Abstract
The haploidy technique is a useful tool for quickly producing pure, fully homozygous lines. Artemisia annua L. is a medicinal plant that produces artemisinin, a widely used antimalarial drug. Because of its extremely small flowers (≤ 3.0 mm), this study conducted microscopic observations to determine the types of flowers of A. annua suitable for microspore embryogenesis, as well as their corresponding microspore development stages, and obtained embryoids (non-zygotic embryos) from isolated microspore cultures. The media for inducing embryoid production were based on Nitsch and Nitsch medium containing 13% or 17% sucrose and the following plant growth regulators: (1) a combination of naphthaleneacetic acid and 6-benzyladenine (MCA medium) and (2) a combination of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (MCAD medium). The results indicated that based on the proportions of uninucleate and binucleate microspores, flowers at the prebloom and early bloom stages contained sufficient late uninucleate to early binucleate microspores suitable for inducing embryogenesis. The production of microspore-derived embryogenic (MDE) structures was faster in MCA13 and MCA17 media than in MCAD13 and MCAD17 media. MCA13 and MCAD13 media induced the production of more callus-like structures than MCA17 and MCAD17 media. Thus, the addition of 2, 4-D to MCAD medium inhibited the growth of MDE structures. Globular embryoids emerged from the multicellular cluster.
Key message
This is the first report of the successful induction of embryoid production in Artemisia annua from an isolated microspore culture, and the induction was closely related to the flower type and microspore developmentstage.
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Abbreviations
- DH:
-
Doubled haploid
- IMC:
-
Isolated microspore culture
- PGRs:
-
Plant growth regulators
- NAA:
-
Naphthaleneacetic acid
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- BA:
-
6-Benzyladenine
- KIN:
-
Kinetin
- EMS:
-
Ethyl methanesulfonate
- NLNL:
-
Nitsch and Nitsch
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Acknowledgements
This work was partially supported by the National Research and Innovation Agency of Indonesia (BRIN) and LPDP through RIIM project grant no. 82/II.7/HK/2022. The authors sincerely thank BBPSI BIOGEN and the Indonesian Ministry of Agriculture for their assistance in providing the facility to implement the project.
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All authors contributed to the study’s conception and design. RP, ISD, DS, AA, and BSP contributed to the design and implementation of the research, analysis of the results, and writing of the manuscript. All authors collaborated in revising the manuscript. All authors have read and approved the final manuscript.
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Purnamaningsih, R., Dewi, I.S., Sukmadjaja, D. et al. Isolated microspore culture for embryoid production in Artemisia annua L.. Plant Cell Tiss Organ Cult 157, 5 (2024). https://doi.org/10.1007/s11240-024-02716-6
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DOI: https://doi.org/10.1007/s11240-024-02716-6