In this study, different seed layers like gold (Au), zinc oxide (ZnO) and aluminum-doped ZnO (AZO) have been associated to ZnO nanowires (NWs) for the development of mechanical energy harvesters. ZnO NWs were grown by using a low temperature hydrothermal method. The morphological properties were investigated using Scanning Electron Microscopy (SEM) and the analysis of crystalline quality and growth orientation was studied using X-ray Diffraction (XRD). The obtained ZnO NWs are found to be highly dense, uniformly distributed and vertically well aligned on the ZnO and AZO seed layers, while ZnO NWs grown on Au possess a low density and follow a non-uniform distribution. Moreover, the NWs exhibited good crystal quality over the seed layers. The piezoelectric nanogenerator (PENG) consists of ZnO NWs grown on the three different seed layers, parylene-C matrix, Ti/Al top electrode and poly(dimethylsiloxane) (PDMS) encapsulated polymer composite. The measurements of the open circuit voltage (VOC) were around 272 mV, 36 mV for ZnO, AZO seed layers while the PENG including Au seed layer presented a short-circuited state. This study is an important step in order to investigate the effect of different seed layers influencing the magnitude of the generated electrical performances under identical growth and measurement conditions. It will also help identify the most suitable seed layers for energy harvesting devices and their future integration in industrial applications.