesh_terms: D000818:Animals; D001343:Autophagy / Q000235:genetics; D002467:Cell Nucleus / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D005670:Fusarium / Q000166:cytology* / Q000235:genetics / Q000254:growth & development* / Q000472:pathogenicity; D017353:Gene Deletion; D005800:Genes, Fungal; D049452:Green Fluorescent Proteins / Q000378:metabolism; D054884:Host-Pathogen Interactions / Q000235:genetics; D025301:Hyphae / Q000166:cytology*; D018551:Solanum lycopersicum / Q000382:microbiology; D009036:Moths / Q000382:microbiology; D010588:Phagosomes / Q000378:metabolism; D013172:Spores, Fungal / Q000378:metabolism; D014774:Virulence / Q000235:genetics
chemical_list: D005656:Fungal Proteins; D049452:Green Fluorescent Proteins
keywords: Atg, autophagy-related; BLAST, basic local alignment search tool; CFW, calcofluor white; ChFP, cherry fluorescent protein; DIC, differential interference contrast; Fusarium oxysporum; GFP, green fluorescent protein; HygR, hygromycin resistant; MDC, monodansylcadaverine; ORF, open reading frame; PCR, polymerase chain reaction; PDA, potato dextrose agar; PDB, potato dextrose broth; PMSF, phenylmethylsulfonyl fluoride; SM, synthetic medium; WT, wild-type; autophagy; filamentous fungi; gDNA, genomic DNA; hyphal fusion; nuclear dynamics; virulence