Background: Nanotechnology, particularly the use of nanoparticles (NPs), is rapidly evolving and has the potential to transform various scientific and technological sectors. Aim: Prepration of the green synthesis and characterization of silver nanoparticles using an extract of the plant Codonopsis pilosula, and the evaluation of its antimicrobial agents. Methodology: This paper used a green synthesis approach to synthesize silver nanoparticles (AgNPs) using an aqueous extract of Codonopsis pilosula roots. It was done by taking the plant extract and mixing it with 2 mM AgNO3 solution and letting the mixture stay at 30 o C for four hours. A change of color proved the formation of AgNPs and was characterized by Scanning Electron Microscopy (SEM), X-ray Diffraction (XRD), and UV-Vis spectroscopy. The FTIR analysis of the plant extract identified important functional groups used in the plant. Results: Successful preparation of spherical sized AgNPs with SEM indicating size ranges of 33.74-81.63 nm. XRD patterns were used to confirm the presence of crystalline silver compounds and a characteristic absorbance peak of 340 nm was exhibited by the UV-Vis spectroscopy. More importantly, these biosynthesized AgNPs demonstrated high levels of antibacterial activity against different pathogenic bacteria such as Acinetobacter baumanii, Escherichia coli, and most importantly, Staphylococcus aureus, by disrupting cell membranes and reactive oxygen species generation. Conclusion: This study demonstrates a greener, more affordable and biocompatible method of making metallic nanoparticles, providing a greener alternative to traditional chemical techniques of making more complex biomedical and pharmaceutical applications.