title: Production of a cell wall-associated endopolygalacturonase by Colletotrichum lindemuthianum and pectin degradation during bean infection.
abstract: The bean pathogen Colletotrichum lindemuthianum expresses two endopolygalacturonase genes, CLPG1 and CLPG2, during interaction with its host plant. However, only CLPG1 was found to be secreted to the extracellular medium during saprophytic growth of the fungus on pectin. To localize CLPG2, a FLAG epitope sequence was inserted in the C-terminal sequence of CLPG2 and the modified gene was introduced into C. lindemuthianum. Western blot analysis using a FLAG monoclonal antibody allowed the detection of CLPG2 in intracellular protein extracts and in the cell wall fraction, but not in the culture medium. Indirect immunofluorescence microscopy was performed to detect CLPG2 during saprophytic or parasitic growth. According to the expression pattern of CLPG2, it was found that CLPG2 accumulates in the fungal cell wall during growth on pectin medium and during appressorium formation, both in vitro and during interaction with the plant. Pectin degradation was not detected around the infection peg using the monoclonal antibody JIM7, specific for methyl-esterified galacturonan. However, extensive pectin dissolution was observed during the development of secondary hyphae.
authors.affiliation: UMR 5546 CNRS-Université Paul Sabatier, Pôle de Biotechnologie Végétale, 24 Chemin de Borde Rouge, BP17 Auzeville, 31326 Castanet-Tolosan, France.
authors.forename: Corentin
authors.initials: C
authors.lastname: Herbert
authors.forename: Richard
authors.initials: R
authors.lastname: O'Connell
authors.forename: Elodie
authors.initials: E
authors.lastname: Gaulin
authors.forename: Vincent
authors.initials: V
authors.lastname: Salesses
authors.forename: Marie Thérèse
authors.initials: MT
authors.lastname: Esquerré-Tugayé
authors.forename: Bernard
authors.initials: B
authors.lastname: Dumas
mesh_terms: D002473:Cell Wall / Q000201:enzymology* / Q000378:metabolism; D020231:Colletotrichum / Q000201:enzymology* / Q000254:growth & development / Q000472:pathogenicity*; D007887:Fabaceae / Q000382:microbiology*; D005656:Fungal Proteins / Q000235:genetics / Q000502:physiology; D015966:Gene Expression Regulation, Fungal; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D010368:Pectins / Q000378:metabolism*; D010935:Plant Diseases / Q000382:microbiology; D011096:Polygalacturonase / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D005656:Fungal Proteins; D010368:Pectins; D011096:Polygalacturonase