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Open Access Methodology article

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

Sang-Wook Han, Chang-Jin Park, Sang-Won Lee and Pamela C Ronald*

Author Affiliations

Department of Plant Pathology, University of California, One Shields Avenue, Davis, CA., 95616, USA

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BMC Microbiology 2008, 8:164  doi:10.1186/1471-2180-8-164

Published: 30 September 2008

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.