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Hot compression tests were performed with strain rates (0.01–10 s−1) and temperatures (850–1150 °C). The power law relationship between the critical stress and critical strain and Zener–Hollomon parameters was determined by θ-σ curves. Microstructure was investigated by electron backscattered diffraction. The results showed that the flow behavior and microstructure of 35CrMo steel was affected by ultrasonic-assisted casting. The activation energy of non-ultrasonic and ultrasonic-assisted 35CrMo steel were 410 ± 9.9 and 386 ± 9.4 kJ/mol, respectively, and the activation energy of ultrasonic-assisted specimens was reduced by 6%. In addition, the ultrasonic-assisted treatment refines the grains to some extent and makes the softening process of ultrasonic-assisted samples progress faster, which promoted the development of dynamic recrystallization and the production of Σ3 boundaries. The discontinuous dynamic recrystallization was the main DRX nucleation mechanism of the 35CrMo steel. [ABSTRACT FROM AUTHOR]