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Introduction: one dimensional nanocrystals especially nano-rods have attracted a great deal of attention due to their unique properties and wide applications in industry. Chemical and physical methods which are currently used to produce zinc oxide nano- rods, often leave toxic chemicals on surface of nanoparticles limiting their applications for health and medical purposes. Therefore, biological synthesis of zinc oxide nanoparticles has been considered as an environmentally friendly process and a potential alternative to chemical and physical methods. Materials and methods: Nano-rods of zinc oxide were produced by Xanthomonas campestris using zinc nitrate hexa hydrate as substrate, in a shaker incubator at 37 ° C and pH 7. The powder produced was then calcined at 600 ° C for 2 hours after drying. The synthesized ZnO nanoparticles were characterized using FTIR, XRD, SEM, EDX and UV–vis spectroscopy. Results: FTIR analysis was used to identify functional groups involved in the biosynthesis of ZnO NPs. The peak observed at 563 cm-1 corresponds to the stretching vibrations of ZnO NPs. XRD analysis revealed that the hexagonal ZnO nanoparticles synthesized were pure and crystalline in nature. The morphology and size of the powder were investigated using SEM analysis and the results showed that ZnO nano-rods have a diameter ranging from 122–200 nm with an average length about 300 nm. EDX analysis was performed for determination of the elemental composition and purity of samples. The recorded EDX spectrum revealed the high purity of the synthesized ZnO nano-rods without detection of any impurities. The absorption peak at 376 nm indicating the presence of zinc oxide nanoparticles was further confirmed by UV-Vis spectroscopy. Discussion and conclusion: The current research work describes a low-cost, unreported, nontoxic, simple, safe and eco-friendly method for the biosynthesis of zinc oxide nanoparticle using xanthomonas campestris as the reducing and capping agent.