Tobacco mosaic virus (TMV) is a major pathogen affecting tomato plants worldwide. The efficacy of silver nanoparticles (Ag-NPs) mediated by Punica granatum biowaste peel extract in mitigating the negative impact of TMV infection on tomato growth and oxidative stress was investigated through scanning electron microscopy (SEM), transmission electron microscopy (TEM), UV-Visible (UV-Vis) spectrophotometer, X-ray Diffraction (XRD), dynamic light scattering (DLS), zeta potential, energy-dispersive X-ray spectroscopy (EDX), and Fourier-transform infrared spectra (FTIR). Results of SEM analysis of green Ag-NPs revealed the presence of condensed spherical or round NPs with diameters ranging between 61 and 97 nm. TEM confirmed the SEM results and showed round-shaped Ag-NPs with an average size of 33.37 ± 12.7 nm. The elemental analysis (EDX) of prepared Ag-NPs revealed the presence of elemental Ag as a major peak (64.43%) at 3–3.5 KeV. The FTIR revealed several functional groups on the prepared Ag-NPs, for which three treatment strategies for Ag-NP applications were evaluated in the greenhouse study and compared to inoculated TMV and control plants: pre-infection treatment (TB), post-infection treatment (TA), and dual treatment (TD). The results showed that the TD strategy is the most effective in improving tomato growth and reducing viral replication, whereas all Ag-NP treatments (TB, TA, and TD) were found to significantly increase expression of the pathogenesis-related (PR) genes PR-1 and PR-2, as well as polyphenolic compounds, HQT, and C4H genes compared to control plants. In contrast, the flavonoid content of tomato plants was not affected by the viral infection, while the phenolic content was significantly reduced in the TMV group. Furthermore, TMV infection led to a significant increase in oxidative stress markers MDA and H₂O₂, as well as a reduction in the enzymatic activity of the antioxidants PPO, SOD, and POX. Our results clearly showed that the application of Ag-NPs on TMV-infected plants reduces virus accumulation, delays viral replication in all treatments, and greatly enhances the expression of the CHS gene involved in flavonoid biosynthesis. Overall, these findings suggest that treatment with Ag-NPs may be an effective strategy to mitigate the negative impact of TMV infection on tomato plants.

Cinnamomum camphora methanolic extract was tested for antifungal activity against three common, isolated, widespread phytopathogens: Alternaria alternata, Fusarium solani, and Fusarium oxysporum, which were molecularly identified and assigned accession numbers ON795987, ON795988, and ON795989, respectively. At 4000 µg/mL, the highest concentration of C. camphora methanolic extract inhibits the fungal mycelia weight of F. oxysporum, A. alternata, and F. solani by 60, 49, and 24%, respectively. The presence of several bioactive metabolites in the C. camphora extract could explain its antifungal activity. The presence of numerous phenolic and flavonoid compounds in the extract was revealed by HPLC analysis, including catechin and gallic acid, which had the highest concentrations of 6.21 and 6.98 µg/mL, respectively. Furthermore, osmoprotectants, total amino acids, and glycine betaine were abundant. Furthermore, total antioxidant activities, as measured by PMA and DPPH, were significant. The most abundant compound in the extract, according to GC-MS analysis, was mono(2-ethylhexyl) ester of 1,2-benzene dicarboxylic acid. Based on its in vitro efficacy in inhibiting mycelial growth weight, the tested extract could be recommended as a safe fungicide instead of a chemical treatment.

Plant diseases significantly reduce crop yields, threatening food security and agricultural sustainability. Fungi are the most destructive type of phytopathogen, and they are responsible for major yield losses in some of the most crucial crops grown across the world. In this study, a fungus isolate was detected from infected tomato plants and molecularly identified as Pythium aphanidermatum (GenBank accession number MW725032). This fungus caused damping-off disease and was shown to be pathogenic. Moreover, the expression of five pathogenesis-related genes, namely PR-1, PR-2, PR-3, PR-4, and PR-5, was quantitatively evaluated under the inoculation of tomato with P. aphanidermatum. The quantitative polymerase chain reaction (qPCR) showed that the expression levels of PR-1, PR-2, and PR-5 genes went up significantly at 5 days post-inoculation (dpi). The expression of the PR-1 gene also increased the variably, which reached its highest value at 20 dpi, with a reported relative expression level 6.34-fold higher than that of the control. At 15 dpi, PR-2 and PR-5 increased the most, while PR-1, PR-3, and PR-5 also increased noticeably at 20 dpi. On the contrary, PR-4 gene expression significantly decreased after inoculation, at all time intervals. Regarding PR-5 gene expression, the data showed a variable change in PR-5 gene expression at a different sample collection period. Still, it was highly expressed at 15 dpi and reached 3.99-fold, followed by 20 dpi, where the increasing percentage reached 3.70-fold, relative to the untreated control. The HPLC analysis indicated that the total concentration of all detected polyphenolic compounds was 3858 µg/g and 3202.2 µg/g in control and infected plant leaves, respectively. Moreover, the HPLC results concluded that Pythium infection decreased phenolic acids, such as chlorogenic and ellagic acids, which correlated with the infection–plant complex process. Based on the results, P. aphanidermatum could be a biotic stress pathogen that causes the expression of pathogen-related genes and stops the regulation of defensin phenolic compounds.

