title: Fluorescent markers of the endocytic pathway in Zymoseptoria tritici.
abstract: Hyphal growth in filamentous fungi is supported by the uptake (endocytosis) and recycling of membranes and associated proteins at the growing tip. An increasing body of published evidence in various fungi demonstrates that this process is of essential importance for fungal growth and pathogenicity. Here, we introduce fluorescent markers to visualize the endocytic pathway in the wheat pathogen Zymoseptoria tritici. We fused enhanced green-fluorescent protein (eGFP) to the actin-binding protein fimbrin (ZtFim1), which is located in actin patches that are formed at the plasma membrane and are participating in endocytic uptake at the cell surface. In addition, we tagged early endosomes by eGFP-labelling a Rab5-homologue (ZtRab5) and late endosomes and vacuoles by expressing eGFP-Rab7 homologue (ZtRab7). Using fluorescent dyes and live cell imaging we confirmed the dynamic behavior and localization of these markers. This set of molecular tools enables an in-depth phenotypic analysis of Z. tritici mutant strains thereby supporting new strategies towards the goal of controlling wheat against Z. tritici.
authors.affiliation: Biosciences, University of Exeter, Exeter EX4 4QD, UK.
authors.forename: S
authors.initials: S
authors.lastname: Kilaru
authors.affiliation: Biosciences, University of Exeter, Exeter EX4 4QD, UK.
authors.forename: M
authors.initials: M
authors.lastname: Schuster
authors.affiliation: Biosciences, University of Exeter, Exeter EX4 4QD, UK.
authors.forename: M
authors.initials: M
authors.lastname: Latz
authors.affiliation: Biosciences, University of Exeter, Exeter EX4 4QD, UK.
authors.forename: M
authors.initials: M
authors.lastname: Guo
authors.affiliation: 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 / Q000235:genetics / Q000502:physiology*; D004705:Endocytosis; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000235:genetics; D008840:Microfilament Proteins / Q000032:analysis* / Q000235:genetics; D061848:Optical Imaging / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; C026742:plastin; D049452:Green Fluorescent Proteins
keywords: Actin patches; Endosomes; Mycosphaerella graminicola; Pathogenic fungi; Septoria tritici blotch