The role of the antimicrobial compound 2,4-diacetylphloroglucinol in the impact of biocontrol Pseudomonas fluorescens F113 on Azospirillum brasilense phytostimulators.

Pseudomonads producing the antimicrobial metabolite 2,4-diacetylphloroglucinol (Phl) can control soil-borne phytopathogens, but their impact on other plant-beneficial bacteria remains poorly documented. Here, the effects of synthetic Phl and Phl(+) Pseudomonas fluorescens F113 on Azospirillum brasilense phytostimulators were investigated. Most A. brasilense strains were moderately sensitive to Phl. In vitro, Phl induced accumulation of carotenoids and poly-ß-hydroxybutyrate-like granules, cytoplasmic membrane damage and growth inhibition in A. brasilense Cd. Experiments with P. fluorescens F113 and a Phl(-) mutant indicated that Phl production ability contributed to in vitro growth inhibition of A. brasilense Cd and Sp245. Under gnotobiotic conditions, each of the three strains, P. fluorescens F113 and A. brasilense Cd and Sp245, stimulated wheat growth. Co-inoculation of A. brasilense Sp245 and Pseudomonas resulted in the same level of phytostimulation as in single inoculations, whereas it abolished phytostimulation when A. brasilense Cd was used. Pseudomonas Phl production ability resulted in lower Azospirillum cell numbers per root system (based on colony counts) and restricted microscale root colonization of neighbouring Azospirillum cells (based on confocal microscopy), regardless of the A. brasilense strain used. Therefore, this work establishes that Phl(+) pseudomonads have the potential to interfere with A. brasilense phytostimulators on roots and with their plant growth promotion capacity.

The Pseudomonas secondary metabolite 2,4-diacetylphloroglucinol is a signal inducing rhizoplane expression of Azospirillum genes involved in plant-growth promotion.

During evolution, plants have become associated with guilds of plant-growth-promoting rhizobacteria (PGPR), which raises the possibility that individual PGPR populations may have developed mechanisms to cointeract with one another on plant roots. We hypothesize that this has resulted in signaling phenomena between different types of PGPR colonizing the same roots. Here, the objective was to determine whether the Pseudomonas secondary metabolite 2,4-diacetylphloroglucinol (DAPG) can act as a signal on Azospirillum PGPR and enhance the phytostimulation effects of the latter. On roots, the DAPG-producing Pseudomonas fluorescens F113 strain but not its phl-negative mutant enhanced the phytostimulatory effect of Azospirillum brasilense Sp245-Rif on wheat. Accordingly, DAPG enhanced Sp245-Rif traits involved in root colonization (cell motility, biofilm formation, and poly-ß-hydroxybutyrate production) and phytostimulation (auxin production). A differential fluorescence induction promoter-trapping approach based on flow cytometry was then used to identify Sp245-Rif genes upregulated by DAPG. DAPG enhanced expression of a wide range of Sp245-Rif genes, including genes involved in phytostimulation. Four of them (i.e., ppdC, flgE, nirK, and nifX-nifB) tended to be upregulated on roots in the presence of P. fluorescens F113 compared with its phl-negative mutant. Our results indicate that DAPG can act as a signal by which some beneficial pseudomonads may stimulate plant-beneficial activities of Azospirillum PGPR.

Biochemical and genomic comparison of inorganic phosphate solubilization in Pseudomonas species.

Mobilization of insoluble soil inorganic phosphate by plant beneficial rhizobacteria is a trait of key importance to the development of microbial biofertilizers. In this study, the ability of several Pseudomonas spp. to solubilize Ca3 (PO4 )2 was compared. While all Pseudomonas spp. were found to facilitate a decrease in pH and solubilize inorganic phosphate by the production of extracellular organic acids, strains varied by producing either gluconic or 2-ketogluconic acid. Furthermore, comparison between the Pseudomonas spp. of the genes involved in oxidative glucose metabolism revealed variations in genomic organization. To further investigate the genetic mechanisms involved in inorganic phosphate solubilization by Pseudomonas spp., a transposon mutant library of P. fluorescens F113 was screened for mutants with reduced Ca3 (PO4 )2 solubilization ability. Mutations in the gcd and pqqE genes greatly reduced the solubilization ability, whereas mutations in the pqqB gene only moderately reduced this ability. The combination of biochemical analysis and genomic comparisons revealed that alterations in the pqq biosynthetic genes, and the presence/absence of the gluconate dehydrogenase (gad) gene, fundamentally affect phosphate solublization in strains of P. fluorescens.