We have synthesized ZnO nanorods by hydrothermal at 95 °C for two hours on different substrates i.e. glass, ZnO, and ITO. Morphologically, the average diameter of ZnO nanorods on glass, ZnO, ITO are 117.6 µm, 133 µm, 98.4 µm, respectively. According to the international center for diffraction data (ICDD) number #01-079-0207, the ZnO nanorods on all substrates possess a polycrystalline wurtzite structure. By full width at half maximum (FWHM) of ZnO NRs in (002) plane, we have determined crystallite size and micro-strain using Scherer’s equation and Stokes-Wilson equation, respectively. The crystallite size and micro-strain of ZnO NRs vary with different kind of substrates. Optically, ZnO nanorods on a glass substrate have the strongest excitonic-related ultraviolet (UV) emission. Further observation, we have determined the UV/GB ratio to confirm the crystalline quality of ZnO NRs on glass, ZnO, and ITO substrates. These UV/GB ratios are 101.35, 68.14, and 22.74. It is indicating the ZnO NRs on glass has a better crystalline quality which indicates fewer native defects concentration compared to the others.