Plant Cell Reports

, Volume 21, Issue 2, pp 118–124 | Cite as

Cryopreservation of embryogenic cultures of Picea mariana (black spruce) using vitrification

  •  D. Touchell
  •  V. Chiang
  •  C.-J. Tsai
Cell Biology and Morphogenesis

Abstract.

This study reports on the first use of a vitrification procedure for the successful cryopreservation of embryogenic cultures of a coniferous species. Using Picea mariana embryogenic cultures, we obtained the highest survival by first preculturing embryogenic masses on semi-solid medium containing 0.8 M sorbitol for 48 h followed by incubation in PVS2 cryoprotective vitrification solution at 0°C for 30 min and direct immersion in liquid nitrogen. The replacement of sorbitol with 1.6 M glycerol also resulted in high survival. When sorbitol and glycerol were used in preculture treatments, 9 of 11 embryogenic lines survived liquid nitrogen treatments. We also demonstrated that 100% post-liquid nitrogen survival of mature somatic embryos could be obtained without pretreatments. A brief desiccation pretreatment, however, increased uniformity of somatic embryo germination.

Cryopreservation Vitrification Picea mariana Somatic embryogenesis 

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Copyright information

© Springer-Verlag 2002

Authors and Affiliations

  •  D. Touchell
    • 1
  •  V. Chiang
    • 1
  •  C.-J. Tsai
    • 1
  1. 1.Plant Biotechnology Research Center, School of Forestry and Wood Products, Michigan Technological University, 1400 Townsend Drive, Houghton, MI 49931, USA

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