title: bcpmr1 encodes a P-type Ca(2+)/Mn(2+)-ATPase mediating cell-wall integrity and virulence in the phytopathogen Botrytis cinerea.
abstract: The cell wall of fungi is generally composed of an inner skeletal layer consisting of various polysaccharides surrounded by a layer of glycoproteins. These usually contain both N- and O-linked oligosaccharides, coupled to the proteins by stepwise addition of mannose residues by mannosyltransferases in the endoplasmic reticulum and the Golgi apparatus. In yeast, an essential luminal cofactor for these mannosyltransferases is Mn(2+) provided by the Ca(2+)/Mn(2+)-ATPase known as Pmr1. In this study, we have identified and characterized the Botrytis cinerea pmr1 gene, the closest homolog of yeast PMR1. We hypothesized that bcpmr1 also encodes a Ca(2+)/Mn(2+)-ATPase that plays an important role in the protein glycosylation pathway. Phenotypic analysis showed that bcpmr1 null mutants displayed a significant reduction in conidial production, radial growth and diameter of sclerotia. Significant alterations in hyphal cell wall composition were observed including a 83% decrease of mannan levels and an increase in the amount of chitin and glucan. These changes were accompanied by a hypersensitivity to cell wall-perturbing agents such as Calcofluor white, Congo red and zymolyase. Importantly, the Δbcpmr1 mutant showed reduced virulence in tomato (leafs and fruits) and apple (fruits) and reduced biofilm formation. Together, our results highlight the importance of bcpmr1 for protein glycosylation, cell wall structure and virulence of B. cinerea.
authors: Laboratorio de Bioquímica y Biología Molecular, Departamento de Biología, Universidad de La Serena, La Serena, Chile; Millennium Nucleus for Fungal Integrative and Synthetic Biology (FISB), Chile.
authors: Verónica
authors: V
authors: Plaza
authors: Laboratorio de Bioquímica y Biología Molecular, Departamento de Biología, Universidad de La Serena, La Serena, Chile.
authors: Yanssuy
authors: Y
authors: Lagües
authors: Laboratorio de Bioquímica y Biología Molecular, Departamento de Biología, Universidad de La Serena, La Serena, Chile.
authors: Mauro
authors: M
authors: Carvajal
authors: Departamento de Biología, División de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta s/n, Col. Noria Alta, C.P. 36050 Guanajuato, Gto., Mexico.
authors: Luis A
authors: LA
authors: Pérez-García
authors: Departamento de Biología, División de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta s/n, Col. Noria Alta, C.P. 36050 Guanajuato, Gto., Mexico.
authors: Hector M
authors: HM
authors: Mora-Montes
authors: Departamento de Genética Molecular y Microbiología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile; Millennium Nucleus for Fungal Integrative and Synthetic Biology (FISB), Chile.
authors: Paulo
authors: P
authors: Canessa
authors: Departamento de Genética Molecular y Microbiología, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile; Millennium Nucleus for Fungal Integrative and Synthetic Biology (FISB), Chile.
authors: Luis F
authors: LF
authors: Larrondo
authors: Laboratorio de Bioquímica y Biología Molecular, Departamento de Biología, Universidad de La Serena, La Serena, Chile; Millennium Nucleus for Fungal Integrative and Synthetic Biology (FISB), Chile. Electronic address: lcastillo@userena.cl.
authors: Luis
authors: L
authors: Castillo
mesh_terms: D020171:Botrytis / Q000254:growth & development / Q000472:pathogenicity / Q000502:physiology*; D002473:Cell Wall / Q000378:metabolism*; D005638:Fruit / Q000382:microbiology; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism*; D018551:Solanum lycopersicum / Q000166:cytology / Q000382:microbiology*; D027845:Malus / Q000166:cytology / Q000382:microbiology*; D009154:Mutation; D018515:Plant Leaves / Q000382:microbiology; D013172:Spores, Fungal / Q000254:growth & development; D014774:Virulence
chemical_list: D005656:Fungal Proteins
keywords: Botrytis cinerea; Cell wall; Glycosylation; PMR1