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A clonal propagation system for Atlantic white cedar (Chamaecyparis thyoides) via somatic embryogenesis without the use of plant growth regulators

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Abstract

Atlantic white cedar (AWC; Chamaecyparis thyoides), an aromatic evergreen conifer native to swamps and bogs along the Atlantic and Gulf coasts of the eastern United States was once an important species for timber production due to its durable wood. However, native populations have declined over the past two centuries. We established an in vitro propagation system for AWC via somatic embryogenesis (SE) without the use of plant growth regulators (PGRs). Whole megagametophytes with zygotic embryos from immature AWC cones were cultured on a modified half-strength embryo maturation (EM) medium with three different PGR treatments, including one devoid of PGRs. Both PGR treatment and cone collection date had significant effects on embryogenesis induction, with EM with no PGRs giving the highest embryogenesis induction, which ranged as high as 27%. We also conducted experiments to determine the effects of activated carbon (AC) and abscisic acid (ABA) in the maturation medium on production of mature somatic embryos. AC significantly affected this variable, with 2 g l−1 producing more embryos than 0 g l−1. Application of exogenous ABA not only failed to improve production of mature somatic embryos, the highest level tested (200 µM), apparently lowered production of mature embryos compared to the 0 ABA control. The highest numbers of mature somatic embryos per ml of plated embryogenic suspension (32–37) were produced on medium with 2 g l−1 AC and levels of ABA at 100 µM or lower. The SE system described here has the potential to contribute the restoration of Atlantic white cedar to its native habitat.

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Abbreviations

ABA:

Abscisic acid

AC:

Activated carbon

AWC:

Atlantic white cedar

BA:

Benzylamino purine

EM:

Embryo maturation medium of Maruyama et al. (2000)

IAA:

Indoleacetic acid

PGR:

Plant growth regulator

2,4-D:

2,4-Dichlorophenoxyacetic acid.

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Acknowledgements

This research was supported by Mclntire-Stennis funds allocated to the Warnell School of Forestry and Natural Resources and the USDA Forest Service, Southern Region National Forest System Genetics Program. We would like to thank Dr. Robert Jetton (Camcore, NC State University) and Bill Lott (Thompson Mills Forest State Arboretum of Georgia) for helping us obtain Atlantic white cedar material for culture initiation. We also thank Heather Gladfelter and Dr. Gerald Pullman for technical assistance and advice.

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  1. Chang-Ho Ahn

    Present address: Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chuncheon, 24341, Republic of Korea

Authors and Affiliations

  1. Warnell School of Forestry and Natural Resources, University of Georgia, Athens, GA, 30602, USA

    Chang-Ho Ahn, A. Ryan Tull, Paul M. Montello & Scott A. Merkle

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  1. Chang-Ho Ahn
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  2. A. Ryan Tull
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Correspondence to Scott A. Merkle.

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Ahn, CH., Tull, A.R., Montello, P.M. et al. A clonal propagation system for Atlantic white cedar (Chamaecyparis thyoides) via somatic embryogenesis without the use of plant growth regulators. Plant Cell Tiss Organ Cult 130, 91–101 (2017). https://doi.org/10.1007/s11240-017-1206-7

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  • DOI: https://doi.org/10.1007/s11240-017-1206-7

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