In the current investigation, we report the biosynthesis of silver nanoparticles (Ag NPs) employing extract of Alternaria alternata, which is an eco-friendly process for the synthesis of metallic nanoparticles. Ag NPs were synthesised through the reduction of aqueous Ag+ ion using the cell extract of fungus A. alternata in the dark conditions. The synthetic process was relatively fast and Ag NPs were formed within 24 h. UV–visible spectrum of the aqueous medium containing silver ion showed a peak at 435 nm corresponding to the plasmon absorbance of Ag NPs and another peak at 280 nm refers to tyrosine amino acid. The nanoparticles were characterised by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). The morphology of nanoparticles is found to be spherical mostly, with ranging size of 27–79 nm; as revealed by SEM. The FTIR spectrum analysis indicated that biomolecules were involved in the synthesis of Ag NPs. The presence of the amino groups is expected to pack differently around the Ag NPs. This in turn will influence the self-assembly of nanoparticles on substrates as well as their stability. The present study demonstrates the possible use of biologically synthesised Ag NPs in the field of agriculture, when A. alternata could be used for simple, nonhazardous and efficient synthesis of Ag NPs.