Antimicrobial activity of experimental formulations of two structurally different nano-zinc oxide materials, plate-like Zinkicide SG4 and particulate Zinkicide SG6, was evaluated against Xanthomonas citri subsp. citri, the cause of citrus canker. In vitro assay demonstrated Zinkicide SG4 had a twofold lower minimum inhibitory concentration (MIC) against Escherichia coli and X. alfalfae subsp. citrumelonis (62.5 to 250 µg/ml) compared with copper sulfate (250 µg/ml), copper hydroxide (250 to 500 µg/ml), or cuprous oxide/zinc oxide (125 to 250 µg/ml). Zinkicide SG6 had a sevenfold to eightfold lower MIC against Escherichia coli and X. alfalfae subsp. citrumelonis (31 to 250 μg/ml). Leaves of sweet orange (Citrus sinensis) and fruit of ‘Ruby Red’ grapefruit (C. paradisi) were evaluated for citrus canker disease control. A greenhouse assay with foliage demonstrated that spray treatment with Zinkicide reduced citrus canker lesion development after injection-infiltration of X. citri subsp. citri into the leaf intercellular space. In field trials conducted in Southeast Florida in 2014 and 2015, Zinkicide SG4 and SG6 reduction of grapefruit canker incidence exceeded that of cuprous oxide and cuprous oxide/zinc oxide bactericides. Zinkicide formulations were also effective against the fungal diseases, citrus scab (Elsinoe fawcetti) and melanose (Diaporthe citri), on grapefruit. No sign of phytotoxicity to the fruit rind was observed during either season. Antimicrobial activity of Zinkicide for protection of leaves and fruit against X. citri subsp. citri was comparable or exceeded that for commercial copper and zinc oxide formulations which may be attributed to translaminar movement of Zinkicide.
Bacterial spot, caused by Xanthomonas spp., is a widespread and damaging bacterial disease of tomato (Solanum lycopersicum). For disease management, growers rely on copper bactericides, which are often ineffective due to the presence of copper-tolerant Xanthomonas strains. This study evaluated the antibacterial activity of the new copper composites core-shell copper (CS-Cu), multivalent copper (MV-Cu), and fixed quaternary ammonium copper (FQ-Cu) as potential alternatives to commercially available micron-sized copper bactericides for controlling copper-tolerant Xanthomonas perforans. In vitro, metallic copper from CS-Cu and FQ-Cu at 100 μg/ml killed the copper-tolerant X. perforans strain within 1 h of exposure. In contrast, none of the micron-sized copper rates (100 to 1,000 μg/ml) from Kocide 3000 significantly reduced copper-tolerant X. perforans populations after 48 h of exposure compared with the water control (P < 0.05). All copper-based treatments killed the copper-sensitive X. perforans strain within 1 h. Greenhouse studies demonstrated that all copper composites significantly reduced bacterial spot disease severity when compared with copper-mancozeb and water controls (P < 0.05). Although there was no significant impact on yield, copper composites significantly reduced disease severity when compared with water controls, using 80% less metallic copper in comparison with copper-mancozeb in field studies (P < 0.05). This study highlights the discovery that copper composites have the potential to manage copper-tolerant X. perforans and tomato bacterial spot.
Copper (Cu) compounds are widely used as antibacterial/antifungal agents for protecting food crops. Prolonged use of Cu biocides would lead to undesirable Cu levels in agricultural soil. In the absence of a suitable alternative, prudent use of Cu biocides is required. This paper reports for the first time a composite material of sol-gel silica host matrix loaded with mixed-valence Cu as an alternative to conventional biocides. In this composite material, Cu is present in different oxidation states. The hydrophilic silica matrix serves as a water-dispersible delivery vehicle for antimicrobial Cu. It is hypothesized that a mixed-valence Cu system, specifically enriched with Cu(0) and Cu(I), will exhibit enhanced antimicrobial efficacy over traditional Cu(II) compounds. Materials were characterized by high-resolution transmission electron microscopy and X-ray photoelectron spectroscopy for the determination of particle size, morphology, crystallinity, and Cu oxidation states. Antimicrobial studies against Xanthomonas alfalfae and Escherichia coli (minimum inhibitory concentration) showed improved efficacy in MV-CuSiNG treatment compared to CuSiNG and other controls. Phytotoxicity studies performed (in Vinca sp. and Hamlin orange) under greenhouse conditions showed that the newly prepared nanocomposite is safe for plants, demonstrating potential usefulness of the material in agricultural biocides.
Copper (Cu) bactericides/fungicides are used extensively for crop protection in agriculture. Concerns for Cu accumulation in soil, Cu leaching into the surrounding ecosystem, and development of Cu resistance in phytopathogenic bacteria are evident. While there is no suitable alternative to Cu available to date for agricultural uses, it is possible to reduce Cu per application by supplementing with Zn and improving Cu bioavailability using nanotechnology. We have prepared a non-phytotoxic composite material consisting of generally recognized as safe ZnO 800 particles and nanocopper-loaded silica gel (ZnO-nCuSi). The morphology of the ZnO-nCuSi material was characterized using scanning electron microscopy, showing ZnO particles dispersed in the silica gel matrix. ZnO-nCuSi demonstrated strong in vitro antimicrobial properties against several model plant bacterial species. Two consecutive year field efficacy results showed that agri-grade ZnO-nCuSi was effective in controlling citrus canker disease at less than half the metallic rate of the commercial cuprous oxide/zinc oxide pesticide.
In this paper, we report a nonphytotoxic bactericide and fungicide formulation containing a composite of silica and quaternary ammonium compound (quat). The composite material was prepared using an acid-catalyzed sol–gel method. Positively charged quat was associated with a negatively charged silica-gel matrix, producing a stable suspension of fixed-quat gel (FQ-G). The morphology of FQ-G and the interaction of quat with silica were characterized using SEM and FTIR, respectively. Silica gel significantly reduced quat phytotoxicity when tested at 500 and 1000 μg/mL foliar-application rates. The in vitro antimicrobial efficacy of FQ-G was evaluated against Xanthomonas alfalfae, Pseudomonas syringae, and Clavibacter michiganensis, showing comparable efficacies to that of quat itself. In field conditions, its efficacy in controlling the bacterial and fungal diseases citrus canker, scab, and melanose on ‘Ray Ruby’ red grapefruit was evaluated. Foliar application rates at 100 and 200 μg/mL provided comparable disease control to those of several copper standards, demonstrating the potential for use as an alternative agricultural biocide.
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