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Halogeton glomeratus (H. glomeratus) is one of the most important halophytes in Asia, and the research on the genes and salt-tolerant mechanisms of this species is limited because of the lack of an optimal and efficient in vitro regeneration system. Here, we developed an efficient plant regeneration protocol using H. glomeratus leaves as explants, which has not been previously reported. High-quality calli were successfully obtained from the H. glomeratus leaves at a frequency of 100% supplemented with 2.00 mg/L 2,4-dichlorophenoxyacetic acid, 0.50 g/L PVP, 5.00 g/L agar, and 30.00 g/L sucrose; the optimum callus subculture medium is 1.00 mg/L 2,4-dichlorophenoxyacetic acid, 0.50 g/L PVP, 5.00 g/L agar, and 30.00 g/L sucrose. Shoots were regenerated on shoot induction medium at a frequency of 100% added with 0.50 mg/L 6-benzylaminopurine, 2.00 mg/L kinetin, 0.20 mg/L naphthaleneacetic acid, 0.50 g/L PVP, 5.00 g/L agar, and 30.00 g/L sucrose; then, shoots grew up on shoot regeneration medium. Finally, roots were regenerated from the shoots on Murashige and Skoog medium with high efficiency. This study firstly offers a rapid and efficient system for plantlet regeneration from leaves of H. glomeratus. Whether our protocol applies well to other tissue regeneration of H. glomeratus needs further confirmation. This work will facilitate basic research and salt-tolerant mechanisms of this important halophyte species. [ABSTRACT FROM AUTHOR]