Green synthesis of nanomaterials is advancing due to its ease of synthesis, inexpensiveness, nontoxicity and renewability. In the present study, an eco-friendly biogenic method was developed for the green synthesis of nickel oxide nanoparticles (NiONPs) using phytochemically rich Berberis balochistanica stem (BBS) extract. The BBS extract was rich in phenolics, flavonoids and berberine. These phytochemicals successfully reduced and stabilised the NiNO₃ (green) into NiONPs (greenish-gray). BBS-NiONPs were confirmed by using UV-visible spectroscopy (peak at 305 nm), X-ray diffraction (size of 31.44 nm), Fourier transform infrared spectroscopy (identified -OH group and Ni-O formation), energy dispersive spectroscopy (showed specified elemental nature) and scanning electron microscopy (showed rhombohedral agglomerated shape). BBS-NiONPs were exposed to multiple in vitro bioactivities to ascertain their beneficial biological applications. They exhibited strong antioxidant activities: total antioxidant capacity (64.77%) and 2, 2-diphenyl-1-picrylhydrazyl (71.48%); and cytotoxic potential: Brine shrimp cytotoxicity assay with IC₅₀ (10.40 µg/mL). BBS-NiONPs restricted the bacterial and fungal pathogenic growths at 1000, 500 and 100 µg/mL. Additionally, BBS-NiONPs showed stimulatory efficacy by enhancing seed germination rate and seedling growth at 31.25 and 62.5 µg/mL. In aggregate, BBS extract has a potent antioxidant activity which makes the green biosynthesis of NiONPs easy, economical and safe. The biochemical potential of BBS-NiONPs can be useful in various biomedical and agricultural fields.

Grapefruit (Citrus paradisi) is a widely grown citrus and its fruit is affected by a variety of biotic and abiotic stress. Keeping in view the hazardous effects of synthetic fungicides, the recent trend is shifting towards safer and eco-friendly control of fruit diseases. The present study was aimed to diagnose the fruit rot disease of grapefruit and its control by using zinc oxide green nanoparticles (ZnO NPs). Fruit rot symptoms were observed in various grapefruit growing sites of Pakistan. Diseased samples were collected, and the disease-causing pathogen was isolated. Following Koch's postulates, the isolated pathogen was identified as Rhizoctonia solani. For eco-friendly control of this disease, ZnO NPs were prepared in the seed extract of Trachyspermum ammi and characterized. Fourier transform infrared spectroscopy (FTIR) of these NPs described the presence of stabilizing and reducing compounds such as phenols, aldehyde and vinyl ether, especially thymol (phenol). X-ray diffraction (XRD) analysis revealed their crystalline nature and size (48.52 nm). Energy dispersive X-ray (EDX) analysis elaborated the presence of major elements in the samples, while scanning electron microscopy (SEM) confirmed the morphology of bio fabricated NPs. ZnO NPs exhibited very good anti-fungal activity and the most significant fungal growth inhibition was observed at 1.0 mg/ml concentration of green NPs, in vitro and in vivo. These findings described that the bioactive constituents of T. ammi seed extract can effectively reduce and stabilize ZnO NPs. It is a cost-effective method to successfully control the fruit rot disease of grapefruit.

