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Regulation of production of the antifungal metabolite 2,4-diacetylphloroglucinol in Pseudomonas fluorescens F113: genetic analysis of phlF as a transcriptional repressor

Biomerit Research Centre, Department of Microbiology, National University of Ireland, Cork, Ireland¹

Author for correspondence: Fergal O’Gara. Tel: +353 21 272098. Fax: +353 21 275934. e-mail: f.ogara{at}ucc.ie

The antifungal metabolite 2,4-diacetylphloroglucinol plays a major role in the biocontrol capabilities of Pseudomonas fluorescens. The phloroglucinol biosynthetic locus of P. fluorescens F113 has been isolated previously. From nucleotide sequence data, a putative regulator gene (phlF) was identified upstream and divergently transcribed from the phlACBD phloroglucinol biosynthetic genes. PhlF shows similarity to various transcriptional repressors in the EMBL database and exhibits a helix–turn–helix motif in its amino acid sequence. phlF was cloned into an expression vector and the PhlF protein product was purified. Gel retardation experiments demonstrated PhlF to be a DNA-binding protein and showed that it binds to the phlA–phlF intergenic region. Introduction of phlF into P. fluorescens F113 in multiple copies resulted in repression of phloroglucinol production in this strain. This effect was mediated at the transcription level since the expression of a phloroglucinol biosynthetic gene fusion in this background was equally repressed. Furthermore, the inactivation of phlF results in derepression of phloroglucinol production in this strain.

Keywords: biocontrol, repressor, transcriptional control, Pseudomonas fluorescens

The GenBank accession number for the sequence reported in this paper is AF129856.

^a Present address: Department of Molecular Biology, IRIS, Chiron, Via Fiorentina 1, 53100 Siena, Italy.

^b Present address: Department of Agronomy, Lethbridge Research Centre, Agriculture and Agri-Food Canada, 5403 1st Avenue South, Lethbridge, Alberta T1J 4B1, Canada.

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