Potato (Solanum tuberosum) is an important crop plant grown around the world for food and nutrition. In Pakistan, potato production averaged a yield of 8 million tons in 1. Potato is affected by many fungal pathogens that cause major economic losses. Potato dry rot is caused by Fusarium that is a devastating fungal disease 2. Fusarium equiseti, Fusarium sambucinum, Fusarium solani complex, Fusarium oxysporum, Fusaium semitectum Fusarium graminearum and Fusarium venenatum are pathogenic Fusarium species on potato worldwide 3,4. Symptoms of dry rot in potato include sunken and wrinkled brown to black patches on tubers, which exhibit reduced dry matter content and shriveled flesh. As storage duration increases, these wrinkled patches may develop cottony white, purple, pink, or brick orange spores and mycelia masses 4. Consequently, internal tissues undergo black or brown rot. The pathogen gains entry through wounds, which serve as primary infection site, entering xylem vessels causing leaf chlorosis followed by necrosis, stunting, wilting and eventually plant death 2. Fusarium species can survive for many years as fungal propagules in the soil, colonize living plants or plant debris, as saprophytes, endophytes or heterophytes. Of different Fusarium species, Fusarium oxysporum produces white color colony with a pink or violet center when grown on potato dextrose agar (PDA) media. Fusarium solani also produces white color colony on PDA but it becomes blue-green or bluish brown. On the underside, this may be pale, tea-with-milk-brown, or reddish brown. Fusarium solani grows rapidly, but not as rapidly as Fusarium oxysporum 2,5. A recent study revealed Fusarium falciforme (from Fusarium soloni species complex) and Fusarium oxysporum were the causal pathogens of fruit rot on postharvest watermelon in Malaysia 6. Fusarium solani species complex (FSSC) is one of the prevalent pathogens of dry rot of potato including F. keratoplasticum, F. falciforme, and F. solani 7. Nearly all species are able to produce mycotoxins that are toxic 2,4. Fusarium mycotoxins are aflatoxins, fumonisins, zearalenone, deoxynivalenol, cyclopiazonic acid and trichothecenes. Trichothecenes possess sesquiterpene isoprenoids that are produced by Fusarium culmorum, Fusarium graminearum and Fusarium sambucinum. Trichothecenes inhibit protein synthesis in the mitochondria that allows reactive oxygen species build up in the cell resulting in oxidative stress and induction of programmed cell death or apoptosis 2,4. Zralenone is another secondary metabolite produced by Fusarium culmorum, Fusarium graminearum, Fusarium cerealis, Fusarium semitectum, Fusarium verticillioides and Fusarium equiseti. Zralenone is thus a potent estrogenic metabolite produced by many Fusarium species. Similarly, Deoxynivalenol is the most widely distributed mycotoxin produced by Fusarium graminearum and Fusarium culmorum 4,8,9. Fumonisins are produced by Fusarium verticillioides and closely related Fusarium proliferatum and Fusarium subglutinans. The fumonisins have a structure similar to sphingolipids. Since Fusarium is of wide occurrence and distribution, a sensitive and rapid surface plasmon resonance imaging assay has detected these mycotoxin such as cyclopiazonic acid in maize and cheese 10. Similarly, nanoparticles-based lateral flow immunoassay is being used for secondary metabolites detection. The rapidly evolving field of nanotechnology has recently gained significant importance due to its management of various medical aliments and agricultural diseases made possible by their strong antimicrobial activity. Nanoparticles typically range from 1 and 100 nm in size and have been implicated into a wide range of profound applications. Nanoparticles, particular silver nanoparticles (Ag NPs) have gained attention due to their unique small size, large surface area to mass ratio that confer nanoparticles distinct physical and chemical properties and rendering them cost-efficient, highly stability and less toxic 11,12. Biologically synthesized Ag NPs are preferred as compared to their counter-parts physical and chemical methods because they encounter problems with scalability and hazardable toxins during mass production. Biologically synthesized Ag NPs are made using plants, fungi and bacteria 11,12. Different fungi-based Ag NPs are reported including Aspergillus, Penicillium and Fusarium species 13,14. These are preferred due to their eco-friendly, bulk production and reproducibility 15. Fusarium oxysporum base Ag NPs were produced with antibacterial activity against Escherichia coli, Staphylococcus aureus and that showed cytotoxic activity against human breast carcinoma MCF-7 cell line 16. Fusarium scirpi Ag NPs had antimicrobial potential against uropathogenic Escherichia coli biofilms 17. Similarly, Fusarium mangiferae Ag NPs also showed antibiofilm potential 18. Ag NPs and Au NPs of Fusarium pseudonygama gave antibiofilm, antioxidant activity and anticancer activity against cancer cell lines 19. Ag NPs were synthesized by using Fusarium pallidoroseum biomass and they showed significant antilarval activity against white grub larvae 20. Extracellular synthesis of Ag NPs by Fusarium acuminatum from ginger showed resistance to Staphylococcus aureus, Salmonella typhi, and Escherichia coli showing simple, eco-friendly method for their production21. Hydrogel with PEG-coated Fusarium verticillioides Ag NPs showed effective wound healing ability with low toxicity and high tolerability with MRSA-infected wounded mice as compared to commercially available silver sulfadiazine cream showing Fusarium coated Ag NPs potential for their wound healing ability 22. Fusarium chlamydosporum and Penicillium chrysogenum Ag NPs showed cytotoxic effect against cancer cells rendering them as potential therapeutic agents 23. Fusarium solani based Ag NPs and Fusarium solani gold (Au) nanoparticles showed significant impact on grain borne fungi and cancer cell lines respectively 24, 25. Thus, Fusarium Ag NPs can be regarded as a source of novel bioactive secondary metabolites having a wide range of biological activities. The objectives of this research was first to identify Fusarium species causing potato dry rot using Elongation Factor (EF) unique sequence followed by construction of phylogenetic tree based on the unique identified sequence. This study is also aimed at investigating Fusarium Ag NPs role against pathogenic bacteria. The synthesized Ag NPs were characterized using UV, SEM, FTIR, XRD and EDS. To the best of our knowledge, this is the first report of fungi that is isolated and used to synthesize and characterize Ag NPs and used against bacterial pathogens.