title: The in planta proteome of wild type strains of the fire blight pathogen, Erwinia amylovora.
abstract: Erwinia amylovora is a Gram-negative plant pathogen that causes fire blight. This disease affects most members of the Rosaceae family including apple and pear. Here, an infection model is introduced to study proteomic changes in a highly virulent E. amylovora strain upon interaction with its host as compared to a lower virulent strain. For this purpose separate shoots of apple rootstocks were wound-infected and when infection became systemic, bacterial cells were isolated and processed for analysis in a proteomics platform combining 2-D fluorescence difference gel electrophoresis and mass spectrometry. Comparing the proteome of the isolates, significant abundance changes were observed in proteins involved in sorbitol metabolism, amylovoran production as well as in protection against plant defense mechanisms. Furthermore several proteins associated with virulence were more abundant in the higher virulent strain. Changes at the proteome level showed good accordance at the transcript level, as was verified by RT-qPCR. In conclusion, this infection model may be a valuable tool to unravel the complexity of plant-pathogen interactions and to gain insight in the molecular mechanisms associated with virulence of E. amylovora, paving the way for the development of plant-protective interventions against this detrimental disease.
  
  
  CONCLUSIONS
  During this research a first time investigation was performed on the proteome of E. amylovora, grown inside a susceptible host plant. This bacterium is the causal agent of fire blight, which can affect most members of the Rosaceae family including apple and pear. To do so, an artificial infection model on shoots of apple rootstocks was optimized and employed. When infection was systemic, bacterial cells were extracted from the plant tissue followed by extraction of the proteins from the bacteria. Further processing of the proteins was done by using a 2-D fluorescence difference gel electrophoresis analysis followed by mass spectrometry. By the use of two strains differing in their virulent ability, we were able to draw conclusions concerning virulence and behavior of different strains inside the host. This research provides a model to investigate plant-pathogen interactions and more importantly, we identified possible new targets for the development of novel control methods against this devastating disease.
authors.affiliation: Molecular and Physical Plant Physiology, Faculty of Sciences, Hasselt University, Belgium. Electronic address: michelle.holtappels@uhasselt.be.
authors.forename: M
authors.initials: M
authors.lastname: Holtappels
authors.affiliation: Zoology department, PCFruit Research Station, Belgium. Electronic address: kristof.vrancken@pcfruit.be.
authors.forename: K
authors.initials: K
authors.lastname: Vrancken
authors.affiliation: Biomedical Research Institute, Hasselt University and Transnational University Limburg, School of Life Sciences, Hasselt University, Belgium. Electronic address: jeanpaul.noben@uhasselt.be.
authors.forename: J P
authors.initials: JP
authors.lastname: Noben
authors.affiliation: Centre for Environmental Sciences, Hasselt University, Belgium. Electronic address: tony.remans@uhasselt.be.
authors.forename: T
authors.initials: T
authors.lastname: Remans
authors.affiliation: Pomology department, PCFruit Research Station, Belgium. Electronic address: hilde.schoofs@pcfruit.be.
authors.forename: H
authors.initials: H
authors.lastname: Schoofs
authors.affiliation: Pomology department, PCFruit Research Station, Belgium. Electronic address: tom.deckers@pcfruit.be.
authors.forename: T
authors.initials: T
authors.lastname: Deckers
authors.affiliation: Molecular and Physical Plant Physiology, Faculty of Sciences, Hasselt University, Belgium. Electronic address: roland.valcke@uhasselt.be.
authors.forename: R
authors.initials: R
authors.lastname: Valcke
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D044084:Erwinia amylovora / Q000378:metabolism*; D027845:Malus / Q000382:microbiology; D010935:Plant Diseases / Q000382:microbiology; D018517:Plant Roots / Q000382:microbiology; D020543:Proteome / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D020543:Proteome
keywords: 2D DIGE; Amylovoran; Erwinia amylovora; In planta; Plant-pathogen interactions; Sorbitol