We reported a new approach for development of lignin bio-oil-based electmspun nanofibers (LENFs) that had high substitution ratio (up to 80 wt%) and good morphology. This approach was particularly unique and translatable as it used small molecule lignin bio-oil with high reactivity and low heterogeneity obtained via lignin depolymerization reaction to produce well-oriented LENFs. Firstly, effects of various blends solutions ratios and electrospinning parameters on the characteristics of the obtained LENFs were analyzed. The results showed that the optimal parameters that resulted in the best electrospun nanofibers were as follows: blend solution ratio, the 20 wt% blend solution containing 80 wt% straw lignin bio-oil (SLB) and 20 wt% polyacrylonitrile (PAN), flow rate, 1 mL/h, voltage, 20 kV, rotational speed, 500 r/min and the distance between needle and collection screen, 20 cm. Secondly, used the best LENFs, we also applied to prepare lignin bio-oil-based carbon nanofibers (LCNFs) and estimated its properties by scanning electron microscope (SEM), X-ray diffraction (XRD) patterns, Raman spectroscopy and tension testing. Our results demonstrated that compared with pure PAN carbon nanofibers (PCNFs), the as-prepared LCNFs had similar smooth surfaces, similar crystallinity and similar mechanical properties. This work can promote the utilization of lignin depolymerization main-products to produce lignin-based materials, while also help to reduce use of high-cost PAN.