Plant Cell, Tissue and Organ Culture

, Volume 27, Issue 1, pp 89–93 | Cite as

Freeze-preservation of buds from Scots pine trees

  • Tero Kuoksa
  • Anja Hohtola


A procedure has been developed for freeze-preservation of buds of the Scots pine (Pinus sylvestris L.). Instead of liquid nitrogen, cold storage in −80°C was used. The partly dormant material used in the experiments was obtained directly from a natural stand in Northern Finland and no prefreezing or cryoprotectants for preconditioning were used. Cooling velocity was 1°C/min up to a terminal freezing temperature of −39°C, after which the buds were immersed in liquid nitrogen at −196°C for 10 minutes. The material was then transferred to a deepfreezer at −80°C and stored up to 6 months. After rapid thawing, the buds were sterilized and their viability was tested by FDA staining and by culturing meristems on 1/2 MS medium for at least two weeks. All the freezing experiments were performed during March and April. The best survival of buds (90–100%) was achieved at the beginning of April, after which a pronounced decline in survival occurred obviously due to a rise in the water content of the buds.

Key words

cryopreservation Pinus sylvestris L. tissue culture 


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Copyright information

© Kluwer Academic Publishers 1991

Authors and Affiliations

  • Tero Kuoksa
    • 1
  • Anja Hohtola
    • 1
  1. 1.Biocenter and Department of BotanyUniversity of OuluOuluFinland

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