title: Proteome analysis of wheat leaf rust fungus, Puccinia triticina, infection structures enriched for haustoria.
abstract: Puccinia triticina (Pt) is a representative of several cereal-infecting rust fungal pathogens of major economic importance world wide. Upon entry through leaf stomata, these fungi establish intracellular haustoria, crucial feeding structures. We report the first proteome of infection structures from parasitized wheat leaves, enriched for haustoria through filtration and sucrose density centrifugation. 2-D PAGE MS/MS and gel-based LC-MS (GeLC-MS) were used to separate proteins. Generated spectra were compared with a partial proteome predicted from a preliminary Pt genome and generated ESTs, to a comprehensive genome-predicted protein complement from the related wheat stem rust fungus, Puccinia graminis f. sp. tritici (Pgt) and to various plant resources. We identified over 260 fungal proteins, 16 of which matched peptides from Pgt. Based on bioinformatic analyses and/or the presence of a signal peptide, at least 50 proteins were predicted to be secreted. Among those, six have effector protein signatures, some are related and the respective genes of several seem to belong to clusters. Many ribosomal structural proteins, proteins involved in energy, general metabolism and transport were detected. Measuring gene expression over several life cycle stages of ten representative candidates using quantitative RT-PCR, all were shown to be strongly upregulated and four expressed solely upon infection.
authors.affiliation: Agriculture & Agri-Food Canada, Pacific Agri-Food Research Centre, Summerland, BC, Canada.
authors.forename: Xiao
authors.initials: X
authors.lastname: Song
authors.forename: Christof
authors.initials: C
authors.lastname: Rampitsch
authors.forename: Bahram
authors.initials: B
authors.lastname: Soltani
authors.forename: Wayne
authors.initials: W
authors.lastname: Mauthe
authors.forename: Rob
authors.initials: R
authors.lastname: Linning
authors.forename: Travis
authors.initials: T
authors.lastname: Banks
authors.forename: Brent
authors.initials: B
authors.lastname: McCallum
authors.forename: Guus
authors.initials: G
authors.lastname: Bakkeren
mesh_terms: D001487:Basidiomycota / Q000737:chemistry* / Q000235:genetics; D002499:Centrifugation, Density Gradient; D016000:Cluster Analysis; D015180:Electrophoresis, Gel, Two-Dimensional; D020224:Expressed Sequence Tags / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D006790:Host-Parasite Interactions; D013058:Mass Spectrometry; D010935:Plant Diseases / Q000382:microbiology; D018515:Plant Leaves / Q000382:microbiology; D020543:Proteome / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017421:Sequence Analysis; D017385:Sequence Homology; D014908:Triticum / Q000382:microbiology; D015854:Up-Regulation
chemical_list: D005656:Fungal Proteins; D020543:Proteome