Potato microtuber induction is the basis of original-original seeds with virus-free production. In this study, the potato cultivar Atlantic and Longshu 3 was selected to form tubers in vitro by solid cultivation of strong seedlings and liquid cultivation induction. Single or matching combinations of CCC, NAA and 6-BA were added in the induction process. The results showed that the use of 10 mg/L hormone CCC or the combination of 0.4 mg/L NAA, 0.5 mg/L 6-BA and 9 mg/L CCC could effectively increase the percentage of large tubers, average weight of large tubers and final yield per plant, and shorten the tuber setting period. The study explored the culture method of mircrotuber induction in vitro, which provided theoretical basis for the development and application of microtuber with virus-free.