Three layer-by-layer (LBL) assembled gold nanoparticles (AuNPs)/lower-generation (Gn <= 3) polyamidoamine dendrimer (PD) with reduced graphene oxide (rGO) as the core/mercaptopropinoic acid (MPA)/Au were successfully fabricated and employed as electrochemical gene nanobiosensing platforms with three-dimensional (3D) fractal nanoarchitecture for fast, ultra-trace determination of label-free DNA hybridization. Three Gn <= 3PD were initially grafted to graphite oxide (GO) via the covalent functionalization between amino terminals of PD and carboxyl terminals of GO where a concomitant reduction of GO, which were covalently linked onto MPA that was self-assembled onto Au substrate, and finally AuNPs were encapsulated onto GG1PD by strong physicochemical interaction between AuNPs and-OH of rGO in GG1PD, Their morphologies, structures, electrochemical properties, and gene nanobiosensing performances were characterized and evaluated. AuNPs/GG2PD-based probe displayed the best excellent structural stability, lowest mobility on solid surface with the increasing charge resistance, widest linear range (1.1 x 10(-6) - 1 x 10(-18)), and the lowest limit of detection (1.87 x 10(-19) M) in comparison with both AuNPs/GG1PD-based and AuNPs/GG3PD-based probes. This work will provide a new candidate for the development of metal nanoparticles functionalized PD with inorganic nonmetallic nanomaterials as cores with 3D fractal nanoarchitecture and promising electrochemical gene nanobiosensing platforms based on dendrimer-nanoinorganic hybrids with 3D nanoarchitectures and LBL assembly for fast and ultra-trace detection of label-free DNA hybridization with potential application in bioanalysis and medical diagnosis of genetic diseases.