title: Erwinia carotovora elicitors and Botrytis cinerea activate defense responses in Physcomitrella patens.
abstract: BACKGROUND
  Vascular plants respond to pathogens by activating a diverse array of defense mechanisms. Studies with these plants have provided a wealth of information on pathogen recognition, signal transduction and the activation of defense responses. However, very little is known about the infection and defense responses of the bryophyte, Physcomitrella patens, to well-studied phytopathogens. The purpose of this study was to determine: i) whether two representative broad host range pathogens, Erwinia carotovora ssp. carotovora (E.c. carotovora) and Botrytis cinerea (B. cinerea), could infect Physcomitrella, and ii) whether B. cinerea, elicitors of a harpin (HrpN) producing E.c. carotovora strain (SCC1) or a HrpN-negative strain (SCC3193), could cause disease symptoms and induce defense responses in Physcomitrella.
  
  
  RESULTS
  B. cinerea and E.c. carotovora were found to readily infect Physcomitrella gametophytic tissues and cause disease symptoms. Treatments with B. cinerea spores or cell-free culture filtrates from E.c. carotovoraSCC1 (CF(SCC1)), resulted in disease development with severe maceration of Physcomitrella tissues, while CF(SCC3193) produced only mild maceration. Although increased cell death was observed with either the CFs or B. cinerea, the occurrence of cytoplasmic shrinkage was only visible in Evans blue stained protonemal cells treated with CF(SCC1) or inoculated with B. cinerea. Most cells showing cytoplasmic shrinkage accumulated autofluorescent compounds and brown chloroplasts were evident in a high proportion of these cells. CF treatments and B. cinerea inoculation induced the expression of the defense-related genes: PR-1, PAL, CHS and LOX.
  
  
  CONCLUSIONS
  B. cinerea and E.c. carotovora elicitors induce a defense response in Physcomitrella, as evidenced by enhanced expression of conserved plant defense-related genes. Since cytoplasmic shrinkage is the most common morphological change observed in plant PCD, and that harpins and B. cinerea induce this type of cell death in vascular plants, our results suggest that E.c. carotovora CFSCC1 containing HrpN and B. cinerea could also induce this type of cell death in Physcomitrella. Our studies thus establish Physcomitrella as an experimental host for investigation of plant-pathogen interactions and B. cinerea and elicitors of E.c. carotovora as promising tools for understanding the mechanisms involved in defense responses and in pathogen-mediated cell death in this simple land plant.
authors.affiliation: Departamento de Biología Molecular, Instituto de Investigaciones Biológicas Clemente Estable, Avenida Italia 3318, CP 11600, Montevideo, Uruguay. iponce@iibce.edu.uy
authors.forename: Inés
authors.initials: I
authors.lastname: Ponce de León
authors.forename: Juan Pablo
authors.initials: JP
authors.lastname: Oliver
authors.forename: Alexandra
authors.initials: A
authors.lastname: Castro
authors.forename: Carina
authors.initials: C
authors.lastname: Gaggero
authors.forename: Marcel
authors.initials: M
authors.lastname: Bentancor
authors.forename: Sabina
authors.initials: S
authors.lastname: Vidal
mesh_terms: D020171:Botrytis / Q000502:physiology*; D019068:Bryopsida / Q000166:cytology / Q000235:genetics / Q000276:immunology* / Q000382:microbiology*; D016923:Cell Death; D002736:Chloroplasts / Q000378:metabolism; D003593:Cytoplasm / Q000382:microbiology; D005453:Fluorescence; D018506:Gene Expression Regulation, Plant; D016973:Pectobacterium carotovorum / Q000502:physiology*; D010935:Plant Diseases / Q000276:immunology / Q000382:microbiology*; D018515:Plant Leaves / Q000382:microbiology; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D010940:Plant Proteins