Lychee (Litchi chinensis Sonn.) is a famous fruit species of tropical and subtropical regions of the world and many biotic and abiotic stresses affect its yield. In this study, lychee fruit rot has been observed and its incidence has been controlled by using zinc oxide nanoparticles (ZnO NPs). Diseased lychee fruits were collected and diagnosed to identify disease-causing pathogens. Morphological appearance, microscopic observation, and sequence analysis of the amplified ITS region identified this isolated pathogen as Aspergillus niger. To control this problem, ZnO NPs were prepared in the leaf extract of Azadirachta indica. Before their antifungal activity, ZnO NPs were characterized using sophisticated approaches. FTIR revealed the presence of reducing and stabilizing molecules on ZnO NPs including alcohol, carboxylic acid, alkyl halide, amine, and alkyl halide. Crystalline nature and average size (29.024 nm) of synthesized ZnO NPs were described by X-ray diffraction. EDX analysis depicted the mass percentage of zinc (30.15%) and oxygen (14.90%). SEM analysis displayed the irregular shape of nanoparticles and confirmed the nano-size of ZnO NPs. Maximum mycelial growth inhibition (70.5%) was observed at 1.0 mg/mL concentration of ZnO NPs in vitro. In in-vivo disease-control analysis, maximum control of lychee fruit rot disease was observed at the same concentration. These results reveal the potential use of these ZnO NPs on a larger scale to replace hazardous chemical fungicides.

Cherry is a fleshy drupe, and it is grown in temperate regions of the world. It is perishable, and several biotic and abiotic factors affect its yield. During April–May 2021, a severe fruit rot of cherry was observed in Swat and adjacent areas. Diseased fruit samples were collected, and the disease-causing pathogen was isolated on PDA. Subsequent morphological, microscopic, and molecular analyses identified the isolated pathogen as Aspergillus flavus. For the control of the fruit rot disease of cherry, iron oxide nanoparticles (Fe₂O₃ NPs) were synthesized in the leaf extract of Calotropis procera and characterized. Fourier transform infrared (FTIR) spectroscopy of synthesized Fe₂O₃ NPs showed the presence of capping and stabilizing agents such as alcohols, aldehydes, and halo compounds. X-ray diffraction (XRD) analysis verified the form and size (32 nm) of Fe₂O₃ NPs. Scanning electron microscopy (SEM) revealed the spinal-shaped morphology of synthesized Fe₂O₃ NPs while X-ray diffraction (EDX) analysis displayed the occurrence of main elements in the samples. After successful preparation and characterization of NPs, their antifungal activity against A. flavus was determined by poison technique. Based on in vitro and in vivo antifungal activity analyses, it was observed that 1.0 mg/mL concentration of Fe₂O₃ can effectively inhibit the growth of fungal mycelia and decrease the incidence of fruit rot of cherry. The results confirmed ecofriendly fungicidal role of Fe₂O₃ and suggested that their large-scale application in the field to replace toxic chemical fungicides.

Heavy metals contaminate soil and adversely affect plant growth. These soils contain different fungi and bacteria which exhibit metal tolerance and work as bioremediation agents to detoxify polluted soils. In the present study, polluted soil samples were collected to estimate the contamination of copper (Cu) and cadmium (Cd). From this contaminated soil, metal tolerant fungi were isolated and characterized. Copper and cadmium were found in a range of 190.2–300.4 mg/kg and 46.8–56.1 mg/kg, respectively. For the isolation of metal tolerant fungi, soil dilutions were made in water and inoculated on potato dextrose agar (PDA) media. Fungal growth was observed on PDA and successive screening resulted in the isolation of four multi-metal tolerant fungal species, including Penicillim oxalicum, Fusarium solani, Aspergillus niger and Trichoderma harzianum. Sequencing of 18S rRNA genes of isolated fungi also efficiently identified them. To reveal minimum inhibitory concentrations (MIC), these fungi were exposed to increasing concentrations of cadmium and copper chlorides (100 to 1000 ppm) and a variable MIC range of 400 ppm to 1000 ppm was estimated. Based on tolerance index analysis, F. solani was found to be resistant at 1 mM copper, while P. oxalicum was the most tolerant species under cadmium stress. F. solani and P. oxalicum demonstrated the highest biosorption capacity of Cu and Cd, respectively. Both metals negatively affected wheat seedlings in a pot experiment, while the treatment of both F. solani and P. oxalicum positively influenced the germination and growth of wheat. Based on these observations, it could be inferred that F. solani and P. oxalicum can be used for the myco-remediation of Cu and Cd, respectively.