Abstract
In plant biotechnology, protoplasts are a versatile tool since they are very helpful for both fundamental biology studies and for genetic improvement and genome editing studies. In many plant species, however, reproducible regeneration from protoplasts continues to be a bottleneck. In the present study, we report the development of an efficient method for protoplast isolation, and plant regeneration in Angelica gigas via indirect somatic embryogenesis. Protoplasts were isolated from embryogenic callus using an enzyme mixture of 1.0% Viscozyme® L + 1% Celluclast® 1.5 L + 0.5% Pectinex® XXL with 7 h treatments. Initially, protoplasts were cultured in MS, modified MS (NH4NO3-free medium), and KM media, and viability and cell division data showed the MS medium was suitable for protoplast culture. Subsequently, the thin alginate layer method was applied to the protoplast culture at an optimal density of 1 × 106 cells per mL− 1 and verified the effect of 2,4-D (0.1, 0.5, and 1.0 mg L− 1) alone, and 2,4-D (0.5, and 1.0 mg L− 1) in combination with BA (0.1 and 0.5 mg L− 1) or KN (0.1 and 0.5 mg L− 1) on cell division, micro-callus formation. MS medium supplemented with 1.0 mg L− 1 2,4-D and 0.1 mg L− 1 KN induced optimal cell division, callus formation, and subsequent induction of somatic embryogenesis from the callus. The somatic embryos germinated and converted into plantlets upon transferring to the MS basal medium. This method of Angelica gigas protoplast regeneration can be used for the genetic improvement of this plant.
Key message
The primary goal of the current study was to develop plant regeneration in Angelica gigas using protoplast culture, which could be valuable for the genetic improvement of this significant medicinal plant.
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Acknowledgements
H.S.L and J.E.H. were supported by the Brain Korea (BK) 21 Plus Program. HNM was supported by the Brain Pool Program of the National Research Foundation of Korea (Grant No. 2022H1D3A2A02056665).
Funding
The work was supported by grants from the National Research Foundation of Korea (No. NRF-2020R1A2C2102401) and the Korea Research Institute of Bioscience and Biotechnology (KGM-5282331).
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H.S.L and J.E.H contributed to experiments and data acquisition. E.Y.J., S.W.K., H.J.K, G.M.L., and H.S.L. in procuring material, interpretation of data, and intellectual content. S.Y.P. contributed to the conceptualization and design of the study. H.N.M. helped in the interpretation of data and writing of the manuscript. All the authors have read and approved the final version of the manuscript.
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Communicated by Francisco de Assis Alves Mourão Filho.
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Lee, HS., Han, JE., Jie, E.Y. et al. Isolation, culture of protoplasts of Angelica gigas Nakai and regeneration of plants via somatic embryogenesis. Plant Cell Tiss Organ Cult 156, 40 (2024). https://doi.org/10.1007/s11240-023-02666-5
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DOI: https://doi.org/10.1007/s11240-023-02666-5