The Ti-6Al-4V alloy is widely used in the manufacture of components that must have low density and high corrosion resistance and fatigue strength. The fatigue strength can be improved by surface modification. The aim of this study was to determine the influence of plasma nitriding on the fatigue behavior of a Ti-6Al-4V alloy with a lamellar microstructure (Widmanstätten type). Nitriding was executed at 720 °C for 4 h in an atmosphere with N₂, Ar, and H₂. Microstructure characterization of the samples was carried out by X-ray diffraction analysis, optical microscopy, and scanning electron microscopy. The average roughness of the specimens was determined, and fatigue tests were executed in a bending⁻rotating machine with reverse tension cycles (R = -1). X-ray diffraction analysis of the nitrided alloy revealed the following matrix phases: α, β, ε-Ti₂N, and δ-TiN. A nitrogen diffusion layer was formed between the substrate and the titanium nitrides. Plasma nitriding resulted in an increase in low-cycle fatigue strength, whereas at high cycles of 200 MPa, both conditions exhibited similar behaviors. The fracture surface of the fatigue-tested specimens clearly revealed the lamellar microstructure. The fracture mechanism in the non-nitrided specimens appears to be due to cracking at the interface of the α and β phases of the lamellar microstructure.