In the present study, silver nanoparticles (Ag-NPs) were synthesized by a chemical and biological method. Further, nanoparticles were characterized for their morphological feature using techniques like UV-Visible, TEM, XRD, and zeta potential. Sharp UV-visible absorption maximum at 410 was observed for biological synthesized silver nanoparticles (Bio-AgNPs), whereas for chemical synthesized silver nanoparticles (CH-AgNPs) peak was observed at 414 nm. TEM micrograph confirmed the formation of spherical nanoparticles dominantly via both protocols with an average size of nanoparticles was 50 nm and 25 nm for CH-AgNPs and Bio-NPs, respectively. Further, the antimicrobial potential of AgNPs was evaluated at different concentrations (25-100 ppm), against three pathogenic plant fungus plant (Alternaria solani, Corynespora cassiicola, and Fusarium spp.), in two different fungal media in term of inhibition of radial growth. Up to 100% inhibition for Alternaria solani and Fusarium spp. and 85% inhibition for Corynespora cassiicola was observed at 100 ppm AgNPs concentration on potato dextrose agar (PDA). Further, exposure of AgNPs on Drosophila melanogaster confirmed that Bio-AgNPs are nontoxic as compared to CH-AgNPS. Hence it can be concluded that Bio-AgNPs are safe to use due to their nontoxic nature.