<p>Biomass-derived filler (bio-filler) shows potential to replace mineral filler against the backdrop of a growing shortage of mineral resources, offering innovative solutions for the sustainable development of road infrastructure. To advance the research and application of bio-filler, this review first outlines the concept, preparation methods, and properties of bio-fillers, followed by a discussion of the interaction between asphalt and bio-filler, while evaluating their economic viability and environmental benefits. Finally, the limitations of bio-filler and future research needs are discussed and pointed out. Relevant studies indicate that the interaction between bio-filler and asphalt is a physical process. The substitution of mineral filler with bio-filler enhances the high temperature performance of mastic and mixture and shows promise in terms of economic and environmental benefits, but the studies on the medium- and low-temperature performances and the water stability of mixture as well as the correlations still need to be further investigated. Future research should begin with preparing bio-fillers and exploring each stage of their application, with the aim of achieving comprehensive breakthroughs in substitution ratio, performance, economic, and environmental benefits. This review highlights the application potential of bio-fillers and provides novel insights into the advancement of asphalt road-use biotechnology.</p> Graphical abstract <p></p>