Abstract
The genome of Pseudomonas fluorescens F113, a model rhizobacterium and a plant growth-promoting agent, encodes three putative type VI secretion systems (T6SSs); F1-, F2- and F3-T6SS. Bioinformatic analysis of the F113 T6SSs has revealed that they belong to group 3, group 1.1, and group 4a, respectively, similar to those previously described in Pseudomonas aeruginosa. In addition, in silico analyses allowed us to identify genes encoding a total of five orphan VgrG proteins and eight putative effectors (Tfe), some with their cognate immunity protein (Tfi) pairs. Genes encoding Tfe and Tfi are found in the proximity of P. fluorescens F113 vgrG, hcp, eagR and tap genes. RNA-Seq analyses in liquid culture and rhizosphere have revealed that F1- and F3-T6SS are expressed under all conditions, indicating that they are active systems, while F2-T6SS did not show any relevant expression under the tested conditions. The analysis of structural mutants in the three T6SSs has shown that the active F1- and F3-T6SSs are involved in interbacterial killing while F2 is not active in these conditions and its role is still unknown.. A rhizosphere colonization analysis of the double mutant affected in the F1- and F3-T6SS clusters showed that the double mutant was severely impaired in persistence in the rhizosphere microbiome, revealing the importance of these two systems for rhizosphere adaption.
Highlights
The genome of Pseudomonas fluorescens F113, a model rhizobacterium and a plant growth-promoting agent, encodes three putative type VI secretion systems (T6SSs); F1, F2- and F3-T6SS
In silico analyses of the genomes of 134 strains belonging to the P. fluorescens species complex revealed that only 20% of them encode T6SS clusters (Table S3)
We identified sixty-two complete T6SS gene clusters distributed mostly in three main phylogenetic clades
Summary
The genome of Pseudomonas fluorescens F113, a model rhizobacterium and a plant growth-promoting agent, encodes three putative type VI secretion systems (T6SSs); F1-, F2- and F3-T6SS. The core genes of the T6SSs are located in genomic clusters commonly comprised of 13 to 15 genes, which encode the structural proteins of the system with well-conserved functions[7,18,19,20]. The encoded-core proteins are named Tss (type six secretion)[18] and include the structural components that form the membrane complex, the baseplate, and the tail. These clusters can encode accessory proteins named Tag (type VI accessories genes) such as TagABFJLPRSTQ involved in regulation or other mechanistic aspects of the T 6SSs7,24,33–35
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