Abstract
A quorum-sensing (QS) locus PcoI/PcoR had been identified previously in the biological control bacterium Pseudomonas fluorescens 2P24. In this paper, we investigated the upstream regulators that influence the transcription of the N-acyl homoserine lactone (AHL) biosynthase gene pcoI using a chromosomal pcoIColon, two colonslacZ fusion reporter strain. Stationary sigma factor RpoS was identified as a negative regulator of QS system using a random mini-Tn5 mutant procedure. Furthermore, deletion mutagenesis and complementation experiments demonstrated that the two-component system GacS/GacA positively regulated the QS system by upregulating pcoI transcription. However, compared with the gacA or gacS mutant, introduction of a second mutation of rpoS in the gacA(-) or gacS(-) background did not lead to further change in the transcriptional expression of the pcoI gene or the synthesis of AHL. Our results suggest that in strain 2P24, RpoS could only play its negative regulatory role on the pcoI gene under a functional GacS/GacA system background.
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