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  1. Dissolution of CuO nanoparticles, releasing Cu ions, is a primary mechanism of Cu interaction in the rooting zone of plants. CuO dissolution is sometimes incorrectly considered negligible at high pH, since complexation of Cu with dissolved organic matter may enhance nanoparticle dissolution. Therefore data on the effects of plant-microbial-soil interactions on nanoparticle dissolution, particularly in alkaline soils, are needed. Dissolution of CuO nanoparticles (100 mg kg −1 Cu) was studied in sand supplemented with factorial combinations of wheat growth, a root-colonizing bacterium, and saturated paste extracts (SPEs) from three alkaline, calcareous soils. In control sand systems with 3.34 mM Ca(NO 3 ) 2 solution, dissolved Cu was low (266 μg L −1 Cu). Addition of dissolved organic matter via wheat root metabolites and/or soil SPEs increased dissolved Cu to 795–6250 μg L −1 Cu. Dissolution was correlated with dissolved organic carbon ( R = 0.916, p < 0.0001). Ligands >3 kDa, presumably fulvic acid from the SPEs, complexed Cu driving solubility; the addition of plant exudates further increased solubility 1.5–3.5×. The root-colonizing bacterium decreased dissolved Cu in sand pore waters from planted systems due to metabolism of root exudates. Batch solubility studies (10 mg L −1 Cu) with the soil SPEs and defined solutions containing bicarbonate or fulvic acid confirmed elevated CuO nanoparticle solubility at >7.5 pH. Nanoparticle dissolution was suppressed in batch experiments compared to sand, via nanoparticle organic matter coating or homoconjugation of dissolved organic matter. Alterations of CuO nanoparticles by soil organic matter, plant exudates, and bacteria will affect dissolution and bioavailability of the CuO nanoparticles in alkaline soils. 
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  2. CuO nanoparticles (NPs) are explored as fungicides and fertilizers, and are increasingly likely to be applied to agricultural soils. Consequently, interactions of CuO NPs with soil pore water (SPW) components, plants, and microbes must be understood. These experiments examined whether dissolved natural organic matter (DNOM) from SPW, or root/bacterial exudates, changed wheat ( Triticum aestivum L. v. Deloris) responses to 100 mg kg −1 (Cu/sand) as CuO NPs. Seedlings were grown in sand with 3.34 mM Ca(NO 3 ) 2 or one of three SPWs, differing in DNOM concentration and composition. At 10 days post-germination, CuO NPs stunted roots by 59% in the 3.34 mM Ca(NO 3 ) 2 and 26–35% in the three SPWs compared to plants grown without NPs. Malate, citrate, gluconate, and 2′-deoxymugineic acid (DMA), were elevated 1.3 to 5-fold in the rhizosphere with CuO NPs present. Cu was bioavailable through metallo-organic complexes, including Cu–DMA and Cu–gluconate. Fulvic acid in SPWs mitigated CuO NP-induced wheat root shortening. Pseudomonas chlororaphis O6 eliminated malate and citrate in the rhizospheres, reduced rhizosphere dissolved Cu ∼18–66%, and reduced root Cu 39% across all SPWs while enhancing root stunting ∼17% more across all SPWs than non-inoculated wheat grown with CuO NPs. Thus, both SPW components and root microbial colonization influenced wheat responses to CuO NPs. These interactions are likely in agricultural soils with additional processes, such as ion sorption, to influence CuO NP phytotoxicity, highlighting the importance of considering not just the target plant, but soil properties and associated microbiomes when evaluating impacts of NPs in agricultural usage. 
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