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Improved germination conditions for Norway spruce somatic cotyledonary embryos increased survival and height growth of emblings

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Abstract

Norway spruce is one of the most cultivated tree species in Nordic countries. However, intermittent shortages of improved seeds occur. As a powerful vegetative propagation technology, somatic embryogenesis (SE) could provide an alternative solution for this problem and also shorten the time required to obtain breeding gains. However, there are still large bottlenecks in SE, e.g. in the germination and acclimatization phases, which greatly affect the final outcome of somatic embryo plants (emblings). In this work, we examined the effects of in vitro embryo storage and germination treatments and ex vitro growing techniques on the survival and growth of emblings. The study comprised 32 genotypes from 18 full-sib families in four experiments, testing two different cold storage methods, three durations of in vitro germination, lower inorganic nitrogen content in the germination medium, and two plant-growing techniques. The best treatment combination—cold storage on filter paper, lower nitrogen content in the germination medium and one-week in vitro germination—resulted in an 88% higher survival and 28% higher growth compared to the poorest, reference treatment in the same test year. These emblings could be planted after a nursery period one year sooner than that of the control emblings. The results indicate that Norway spruce emblings germinated for one week in vitro can be transplanted and grown in nurseries without any additional treatments or environmental control differing from seedlings, which is a prerequisite to reach standards for forest regeneration material.

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Acknowledgements

The authors would like to acknowledge the editor and referees for their dedication to improve the manuscript with valuable comments.

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Authors and Affiliations

  1. Natural Resources Institute Finland (Luke), Production Systems, Finlandiantie 18, 58450, Punkaharju, Finland

    Mikko Tikkinen, Saila Varis & Tuija Aronen

  2. Faculty of Science and Forestry, School of Forest Sciences, University of Eastern Finland (UEF), Joensuu Campus (Yliopistokatu 7), PO Box 111, 80101, Joensuu, Finland

    Heli Peltola

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  1. Mikko Tikkinen
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  2. Saila Varis
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468_2018_1728_MOESM1_ESM.pdf

Appendix 1 Average length of shoot (positive scale) and root (negative scale) after in vitro germination treatments (1w-gel, 3w-gel and 5w-gel) in different genotypes in Experiment II. Average values for root and shoot length after in vitro germination are presented with standard error. The genotypes are organized in the same order as in Fig. 3 except the three additional genotypes included in Experiment II are added in the right (PDF 42 KB)

468_2018_1728_MOESM2_ESM.pdf

Appendix 2 Logistic regression models used for analyzing binary response (living or dead) in the nursery testing of emblings after first growth season. In the models, e1 is a design variable for embryo length before germination, g1 to g5 are design variables for germination treatments, c1 is a design variable for preservation method of embryogenic cell mass, and a1 – a21 are design variables for genotypes (PDF 20 KB)

468_2018_1728_MOESM3_ESM.pdf

Appendix 3 Mean height and survival of genotypes in different treatments in Experiment III. Average values are presented with standard error (PDF 27 KB)

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Tikkinen, M., Varis, S., Peltola, H. et al. Improved germination conditions for Norway spruce somatic cotyledonary embryos increased survival and height growth of emblings. Trees 32, 1489–1504 (2018). https://doi.org/10.1007/s00468-018-1728-6

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