The eco-friendly synthesis of nanoparticles through various biological means helps to explore various plants for their ability to synthesize silver nanoparticles (AgNPs). Here we have synthesized AgNPs by using rhizome extract of Dioscorea batatas at 80∘C as well as room temperature (25∘C). AgNPs were characterized under UV-vis spectrophotometer, SEM, FTIR, XRD, and EDX. The antimicrobial activity of AgNPs was evaluated on gram positive (B. substilis and S. aureus), gram negative (E. coli), and fungi (S. cerivisae and C. albicans). At room temperature, S. cerivisae and C. albicans were found to be more susceptible to AgNPs than at 80∘C.