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Plant extract-based green synthesis of nanoparticles is an emerging class of nanotechnology that has revolutionized the entire field of biological sciences. Green synthesized nanoparticles are used as super-growth promoters and antifungal agents. In this study, selenium nanoparticles (SeNPs) were synthesized using Melia azedarach leaves extract as the main reducing and stabilizing agent and characterized by UV–visible spectroscopy, scanning electron microscopy (SEM), X-ray diffraction (XRD), energy-dispersive X-ray (EDX), and fourier transform infrared spectrometer (FTIR). The green synthesized SeNPs were exogenously applied on Mangifera indica infected with mango malformation disease. The SeNPs at a concentration of 30 μ g/mL were found to be the best concentration which enhanced the physiological (chlorophyll and membrane stability index), and biochemical (proline and soluble sugar) parameters. The antioxidant defense system was also explored, and it was reported that green synthesized SeNPs significantly reduced the biotic stress by enhancing enzymatic and non-enzymatic activities. In vitro antifungal activity of SeNPs reported that 300 μ g/mL concentration inhibited the Fusarium mangiferae the most. This study is considered the first biocompatible approach to evaluate the potential of green synthesized SeNPs to improve the health of mango malformation-infected plants and effective management strategy to inhibit the growth of F . mangifera .

The current study involves the biogenesis of titanium dioxide nanoparticles (TiO 2 NPs) by using Moringa oleifera Lam. aqueous leaf extract for the reduction of titanium dioxide salt into TiO 2 nanoparticles. The biosynthesized TiO 2 nanoparticles were observed by using the UV-visible spectrophotometry, SEM, EDX and XRD analytical methods. It was confirmed that the nanoparticles are crystalline and exist in the size range of 10–100 nm. The FTIR analysis confirmed the presence of O-H (hydrogen bonding), N-H (amide), C-C (alkanes) and C-I (Iodo-stretch) functional groups responsible for the stabilization of nanoparticles. Various concentrations (20, 40, 60 and 80 mg/L) of TiO 2 NPs were applied exogenously on wheat plants infected with a fungus Bipolaris sorokiniana responsible to cause spot blotch disease at different time intervals. The measurement of disease incidence and percent disease index showed the time-dependent response and 40 mg/L was reported a stable concentration of TiO 2 NPs to reduce the disease severity. The effects of biosynthesized TiO 2 NPs were also evaluated for agro-morphological (leaf and root surface area, plant fresh and dry weight and yield parameters), physiological (relative water content, membrane stability index and chlorophyll content) and non-enzymatic metabolites (soluble sugar, protein, soluble phenol and flavonoid content) in wheat plants under biotic stress and 40 mg/L concentration of TiO 2 NPs was found to be effective to elicit modifications to reduce biotic stress. The current study highlights the significant role of biosynthesized TiO 2 NPs in controlling fungal diseases of wheat plants and thus ultimately improving the quality and yield of wheat plants.

Aflatoxins (AFs) are the most important toxic, mutagenic, and carcinogenic fungal toxins that routinely contaminate food and feed. While more than 20 AFs have been identified to date, aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1), G2 (AFG2), and M1 (AFM1) are the most common. Over 25 species of Aspergillus have been shown to produce AFs, with Aspergillus flavus, Aspergillus parasiticus, and Aspergillus nomius being the most important and well-known AF-producing fungi. These ubiquitous molds can propagate on agricultural commodities to produce AFs in fields and during harvesting, processing, transportation, and storage. Countries with warmer climates and that produce foods susceptible to AF contamination shoulder a substantial portion of the global AF burden. Pakistan’s warm climate promotes the growth of toxigenic fungi, resulting in frequent AF contamination of human foods and animal feeds. The potential for contamination in Pakistan is exacerbated by improper storage conditions and a lack of regulatory limits and enforcement mechanisms. High levels of AFs in common commodities produced in Pakistan are a major food safety problem, posing serious health risks to the population. Furthermore, aflatoxin contamination contributes to economic losses by limiting exports of these commodities. In this review, recent information regarding the fungal producers of AFs, prevalence of AF contamination of foods and feed, current regulations, and AF prevention and removal strategies are summarized, with a major focus on Pakistan.

