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Cryopreservation of embryogenic suspension cultures of Cyclamen persicum Mill.

  • Cell Biology and Morphogenesis
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Abstract

We have developed a simple protocol for the cryopreservation of embryogenic suspension cultures of Cyclamen persicum. Embryogenic suspension cultures in the linear growth phase 7–10 days after subculture were used for cryopreservation. Of the different cryoprotectants tested during a 2-day pre-culture, 0.6 M sucrose resulted in the highest re-growth rates of 75%. An additional pre-treatment with 0.6 M sucrose and 10% DMSO (dimethylsulfoxide) for 1 h also positively affected re-growth. Microscopic studies on viability revealed that only few small embryogenic cells survived cryopreservation, while vacuolated single cells died. Experiments in which the duration of the pre-culture period—i.e. the length of time the embryogenic suspension cells were exposed to 0.6 M sucrose—was varied showed that 2–4 days was the most optimal exposure time to 0.6 M sucrose. Callus re-grown after cryopreservation showed growth rates similar to that of unfrozen callus and regenerated even higher numbers of somatic embryos than unfrozen callus.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

DM:

Dry mass

DMSO:

Dimethylsulfoxide

FDA:

Fluorescein diacetate

FM:

Fresh mass

2IP:

6-(γ,γ-Dimethylallylamino)purine

LN:

Liquid nitrogen

rpm:

Rounds per minute

SCV:

Sedimented cell volume

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Acknowledgement

The authors would like to thank Dr. Gunda Mix-Wagner for the introduction to cryopreservation techniques.

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Authors and Affiliations

  1. Section of Floriculture, Institute of Floriculture, Tree Nursery Science and Plant Breeding, University of Hannover, Herrenhaeuser Str. 2, 30419, Hannover, Germany

    Traud Winkelmann, Viola Mußmann & Margrethe Serek

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  1. Traud Winkelmann
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  2. Viola Mußmann
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  3. Margrethe Serek
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Correspondence to Traud Winkelmann.

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Communicated by H. Lörz

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Winkelmann, T., Mußmann, V. & Serek, M. Cryopreservation of embryogenic suspension cultures of Cyclamen persicum Mill.. Plant Cell Rep 23, 1–8 (2004). https://doi.org/10.1007/s00299-004-0783-1

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  • DOI: https://doi.org/10.1007/s00299-004-0783-1

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