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Development and Application of a dapB-Based In Vivo Expression Technology System To Study Colonization of Rice by the Endophytic Nitrogen-Fixing Bacterium Pseudomonas stutzeri A15

Hans Rediers,¹ Victoria Bonnecarrère,^1, Paul B. Rainey,^2,3 Kelly Hamonts,¹ Jos Vanderleyden,¹ and René De Mot^1*

Centre of Microbial and Plant Genetics, Katholieke Universiteit Leuven, B-3001 Heverlee, Belgium,¹ Department of Plant Sciences, University of Oxford, Oxford OX1 3RB, United Kingdom,² School of Biological Sciences, University of Auckland, Auckland, New Zealand³

Received 14 May 2003/ Accepted 5 August 2003

Pseudomonas stutzeri A15 is a nitrogen-fixing bacterium isolated from paddy rice. Strain A15 is able to colonize and infect rice roots. This strain may provide rice plants with fixed nitrogen and hence promote plant growth. In this article, we describe the use of dapB-based in vivo expression technology to identify P. stutzeri A15 genes that are specifically induced during colonization and infection (cii). We focused on the identification of P. stutzeri A15 genes that are switched on during rice root colonization and are switched off during free-living growth on synthetic medium. Several transcriptional fusions induced in the rice rhizosphere were isolated. Some of the corresponding genes are involved in the stress response, chemotaxis, metabolism, and global regulation, while others encode putative proteins with unknown functions or without significant homology to known proteins.

Present address: Biotechnology Unit, Instituto Nacional de Investigación Agropecuaria, Canelones, Uruguay.

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