In this study, selenium nanoparticles (SeNPs) and cerium oxide nanoparticles (CeONPs) were synthesized by using the extract of Melia azedarach leaves, and Acorus calamusas rhizomes, respectively, and investigated for the biological and sustainable control of yellow, or stripe rust, disease in wheat. The green synthesized NPs were characterized by UV-Visible spectroscopy, scanning electron microscopy (SEM), energy-dispersive X-ray (EDX), and X-ray diffraction (XRD). The SeNPs and CeONPs, with different concentrations (i.e., 10, 20, 30, and 40 mg/L), were exogenously applied to wheat infected with Puccinia striformis. SeNPs and CeONPs, at a concentration of 30 mg/L, were found to be the most suitable concentrations, which reduced the disease severity and enhanced the morphological (plant height, root length, shoot length, leaf length, and ear length), physiological (chlorophyll and membrane stability index), biochemical (proline, phenolics and flavonoids) and antioxidant (SOD and POD) parameters. The antioxidant activity of SeNPs and CeONPs was also measured. For this purpose, different concentrations (50, 100, 150, 200 and 400 ppm) of both SeNPs and CeONPs were used. The concentration of 400 ppm most promoted the DPPH, ABTS and reducing power activity of both SeNPs and CeONPs. This study is considered the first biocompatible approach to evaluate the potential of green synthesized SeNPs and CeONPs to improve the health of yellow, or stripe rust, infected wheat plants and to provide an effective management strategy to inhibit the growth of Puccinia striformis.

Citrus production is harmed worldwide by yellow dragon disease, also known as Huanglongbing (HLB), or citrus greening. As a result, it has negative effects and a significant impact on the agro-industrial sector. There is still no viable biocompatible treatment for Huanglongbing, despite enormous efforts to combat this disease and decrease its detrimental effects on citrus production. Nowadays, green-synthesized nanoparticles are gaining attention for their use in controlling various crop diseases. This research is the first scientific approach to examine the potential of phylogenic silver nanoparticles (AgNPs) to restore the health of Huanglongbing-diseased ‘Kinnow’ mandarin plants in a biocompatible manner. AgNPs were synthesized using Moringa oleifera as a reducing, capping, and stabilizing agent and characterized using different characterization techniques, i.e., UV–visible spectroscopy with a maximum average peak at 418 nm, scanning electron microscopy (SEM) with a size of 74 nm, and energy-dispersive spectroscopy (EDX), which confirmed the presence of silver ions along with different elements, and Fourier transform infrared spectroscopy served to confirm different functional groups of elements. Exogenously, AgNPs at various concentrations, i.e., 25, 50, 75, and 100 mgL−1, were applied against Huanglongbing-diseased plants to evaluate the physiological, biochemical, and fruit parameters. The findings of the current study revealed that 75 mgL−1 AgNPs were most effective in boosting the plants’ physiological profiles, i.e., chl a, chl b, total chl, carotenoid content, MSI, and RWC up to 92.87%, 93.36%, 66.72%, 80.95%, 59.61%, and 79.55%, respectively; biochemical parameters, i.e., 75 mgL−1 concentration decreased the proline content by up to 40.98%, and increased the SSC, SOD, POD, CAT, TPC, and TFC content by 74.75%, 72.86%, 93.76%, 76.41%, 73.98%, and 92.85%, respectively; and fruit parameters, i.e., 75 mgL−1 concentration increased the average fruit weight, peel diameter, peel weight, juice weight, rag weight, juice pH, total soluble solids, and total sugarby up to 90.78%, 8.65%, 68.06%, 84.74%, 74.66%, 52.58%, 72.94%, and 69.69%, respectively. These findings enable us to develop the AgNP formulation as a potential citrus Huanglongbing disease management method.

