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Plant probiotic bacteria are being increasingly used to maximize both the productivity and quality of field crops. Pseudomonas chlororaphis O6 (PcO6) is a plant root colonizer with probiotic activities. This bacterium produces an array of metabolites, including a group of phenazines that are functional in plant protection. The paper reports responses of PcO6 to a nonionic triblock copolymer surfactant, Pluronic F68. This Pluronic exhibits membrane “healing” activity and improves cryopreservation recovery in eukaryotic cells. The product is FDA-approved and is applied as an adjuvant in formulations used in agriculture, medicine, and biotechnology. Growth of PcO6 on lysogeny broth at 25 °C was unhindered by 0.1 and 1.0 g/L F68, reduced at 10 g/L, and with significant inhibition at 100 g/L F68; micelle formation could account for inhibited growth at higher doses. Phenazine production was not changed by F68, whereas the surfactant activity of F68 induced the spread of bacterial colonization on 0.5% agar. Exposure of cells to fluorescein-labeled F68 resulted in intense fluorescence, stable to washing, showing a direct association of the Pluronic with the bacterium. However, neither protection nor harm was found for PcO6 cells suspended in either 0.1% or 1% F68 after three freeze (−20 °C)/thaw cycles. These findings suggest that F68 could be compatible for use in agricultural formulations with little effect on probiotics such as PcO6. 

Abstract Outer membrane vesicles (OMVs) produced by Gram-negative bacteria have roles in cell-to-cell signaling, biofilm formation, and stress responses. Here, the effects of abiotic stressors on OMV contents and composition from biofilm cells of the plant health-promoting bacteriumPseudomonas chlororaphisO6 (PcO6) are examined. Two stressors relevant to this root-colonizing bacterium were examined: CuO nanoparticles (NPs)-a potential fertilizer and fungicide- and H₂O₂-released from roots during plant stress responses. Atomic force microscopy revealed 40–300 nm diameter OMVs from control and stressed biofilm cells. Raman spectroscopy with linear discriminant analysis (LDA) was used to identify changes in chemical profiles ofPcO6 cells and resultant OMVs according to the cellular stressor with 84.7% and 83.3% accuracies, respectively. All OMVs had higher relative concentrations of proteins, lipids, and nucleic acids thanPcO6 cells. The nucleic acid concentration in OMVs exhibited a cellular stressor-dependent increase: CuO NP-induced OMVs > H₂O₂-induced OMVs > control OMVs. Biochemical assays confirmed the presence of lipopolysaccharides, nucleic acids, and protein in OMVs; however, these assays did not discriminate OMV composition according to the cellular stressor. These results demonstrate the sensitivity of Raman spectroscopy using LDA to characterize and distinguish cellular stress effects on OMVs composition and contents. 

Strategies to reduce crop losses due to drought are needed as climate variability affects agricultural productivity. Wheat (Triticum aestivum var. Juniper) growth in a nutrient-sufficient, solid growth matrix containing varied doses of CuO, ZnO, and SiO2 nanoparticles (NPs) was used to evaluate NP mitigation of drought stress. NP amendments were at fertilizer levels, with maxima of 30 Cu, 20 Zn, and 200 Si (mg metal/kg matrix). Seeds of this drought-tolerant cultivar were inoculated with Pseudomonas chlororaphis O6 (PcO6) to provide a protective root microbiome. An 8 day drought imposed on 14 day-old wheat seedlings decreased shoot and root mass, shoot water content, and the quantum yield of photosystem II when compared to watered plants. PcO6 root colonization was not impaired by drought or NPs. A dose-dependent increase in the Cu, Zn, and Si from the NPs was observed from analysis of the rhizosphere solution, and this process was not affected by drought. Consequently, fertilizer concentrations of the NPs did not further improve drought tolerance in wheat seedlings under the growth conditions of adequate mineral nutrition and the presence of a beneficial microbiome. These findings suggest that potential NP benefits in promoting plant drought tolerance occur only under certain environmental conditions. 

Coatings offer a means to control nanoparticle (NP) size, regulate dissolution, and mitigate runoff when added to crops through soil. Simultaneously, coatings can enhance particle binding to plants and provide an additional source of nutrients, making them a valuable component to existing nanoparticle delivery systems. Here, the surface functionalization of metal and metal-oxide nanoparticles to inhibit aggregation and preserve smaller agglomerate sizes for enhanced transport to the rooting zone and improved uptake in plants is reviewed. Coatings are classified by type and by their efficacy to mitigate agglomeration in soils with variable pH, ionic concentration, and natural organic matter profiles. Varying degrees of success have been reported using a range of different polymers, biomolecules, and inorganic surface coatings. Advances in zwitterionic coatings show the best results for maintaining nanoparticle stability in solutions even under high salinity and temperature conditions, whereas coating by the soil component humic acid may show additional benefits such as promoting dissolution and enhancing bioavailability in soils. Pre-tuning of NP surface properties through exposure to select natural organic matter, microbial products, and other biopolymers may yield more cost-effective nonagglomerating metal/metal-oxide NPs for soil applications in agriculture. 

