One-dimensional electrospun nanofibers of p-type NiO/n-type ZnO heterojunctions with different molar ratios of Ni to Zn were successfully synthesized using a facile electrospinning technique. X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) spectroscopy, transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), UV-vis diffuse reflectance (DR) spectroscopy, resonant Raman spectroscopy, photoluminescence (PL) spectroscopy, and surface photovoltage spectroscopy (SPS) were used to characterize the as-synthesized nanofibers. The results indicated that the p-n heterojunctions formed between the cubic structure NiO and hexangular structure ZnO in the NiO/ZnO nanofibers. Furthermore, the photocatalytic activity of the as-electrospun NiO/ZnO nanofibers for the degradation of rhodamine B (RB) was much higher than that of electrospun NiO and ZnO nanofibers, which could be ascribed to the formation of p-n heterojunctions in the NiO/ZnO nanofibers. In particular, the p-type NiO/n-type ZnO heterojunction nanofibers with the original Ni/Zn molar ratio of 1 exhibited the best catalytic activity, which might be attributed to their high separation efficiency of photogenerated electrons and holes. Notably, the electrospun nanofibers of p-type NiO/n-type ZnO heterojunctions could be easily recycled without a decrease of the photocatalytic activity due to their one-dimensional nanostructural property.