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Investigating a new cryopreservation protocol for yams (Dioscorea spp.)

  • Cell Biology and Morphogenesis
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Abstract

Experiments were initiated to develop protocols for four species of Dioscorea, namely D. bulbifera L., D. oppositifolia L., D. alata L. and D. cayenensis Lam., and to assess the effects of various factors on the survival and regrowth of explants after rewarming. The treatments consisted of three different methods—vitrification, droplet and a modified droplet method—as well as the following variables—cryoprotective solution, sucrose concentration in the preadaptation medium and cold acclimation. While most of the protocols resulted in low to zero survival and regrowth rates, maximum survival rates of 100% and 75% were obtained for D. oppositifolia L. and D. cayenensis Lam., respectively, using two different protocols of the modified droplet method. A higher average survival rate was obtained using the droplet and modified droplet techniques than for the original vitrification methods. When the average of all the experiments was taken, more than one-half (52%) of the surviving explants developed further within 1 month with the modified droplet method compared to zero regrowth with the two other methods. The optimum protocol for cryopreservation is specific for each genotype.

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Fig. 1a–d.
Fig. 2a–f.
Fig. 3.

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Abbreviations

AC :

Activated charcoal

BAP :

6-Benzylaminopurine

DMSO :

Dimethylsulfoxide

EG :

Ethylene glycol

GA 3 :

Gibberellic acid

Gly :

Glycerol

IAA :

Indole-3-acetic acid

Kin :

Kinetin

NAA :

α-Naphthaleneacetic acid

PVS :

Plant vitrification solution

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Acknowledgements

The authors thank Mrs. Doris Büchner and Mrs. Marion Dreiling for their skilful technical assistance in the experiments and micropropagation of yams. S.L. thanks the German Academic Exchange Service (DAAD) for funding his studies.

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  1. S. Leunufna

    Present address: Genebank Department, Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, Germany

Authors and Affiliations

  1. Pattimura University, Ambon, Indonesia

    S. Leunufna

  2. Genebank Department, Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, Germany

    E. R. J. Keller

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  1. S. Leunufna
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Correspondence to E. R. J. Keller.

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Communicat by H. Lörz

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Leunufna, S., Keller, E.R.J. Investigating a new cryopreservation protocol for yams (Dioscorea spp.). Plant Cell Rep 21, 1159–1166 (2003). https://doi.org/10.1007/s00299-003-0652-3

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  • DOI: https://doi.org/10.1007/s00299-003-0652-3

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