A new approach has been developed to improve sensing performances of electrochemically grown Au nanostructures (AuNSs) based on the pre-seeding of the electrode. The pre-seeding modification is simply carried out by vacuum thermal deposition of 5 nm thin film of Au on the substrate followed by thermal annealing at 500 °C. The electrochemical growth of AuNSs on the pre-seeded substrates leads to impressive electrochemical responses of the electrode owing to the seeding modification. The dependence of the morphology and the electrochemical properties of the AunSs on various deposition potentials and times have been investigated. for the positive potentials, the pre-seeding leads to the growth of porous and hole-possess networks of AunSs on the surface. for the negative potentials, AunSs with carved stone ball shapes are produced. the superior electrode was achieved from AunSs developed at 0.1 V for 900 s with pre-seeding modification. The sensing properties of the superior electrode toward glucose detection show a high sensitivity of 184.9 µA mM −1 cm −2 , with a remarkable detection limit of 0.32 µM and a wide range of linearity. the excellent selectivity and reproducibility of the sensors propose the current approach as a large-scale production route for non-enzymatic glucose detection. During past decades, electrochemical sensing has attracted many attentions due to their considerable advantages such as high sensitivity, low cost, fast response time, wide linear range and portability and widespread applications 1-9. Up to now various types of nanomaterial such as carbon nanotubes 10 , graphene 11,12 , noble metals 13,14 , metal oxides 15 , hybrid 16 and composite nanostructures 17,18 have been developed for electrochemical sensing. Among different nanomaterials, gold nanostructures (AuNSs) have attracted significant interests because of their remarkable electrical and catalytic properties, easy functionalization and excellent biocompatibility. Since the electron transfer and catalytic activity of AuNSs strongly depend on their morphology and their size, extensive researches have been proposed in literatures to develop optimized AuNSs for sensing applications 19,20. The notable approaches between them are based on the electrodeposition 19 , direct electrostatic self-assembly 21 , self-assembly with polymer 22-24 and seed-mediated growth 25-28. In the cases of self-assembly and seed-mediated strategies, the electron transfer and catalytic activity of AuNSs are hindered due to the existence of surfactant and linker molecules surrounding the nanostructures. The electrodeposition is the most reliable approach for shape and size controlled construction of AuNSs due to the possibility of controlling the nucleation and the growth processes. However, in this method the well control of morphology is provided by using template, surfactant or pre-treatment of the substrate that limit the applicability of the method 20,29,30 .