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Pine somatic embryogenesis: analyses of seed tissue and medium to improve protocol development

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Abstract

The shift from vegetative to embryogenic growth requires tissue to enter a radically different program of development and can be studied in vitro through the development of somatic embryos. From an applied perspective somatic embryogenesis (SE) is expected to play an important role in increasing productivity, sustainability, and uniformity of future forests. For commercial use, SE technology must work with a variety of genetically diverse trees. Since the first reports of SE in Picea abies and Larix decidua in 1985, many different coniferous species have shown the ability to produce embryogenic tissue. However, initiation frequency is often low, many desired seed sources are recalcitrant, and culture survival is often poor, raising costs of somatic seedlings produced from successful genotypes. A number of tools are now available to improve embryogenic tissue initiation and somatic embryo development in vitro that have resulted from analytical studies of seed tissues, the seed environment and gene expression in megagametophyte, zygotic embryos and somatic embryos. Benefits have occurred from medium supplementation with hormones, plant growth regulators, hormone inhibitors and polyamines. Somatic embryo growth has been enhanced with medium supplementation of nutritional components including specific sugar types, vitamins, organic acids, and redox potential modifiers. Control of environmental factors including, water potential, pH, adsorption of medium components by activated carbon and liquid versus gelled medium have also led to SE protocol improvements. The use of analytical studies to duplicate the seed environment in vitro is improving protocol development resulting in increased initiation, improved yields and higher-quality somatic embryos.

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Acknowledgments

The authors thank the member companies of the Institute of Paper Science and Technology and IPST at Georgia Tech, State of Georgia TIP3 Program, Consortium for Plant Biotechnology Research by the DOE Prime Agreement No. DEFG36-02GO12026 and USEPA grant EM-83438801, and Arborgen, Monsanto Company and Weyerhaeuser Company as members of CPBR, for financial support. We thank Arborgen, MeadWestvaco Corporation and Weyerhaeuser NR Company for cone collections. In addition, the authors are grateful for the valuable assistance of Michael Buchanan, Kylie Bucalo, John Cairney, Erin Clark, Molly Clark, Brandi Copeland, Kelly-Marie Chase, Xiaorong Feng, Jennifer Grabowski, Shannon Johnson, Sheldon W. May, Jonathan Mein, Paul Montello, Kavita Namjoshi, Anna Skryabina, Xiaoyan Zeng and Yalin Zhang and PK Gupta for critical reading of the early draft.

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  1. Forest Biology, Institute of Paper Science and Technology at Georgia Tech, Georgia Institute of Technology, Atlanta, GA, 30332-0620, USA

    Gerald S. Pullman & Kylie Bucalo

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

    Gerald S. Pullman

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

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Pullman, G.S., Bucalo, K. Pine somatic embryogenesis: analyses of seed tissue and medium to improve protocol development. New Forests 45, 353–377 (2014). https://doi.org/10.1007/s11056-014-9407-y

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