Abstract
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.
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Funding
This work was supported by the Open Funds of Laboratory of Gansu Agricultural University (Grant GSCS-2019-07), National Natural Science Foundation of China (Grant 31960072, 32001514), Innovation Capacity Enhancement Project of Gansu Education Department (Grant 2019A-053), China Agriculture Research System (Grant CARS-05-04B-2), Key Projects of Natural Science Foundation of Gansu Province (20JR10RA507), National Basic Research Program of China (973 program, 2014CB160313), National University Innovation and Entrepreneurship Training Program (202010733004, 202010733003), and Scientific Research Start-Up Funds for Openly-Recruited Doctors of Gansu Agricultural University (Grant GAU-KYQD-2018-22, 02).
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HW and XS designed the experiments. LY, JW, BL, YM, ES, and KY performed the experiments. LY, JW, XM, YL, and PR observed the morphologic changes of explants and analyzed the data. LY wrote the manuscript. All authors read and approved the final manuscript.
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Editor: Yong Eui Choi
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Yao, L., Wang, J., Yang, K. et al. In vitro regeneration system of Halogeton glomeratus: an important halophyte. In Vitro Cell.Dev.Biol.-Plant 57, 332–340 (2021). https://doi.org/10.1007/s11627-021-10169-1
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DOI: https://doi.org/10.1007/s11627-021-10169-1