RESUMEN

The present study evaluated the priming efficacy of chitosan and chitosan-derived nanoparticles (CNPs) against bacterial wilt of tomato. In the current study, seed-treated CNPs plus pathogen-inoculated tomato seedlings recorded significant protection of 62 % against pathogen-induced wilt disease and subsequently better growth. The induced resistance was witnessed by a prominent increase in lignin, callose and H2O2 deposition, followed by superoxide radical accumulation in leaves. Additionally, chitosan and CNPs-treated tomato plants recorded a remarkable increase in the upregulation of phenylalanine ammonia-lyase (PAL), peroxidase (POX), polyphenol oxidase (PPO), catalase (CAT) and ß-1, 3 glucanase (GLU) in comparison with untreated plants. The chitosan and CNPs-induced antioxidant enzymes were positively correlated with the stimulation of corresponding gene expression in CNPs treated plants related to pathogen-inoculated ones. The results of this study describe that how the application of chitosan and CNPs elicit defense responses at the cellular, biochemical and gene expression in tomato plants against bacterial wilt disease, thereby improve growth and yield.

Asunto(s)

Quitosano , Nanopartículas , Solanum lycopersicum , Antioxidantes/metabolismo , Catalasa/genética , Catalasa/metabolismo , Catecol Oxidasa/metabolismo , Quitosano/metabolismo , Quitosano/farmacología , Peróxido de Hidrógeno/metabolismo , Inmunidad , Lignina/metabolismo , Solanum lycopersicum/microbiología , Fenilanina Amoníaco-Liasa/metabolismo , Enfermedades de las Plantas/microbiología , Superóxidos/metabolismo

RESUMEN

PURPOSE: The study aimed to find an effective method for fungal-mediated synthesis of zinc oxide nanoparticles using endophytic fungal extracts and to evaluate the efficiency of synthesized ZnO NPs as antimicrobial and anticancerous agents. METHODS: Zinc oxide nanoparticles (ZnO NPs) were produced from zinc nitrate hexahydrate with fungal filtrate by the combustion method. The spectroscopy and microscopy techniques, such as ultraviolet-visible spectroscopy, Fourier transform infrared spectroscopy (FT-IR), powder X-ray diffraction (PXRD), scanning electron microscopy (SEM) with energy-dispersive X-ray spectroscopy (EDX), dynamic light scattering (DLS), and transmission electron microscopy (TEM) with selected area electron diffraction (SAED), were used to characterize the obtained product. Antibacterial activity on Gram-positive (Staphylococcus aureus and Bacillus cereus) and Gram-negative (Pseudomonas aeruginosa and Escherichia coli) samples was tested by broth microplate dilution technique. ZnO NPs antifungal activity was determined against plant pathogenic and regular contaminating fungi using the food-poison method. The anticancerous assay of the synthesized ZnO NPs was also investigated by cell uptake, MTT assay, and apoptosis assay. RESULTS: The fungal synthesized ZnO NPs were pure, mainly hexagonal in shape and size range of 34-55 nm. The biosynthesized ZnO NPs could proficiently inhibit both Gram-positive and Gram-negative bacteria. ZnO NPs synthesized from fungal extract exhibited antifungal activity in a dose-dependent manner with a high percentage of mycelial inhibition. The cell uptake analysis of ZnO NPs suggests that a significant amount of ZnO NPs (1 µg/mL) was internalized without disturbing cancer cells' morphology. As a result, the synthesized ZnO NPs showed significant anticancer activity against cancer cells at 1 µg/mL concentration. CONCLUSION: This fungus-mediated synthesis of ZnO NPs is a simple, eco-friendly, and non-toxic method. Our results show that the synthesized ZnO NPs are an excellent novel antimicrobial and anticancer agent. Further studies are required to understand the mechanism of the antimicrobial, anticancerous action of ZnO NPs and their possible genotoxicity.

Asunto(s)

Antibacterianos/farmacología , Antifúngicos/farmacología , Ascomicetos/metabolismo , Nanopartículas del Metal/química , Óxido de Zinc/metabolismo , Antibacterianos/química , Antifúngicos/química , Antifúngicos/metabolismo , Antineoplásicos/química , Antineoplásicos/farmacología , Ascomicetos/aislamiento & purificación , Línea Celular Tumoral , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Grampositivas/efectos de los fármacos , Humanos , Lamiales/microbiología , Pruebas de Sensibilidad Microbiana , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Hojas de la Planta/microbiología , Espectrometría por Rayos X , Espectroscopía Infrarroja por Transformada de Fourier , Staphylococcus aureus/efectos de los fármacos , Difracción de Rayos X , Óxido de Zinc/química , Óxido de Zinc/farmacología

RESUMEN

In the present study, oosporein, a fungal toxic secondary metabolite known to be a toxic agent causing chronic disorders in animals, was isolated from fungus Cochliobolus kusanoi of Nerium oleander L. Toxic effects of oosporein and the possible mechanisms of cytotoxicity as well as the role of oxidative stress in cytotoxicity to Madin-Darby canine kidney kidney cells and RAW 264.7 splene cells were evaluated in vitro. Also to know the possible in vivo toxic effects of oosporein on kidney and spleen, Balb/C mouse were treated with different concentrations of oosporein ranging from 20 to 200 µM). After 24 h of exposure histopathological observations were made to know the effects of oosporein on target organs. Oosporein induced elevated levels of reactive oxygen species (ROS) generation and high levels of malondialdehyde, loss of mitochondrial membrane potential, induced glutathione hydroxylase (GSH) production was observed in a dose depended manner. Effects oosporein on chromosomal DNA damage was assessed by Comet assay, and increase in DNA damage were observed in both the studied cell lines by increasing the oosporein concentration. Further, oosporein treatment to studied cell lines indicated significant suppression of oxidative stress related gene (Superoxide dismutase1 and Catalase ) expression, and increased levels of mRNA expression in apoptosis or oxidative stress inducing genes HSP70, Caspase3, Caspase6, and Caspase9 as measured by quantitative real time-PCR assay. Histopathological examination of oosporein treated mouse kidney and splenocytes further revealed that, oosporein treated target mouse tissues were significantly damaged with that of untreated sam control mice and these effects were in directly proportional to the the toxin dose. Results of the present study reveals that, ROS is the principle event prompting increased oosporein toxicity in studied in vivio and in vitro animal models. The high previlance of these fungi in temperate climates further warrants the need of safe food grain storage and processing practices to control the toxic effects of oosporein to humans and live stock.