Unbalanced vibrations of axial threshing cylinders on a combine harvester were coupled to each other through a frame. The intensified unbalanced vibration will shorten the working life of the axial threshing cylinder. In this paper, the theoretical modes of the axial threshing cylinder were carried out by using finite element analysis software ANSYS. The axis trajectory and speed fluctuation of the axial threshing cylinder under no-load state and threshing state were tested and analyzed. According to the amplitude and axis trajectory of the transmission shaft, as well as the variation law of the axial threshing cylinder speed, the influence of rice straw on the vibration under the threshing state was revealed. The rotation speeds of cylinder I and cylinder III were adjusted, and the amplitude of cylinder II transmission shaft was analyzed when cylinder II was under stable condition. Then the disturbance and coupling relationships among the unbalanced vibration of axial threshing cylinders were compared. Test results showed that the rotational frequency of the axial threshing cylinder was not in its resonance interval at rated speed. When the axial threshing cylinder was threshing, the horizontal amplitude increased by 0.366 mm. The vertical amplitude increased by 0.697 mm. The speed decreased from 763 rpm to about 750 rpm. The rotational frequency of the axial threshing cylinder would not cause the resonance. With the feeding of rice, the amplitude of the axial threshing cylinder increased slightly and the operating speed was lower than the rated speed. The unbalanced vibration of the axial threshing cylinder transmitted along the frame and coupled with each other, causing the vibration of the axial threshing cylinder to be intensified.