Malvaceae and Brassicaceae family crops are economically important; however, their production has been markedly decreased in recent years due to various plant pests. Hence, the search for novel classes of efficient biological approaches continues due to unavailability of precise pesticides. The present study was designed to synthesize, characterize and evaluate the efficacy of silver nanoparticles (AgNPs) obtained using stem extract of Gossypium hirsutum (cotton plant) against plant pathogens Xanthomonas axonopodis pv.malvacearum and Xanthomonas campestris pv. campestris. Biosynthesized AgNPs were characterized using UV-visible spectrophotometry, Dynamic Light Scattering, Scanning Electron Microscopy combined with energydispersive X-ray analysis and Fourier transform infrared spectroscopy. The synthesized AgNPs were spherical in shape with size ranging from 20 to 100 nm.The characterized AgNPs were investigated for their efficacy against bacterial plant pathogens using the paper disc method. In vitro studies with two concentrations of AgNPs (50 and 100 μg mL −1 ) showed zone of inhibition 11.0 ± 1.0 and 12.3 ± 0.5 mm for X. axonopodis pv. malvacearum and 9.7 ± 0.6 and 15.33 ± 1.0 mm for X. campestris pv. campestris. Furthermore, the AgNPs exhibited strong antioxidant activity, and a phytotoxicity study on Vigna unguiculata (cowpea plant) showed no toxicity. Overall, the findings suggest that G. hirsutum stem extract could be efficiently used in the synthesis of AgNPs and showed antimicrobial activity against plant pathogens. Hence, the synthesized nanoparticles could be used to combat plant pathogens in the agriculture sector.