When introduced into autoclaved soil, the population of the hfq mutant PM107 colonized on the wheat rhizosphere was 11-fold lower than that of the wild-type strain 2P24 and its complemented strain PM107/p415-hfq (Fig. 5a). A similar tendency was also observed in the natural soil that was not autoclaved (Fig. 5b). Determinations of population densities on the wheat tips in the same experiments yielded similar results, except that the overall recovered populations of the inoculated strains on the wheat tips were lower than in the wheat rhizospheres (Fig. 5c and d). These results indicated that rhizosphere colonization of P. fluorescens 2P24
in wheat is strongly influenced by the hfq gene. Our study provides PI3K inhibitor evidence that the hfq gene selleck chemicals significantly regulates the transcription of the 2,4-DAPG biosynthetic gene
phlA and the AHL synthase gene pcoI in P. fluorescens 2P24, and consequently affects the production of 2,4-DAPG and AHL, respectively (Figs 2 and 3). Hfq was first identified in E. coli as a factor required for the replication of phage Qβ RNA and subsequently as an important regulator of bacterial gene expression participating in numerous regulatory pathways (Tsui et al., 1994; Valentin-Hansen et al., 2004). Previous studies have shown that Hfq modulates the activity of small regulatory RNAs (sRNAs) by stimulating the pairing between sRNAs and their target mRNAs, thereby facilitating sRNA–mRNA interactions. In Vibrio harveyi and Vibrio cholerae, Hfq Thymidylate synthase mediates interactions between multiple sRNAs and luxR and hapR mRNA targets, which may regulate virulence
gene expression (Lenz et al., 2004). Interaction between Hfq and sRNAs has been described in Pseudomonas spp., and it has been suggested that Hfq may bind to sRNA RsmY and protect RsmY from endonucleolytic cleavage (Sonnleitner et al., 2006; Sorger-Domenigg et al., 2007). In the pathogen P. aeruginosa, sRNAs RsmZ and RsmY were reported to be necessary for the production of AHL and extracellular virulence factors (Heurlier et al., 2004; Kay et al., 2006). Moreover, in plant-beneficial bacterium P. fluorescens CHA0, sRNAs RsmZ, RsmY and RsmX positively regulate the production of the antibiotic 2,4-DAPG and other secondary metabolites by repression of the RsmA and RsmE proteins (Heeb et al., 2002; Valverde et al., 2003; Kay et al., 2005). In strain 2P24, sRNA RsmZ was identified as a positive factor influencing the production of 2,4-DAPG (Jiang et al., 2008) and AHL (unpublished data). Sequence analyses of the P. fluorescens 2P24 genome draft map revealed two homologues of sRNAs, RsmY and RsmX, and the nucleotide sequence of the rsmY gene has 92% and 68% identities with the corresponding gene in P. fluorescens CHA0 and P. aeruginosa PAO1, respectively (data not shown).