ABSTRACT
Nitrogen is the second most critical factor for crop production after water. In this study, the beneficial rhizobacterium Pseudomonas protegens Pf-5 was genetically modified to fix nitrogen using the genes encoding the nitrogenase of Pseudomonas stutzeri A1501 via the X940 cosmid. Pf-5 X940 was able to grow in L medium without nitrogen, displayed high nitrogenase activity and released significant quantities of ammonium to the medium. Pf-5 X940 also showed constitutive expression and enzymatic activity of nitrogenase in ammonium medium or in nitrogen-free medium, suggesting a constitutive nitrogen fixation. Similar to Pseudomonas protegens Pf-5, Pseudomonas putida, Pseudomonas veronii and Pseudomonas taetrolens but not Pseudomonas balearica and Pseudomonas stutzeri transformed with cosmid X940 showed constitutive nitrogenase activity and high ammonium production, suggesting that this phenotype depends on the genome context and that this technology to obtain nitrogen-fixing bacteria is not restricted to Pf-5. Interestingly, inoculation of Arabidopsis, alfalfa, tall fescue and maize with Pf-5 X940 increased the ammonium concentration in soil and plant productivity under nitrogen-deficient conditions. In conclusion, these results open the way to the production of effective recombinant inoculants for nitrogen fixation on a wide range of crops.
Subject(s)
Bacterial Proteins/metabolism , Genome, Bacterial , Nitrogen Fixation/genetics , Nitrogenase/metabolism , Pseudomonas/genetics , Soil Microbiology , Arabidopsis/growth & development , Bacterial Proteins/genetics , Genetic Vectors , Medicago sativa/growth & development , Nitrogen/metabolism , Nitrogenase/genetics , Pseudomonas/enzymology , Pseudomonas putida/enzymology , Pseudomonas putida/genetics , Pseudomonas stutzeri/enzymology , Pseudomonas stutzeri/genetics , Transformation, Bacterial , Zea mays/growth & developmentABSTRACT
Polyhydroxybutyrate (PHB), a typical carbon and energy storage compound, is widely found in Bacteria and Archae domains. This polymer is produced in response to conditions of physiological stress. PHB is composed of repeating units of ß-hydroxybutyrate (R-3HB). It has been previously shown that R-3HB functions as an osmolyte in extremophile strains. In this study, Pseudomonas sp. CT13, a halotolerant bacterium, and its PHB synthase-minus mutant (phaC) were used to analyze the chaperone role of R-3HB. The production of this compound was found to be essential to salt stress resistance and positively correlated with salt concentration, suggesting that PHB monomer acts as a compatible solute in Pseudomonas sp. CT13. R-3HB accumulation was also associated with the prevention of protein aggregation under combined salt and thermal stresses in Pseudomonas sp. CT13. Physiological concentrations of R-3HB efficiently reduced citrate synthase (CS) aggregation and stabilized the enzymatic activities of CS during thermal stress. Docking analysis of the CS/R-3HB interaction predicted the stability of this complex under physiological concentrations of R-3HB. Thus, in vivo, in vitro and in silico analyses suggest that R-3HB can act as a chemical chaperone.
Subject(s)
Bacterial Proteins/metabolism , Citrate (si)-Synthase/metabolism , Heat-Shock Response/drug effects , Hydroxybutyrates/pharmacology , Pseudomonas/enzymology , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Citrate (si)-Synthase/chemistry , Citrate (si)-Synthase/genetics , Enzyme Stability/drug effects , Heat-Shock Response/physiology , Oxidation-ReductionABSTRACT
During the last few years, RIMAPS technique has been used to characterize the micro-relief of metallic surfaces and recently also applied to biological surfaces. RIMAPS is an image analysis technique which uses the rotation of an image and calculates its average power spectrum. Here, it is presented as a tool for describing the morphology of the trichodium net found in some grasses, which is developed on the epidermal cells of the lemma. Three different species of grasses (herbarium samples) are analyzed: Podagrostis aequivalvis (Trin.) Scribn. & Merr., Bromidium hygrometricum (Nees) Nees & Meyen and Bromidium ramboi (Parodi) Rúgolo. Simple schemes representing the real microstructure of the lemma are proposed and studied. RIMAPS spectra of both the schemes and the real microstructures are compared. These results allow inferring how similar the proposed geometrical schemes are to the real microstructures. Each geometrical pattern could be used as a reference for classifying other species. Finally, this kind of analysis is used to determine the morphology of the trichodium net of Agrostis breviculmis Hitchc. As the dried sample had shrunk and the microstructure was not clear, two kinds of morphology are proposed for the trichodium net of Agrostis L., one elliptical and the other rectilinear, the former being the most suitable.
