Due to the complicated working environment in the field, the GPS data cannot accurately reflect the position of the seedlings during the automatic navigation of the rice transplanter, and may not meet the corresponding operational requirements. Therefore, in order to verify the accuracy of the satellite navigation operation of the rice transplanter, a method of using the total station to test the seedling path was proposed to verify the accuracy of the transplanter. By studying the data of the seedling point obtained by the singlepoint static measurement of the total station, the straightness of the seedling row and the parallel accuracy of the seedling rows were obtained, and the operation precision of the transplanter was evaluated. From the analysis of relevant data, the minimum and maximum values of the straightness of the seedling row were 3.08cm and 4.93cm, respectively, and the minimum and maximum root mean square error were 4.299cm and 6.263cm, respectively; the minimum and maximum values of parallel accuracy of seedling rows were 5.17cm and 15.53cm, respectively, and the minimum and maximum root mean square errors were 4.29cm and 5.43cm, respectively. The results showed that the method can achieve quantitative evaluation of the accuracy of automatic navigation operations in agricultural machinery fields.