1. Instituto de Biología Molecular y Celular de Plantas (Consejo Superior de Investigaciones Científicas—Universitat Politècnica de València), Avenida de los Naranjos, 46022 Valencia, Spain
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Dr. Mohamed A. Mohamed
1. Instituto de Biología Molecular y Celular de Plantas (Consejo Superior de Investigaciones Científicas—Universitat Politècnica de València), Avenida de los Naranjos, 46022 Valencia, Spain
Sustainable use of nanotechnology in crop protection requires an understanding of the plant’s life cycle, potential toxicological impacts of nanomaterials and their mechanism of action against the target pathogens. Herein, we show some properties of a candidate antifungal nanocomposite made from copper oxide (CuO; otherwise an essential soil nutrient) nanoparticles (NPs), with definite size and shape, decorating the surface of reduced graphene oxide (rGO) nanosheets. The successful preparation of the rGO-CuO NPs was confirmed by spectroscopic and microscopic analyses, and its antifungal activity against wild strains of Fusarium oxysporum affecting tomato and pepper plants was successfully confirmed. A comparative analysis in vitro indicated that this nanocomposite had higher antifungal activity at only 1 mg/L than the conventional fungicide Kocide 2000 at 2.5 g/L. Further investigation suggested that rGO-CuO NPs creates pits and pores on the fungal cell membranes inducing cell death. In planta results indicated that only 1 mg/L from the nanocomposite is required to reduce Fusarium wilt and root rot diseases severity below 5% for tomato and pepper plants without any phytotoxicity for about 70 days. Comparatively, 2.5 g/L of Kocide 2000 are required to achieve about 30% disease reduction in both plants. The present study contributes to the concept of agro-nanotechnology, showing the properties of a novel ecofriendly and economic nanopesticide for sustainable plant protection.
Mycogenic synthesis of silver nanoparticles (AgNPs) was carried out in the present investigation using an aqueous extract of endophytic non-pathogenic Alternaria solani F10 (KT721914). The mycosynthesized AgNPs were characterized by means of spectroscopic and microscopic techniques. The surface plasmon resonance found at 430 nm confirmed the formation of stable AgNPs for several weeks at room temperature. Also, the results revealed the formation of spherical and monodispersed AgNPs with an average size of 14.8 ± 1.2 nm. The FT-IR spectrum suggested that the fungal extracellular proteins and secondary metabolites had the role in Ag reduction and AgNPs capping of which protein Ag nanoconjugates were formed. Furthermore, the mycosynthesized AgNPs exhibited potent antifungal activity against different pathogenic isolates of the same Alternaria solani fungus, the causal pathogen of tomato early blight disease. The antifungal efficiency of the AgNPs at 1, 5 and 10 ppm were evaluated for 8 days after incubation by measuring the inhibition rate of fungal radial growth. The results were further supported by investigating fungal hyphae morphology alteration by scanning and transmission electron microscopy. Treated fungal hyphae showed formation of pits and pores. Also, the mycosynthesized AgNPs were able to pass and distribute throughout the fungal cell area and interact with the cell components.
Nature produces large quantities of superbly complex and highly reliable microcapsules. The micrometre-sized Lycopodium clavatum spores are one example of these robust capsules. The encapsulation of erythromycin (EM) and bacitracin (BAC) antibiotics into the Lycopodium clavatum sporopollenin (LCS) extracted from these spore species is explored for the first time. The LCS microparticles are extensively characterised before and after loading using SEM, CLSM, TGA and FTIR techniques. The loading capacity and entrapping efficiency of EM were 16.2 and 32.4%, respectively. The antibacterial activities of pure antibiotics, empty LCS and the antibiotic-loaded LCS were evaluated against Staphylococcus aureus (Gram-positive), Pseudomonas aeruginosa (Gram-negative), and Klebsiella pneumoniae (Gram-negative) human pathogenic bacterial strains. A remarkable increase in the antibacterial fold activity of both EM- and BAC-loaded LCS compared to that of the pure antibiotics is observed. Crucial for drug delivery applications, empty LCS, EM- and BAC-loaded LCS were found to be nontoxic against human epithelial colorectal adenocarcinoma cells Caco-2 as revealed by the cytotoxicity evaluation. The in vitro release mechanism of EM in pH 7.4 showed a deviation from Fick's law. In vivo release of EM from EM-loaded LCS (an oral dose of 50 mg kg−1) revealed high values of the area under the plasma concentration–time curve (AUC0–6 h and AUC0–∞ were 1620 and 2147 μg h L−1, respectively) indicative of the enhanced EM bioavailability. The successful loading of antibiotics into the nontoxic LCS and the enhanced bioavailability can open up intriguing applications in oral and topical drug delivery strategies.
The extensive use of agrochemicals for crop protection is increasing their environmental risks. Due to the incredible antimicrobial potential of nanomaterials, research into their potential use in sustainable agriculture as alternatives to chemical fungicides is advancing rapidly. In this study, we evaluated the possible antifungal properties of copper oxide nanocomposite (CH@CuO NPs) coated with chitosan in order to fend off Fusarium wilt diseases in tomato plants caused by F. oxysporum f. sp. lycopersici (FOL) throughout in vitro and in vivo experiments. Here, we demonstrate some of the characteristics of a potential antifungal nanocomposite composed of copper oxide nanoparticles (CuO NPs), firmly immobilized on chitosan nanoparticle (CH) surfaces as dark spots, with an irregular shape and 54.22 nm in size, as indicated by Transmission electron microscope (TEM) analysis. Spectroscopic and microscopic investigations, as well as its antifungal efficacy, verified that the successful synthesis of the CH@CuO NPs at three different concentrations (1, 25, and 50) mg/L against three different wild isolates of the pathogenic Fusarium oxysporum that infect tomatoes was successfully proven to be effective. In vitro comparisons revealed that CH@CuO NPs showed stronger antifungal activity at only 1 mg/L (96.22 ± 1.35) than the classical chemical fungicide “Kocide 2000” at conc. 2.5 g/L (77.34 ± 0.33), for example, in the case of FOL1 isolate. In accordance with the in vivo data, tomato plants can be treated with only 1 mg/L of CH@CuO NPs for up to 75 days, by which time Fusarium wilt disease severity is reduced by 91.5% In contrast, 2.5 g/L of Kocide 2000 is required to reduce disease in tomato plants by about 90%. This research expands our understanding of agro-nanotechnology by outlining the characteristics of a unique, environmentally friendly, and economically viable nanopesticide for long-term plant protection.