Ti2AlNb-based alloys, spark plasma sintered from ball-milled octahedral Ti-22Al-25Nb powder, are aged for 0–3 h in this study. Ball milling tunes the morphology of primary orthorhombic (O) phase to be columnar, and improves the mechanical properties of the alloys. The evolution of the Widmanstätten O phase, which brought main precipitation strengthening effect, is then investigated during the ageing process. The hardness of the alloys is improved by an increasing content of O phase and the reduced width of the Widmanstätten O phase; in addition, the length of Widmanstätten O phase also brings positive effect on the hardness, which indicates that the hardness is strongly coupled with the interface of the Widmanstätten O precipitates and B2 matrix. By means of transmission electron microscopy observation, the precipitation of O initials with an amorphous state, followed by a coherent interface between (110)B2 and (002)O, which is destroyed to be semi-coherent and incoherent during the next ageing stage.