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An assessment of somatic embryogenesis and cryo-preservation methods with a wide range of Sitka spruce breeding material from the UK

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Abstract

A wide range of Sitka spruce breeding material (Picea sitchensis (Bong.) Carr.) was assessed for its ability to respond to somatic embryogenesis (SE) and cryo-preservation methods, with 765 cell lines being established from seeds taken from the UK’s breeding population for testing with the relevant methodologies. Cell lines were most readily established from immature seeds (63%) on a proliferation medium originally developed for white spruce (P. glauca) with agar as the gelling agent. However, it was also found that 48% of the zygotic embryos taken from freshly harvested mature seed were also able to establish cell lines on the same medium, as were 20% of seeds that had been stored at −20 °C for up to 8 years. Most of the cell lines tested (57%) produced significant numbers of somatic embryos, but the rate at which they could be germinated into plants capable of surviving the transfer to nursery conditions varied considerably from 67 to 8% between years for reasons that were not clear. The ability of the cell lines to survive cryo-preservation was also assessed, and out of 199 cell lines tested, 45 cell lines from 21 breeding families were successfully established in a cryo-storage facility. The best rates of recovery were obtained when Phytagel™ was used in the medium for growing the samples before and after they were cryo-preserved.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

ABA:

Abscisic acid

BAP:

Benzyl amino purine

DMSO:

Di-methyl sulfoxide

FR:

Forest research

MVF:

Multi-varietal forestry

SE:

Somatic embryogenesis

UK:

United Kingdom

WSGM:

White spruce germination medium

WSMM:

White spruce maturation medium

WSPM:

White spruce proliferation medium

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Acknowledgements

We gratefully acknowledge Steve Lee of FR and Andy Leitch of the UK Forestry Commission for their support of this work, and thank Yill-Sung Park and Ian MacEacheron of the Canadian Forest Service; David Thompson and Stuart Kennedy of Coillte, Ireland; Cathy Hargreaves of Scion Research, New Zealand for their many helpful comments for the execution of this work; and we also thank Richard Jinks and Glenn Brearley of FR for their help in preparing the manuscript.

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Authors and Affiliations

  1. Forest Research, Northern Research Station, Roslin, Edinburgh, EH25 9SY, Midlothian, Scotland, UK

    Trevor Fenning, Margaret O’Donnell & Thomas Connolly

  2. School of Environment, Science and Engineering, Southern Cross University, Lismore, NSW, Australia

    Trevor Fenning

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  1. Trevor Fenning
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  2. Margaret O’Donnell
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  3. Thomas Connolly
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Correspondence to Trevor Fenning.

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Communicated by Pamela J. Weathers.

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Fenning, T., O’Donnell, M. & Connolly, T. An assessment of somatic embryogenesis and cryo-preservation methods with a wide range of Sitka spruce breeding material from the UK. Plant Cell Tiss Organ Cult 131, 483–497 (2017). https://doi.org/10.1007/s11240-017-1299-z

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  • DOI: https://doi.org/10.1007/s11240-017-1299-z

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