This study reports a protocol for leaf callus induction and suspension culture establishment in lychee cv. Huaizhi. The results showed that 12-day-old leaf explants cultured under a photoperiod of 16/8 h with their adaxial side touching the medium were the optimum conditions for leaf callus induction. Globular embryos were formed when the induced calli were kept on the callus induction medium without 2,4-D for 24 weeks. Friable calli were induced after 2–3 subcultures at 4 weeks intervals on the Murashige and Skoog medium supplemented with 3 mg/L IAA and 2 mg/L BAP. Suspension culture was established when these friable calli were subcultured six times in liquid callus induction medium.