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KMID : 0903619950360040540
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Journal of the Korean Society for Horticultural Science
1995 Volume.36 No. 4 p.540 ~ p.547
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Cryopreservation of Axillary Buds of Dendranthema grandiflorum Ramat . and Subsequent Plant Regeneration
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Abstract
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Excised axillary buds taken from chrysanthemum (Dendranthema grandiflorum) were successfully cryopreserved. The segments were precultured on solidified Murashige-Skoog medium containing 5% dimethyl sulfoxide (DMSO) and 0.5M sucrose at 25℃ in light for 24 hrs. Thereafter, these precultured segments were treated with various cryoprotector solution. The highest survival rate was observed in combinaton with 10% DMSO and 0.5M glucose as a cryoprotectant. Cryoprotected axillary buds were then sufficiently dehydrated with a slow prefreezing treatment prior to a plunge into liquid nitrogen (LN). The highest cell viability was observed when axillary buds were frozen in a cryotube as far as -40℃. In this case, the survival rate of samples prefrozen to -40℃ was about 80%. In prefreezing -40℃, the extremely slow cooling velocity of 0.3℃/min showed the highest survival rate. Ice seeding increased the survival rate of samples by the prevention of cellular supercooling. Survival rates of cryopreserved axillary buds using the same pretreatment procedure were compared for 9 other cultivars of chrysanthemum. Among 9 cultivars tested, 8 showed high survival rate. After the cryopreservation in LN, the survived axillary buds produced shoots and regenerated into plantlets. These results demonstrate the feasibility of cryopreservation for chrysanthemum axillary buds.
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