[Purposes] This work aims to improve the deficient high-temperature performance and applicability of bio-oil, a new environmental material for pavement and determine the performance differences of polyphosphoric acid (PPA) and bio-oil composite modified asphalt, which bio-oil from different sources.[Methods] The bio-oils, corn straw oil (CSO), kitchen waste oil (KWO) and vegetable bio-oil (VBO) were used to prepare PPA modified bio-asphalt. The conventional properties, rheological behavior and SEM tests were conducted to study its high and low-temperature behavior and microstructure.[Findings] The results showed that PPA has a positive effect on the high-temperature rutting resistance of asphalt, and the three kinds of bio-oils improved the low-temperature cracking resistance. The type of bio-oil influenced the temperature sensitivity, high and low-temperature performance of asphalt. The high-temperature performance of the three PPA-modified biological bitumen was in order from high to low: PPA/KWO modified bitumen, PPA/CSO modified bitumen, PPA/VBO modified bitumen; The low-temperature rheological properties of PPA/VBO modified asphalt, PPA/CSO modified asphalt and PPA/KWO modified asphalt were in descending order.[Conclusions] The combination of PPA and bio-oil can significantly improved the high and low-temperature performance of asphalt. This study could provide a reference for applying and promoting bio-oil and make road engineering more environmentally friendly.