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Planta

, Volume 233, Issue 3, pp 635–647 | Cite as

Initiation of somatic embryos and regeneration of plants from primordial shoots of 10-year-old somatic white spruce and expression profiles of 11 genes followed during the tissue culture process

  • Krystyna Klimaszewska
  • Catherine Overton
  • Don Stewart
  • Robert G. Rutledge
Original Article

Abstract

Adult conifers are notoriously recalcitrant in vegetative propagation and micropropagation that would result in the regeneration of juvenile propagules through somatic embryogenesis (SE) has not been demonstrated to date. Because SE-derived material is more amenable in subsequent tissue culture experiments compared with seed-derived material, a multi-year study was conducted to investigate induction of SE from primordial shoot (PS) explants that were excised from shoot buds of somatic embryo-derived white spruce. The SE induction experiments were carried out first with greenhouse-grown and later with field-grown trees each year from 2002 (2-year-old) to 2010 (10-year-old). Of the four genotypes tested, 893-2 and 893-12 never responded, 893-1 responded up to year 4 and 893-6 consistently responded every year. In 2010, for the first time, three of the 17 893-6 clonal trees produced male strobili as well as SE from cultured PS explants. SE induction was associated with formation of a nodule on the surface of an elongated needle primordium or in callus. Early somatic embryos were detectable after about 3 weeks of culture. Of 11 genes whose expression profiles were followed during the PS cultures, CHAP3A, VP1, WOX2 and SAP2C were expressed exclusively in the early stages of SE, and could potentially be used as markers of embryogenecity. Mature somatic embryos and plants were produced from the explants of responding genotype. Implication of these results for future research on adult conifer recalcitrance in micropropagation is discussed.

Keywords

Absolute quantitative real-time PCR Adult tree Conifer Picea glauca Shoot buds Transcription factors 

Abbreviations

BA

6-Benzyladenine

2,4-D

2,4-Dichlorophenoxyacetic acid

EM

Embryonal mass

HP

Hypothetical protein

LEC1

LEAFY COTYLEDON1

PS

Primordial shoot

PTSR

Peroxisomal targeting signal receptor

PVP

Polyvinylpyrrolidone

Qpcr

Quantitative polymerase chain reaction

SE

Somatic embryogenesis

SERK1

Somatic embryogenesis receptor-like kinase 1

WOX2

WUSCHEL

Notes

Acknowledgments

We thank Dr. J. Bonga (Natural Resources Canada, Canadian Forest Service, Atlantic Forestry Centre) for his critical reading of the manuscript and valuable suggestions. Ms. P. Cheers (CFS-Laurentian Forestry Centre) is gratefully acknowledged for English editing. This work was financially supported by the Natural Resources Canada, Canadian Forest Service.

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Krystyna Klimaszewska
    • 1
  • Catherine Overton
    • 1
  • Don Stewart
    • 1
  • Robert G. Rutledge
    • 1
  1. 1.Natural Resources CanadaCanadian Forest Service, Laurentian Forestry Centre, 1055 du P.E.P.SQuebecCanada

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