An efficient method for visualization and growth of fluorescent Xanthomonas oryzae pv. oryzae in planta

BMC Microbiol. 2008 Sep 30:8:164. doi: 10.1186/1471-2180-8-164.

Abstract

Background: Xanthomonas oryzae pv. oryzae, the causal agent of bacterial blight disease, is a serious pathogen of rice. Here we describe a fluorescent marker system to study virulence and pathogenicity of X. oryzae pv. oryzae.

Results: A fluorescent X. oryzae pv. oryzae Philippine race 6 strain expressing green fluorescent protein (GFP) (PXO99GFP) was generated using the gfp gene under the control of the neomycin promoter in the vector, pPneo-gfp. The PXO99GFPstrain displayed identical virulence and avirulence properties as the wild type control strain, PXO99. Using fluorescent microscopy, bacterial multiplication and colonization were directly observed in rice xylem vessels. Accurate and rapid determination of bacterial growth was assessed using fluoremetry and an Enzyme-Linked ImmunoSorbant Assay (ELISA).

Conclusion: Our results indicate that the fluorescent marker system is useful for assessing bacterial infection and monitoring bacterial multiplication in planta.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Fluorescence
  • Gene Expression
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Microscopy, Fluorescence
  • Oryza / microbiology*
  • Plasmids
  • Promoter Regions, Genetic
  • Staining and Labeling / methods
  • Virulence
  • Xanthomonas / genetics
  • Xanthomonas / growth & development*
  • Xanthomonas / pathogenicity*
  • Xylem / microbiology

Substances

  • enhanced green fluorescent protein
  • Green Fluorescent Proteins