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In vitro clonal propagation and stable cryopreservation system for Platycladus orientalis via somatic embryogenesis

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An Erratum to this article was published on 22 September 2017

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Abstract

Platycladus orientalis is a widespread conifer, which is native in eastern Asia, and has recently attracted much attention due to its ornamental value for landscape and gardens. However, native P. orientalis populations have been in decline over the past century. Here, we established an in vitro propagation and cryopreservation system for P. orientalis via somatic embryogenesis (SE). Whole megagametophytes with four development stages (Early embryogeny: E1 and late embryogeny: L1, L2, and L3) of zygotic embryos from immature P. orientalis cones were used as initial explants and cultured on three different basal media such as initiation medium (IM), Litvay (LV), and Schenk and Hildebrandt (SH). Both the developmental stage of zygotic embryos and kind of basal medium had a significant effect on embryogenesis induction with IM (P < 0.001, respectively). The highest frequency of embryogenic callus induction was obtained in megagametophytes with zygotic embryos at L2 stage, which ranged as high as 30%. The maturation medium containing IM basal salts, vitamins and amino acids, 15 g l−1 abscisic acid (ABA), 50 g l−1 maltose, and 100 g l−1 polyethylene glycol 4000 (PEG) was found to be the suitable medium for production of somatic embryos. The frequency of somatic embryo formation from both non-cryopreserved and cryopreserved cell lines was also tested. There were no statistical differences on the production of somatic embryos between non-cryopreserved and cryopreserved cells (P = 0.523). Genetic fidelity of the plantlets regenerated from non-cryopreserved and cryopreserved embryogenic cell lines was assessed by both random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analysis. There was no genetic instability in the regenerated plantlets from cryopreserved embryogenic cell lines. Both the SE protocol and cryopreservation protocols described here have the potential to contribute the conservation and clonal propagation of P. orientalis germplasm.

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Change history

  • 22 September 2017

    In the original publication, the concentration of abscisic acid mentioned in the Abstract and in the Materials and Methods was incorrect. It should have read 15 mg l−1.

Abbreviations

ABA:

Abscisic acid

AC:

Activated carbon

BA:

Benzylamino purine

IM:

Initiation medium

MIM:

Maturation medium

GIM:

Germination medium

PGR:

Plant growth regulator

2,4-D:

2,4-Dichlorophenoxyacetic acid

RAPD:

Random amplified polymorphic DNA

ISSR:

Inter simple sequence repeat

CP:

Cryoprotectant

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Acknowledgements

This work was financially supported by grants from the Post-Genome Multi-Ministry Genome Project (S111414L070110), and a 2014 research grant from Kangwon National University (C1010841-01-01).

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Authors and Affiliations

  1. Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chuncheon, 24341, Republic of Korea

    Chang-Ho Ahn & Yong-Eui Choi

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  1. Chang-Ho Ahn
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  2. Yong-Eui Choi
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Contributions

YEC designed all experiments and wrote drafts of the manuscript. CHA conducted the experiments and data analysis, took photos and wrote the manuscript.

Corresponding author

Correspondence to Yong-Eui Choi.

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The authors declare no conflict of interest.

Additional information

Communicated by Qiao-Chun Wang.

An erratum to this article is available at https://doi.org/10.1007/s11240-017-1307-3.

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Ahn, CH., Choi, YE. In vitro clonal propagation and stable cryopreservation system for Platycladus orientalis via somatic embryogenesis. Plant Cell Tiss Organ Cult 131, 513–523 (2017). https://doi.org/10.1007/s11240-017-1301-9

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  • DOI: https://doi.org/10.1007/s11240-017-1301-9

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