Gradient structure (GS) is commonly designed and processed in engineering materials to improve mechanical properties especially fatigue performance by taking advantage of the strengthened surface. However, whether the very-high-cycle fatigue (VHCF) property can be improved by GS is questioning due to the different crack initiation mechanisms between low-, high-cycle and VHCF. In this paper, GS of a Ti-6Al-4V alloy is generated by pre-torsion and characterized by electron backscatter diffraction. Then the VHCF behavior of the GS specimen is studied. The fractography and synchrotron radiation X-ray microtomography presented detailed characteristics of the internal crack initiation region in VHCF of the titanium alloy with GS. The results indicated that, in contrast to the low-and high-cycle regimes, the VHCF strength is reduced for the specimens with GS. Thus, the GS induced by pre-torsion cannot enhance the VHCF strength of the titanium alloy. This implies that VHCF test (property) is an important consideration for the microstructural designed materials. The graphical abstract is available in Supplementary information.Fatigue damage causes the failure of more than 90% engineering materials and structures. According to the traditional knowledge, fatigue crack initiates from the surface of material 1 . Therefore, many surface strengthening methods, such as surface mechanical attrition or grinding treatment, surface quenching, shot peening, etc., have been widely used as counter strategies to improve the fatigue resistance of the components by introducing the refined microstructure with gradient distribution of grain size and residual compressive stress 2,3 . It is known that crack is hard to initiate from small grains, and the residual compressive stress may enhance the fatigue strength 1,2 . In recent years, pre-torsion method was used to produce gradient structure (GS) in metallic materials owing to its low cost and easy processing 4-7 . During the pre-torsion process, grain size can be substantially refined due to severe plastic deformation (SPD) 8 , and the GS is generated by the gradient strain. A number of experimental results 9,10 confirmed that the layer of GS improves fatigue property in low-cycle-fatigue (LCF) and high-cycle-fatigue (HCF) regimes, i.e. for the cases that the number of fatigue failure cycles is lower than 10 7 . However, with the development of modern civilization, there is practical demand of engineering structures for more than 10 7 loading cycles of safe performance. Thus, very-high-cycle fatigue (VHCF) has become an important topic, and has received increasing attention [11][12][13] . However, VHCF test is very time consuming, e.g. it takes 4 days to reach 10 7 cycles and more than 1 year for 10 9 cycles with conventional frequency of 30 Hz via a servohydraulic system. This limits the VHCF research and hence the VHCF behavior is still rarely studied for metallic materials with a designed microstructure, such as GS, heterogeneous, hierarchical, etc. Thus, whether the VHCF fatigue propert...