The roles of microstructure in plastic deformation and crack growth mechanisms of a titanium alloy with a trimodal microstructure have been systematically investigated. The results show that thick intragranular α lath and a small number of equiaxed α phases avoid the nucleation of cracks at the grain boundary, resulting in branching and fluctuation of cracks. Based on electron back-scattered diffraction, the strain partition and plastic deformation ahead of the crack tip were observed and analyzed in detail. Due to the toughening effect of the softer equiaxed α phase at the grain boundary, crack arresting and blunting are prevalent, improving the crack growth resistance and generating a relatively superior fracture toughness performance. These results indicate that a small amount of large globular α phases is beneficial to increase the crack propagation resistance and, thus, a good combination of mechanical property is obtained in the trimodal microstructure.