title: Identification of Bremia lactucae and Oidium neolycopersici proteins extracted for intact spore MALDI mass spectrometric biotyping.
abstract: Several proteomic approaches were applied to identify protein markers providing typical signals during intact cell/spore (IC/IS) MALDI-TOF MS of two plant pathogens, namely Bremia lactucae (a downy mildew) and Oidium neolycopersici (a powdery mildew). First, proteins were extracted from intact spores of the microorganisms under conditions simulating their treatment prior to the mass spectrometric analysis. After a separation by electrophoresis and tryptic digestion, 198 and 140 proteins were identified in the B. lactucae and O. neolycopersici extracts, respectively. A large portion of them were found to be involved in the process of protein biosynthesis. For the first time, some proteins were assigned to characteristic signals in MS profiles of the investigated pathogens based on an agreement in the molecular mass. There were 9 and 10 proteins recognized, respectively, which could contribute significantly to the spectral patterns. These proteins were assigned tentatively to the following peaks in the MS profiles: (i) m/z 7828; 8593; 10 456; 11 312; 12 450; 12 763; 14 756 and 16 854 for B. lactucae; (ii) m/z 7709; 8895; 9504; 9952; 11 317; 14 082 and 14 839 for O. neolycopersici. We demonstrated the presence of ribosomal proteins and histones, which could be employed as markers in biotyping analyses for pathogen identification.
authors.affiliation: Department of Protein Biochemistry and Proteomics, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Olomouc, Czech Republic.
authors.forename: Jana
authors.initials: J
authors.lastname: Beinhauer
authors.affiliation: Department of Protein Biochemistry and Proteomics, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Olomouc, Czech Republic.
authors.forename: René
authors.initials: R
authors.lastname: Lenobel
authors.affiliation: Department of Protein Biochemistry and Proteomics, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Olomouc, Czech Republic.
authors.forename: Dmitry
authors.initials: D
authors.lastname: Loginov
authors.affiliation: Department of Protein Biochemistry and Proteomics, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Olomouc, Czech Republic.
authors.forename: Ivo
authors.initials: I
authors.lastname: Chamrád
authors.affiliation: Institute of Molecular Pathology, Faculty of Military Health Sciences, University of Defence, Hradec Králové, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Řehulka
authors.affiliation: Department of Botany, Faculty of Science, Palacký University, Olomouc, Czech Republic.
authors.forename: Michaela
authors.initials: M
authors.lastname: Sedlářová
authors.affiliation: Institute of Chemical Technologies and Analytics, Vienna University of Technology (TU Wien), Vienna, Austria.
authors.forename: Martina
authors.initials: M
authors.lastname: Marchetti-Deschmann
authors.affiliation: Institute of Chemical Technologies and Analytics, Vienna University of Technology (TU Wien), Vienna, Austria.
authors.forename: Günter
authors.initials: G
authors.lastname: Allmaier
authors.affiliation: Department of Protein Biochemistry and Proteomics, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Olomouc, Czech Republic.
authors.forename: Marek
authors.initials: M
authors.lastname: Šebela
mesh_terms: D001203:Ascomycota / Q000737:chemistry*; D005656:Fungal Proteins / Q000032:analysis* / Q000737:chemistry; D016533:Mycological Typing Techniques / Q000379:methods*; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D010935:Plant Diseases / Q000382:microbiology; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013172:Spores, Fungal / Q000737:chemistry*
chemical_list: D005656:Fungal Proteins; D010455:Peptides
keywords: Biotyping; Marker; Mass spectrometry; Protein extraction; Protein identification