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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:Tomato (Solanum lycopersicum) is among the most important commercial horticultural crops worldwide. The crop quality and production is largely hampered due to the fungal pathogen Alternaria solani causing necrotrophic foliage early blight disease. Crop plants usually respond to the biotic challenges with altered metabolic composition and physiological perturbations. We have deciphered altered metabolite composition, modulated metabolic pathways and identified metabolite biomarkers in A. solani-challenged susceptible tomato variety Kashi Aman using Liquid Chromatography-Mass Spectrometry (LC-MS) based metabolomics. Alteration in the metabolite feature composition of pathogen-challenged (m/z 9405) and non-challenged (m/z 9667) plant leaves including 8487 infection-exclusive and 8742 non-infection exclusive features was observed. Functional annotation revealed putatively annotated metabolites and pathway mapping indicated their enrichment in metabolic pathways, biosynthesis of secondary metabolites, ubiquinone and terpenoid-quinones, brassinosteroids, steroids, terpenoids, phenylpropanoids, carotenoids, oxy/sphingolipids and metabolism of biotin and porphyrin. PCA, multivariate PLS-DA and OPLS-DA analysis showed sample discrimination. Significantly up regulated 481 and down regulated 548 metabolite features were identified based on the fold change (threshold ≥ 2.0). OPLS-DA model based on variable importance in projection (VIP scores) and FC threshold (> 2.0) revealed 41 up regulated discriminant metabolite features annotated as sphingosine, fecosterol, melatonin, serotonin, glucose 6-phosphate, zeatin, dihydrozeatin and zeatin-β-D-glucoside. Similarly, 23 down regulated discriminant metabolites included histidinol, 4-aminobutyraldehyde, propanoate, tyramine and linalool. Melatonin and serotonin in the leaves were the two indoleamines being reported for the first time in tomato in response to the early blight pathogen. Receiver operating characteristic (ROC)-based biomarker analysis identified apigenin-7-glucoside, uridine, adenosyl-homocysteine, cGMP, tyrosine, pantothenic acid, riboflavin (as up regulated) and adenosine, homocyctine and azmaline (as down regulated) biomarkers. These results could aid in the development of metabolite-quantitative trait loci (mQTL). Furthermore, stress-induced biosynthetic pathways may be the potential targets for modifications through breeding programs or genetic engineering for improving crop performance in the fields.

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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