Vertically aligned, single crystalline ZnO nanorods with a high packing density and diameter of B60 nm have been successfully synthesized via a low-temperature hydrothermal route on glass substrates pre-deposited with a ZnO seeding layer. The seeding layer exhibits an epitaxial effect on the growth and alignment of the ZnO nanorods. This epitaxial effect can arise from two considerations, namely the crystalline orientation and surface roughness of the seeding layer, which can be controlled by the curing temperature. The ZnO seeding layer that was cured at 3501C exhibited a preferred (0002) crystalline orientation of wurtzite hexagonal structure and a low surface roughness. It was demonstrated to promote the vertical growth of ZnO nanorods. The ZnO nanorods grew in an almost linear relationship with hydrothermal time up to 8 h, but thereafter started to dissolve as the reaction time extended beyond 8 h, due to competition from the homogeneous nucleation of ZnO microparticles in the solution.