Background: Nanoparticles, due to their small size and large surface area, have unique physicochemical properties that make them attractive for a wide range of industrial, medical, agricultural application. Objective: To prepare a novel nanoparticles from olive leaf extract and copper oxide and test its physiological effect of strawberry plants. Materials and Methods: Olive leaves extract was prepared according to a certain protocol, and the green nanoparticles were synthesized by coordination this extract with copper oxide. Characterization was done using many techniques such as FTIR, UV-Vis, XRD to identify the new copper nanoparticles. Different concentrations were prepared of CuONPs (10%, 20%, and 30%). Results: The FTIR spectrum shows peaks which confirm the coordination of olive leaves extract with the copper ion, while UV-Vis spectrum shows peaksthat confirm the formation of copper nanoparticles, the SEM shows the homogeneity of the nanoparticle that was confirmed by XRD spectrum and dataUsing30% (C3) give best seedling leaf length, and number of fruitswhile number of leaves are best in 20% (C2) compared to other concentrations. While the highest value in the number of flowers in treatmentof bio fertilizer. Best concentration of DNA appears in C3 (134.7) while best DNA purity is found on C2 (20%). The chlorophyll was also measured using SPADwhich show optimum with C3 (30%). Conclusions: Using30% (C3) give best seedling leaf length, number of fruits and chlorophyll while number of leaves are best in 20% (C2).