In this present work, we have successfully synthesized vertical aligned ZnO nanorod arrays (ZnO NRs) on conductive stainless-steel substrate. These ZnO NRs were obtained by using a low temperature, low cost, and eco-friendly hydrothermal approach. The effect of the sol-gel deposited ZnO seed layer on the vertical alignment ZnO NRs has been thoroughly studied. Characterizations of the seed layer and the as-obtained nanorods were investigated using scanning electron microscopy (SEM), X-ray diffraction (XRD), atomic force microscopy (AFM), high-resolution transmission electron microscopy (HR-TEM), and water contact angles (WCA) measurement. SEM analysis shows that these nanorods were grown with a high uniformity and density over the entire substrate. TEM and XRD studies confirmed the hexagonal wurtzite structure of ZnO nanorods. Finally, field immersion tests using the optimized samples were conducted in a coastal sea environment to study biofouling settlement. To the best of our knowledge, there are few articles that investigate the influence of the seed layer deposited on stainless steel on the formation of vertically aligned nanorods for antifouling applications. The novelty of this work is manifested by optimizing the ZnO seed layer to obtain either thick or fine ZnO nanorods (NRs). Moreover, another novelty of this study is the use of the hydrothermal deposition of ZnO NRs on stainless steel for biofouling to replace paints in marine applications.