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Green Synthesis of Silver Nanoparticles Using Pomegranate and Orange Peel Extracts and Their Antifungal Activity against Alternaria solani, the Causal Agent of Early Blight Disease of Tomato.

ABSTRACT: This study aimed to synthesize silver nanoparticles (AgNPs) by pomegranate and orange peel extracts using a low concentration of AgNO₃ solution to controlearly blight of tomato caused by Alternaria solani. The pathogen was isolated from infected tomato plants growing in different areas of Saudi Arabia. The isolates of this pathogen were morphologically and molecularly identified. Extracts from peels of pomegranate and orange fruits effectively developed a simple, quick, eco-friendly and economical method through a synthesis of AgNPs as antifungal agents against A. solani. Phenolic content in the pomegranate peel extract was greater than orange peel extract. Phenolic compounds showed a variation of both peel extracts as identified and quantified by High-Performance Liquid Chromatography. The phenolic composition displayed variability as the pomegranate peel extract exhibited an exorbitant amount of Quercitrin (23.62 mg/g DW), while orange peel extract recorded a high amount of Chlorogenic acid (5.92 mg/g DW). Biosynthesized AgNPs were characterized using UV- visible spectroscopy which recorded an average wavelength of 437 nm and 450 nm for pomegranate and orange peels, respectively. Fourier-transform infrared spectroscopy exhibited 32x73.24, 2223.71, 2047.29 and 1972.46 cm^-1, and 3260.70, 1634.62, 1376.62 and 1243.76 cm^-1 for pomegranate and orange peels, respectively. Transmission electron microscopy showed spherical shape of nanoparticles. Zetasizer analysis presented negative charge values; -16.9 and -19.5 mV with average particle sizes 8 and 14 nm fin case of pomegranate and orange peels, respectively. In vitro, antifungal assay was done to estimate the possibility of biosynthesized AgNPs and crude extracts of fruit peels to reduce the mycelial growth of A. solani. AgNPs displayed more fungal mycelial inhibition than crude extracts of two peels and AgNO₃. We recommend the use of AgNPs synthesized from fruit peels for controlling fungal plant pathogens and may be applied broadly and safely in place by using the chemical fungicides, which display high toxicity for humans.

SUBMITTER: Mostafa YS

PROVIDER: S-EPMC8619153 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundMultidrug-resistant (MDR) bacteria are acknowledged as one of the main factors contributing to chronic illnesses and fatalities globally. Numerous diseases, including bloodstream infections, pneumonia, urinary tract infections, and surgical site infections, can be brought on by MDR bacteria. Therefore, a crucial topic of continuing research is the development of a novel and different treatment for MDR microbial pathogens. This work is introduce an alternative method for elimination of MDR bacterial isolates which are causative agents of urinary tract infection among people in Egypt. In our study, we need a novel strategy to combat MDR bacteria by green-synthesized metal nanoparticles (MNPs). That is due to the ability of MNPs to penetrate the cell wall and the cell membrane of gram-positive and gram-negative bacteria.MethodsClinical isolates of MDR bacteria had their antibiotic susceptibility assessed before being molecularly identified using 16 s rRNA, sequencing, and phylogenetic analysis. Also, genetic profiles of isolated strains were performed using ISSR and SDS-PAGE. Finally, characterized plant-mediated silver nanoparticles derived from lemon and pomegranate peel extracts were evaluated against isolated multidrug-resistant bacterial stains.ResultsIn our present trial, one-hundred urine samples were collected from 71 females and 29 males complaining of UTI (urinary tract infection) symptoms. One-hundred microbial isolates were isolated, including 88-g negative and only 8-g positive bacteria in addition to four yeast isolates (Candida species). A total of 72% of the isolated bacteria showed MDR activity. The most prevalent MDR bacterial isolates (Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterococcus faecalis, and Klebsiella pneumoniae) were identified through 16S rDNA PCR sequencing as with accession numbers OP741103, OP741104, OP741105, OP741106, and OP741107, respectively. Lemon and pomegranate-mediated silver nanoparticles [Ag-NPs] were characterized by UV spectroscopy, FTIR, XRD, and TEM with average size 32 and 28 nm, respectively. Lemon and pomegranate-mediated silver nanoparticles [Ag-NPs] showed an inhibitory effect on the selected five MDR isolates at MIC 50 and 30 µg/mL, respectively. These common bacterial isolates were also genetically examined using ISSR PCR, and their total protein level was evaluated using SDS-PAGE, showing the presence of distinct genetic and protein bands for each bacterial species and emphasizing their general and protein composition as a crucial and essential tool in understanding and overcoming MDR behavior in UTI patients.ConclusionsLemon and pomegranate-mediated silver nanoparticles [Ag-NPs] were found to have an inhibitory effect on MDR isolates. Therefore, the study suggests that [Ag-NPs] could be a potential treatment for MDR UTI infections caused by the identified bacterial species.

Project description:Antifungal activities of plant-beneficial Bacillus have been widely studied in recent years. Numerous studies have studied the antifungal mechanisms of soluble non-volatile bioactive compounds such as lipopeptides and proteins produced by Bacillus against soil-borne diseases. However, the antagonistic mechanisms of volatile organic compounds (VOCs) from Bacillus against airborne phytopathogens are still largely unknown, and the function of Alternaria solani pathogenic genes has not been well identified. Here, we first isolated a Bacillus strain with strong antifungal activity and finally identified it as B. subtilis ZD01. Then, the antagonistic mechanisms of VOCs produced by strain ZD01, against A. solani, an airborne fungal pathogen that can cause early blight diseases of potato, were studied. We showed that VOCs produced by strain ZD01 can reduce the colony size and mycelial penetration and can cause serious morphological changes of A. solani. Scanning electron microscope (SEM) observation showed that VOCs released by ZD01 could cause more flaccid and gapped hyphae of A. solani. Also, we found that VOCs produced by ZD01 can inhibit the conidia germination and reduce the lesion areas and number of A. solani in vivo significantly. Meanwhile, based on gas chromatography/mass spectrometry (GC/MS) analysis, 29 volatile compounds produced by strain ZD01 were identified. Out of 29 identified VOCs, 9 VOCs showed complete growth inhibition activities against A. solani. Moreover, we identified two virulence-associated genes (slt2 and sod) in A. solani. slt2 is a key gene that regulates the mycelial growth, penetration, sporulation, and virulence in vivo in A. solani. In addition, sod plays a significant role in the SOD synthetic pathway in A. solani. Results from qRT-PCR showed that the transcriptional expression of these two genes was down-regulated after being treated by VOCs produced by ZD01. These results are useful for a better understanding of the biocontrol mechanism of Bacillus and offer a potential method for potato early blight disease control.

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Green Synthesis of Silver Nanoparticles Using Pomegranate and Orange Peel Extracts and Their Antifungal Activity against Alternaria solani, the Causal Agent of Early Blight Disease of Tomato.

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