Identification of N-3-hydroxyoctanoyl-homoserine lactone production in Pseudomonas fluorescens 5064, pathogenic to broccoli, and controlling biosurfactant production by quorum sensing

Quorum sensing controls a number of key processes in growth, reproduction and virulence of many gram-negative bacteria via signalling molecules or autoinducers. It can control, for instance, the production of pectic enzymes which are virulence factors in disease. Pseudomonas fluorescens 5064 produces biosurfactants which are important for bacterial establishment on the plant surface prior to causing disease in broccoli. The aim of this paper was to determine if biosurfactant production in this bacterium is controlled via quorum sensing. To do this, 35 surfactant-minus Tn5 mutants of P. fluorescens 5064 were screened for their abilities to produce a quorum sensing signal. Six of these biosurfactant-deficient mutants showed a large reduction in quorum sensing signal production and varied in their abilities to cause disease. In one mutant, 6423, which contains a single Tn5 insertion, the production of the signal was almost eliminated. Synthetic quorum sensing signal and quorum sensing signal extracted from wild type P. fluorescens 5064 restored biosurfactant production by addition to the culture in mutant 6423. The quorum sensing signal in wild type P. fluorescens 5064 was identified by high pressure liquid chromatography and mass spectrometry as N -3-hydroxyoctanoyl-homoserine lactone.