Characterization the role of GacA-dependent small RNAs and RsmA family proteins on 2,4-diacetylphloroglucinol production in Pseudomonas fluorescens 2P24

Pseudomonas fluorescens 2P24 is a plant-beneficial rhizobacteria that controls many root diseases caused by soilborne pathogens, and the production of the antibiotic compound 2,4-diacetylphloroglucinol (2,4-DAPG) is essential for its biocontrol ability. In the present study, we investigated the regulatory mechanism acting on the production of 2,4-DAPG by the GacA-dependent small non-coding RNAs (sRNAs) and RsmA/E proteins in strain 2P24. Our results showed that the GacS-GacA system regulates the expression of the phlACBD locus, which is responsible for 2,4-DAPG production, by inducing the expression of rsmX, rsmX1, rsmY, and rsmZ. A novel GacA-regulated sRNA, RgsA, was found to negatively regulate 2,4-DAPG production. Activation of the phlACBD locus by the GacS-GacA system is mediated through RsmA and RsmE proteins (but not RsmI), which inhibit phlACBD translation by binding to the putative RsmA/E recognition element in the phlACBD leader. Taken together, our results suggested that in P. fluorescens 2P24, the GacS-GacA system controls the cellular 2,4-DAPG levels in the cell by fine-tuning the function of sRNAs in P. fluorescens.