title: Fluorescent markers for the Spitzenkörper and exocytosis in Zymoseptoria tritici.
abstract: Fungal hyphae are highly polarized cells that invade their substrate by tip growth. In plant pathogenic fungi, hyphal growth is essential for host invasion. This makes polarity factors and secretion regulators potential new targets for novel fungicides. Polarization requires delivery of secretory vesicles to the apical Spitzenkörper, followed by polarized exocytosis at the expanding cell tip. Here, we introduce fluorescent markers to visualize the apical Spitzenkörper and the apical site of exocytosis in hyphae of the wheat pathogen Zymoseptoria tritici. We fused green fluorescent protein to the small GTPase ZtSec4, the myosin light chain ZtMlc1 and the small GTPase ZtRab11 and co-localize the fusion proteins with the dye FM4-64 in the hyphal apex, suggesting that the markers label the hyphal Spitzenkörper in Z. tritici. In addition, we localize GFP-fusions to the exocyst protein ZtExo70, the polarisome protein ZtSpa2. Consistent with results in the ascomycete Neurospora crassa, these markers did localize near the plasma membrane at the hyphal tip and only partially co-localize with FM4-64. Thus, these fluorescent markers are useful molecular tools that allow phenotypic analysis of mutants in Z. tritici. These tools will help develop new avenues of research in our quest to control STB infection in wheat.
authors.affiliation: School of Biosciences, University of Exeter, Exeter EX4 4QD, UK.
authors.forename: M
authors.initials: M
authors.lastname: Guo
authors.affiliation: School of Biosciences, University of Exeter, Exeter EX4 4QD, UK.
authors.forename: S
authors.initials: S
authors.lastname: Kilaru
authors.affiliation: School of Biosciences, University of Exeter, Exeter EX4 4QD, UK.
authors.forename: M
authors.initials: M
authors.lastname: Schuster
authors.affiliation: School of Biosciences, University of Exeter, Exeter EX4 4QD, UK.
authors.forename: M
authors.initials: M
authors.lastname: Latz
authors.affiliation: School of Biosciences, University of Exeter, Exeter EX4 4QD, UK. Electronic address: G.Steinberg@exeter.ac.uk.
authors.forename: G
authors.initials: G
authors.lastname: Steinberg
mesh_terms: D001203:Ascomycota / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D005089:Exocytosis; D005656:Fungal Proteins / Q000032:analysis* / Q000235:genetics; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics; D025301:Hyphae / Q000737:chemistry / Q000235:genetics / Q000502:physiology; D061848:Optical Imaging / Q000379:methods; D015388:Organelles / Q000737:chemistry*; D010935:Plant Diseases / Q000382:microbiology; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics; D013194:Staining and Labeling / Q000379:methods*; D014908:Triticum / Q000382:microbiology
chemical_list: D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: Exocytosis; Hyphal tip; Mycosphaerella graminicola; Pathogenic fungi; Secretion; Septoria tritici blotch