Biosynthesis of metallic nanoparticles is simple, effective, eco-friendly and inexpensive way. It is a non-toxic method. Biosynthesized nanoparticles are primarily biocompatible and can be used in biomedical applications. Silver nanoparticles (AgNPs) are known to have strong antibacterial effects against pathogenic bacteria and have shown antifungal, antiviral activities so they can be helpful not only for fighting against bacteria. In given study silver nanoparticles were biosynthesized using methanolic extracts of Gentiana septemfida, Erysimum contractum and Chelidonium majus. Obtained nanoparticles were evaluated for antibacterial and fungicidal activities against Gram-negative Escherichia coli, Gram-positive Staphylococcus aureus and Candida albicans. Also, their cytotoxic effects were evaluated on human lung carcinoma A-549, colon adenocarcinoma DLD-1, and healthy human skin fibroblasts WS1 cell lines. The results show antibacterial, antifungal and cytotoxic effects at different level for each type. Biosynthesis of silver nanoparticles is simple, cost effective, eco-friendly method and obtained nanoparticles have potential to be used as antibacterial, antifungal and cytotoxic agents.