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Biosynthesized silver nanoparticles (AgNPs) are environment friendly, cost-effective, bio-compatible and expanding research area due to their potential applications in medical domain. The present study focuses on biologically synthesized of AgNPs using aqueous root extract of  Iris germanica as reducing agent as well as capping agent and examined their potential antibacterial efficacy. In reduction reaction it was observed that silver (Ag+1) ions associate in root extract and reduced in solution (Ag0) leads to formation of stable formation of spherical AgNPs. Biosynthesized AgNPs showed an effective and rapid antibacterial activity against both bacterial strain (gram+ve and gram-ve). Results exhibited that AgNPs were extremely toxic towards Bacillus subtilis and Escherichia coli pathogenic bacterial strains and can be utilized for the applications in biomedical science.