In the present study, SeNPs were synthesized using Melia azedarach leaf extracts and investigated for growth promotion in wheat under the biotic stress of spot blotch disease. The phytosynthesized SeNPs were characterized using UV-visible spectroscopy, scanning electron microscopy (SEM), energy-dispersive X-ray (EDX), and Fourier-transformed infrared spectroscopy (FTIR). The in vitro efficacy of different concentrations of phytosynthesized SeNPs (i.e., 100 μg/mL, 150 μg/mL, 200 μg/mL, 250 μg/mL, and 300 μg/mL) was evaluated using the well diffusion method, which reported that 300 μg/mL showed maximum fungus growth inhibition. For in vivo study, different concentrations (10, 20, 30, and 40 mg/L) of SeNPs were applied exogenously to evaluate the morphological, physiological, and biochemical parameters under control conditions and determine when infection was induced. Among all treatments, 30 mg/L of SeNPs performed well and increased the plant height by 2.34% compared to the control and 30.7% more than fungus-inoculated wheat. Similarly, fresh plant weight and dry weight increased by 17.35% and 13.43% over the control and 20.34% and 52.48% over the fungus-treated wheat, respectively. In leaf surface area and root length, our findings were 50.11% and 10.37% higher than the control and 40% and 71% higher than diseased wheat, respectively. Plant physiological parameters i.e., chlorophyll a, chlorophyll b, and total chlorophyll content, were increased 14, 133, and 16.1 times over the control and 157, 253, and 42 times over the pathogen-inoculated wheat, respectively. Our findings regarding carotenoid content, relative water content, and the membrane stability index were 29-, 49-, and 81-fold higher than the control and 187-, 63-, and 48-fold higher than the negative control, respectively. In the case of plant biochemical parameters, proline, sugar, flavonoids, and phenolic contents were recorded at 6, 287, 11, and 34 times higher than the control and 32, 107, 33, and 4 times more than fungus-inoculated wheat, respectively. This study is considered the first biocompatible approach to evaluate the potential of green-synthesized SeNPs as growth-promoting substances in wheat under the spot blotch stress and effective management strategy to inhibit fungal growth.

Sesame Sesamum indicum L. is a major oil-based seed crop that has been widely cultivated and consumed in Pakistan. Unfortunately, sesame is highly prone to Aspergillus fungal growth in the field, and under inappropriate storage conditions can become contaminated with aflatoxins, the most potent carcinogen found in nature. Here, we have isolated a high number of Aspergillus isolates from sesame seeds in fresh and stored conditions obtained from rainfed and irrigated zones of Punjab, Pakistan, and characterized them for aflatoxigenic potentials. Using morphological identification techniques, 260 isolates were grouped as potential Aspergillus section Flavi, with 126 and 134 originating from the rainfed and irrigated zones, respectively. Out of 260 in total, 188 isolates were confirmed to produce aflatoxins. There were no significant differences in potential aflatoxigenic isolates with respect to the rainfed and irrigated zones. However, the number of potential aflatoxigenic isolates was significantly higher (p < 0.05) in stored samples than that of those from fresh sesame seeds in the rainfed and irrigated zone. Whole genome sequencing and comparative analyses of 12 select isolates have revealed that one of the A. flavus isolates, which produced very low aflatoxins (AFP10), has an elevated missense variant rate, numerous high impact mutations, and a 600 base pair deletion in the norB gene. In summary, our study provides insights into aflatoxigenic potential and the associated genetic diversity of indigenous Aspergillus section Flavi isolates and potential management strategies for reducing aflatoxin contamination levels in a major crop consumed in Punjab, Pakistan.

