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Pseudomonas fluorescens  2P24 is a plant growth-promoting rhizobacterium (PGPR) isolated from wheat take-all decline soil. Genomic analysis of strain 2P24 revealed the presence of a complete SPI-1 type III secretion system (T3SS) gene cluster on the chromosome with an organization and orientation similar to the SPI-1 T3SS gene clusters of  Salmonella enterica  and  P. kilonensis  F113. Phylogenetic analysis revealed that the SPI-1 T3SS gene cluster of strain 2P24 might be obtained from  Salmonella  and  Shigella  by horizontal gene transfer. Two transcriptional regulator homologs of HilA and InvF were found from the SPI-1 T3SS gene cluster of strain 2P24. HilA regulated the expression of the structural genes positively, such as  invG ,  sipB ,  sipD ,  prgI , and  prgK . Prediction of transcriptional binding sites and RNA-seq analysis revealed 14 genes were up-regulated by InvF in strain 2P24. Exploring potential roles of SPI-1 T3SS revealed that it was not associated with motility. However, 2P24Δ invF  reduced resistance against  Fusarium graminearum  significantly. 2P24Δ hilA  enhanced formation of biofilm significantly at 48 h. All three mutants 2P24Δ hilA , 2P24Δ invF , and 2P24Δ invE-C  reduced the chemotactic responses to glucose significantly. Finally, the determination of SPI-1 mutants to trigger innate immunity in  Nicotiana benthamiana  showed that 2P24Δ invE-C  reduced the ability to induce the production of reactive oxygen species compared with the wild type strain 2P24.

Characterization of the SPI-1 Type III Secretion System in Pseudomonas fluorescens 2P24