The dynamic performance of radio frequency identification (RFID) under transportation line based on photoelectric technology is investigated. By the laser ranging technology, a testing system is designed to measure the dynamic reading distance of RFID. The reading distance of single-tag or multi-tag system can be measured indirectly, and the automatic measurement to the reading distance and literacy rate of RFID tags is realized. The single-tag and the multi-tag systems are respectively tested for their dynamic performances and the testing results are compared and analyzed. The results show that, in the process of testing the single-tag system, the dynamic reading performance of RFID tags is directly influenced by the material quality of conveyor belts. In contrast, in the process of testing the multi-tag system, the dynamic reading range of RFID tags decreases with the increase of the number of tags, and the collision tag can be detected accurately.