The plastic deformation processes and fracture behavior of a Ti–5Al–5Mo–5V–1Cr–1Fe alloy with bimodal and lamellar microstructures were studied by room-temperature tensile tests with in situ scanning electron microscopy (SEM) observations. The results indicate that a bimodal microstructure has a lower strength but higher ductility than a lamellar microstructure. For the bimodal microstructure, parallel, deep slip bands (SBs) are first noticed in the primary α (αp) phase lying at an angle of about 45° to the direction of the applied tension, while they are first observed in the coarse lath α (αL) phase or its interface at grain boundaries (GBs) for the lamellar microstructure. The β matrix undergoes larger plastic deformation than the αL phase in the bimodal microstructure before fracture. Microcracks are prone to nucleate at the αp/β interface and interconnect, finally causing the fracture of the bimodal microstructure. The plastic deformation is mainly restricted to within the coarse αL phase at GBs, which promotes the formation of microcracks and the intergranular fracture of the lamellar microstructure.