title: Candidate Effector Proteins of the Rust Pathogen Melampsora larici-populina Target Diverse Plant Cell Compartments.
abstract: Rust fungi are devastating crop pathogens that deliver effector proteins into infected tissues to modulate plant functions and promote parasitic growth. The genome of the poplar leaf rust fungus Melampsora larici-populina revealed a large catalog of secreted proteins, some of which have been considered candidate effectors. Unraveling how these proteins function in host cells is a key to understanding pathogenicity mechanisms and developing resistant plants. In this study, we used an effectoromics pipeline to select, clone, and express 20 candidate effectors in Nicotiana benthamiana leaf cells to determine their subcellular localization and identify the plant proteins they interact with. Confocal microscopy revealed that six candidate effectors target the nucleus, nucleoli, chloroplasts, mitochondria, and discrete cellular bodies. We also used coimmunoprecipitation (coIP) and mass spectrometry to identify 606 N. benthamiana proteins that associate with the candidate effectors. Five candidate effectors specifically associated with a small set of plant proteins that may represent biologically relevant interactors. We confirmed the interaction between the candidate effector MLP124017 and TOPLESS-related protein 4 from poplar by in planta coIP. Altogether, our data enable us to validate effector proteins from M. larici-populina and reveal that these proteins may target multiple compartments and processes in plant cells. It also shows that N. benthamiana can be a powerful heterologous system to study effectors of obligate biotrophic pathogens.
authors: 1 The Sainsbury Laboratory, Norwich Research Park, NR4 7UH Norwich, U.K.;|2 INRA, UMR 1136 Interactions Arbres/Microorganismes, Centre INRA Nancy Lorraine, 54280 Champenoux, France;|3 Université de Lorraine, UMR 1136 Interactions Arbres/Microorganismes, Faculté des Sciences et Technologies, 54506 Vandoeuvre-lès-Nancy, France;
authors: Benjamin
authors: B
authors: Petre
authors: 1 The Sainsbury Laboratory, Norwich Research Park, NR4 7UH Norwich, U.K.;|4 The Genome Analysis Centre, Norwich Research Park, NR4 7UH Norwich, U.K.;|5 The John Innes Centre, Norwich Research Park, NR4 7UH Norwich, U.K.
authors: Diane G O
authors: DG
authors: Saunders
authors: 1 The Sainsbury Laboratory, Norwich Research Park, NR4 7UH Norwich, U.K.;
authors: Jan
authors: J
authors: Sklenar
authors: 1 The Sainsbury Laboratory, Norwich Research Park, NR4 7UH Norwich, U.K.;|2 INRA, UMR 1136 Interactions Arbres/Microorganismes, Centre INRA Nancy Lorraine, 54280 Champenoux, France;|3 Université de Lorraine, UMR 1136 Interactions Arbres/Microorganismes, Faculté des Sciences et Technologies, 54506 Vandoeuvre-lès-Nancy, France;
authors: Cécile
authors: C
authors: Lorrain
authors: 1 The Sainsbury Laboratory, Norwich Research Park, NR4 7UH Norwich, U.K.;
authors: Joe
authors: J
authors: Win
authors: 2 INRA, UMR 1136 Interactions Arbres/Microorganismes, Centre INRA Nancy Lorraine, 54280 Champenoux, France;|3 Université de Lorraine, UMR 1136 Interactions Arbres/Microorganismes, Faculté des Sciences et Technologies, 54506 Vandoeuvre-lès-Nancy, France;
authors: Sébastien
authors: S
authors: Duplessis
authors: 1 The Sainsbury Laboratory, Norwich Research Park, NR4 7UH Norwich, U.K.;
authors: Sophien
authors: S
authors: Kamoun
mesh_terms: D001487:Basidiomycota / Q000235:genetics / Q000502:physiology; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D016681:Genome, Fungal / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D010935:Plant Diseases / Q000382:microbiology; D018515:Plant Leaves / Q000382:microbiology; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D032107:Populus / Q000382:microbiology*; D021381:Protein Transport; D011993:Recombinant Fusion Proteins; D014026:Nicotiana / Q000166:cytology / Q000382:microbiology*; D019076:Transgenes
chemical_list: D005656:Fungal Proteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins