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Fraser fir somatic embryogenesis: high frequency initiation, maintenance, embryo development, germination and cryopreservation

New Forests volume 47, pages 453–480 (2016)Cite this article

Abstract

Fraser fir (Abies fraseri [Pursh] Poir.) is a coniferous species native to the Southern Appalachian Mountains of the eastern United States. The species has high economic and recreational value but is vulnerable to extinction due to introduced pests and global warming. Somatic embryogenesis technology may assist in the clonal production of desired lines of Christmas trees and safekeeping of rare and valuable germplasm via cryopreservation. We have developed a highly effective medium for initiation of embryogenic tissue from immature or mature seeds of Fraser fir that contains AL salts (Kvaalen et al., in Can J For Res 35:1053–1060, 2005), brassinolide, paclobutrazol and abscisic acid. Using dominant embryos attached to the female gametophyte placed on medium, the highest initiation percentages occurred with precotyledonary stage 3 embryos. When tested with 11 high-value open-pollinated families over 5 years, initiation tests for medium containing brassinolide and paclobutrazol averaged 6–62 % initiation. A maintenance medium was developed that contained AL salts and 1.1 mg L−1 BAP and was able to capture approximately 50 % of the initiations. A maturation medium was developed containing AL salts, maltose, polyethylene glycol 8000 and abscisic acid that produced cotyledonary embryos capable of germination to produce a root and shoot. Culture cryopreservation and retrieval was also demonstrated.

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Acknowledgments

We thank Jerry Moody, Avery County Extension Agent, NCSU, Ken Roeder, Forest Geneticist, North Carolina Division of Forest Resources, and Bryan Davis, formerly NCSU Cooperative Extension Technician, for cone collections. We also thank the NC Christmas Tree Association for financial support and the Institute of Paper Science and Technology (Renewable Bioproducts Institute) and Georgia Institute of Technology for providing materials and supplies for undergraduate research in support of this project. We are grateful for the help of K-M Chase, E. Clark, J. Dregar, J. Grabowski, R. Gray, R. Gupta, C. Umejiego, and Y. Zhang.

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  1. Kylie Bucalo

    Present address: Department of Conservation Research, Atlanta Botanical Garden, Atlanta, GA, 30309, USA

Authors and Affiliations

  1. School of Biology, Renewable Bioproducts Institute (Formerly the Institute of Paper Science and Technology), Georgia Institute of Technology, 500 10th Street N.W., Atlanta, GA, 30332-0620, USA

    Gerald S. Pullman

  2. Institute of Paper Science and Technology, Georgia Institute of Technology, 500 10th Street N.W., Atlanta, GA, 30332, USA

    Katie Olson & Kylie Bucalo

  3. School of Biology, Georgia Institute of Technology, Atlanta, GA, USA

    Taylor Fischer

  4. G. W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, 500 10th Street N.W., Atlanta, GA, 30332, USA

    Ulrika Egertsdotter

  5. Umea Plant Science Center, Department of Forest Genetics and Plant Physiology, Swedish Agricultural University, 901 83, Umeå, Sweden

    Ulrika Egertsdotter

  6. Department of Forestry and Environmental Resources, N.C. State University, 3219 Jordan Hall, Raleigh, NC, 27695-8008, USA

    John Frampton

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  1. Gerald S. Pullman
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Correspondence to Gerald S. Pullman.

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Pullman, G.S., Olson, K., Fischer, T. et al. Fraser fir somatic embryogenesis: high frequency initiation, maintenance, embryo development, germination and cryopreservation. New Forests 47, 453–480 (2016). https://doi.org/10.1007/s11056-016-9525-9

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Keywords

  • Abies fraseri
  • Embryogenic tissue initiation
  • Conifer
  • Fraser fir
  • Somatic embryogenesis