Chickpea (Cicer arietinum L.) is one of the main pulse crops of Pakistan. The yield of chickpea is affected by a variety of biotic and abiotic factors. Due to their environmentally friendly nature, different nanoparticles are being synthesized and applied to economically important crops. In the present study, Trichoderma harzianum has been used as a stabilizing and reducing agent for the mycosynthesis of zinc oxide nanoparticles (ZnO NPs). Before their application to control Fusarium wilt of chickpea, synthesized ZnO NPs were characterized. X-ray diffraction (XRD) analysis revealed the average size (13 nm) of ZnO NPs. Scanning electron microscopy (SEM) indicated their spherical structure, and energy dispersive X-ray analysis (EDX) confirmed the oxide formation of ZnO NPs. Transmission electron microscopy (TEM) described the size and shape of nanoparticles, and Fourier transform infrared (FTIR) spectroscopy displayed the presence of reducing and stabilizing chemical compounds (alcohol, carboxylic acid, amines, and alkyl halide). Successfully characterized ZnO NPs exhibited significant mycelial growth inhibition of Fusarium oxysporum, in vitro. In a greenhouse pot experiment, the priming of chickpea seeds with ZnO NPs significantly increased the antioxidant activity of germinated plants and they displayed 90% less disease incidence than the control. Seed priming with ZnO NPs helped plants to accumulate higher quantities of sugars, phenol, total proteins, and superoxide dismutase (SOD) to create resistance against wilt pathogen. These nanofungicides were produced in powder form and they can easily be transferred and used in the field to control Fusarium wilt of chickpea.

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.