Background Green synthesis of metallic nanoparticles has gained significant attention in the field of nano-medicine due to environment-friendly and cost-effective alternative in comparison to other physical and chemical methods. Several metals such as silver, gold, iron, titanium, zinc, magnesium, and copper have been subjected to nano-formulation for a wide range of useful applications. Silver nanoparticles (AgNPs) are playing a major role in the field of nano-medicine and nanotechnology. They are widely used in diagnostics, therapeutic and pharmaceutical industries. Studies have shown potential inhibitory antimicrobial, anti-inflammatory and anti-angiogenesis activities of AgNPs. Methods AgNPs have been synthesized using silver nitrate (AgNO3) and methanolic root extract of Rhazya stricta that belongs to the Apocynaceae family. Stability and dispersion of nanoparticles were improved by adding xylitol. Synthesized nanoparticles were characterized by UV-vis spectroscopy, scanning electron microscopy (SEM), Energy dispersive spectroscopy (EDS), X-ray diffractometer (XRD), and Fourier transforms infrared spectroscopy (FTIR). Furthermore, the antibacterial effect of the plant extract and the nanoparticles were evaluated against gram-positive (Bacillus subtilis) and gram-negative (Escherichia coli) bacteria. Results The Average size of AgNPs synthesized, was 20 nm with the spherical shape. Rhazya stricta based nanoparticles exhibited improved antibacterial activity against both gram-positive and negative strains.