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In vitro propagation and cryopreservation of Thuja koraiensis Nakai via somatic embryogenesis

  • Chang Ho Ahn
  • Kweon Heo
  • Hyeong Soo Park
  • Yong Eui ChoiEmail author
Plant Tissue Culture
  • 57 Downloads

Abstract

Korean arbor vitae (KAV; Thuja koraiensis Nakai) is a critically endangered coniferous tree in Korea. Here, we report the somatic embryogenesis (SE) and cryopreservation system that can be used for micropropagation of KAV and long-term storage of KAV cultures. To induce SE in KAV, the influence of the developmental stage of zygotic embryos and the effect of basal medium on embryogenesis induction were examined. The developmental stage of zygotic embryos had a significant effect on the embryogenesis induction (P < 0.0001). The highest frequency of embryogenesis induction occurred in megagametophytes with zygotic embryos at precotyledonary (P) and late embryogeny (L1) stage (36%). The highest frequency of embryogenesis induction was obtained on initiation medium containing IM basal salts with 2.2 μM 6-benzylaminopurine and 4.5 μM 2,4-dichlorophenoxyacetic acid (35%). The effect of abscisic acid (ABA) on production of somatic embryos was tested. The highest number of somatic embryos per 50 mg of embryogenic tissue was achieved on maturation medium with levels of 100 μM ABA (24.0 ± 2.4). The effect of cryopreservation treatment to embryogenic tissues on the maturation capacity of somatic embryos was also tested. No significant differences between noncryopreservation and cryopreservation treatment were observed (P = 0.1896), and the highest mean number of somatic embryo per 50 mg of embryogenic tissues was obtained in noncryopreserved cell line (28.17 ± 5.66). Finally, the genetic identities of the plantlets regenerated from non- and cryopreserved embryogenic cell lines were verified and there was no genetic variation in the regenerated plantlets from cryostored embryogenic cell lines. This study is the first report on SE and the successful cryopreservation of embryogenic culture of the genus Thuja.

Keywords

Cupressaceae Endangered conifer Embryogenic tissue Cryostorage Genetic fidelity 

Notes

Acknowledgements

This research was carried out with the support of ‘R&D Program for Forest Science Technology (Project No. FTIS 2018131B10-1820-BB01) provided by Korea Forest Service (Korea Forestry Promotion Institute). We would like to thank the National Park Research Institute of Korea and Korea National Arboretum. We also thank to Dr. Hyo-In Lim (Korea Forest Research Institute) for the assistance.

Author contributions

YEC and CHA designed all experiments and wrote all drafts of the manuscript. CHA conducted the experiments and data analysis. KH carried out cytological analysis. HSP collected KAV cones for culture initiation and provided some photos.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11627_2019_9989_MOESM1_ESM.docx (15 kb)
Table S1 (DOCX 14 kb)
11627_2019_9989_MOESM2_ESM.docx (15 kb)
Table S2 (DOCX 15 kb)

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Copyright information

© The Society for In Vitro Biology 2019

Authors and Affiliations

  • Chang Ho Ahn
    • 1
  • Kweon Heo
    • 2
  • Hyeong Soo Park
    • 3
  • Yong Eui Choi
    • 1
    Email author
  1. 1.Devision of Forest Resources, College of Forest and Environmental SciencesKangwon National UniversityChuncheonRepublic of Korea
  2. 2.Department of Applied Plant ScienceKangwon National UniversityChuncheonRepublic of Korea
  3. 3.Seoraksan National Park Office, Korea National Park ServiceSokchoRepublic of Korea

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