Abstract. Alpha phase exhibits equiaxed or lamellar morphologies with size from submicron to microns in an alpha-beta titanium alloy. Cyclic deformation, slip characteristics and crack nucleation during fatigue in different microstructures of TC21 alloy (Ti-6Al-2Sn-2Zr-3Mo-1Cr-2Nb-0.1Si) were systematically investigated and analyzed. During low-cycle fatigue, equiaxed microstructure (EM) in TC21 alloy exhibits higher strength, ductility and longer low-cycle fatigue life than those of the lamellar microstructure (LM). There are more voids in the single lamellar alpha than the equiaxed alpha grains. As a result, voids more easily link up to form crack in the lamellar alpha phase than the equiaxed alpha phase. However, during high-cycle fatigue, the fine lamellar microstructure (FLM) shows higher fatigue limit than bimodal microstructure (BM). The localized plastic deformation can be induced during high-cycle fatigue. The slip bands or twins are observed in the equiaxed and lamellar alpha phases( >1micron), which tends to form strain concentration and initiate fatigue crack. The localized slip within nanoscale alpha plates is seldom observed and extrusion/intrusion dispersedly distributed on the sample surface in FLM. This indicates that FLM show super resistance to fatigue crack which bring about higher fatigue limit than BM.