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Shoot organogenesis and somatic embryogenesis in leaf tissue of Pulsatilla tongkangensis Y.N. Lee & T.C. Lee

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Abstract

Leaf material explants of Pulsatilla tongkangensis Y. N. Lee & T. C. Lee were used to regenerate plants of this endangered species by somatic embryogenesis and organogenesis from meristematic nodules, induced by MS medium supplemented with zeatin (Zn) and indole-3-acetic acid (IAA). Globular structures were induced on the surface of the explants after 2 weeks and after 6–7 weeks of culture, multiple shoots developed from the nodules. Morpho-histological analysis of light green globular, heart-shaped structures resembling somatic embryos revealed, however, that these were organogenic, with strongly vacuolated parenchymatous cells surrounded by a single layer of epithermal cells, and tracheid elements, but no root pole. Milky-white callus also developed around the nodules after 4–6 weeks. Morpho-histological analysis of the globular, heart-, and torpedo-shaped stages of regenerants present in this callus confirmed the development of somatic embryos in the milky-white structures, characterized by deeply staining, small cells with rich cytoplasm, very little vascular tissue in the developing embryos, and no vascular connection with the surrounding callus. The highest rooting frequency (93.33%) was achieved on MS medium containing 1.5 mg/l NAA. Plantlets were acclimatized and successfully transferred to pots. Our results provide a plant regeneration system with potential for germplasm conservation of endangered plants and the rapid propagation and molecular breeding of P. tongkangensis.

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Acknowledgements

The authors wish to thank Prof. Sun-Kuan Hong for providing the plant materials for the experiments.

Funding

This research was funded by Subsidy for enhancement of medical service and security capability (Shan dong Province 2019, 39) and horizontal projects of Linyi University (HX160402).

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Authors and Affiliations

  1. Key Lab of Plant Stress Research, College of Life Sciences, Shandong Normal University, No. 88 Wenhuadong Road, Jinan, 250014, China

    Xiao-mei Zhao, Xue-jie Zhang & Shou-jin Fan

  2. Laboratory of Plant Molecular Genetics and Crop Gene Editing, College of Life Sciences, Linyi University, Shuangling Road, Linyi, 276005, Shandong, China

    Xiao-mei Zhao, Yu-ji Lian & Ze-lin Jin

  3. Jinan Vocational College, No.12 Shungeng Road, Lixia District, Jinan, 250014, Shandong, China

    Yan Y

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  1. Xiao-mei Zhao
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  2. Yu-ji Lian
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  3. Ze-lin Jin
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  4. Xue-jie Zhang
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  5. Yan Y
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Zhao, Xm., Lian, Yj., Jin, Zl. et al. Shoot organogenesis and somatic embryogenesis in leaf tissue of Pulsatilla tongkangensis Y.N. Lee & T.C. Lee. Plant Biotechnol Rep 16, 389–400 (2022). https://doi.org/10.1007/s11816-021-00727-9

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