Organosilanes (e.g., R’-SiOR3) provide hydrophobic functionality in thin-film coatings, porous gels, and particles. Compared with tetraalkoxysilanes (SiOR4), organosilanes exhibit distinct reaction kinetics and assembly mechanisms arising from steric and electronic properties of the R’ group on the silicon atom. Here, the hydrolysis and condensation pathways of n-propyltrimethoxy silane (nPM) and a tri-fluorinated analog of nPM, 3,3,3-trifluoropropyl trimethoxy silane (3F), were investigated under aqueous conditions at pH 1.7, 2.0, 3.0, and 4.0. Prior to hydrolysis, 3F and nPM are insoluble in water and form a lens at the bottom (3F) or top (nPM) of the solutions. This phase separation was employed to follow reaction kinetics using a Turbiscan instrument to monitor hydrolysis through solubilization of the neat silane lens while simultaneously tracking condensation-induced turbidity throughout the bulk solution. Dynamic light scattering confirmed the silane condensation and particle aggregation processes reported by the turbidity scanning. Employing macroscopic phase separation of the starting reactants from the solvent further allows for control over the reaction kinetics, as the interfacial area can be readily controlled by reaction vessel geometry, namely by controlling the surface area to volume. In-situ turbidity scanning and dynamic light scattering revealed distinct reaction kinetics for nPM and 3F, attributable to the electron withdrawing and donating nature of the fluoro- and organo-side chains of 3F and nPM, respectively. 

Nanofertilizer application is becoming a sustainable alternative for plants micronutrients supply. Seed nutrient priming before seeding reduces non- target dispersion; although, applying nanofertilizer in correct concentration must be narrowly chosen to prevent germination and development issues. Here, we evaluated corn seedlings development and germination after seed priming with Mn3O4 nanoparticle (NP), Mn3O4 bulk and MnCl2. Sterile seeds were soaked for 8hours in priming solutions of 0, 20, 40, 80 and 160mg L
1 for each Mn sources. The seeds vigor and germination were evaluated after 7 days on germination paper. Root, shoot and total lengths were measured as well as root, shoot and total dry biomass. Compared to the control, the Mn3O4 NP and Mn3O4 bulk promoted beneficial effects. Mn3O4 NP seed-priming exhibited a concentration dependent profile in improving seedling growth, with greatest benefit around 20mg L
1, pro- viding higher germination, vigor, dry biomass and length than control and the other source tested. Particle size plays an important role in the reactiv- ity of Mn3O4 NP. On the other hand, seeds primed with soluble source did not differ from the control. These findings support NP-seed priming as an alternative to delivery micronutrients. 

This study introduces a simple and environmentally friendly method to synthesize silica-protein nanocomposite materials using microwave energy to solubilize hydrophobic protein in an aqueous solution of pre-hydrolyzed organo- or fluoro-silane. Sol-gel functionality can be enhanced through biomacromolecule incorporation to tune mechanical properties, surface energy, and biocompatibility. Here, synthetic spider silk protein and organo- and fluoro-silane precursors were dissolved and mixed in weakly acidic aqueous solution using microwave technology. Scanning electron microscopy (SEM) and Atomic force microscopy (AFM) images revealed the formation of spherical nanoparticles with sizes ranging from 100 to 500 nm depending, in part, on silane fluoro- or organo-side chain chemistry. The silane-protein interaction in the nanocomposite was assessed through infrared spectroscopy. Deconvoluted ATR-FTIR (Attenuated total reflectance Fourier-transform infrared spectroscopy) spectra revealed silane chemistry-specific conformational changes in the protein-silane nanocomposites. Relative to microwave-solubilized spider silk protein, the β structure content increased by 14% in the spider silk-organo-silica nanocomposites, but decreased by a net 20% in the spider silk-fluoro-silica nanocomposites. Methods of tuning the secondary structures, and in particular β-sheets that are the cross-linking moieties in spider silks and other self-assembling fibrillar proteins, may provide a unique means to promote protein interactions, favor subsequent epitaxial growth process, and enhance the properties of the protein-silane nanocomposites.