Silver (Ag) nanoparticles (NPs) were green synthesized at room temperature using different concentrations of the Thymbra spicata L. var. spicata (Zahter) aqueous leaf extracts for the first time. With the synthesis of AgNPs using the leaf extract of Cynara scolymus (Artichoke) and Mentha piperita (Peppermint), the biological activities of the nanoparticles synthesized using leaf extract of three economically significant plants have been studied comparatively. Nanoparticles were characterized by different spectroscopic and microscopic analysis. TEM analysis of the biosynthesized AgNPs revealed that the size and shape of the AgNPs were changed with the plant extract concentration. Biologically synthesized AgNPs from leaf extracts of the three different plants displayed significant differences in antibacterial activity against two different gram-negative and gram-positive bacteria. Also, the results from this study show the shape dependence of the antibacterial and cytotoxic activity of silver nanoparticles synthesized using T. spicata leaf extract. The nanoparticles with different shapes exhibited the strongest antibacterial and cytotoxic activity compared to mostly spherical nanoparticles. Present results clearly indicate that biological activities of silver nanoparticles were affected by nanoparticle shape and the source of the plant extract used in the synthesis.