title: In-depth analysis of the Magnaporthe oryzae conidial proteome.
abstract: The filamentous fungus Magnaporthe oryzae (M. oryzae) is the causative agent of rice blast disease and presents a significant threat to worldwide rice production. To establish the groundwork for future research on the pathogenic development of M. oryzae, a global proteomic study of conidia was performed. The filter aided sample preparation method (FASP) and anion StageTip fractionation combined with long, optimized shallow 210 min nanoLC gradients prior to mass spectrometry analysis on an Orbitrap XL was applied, which resulted in a doubling of protein identifications in comparison to our previous GeLC analysis. Herein, we report the identification of 2912 conidial proteins at a 1% protein false discovery rate (FDR) and we present the most extensive study performed on M. oryzae conidia to date. A similar distribution between identified proteins and the predicted proteome was observed when subcellular localization analysis was performed, suggesting the detected proteins build a representative portion of the predicted proteome. A higher percentage of cytoplasmic proteins (associated with translation, energy, and metabolism) were observed in the conidial proteome relative to the whole predicted proteome. Conversely, nuclear and extracellular proteins were less well represented in the conidial proteome. Further analysis by gene ontology revealed biological insights into identified proteins important for central metabolic processes and the physiology of conidia.
authors.affiliation: W.M. Keck FT-ICR Mass Spectrometry Laboratory, Department of Chemistry, and Center for Integrated Fungal Research, North Carolina State University, Raleigh, North Carolina 27695, United States.
authors.forename: Emine
authors.initials: E
authors.lastname: Gokce
authors.forename: William L
authors.initials: WL
authors.lastname: Franck
authors.forename: Yeonyee
authors.initials: Y
authors.lastname: Oh
authors.forename: Ralph A
authors.initials: RA
authors.lastname: Dean
authors.forename: David C
authors.initials: DC
authors.lastname: Muddiman
mesh_terms: D002467:Cell Nucleus / Q000737:chemistry; D005591:Chemical Fractionation / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods; D003593:Cytoplasm / Q000737:chemistry; D005656:Fungal Proteins / Q000032:analysis* / Q000737:chemistry; D020082:Magnaporthe / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods; D036103:Nanotechnology; D009687:Nuclear Proteins / Q000032:analysis / Q000737:chemistry; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013172:Spores, Fungal / Q000737:chemistry*; D013997:Time Factors
chemical_list: D005656:Fungal Proteins; D009687:Nuclear Proteins; D010455:Peptides; D020543:Proteome