This study was carried out to evaluate the prevalence of mycobiota and aflatoxin (AF) B1 contamination in sesame seeds from rainfed and irrigated zones of the Punjab, Pakistan. For this purpose, 100 sesame seed samples were collected directly from the fields of major sesame-producing areas in rainfed and irrigated zones. The agar plate method was used for isolation of mycobiota, and thin-layer chromatography was used to determine AFB1 concentrations. Seed samples were then stored for 12 months. After 12 months, the seeds were again analyzed for mycobiota and AFB1 for comparison. All samples were positive for fungal growth under fresh and stored conditions. Twenty-one fungal species of 10 genera were isolated. Aspergillus flavus was the most prevalent contaminant found in fresh and stored sesame seeds from rainfed and irrigated zones, followed by Aspergillus niger, Alternaria alternata, and Fusarium oxysporum. The least prevalent fungi were Aspergillus ochraceus and Cladosporium oxysporum. Analysis revealed that 92% of fresh and 99% of stored seed samples were contaminated with AFB1. In the rainfed zone, 88% of fresh and 100% of stored seed samples were contaminated with AFB1, with mean concentrations of 15.74 and 33.8 ppb, respectively. In the irrigated zone, 96% of fresh and 98% of stored seed samples were contaminated with AFB1, with mean concentrations of 20.5 and 27.56 ppb, respectively. AFB1 concentrations >20 ppb were found in 20% of fresh and 100% of stored seeds samples from the rainfed zone and in 28% of fresh and 60% of stored samples from the irrigated zone and thus were not fit for human consumption as per the maximum limit (20 ppb) assigned by the U.S. Food and Drug Administration and the Food and Agriculture Organization of the United Nations. This report is the first on the mycobiota and AFB1 contamination in sesame seeds from rainfed and irrigated zones of the Punjab, Pakistan. These baseline data are an initial step in the effort to deal with this significant food safety issue.

Plant diseases caused by fungal and bacterial pathogens are threatening the agriculture and human health all over the world. The efficacies of some antifungal and antimicrobial agents, which are currently used to inhibit disease-causing microorganisms have been weakened by microbial resistance. Therefore, new antimicrobial agents that could overcome this resistance need to be discovered. The present study was carried out to assess the antifungal and antibacterial potential of oilseed cakes against soil-borne pathogenic fungi (Fusarium oxysporum and Rhizoctonia solani) and virulent strains of bacteria (Bacillus cereus and Rhizobium sp). Ethanolic extracts of oilseed cakes of Prunus dulcis, Gossypium hirsutum, Jatropha curcas, Simmondsia chinensis, Azadirachta indica with different concentrations were evaluated for antifungal and antibacterial activity by disk diffusion method. The overall result indicated that that ethanolic extracts of all seed cakes possessed significant antifungal and antibacterial activity at 1000ppm. Gossypium hirsutum seed cake showed a maximum zone of inhibition (ZOI=1.00±0.05) against Fusarium oxysporum. Prunus dulcis seed cake showed maximum inhibition of Rhizoctonia solani (Z0I=1.13±0.18) and Bacillus cereus (Z0I=0.12±0.02). Simmondsia chinensis was found most effective against Rhizobium sp (Z0I=0.30±0.05). These findings suggested that tested oilseed cakes could be utilized for fungal and bacterial disease management and can be further developed to be a natural fungicide and antibacterial for sustainable agriculture. The use of oilseed cakes as biocontrol agents would benefit horticulture crops, as well as the management of industrial waste and the preservation of soil fertility.

Various cultivars of red chilli were collected from a small town named Kunri, located in the province Sindh, Pakistan. This town is a hub of red chilli production in Asia. A total of 69 samples belonging to 6 cultivars were obtained and analysed for the occurrence of aflatoxins and Aspergillus flavus , to explore the potential of resistant and susceptible germplasm. Aflatoxins were detected by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC), while A. flavus was isolated and identified using agar plate, blotter paper, deep freezing and dilution techniques. Molecular characterization using internal transcribed spacer (ITS) 1/4 and A. flavus specific FL1-F/R primers confirmed the identity of A. flavus . The data revealed that 67 and 75% samples contaminated with aflatoxin B 1 (AFB 1 ) and with A. flavus , respectively. A highly susceptible chilli cultivar was ‘Nagina’, showing 78.8% frequency of total aflatoxins (1.2–600 μg/kg) and a mean of 87.7 μg/kg for AFB 1 and 121.9 μg/kg for total aflatoxins. A. flavus was detected with 93% frequency and 2.14 × 10 4 colony forming units. In contrast, cultivars ‘Kunri’ and ‘Drooping Type’ were found to be resistant, with low levels of aflatoxins and fungal counts. The study was conducted for the first time to explore two potential cultivars that were less susceptible towards A. flavus and aflatoxin contamination. These cultivars could be preferably cultivated and thereby boost Pakistan’s chilli production.