Abstract
A fast and efficient genetic transformation pathway is needed for sweet potato transgenics and acclimatization of regenerated plants also needs to be studied. In this experiment, purple sweet potato leaves were used as explants. By adding different hormones at different stages, callus was induced first and then adventitious roots and buds were induced. An acclimatization method with a high survival rate was also found. A new efficient in vitro plant regeneration system characterized by rapid sequential organogenesis using leaf explants has been developed in sweet potato. Optimal plant regeneration was obtained in the variety Ning Purple Potato No. 1 with a four-step protocol. The first stage was callus induction of leaf explants for 40 days on Murashige and Skoog medium containing 0.8 mg/L 2,4-D. The second stage was adventitious root induction from callus on solid MS medium with different combinations of 6-BA and NAA; 0.6 mg/L 6-BA and 0.3 mg/L NAA was the best combination to induce adventitious roots. The third stage was adventitious bud induction from adventitious roots; solid MS medium with 2.0 mg/L ZT, 1.0 mg/L KIN and 1.0 mg/L GA3 produced the maximum of number of adventitious buds after 12 weeks. The fourth stage was acclimatization to hydroponics. The best acclimatization program was floating culture in Hoagland nutrient salts solution for 7 days. We have developed an efficient protocol to generate plants and the regenerated plants survived acclimatization and transplantation. As the method is simple, rapid and reproducible, it may be valuable for transgenic genetic improvement.
Key message
The present study developed a protocol for bud induction from adventitious roots of purple sweet potato and a new method for acclimatizing regenerated seedlings by hydroponics.
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Abbreviations
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- 6-BA:
-
6-benzyladenine
- GA3 :
-
Gibberellic acid
- KIN:
-
Kinetin
- MS:
-
Murashige & Skoog
- NAA:
-
1-Napthyl Acetic Acid
- ZT:
-
Zeatin
- mg/L:
-
Milligrams/Liter
- g/L:
-
Grams/Liter
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Acknowledgements
The authors gratefully acknowledge the management of Henan Institute of Science and Technology, China for laboratory facilities. The authors also acknowledge the financial support of the Science and Technology Project of Henan Provincial Department of Science and Technology (Important gene regulation mechanism and new material creation of anthocyanin synthesis in sweet potato, Item no.222102110282). We thank Robbie Lewis, MSc, from Liwen Bianji (Edanz) (www.liwenbianji.cn/) for editing a draft of this manuscript.
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Genhai Hu designed the experiment. Genhai Hu and Xiaohong Zhang collected the experimental materials and performed callus induction. Genhai Hu and Maoni Chao performed induction of adventitious roots and shoots. Yuanzhi Fu conducted the acclimatization process. All authors wrote the manuscript. Genhai Hu monitored the research. All authors read and approved the manuscript.
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Communicated by Yan Liu.
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Hu, G., Chao, M., Fu, Y. et al. In vitro protocol for bud induction from adventitious roots and hydroponic acclimatization of purple sweet potato (Ipomoea batatas (L.) Lam). Plant Cell Tiss Organ Cult 151, 207–213 (2022). https://doi.org/10.1007/s11240-022-02335-z
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DOI: https://doi.org/10.1007/s11240-022-02335-z