The application of green methods in the synthesis of nanoparticles using plants is a cost-effective and eco-friendly approach. Zinc oxide nanoparticles are of great importance due to their versatile properties. The conditions of synthesis strongly influence the characteristics and functionality of the nanoparticles. The present work studied the biological, green synthesis of zinc oxide nanoparticles (ZnONPs) in the presence of different concentrations of ethanolic extract of Larrea tridentata (10, 20, and 30 mg/mL). The time of the formation of nanoparticles was evaluated at different temperatures and pH values of the reaction medium. The formation of ZnONPs was confirmed by ultraviolet-visible (UV-Vis) and Fourier transform infrared spectroscopies (FT-IR), as well as scanning electron microscopy (SEM). X-ray diffraction analysis (XDR) determined the crystallographic structure of the nanoparticles. Obtained ZnONPs had a size range of 18 to 40 nm. The temperature, reaction time, and pH significantly influenced the nanoparticles’ morphology, size, and aggregation. The impact of chosen ZnONPs was tested on the germination of serrano chili seeds (Capsicum annuum). At 100 ppm, the nanoparticles improved germination percentage, vigor, and seedlings’ growth parameters.