RESUMO
Seed treatment with plant growth-promoting bacteria represents the primary strategy to incorporate them into agricultural ecosystems, particularly for crops under extensive management, such as maize. In this study, we evaluated the seed bacterization levels, root colonization patterns, and root competitiveness of a collection of autochthonous Pseudomonas isolates that have demonstrated several plant-probiotic abilities in vitro. Our findings indicate that the seed bacterization level, both with and without the addition of various protectants, is specific to each Pseudomonas strain, including their response to seed pre-hydration. Bacterization kinetics revealed that while certain isolates persisted on seed surfaces for up to 4 days post-inoculation (dpi), others experienced a rapid decline in viability after 1 or 2 dpi. The observed differences in seed bacterization levels were consistent with the root colonization densities observed through confocal microscopy analysis, and with root competitiveness quantified via selective plate counts. Notably, isolates P. protegens RBAN4 and P. chlororaphis subsp. aurantiaca SMMP3 demonstrated effective competition with the natural microflora for colonizing the maize rhizosphere and both promoted shoot and root biomass production in maize assessed at the V3 grown stage. Conversely, P. donghuensis SVBP6 was detected at very low levels in the maize rhizosphere, but still exhibited a positive effect on plant parameters, suggesting a growth-stimulatory effect during the early stages of plant development. In conclusion, there is a considerable strain-specific variability in the maize seed bacterization and survival capacities of Pseudomonas isolates with plant-probiotic traits, with a correlation in their root competitiveness under natural conditions. This variability must be understood to optimize their adoption as inputs for the agricultural system. Our experimental approach emphasizes the critical importance of tailoring seed bacterization treatments for each inoculant candidate, including the selection and incorporation of protective substances. It should not be assumed that all bacterial cells exhibit a similar performance.
RESUMO
Pseudomonas donghuensis strain SVBP6, an isolate from an agricultural plot in Argentina, displays a broad-spectrum and diffusible antifungal activity, which requires a functional gacS gene but could not be ascribed yet to known secondary metabolites typical of Pseudomonas biocontrol species. Here, we report that Tn5 mutagenesis allowed the identification of a gene cluster involved in both the fungal antagonism and the production of a soluble tropolonoid compound. The ethyl acetate extract from culture supernatant showed a dose-dependent inhibitory effect against the phytopathogenic fungus Macrophomina phaseolina. The main compound present in the organic extract was identified by spectroscopic and X-ray analyses as 7-hydroxytropolone (7HT). Its structure and tautomerism was confirmed by preparing the two key derivatives 2,3-dimethoxy- and 2,7-dimethoxy-tropone. 7HT, but not 2,3- or 2,7-dimethoxy-tropone, mimicked the fungal inhibitory activity of the ethyl acetate extract from culture supernatant. The activity of 7HT, as well as its production, was barely affected by the presence of up to 50 µM added iron (Fe+2 ). To summarize, P. donghuensis SVBP6 produces 7HT under the positive control of the Gac-Rsm cascade and is the main active metabolite responsible for the broad-spectrum inhibition of different phytopathogenic fungi.