In this paper, the characteristics of underactuation are analyzed according to the working principle of underwater glider, a complete 6DOF model of underwater glider is established, and the error equation of 3D path tracking is given. Based on the line-of-sight navigation method in three-dimensional space, the nonlinear adaptive control strategy is designed, and the roll angle control law, pitch angle control law, and depth control law of underwater glider are given. The tracking error of underwater glider is proved to be convergent by Lyapunov stability theorem. Finally, the results of marine experiments verify that the nonlinear controller designed in this paper can overcome the influence of constant current disturbance under certain conditions and has good robustness and good tracking effect under different ocean current sizes and course angles. Of course, one control method cannot be applicable to all sea conditions. When the angle between the size and direction of the ocean current exceeds a certain value, the tracking control of the underwater glider will show obvious divergence. Therefore, the stability range against the interference of the sea current is given in this paper.