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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. 

Pseudomonas chlororaphis isolates have been studied intensively for their beneficial traits. P. chlororaphis species function as probiotics in plants and fish, offering plants protection against microbes, nematodes and insects. In this review, we discuss the classification of P. chlororaphis isolates within four subspecies; the shared traits include the production of coloured antimicrobial phenazines, high sequence identity between housekeeping genes and similar cellular fatty acid composition. The direct antimicrobial, insecticidal and nematocidal effects of P. chlororaphis isolates are correlated with known metabolites. Other metabolites prime the plants for stress tolerance and participate in microbial cell signalling events and biofilm formation among other things. Formulations of P. chlororaphis isolates and their metabolites are currently being commercialized for agricultural use. 

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.