In this study, we used RT-qPCR to examine how PR genes were expressed in model tomato (Solanum lycopersicum L.) plants that had been infected with TMV or CMV. Under greenhouse conditions, the indirect ELISA data showed that both viruses were detected for the first time at 6 dpi. Then, the levels of accumulation increased very quickly, reaching a peak of 15 dpi. During the course of the study (1–15 dpi), the Delta C_T, NormFinder, BestKeeper, and GeNorm software tools revealed that the β-actin gene was the most informative reference gene in the virally infected tomato tissues. For both the TMV- and CMV-infected tomato plants, the transcriptional expression levels of most tested genes changed between activation and repression, especially in the first 12 dpi. Compared to mock-inoculated plants, the expression levels of PR-1 were induced at all time intervals except at 8 dpi for CMV and at 6, 7, and 8 dpi for TMV infection. Conversely, the greater activation and accumulation of both viruses were associated with the greater up-regulation of PR-2 at 8 dpi, with relative expression levels of 7.28- and 5.84-fold for TMV and CMV, respectively. The up-regulated expression of PR-3, PR-4, and PR-7 was shown at 4 dpi. In contrast, the PR-5 gene was inhibited in TMV at 1 dpi until 9 dpi, and the induction of this gene at 10 dpi increased by 1.72-fold, but PR-5 was observed to up-regulate the expression of CMV at 1 dpi. This study provides the first valuable information on the comparative transcriptional levels of these tomato genes between TMV and CMV infections.

Three different concentrations of four (ethanol, acetone, methanol, and diethyl ether) extracts of licorice, doum, and banana peel were evaluated for antifungal and antimycotoxigenic efficiency against a maize aflatoxigenic fungus, Aspergillus flavus. Among them, the licorice diethyl ether 75% extract was intensely active, showing the best wet and dry weight inhibition and exhibiting the highest efficacy ratio (91%). Regarding aflatoxin B1 (AFB1) production, all the plant extracts tested were effective against AFB1 production after one month of maize storage, with average efficacy ratios ranging from 74.1% to 97.5%. At the same time, Thiram fungicide exhibited an efficacy ratio of 20.14%. The relative expression levels of three structural genes (aflD, aflP, and aflQ) and two regulatory genes (aflR and aflS) were significantly downregulated when compared to untreated maize grains or Thiram-treated maize grains. The doum diethyl ether 75% peel extract showed the highest total phenolic content (60.48 mg GAE/g dry extract wt.) and antioxidant activity (84.71 μg/mL). GC–MS analysis revealed that dimethoxycinnamic acid, aspartic acid, valproic acid, and linoleic acid might imbue the extracts with antioxidant capacities in relation to fungal growth and aflatoxin biosynthesis. Finally, the results suggest that the three plant extracts can be considered a promising source for developing potentially effective and environmentally safer alternative ways to control aflatoxin formation, thus creating a potentially protective method for grain storage.

This study aims to detect Clostridium botulinum and its control using natural leaf extracts of Citrus limon, Citrus sinensis, and Citrus unshiu in Egyptian fish products, e.g., canned tuna, canned sardine, canned mackerel, fesikh, moloha, and renga, as well the application of C. limon in tuna. Moreover, the antibacterial activity of the C. limon leaf extract was also estimated. In the water extract, ascorbic acid, total flavonoid content (TFC), and total phenolic content (TPC) were determined by volumetric, aluminum chloride, and Folin–Ciocalteu approaches, respectively. The antioxidant ability of the extract was analyzed in vitro via free radical scavenging (DPPH) and Ferric reducing assays. The results showed variability in the distribution of the total number of positive C. botulinum in fish samples from three different governorates under study, which were (24) Alexandria, (16) Beheira, and (17) Gharbia, out of the 120 tested samples in each governorate. Additionally, the findings revealed that all three Citrus extracts contain an appropriate number of secondary metabolites, with a sustainable presence of saponin and tannins in the C. limon extract. Furthermore, all Citrus extracts inhibited bacterial growth by increasing the inhibition zone, with C. limon being the best extract (25 mm) compared to C. sinensis and C. unshiu. The overall results showed the high antioxidant and anti-Clostridium powers (p < 0.05) of C. limon leaf extract, indicating its preservative activity in fishery products during storage. Finally, C. limon leaf extract can fight off C. botulinum and is considered a promising natural preservation candidate in ensuring safe and fresh fishery products.