Survival of carnation (Dianthus caryophyllus L.) shoot apices frozen to the temperature of liquid nitrogen

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Effects of cooling and rewarming rates on the survival of carnation shoot apices frozen to the temperature of liquid nitrogen were investigated. Ten percent dimethyl sulfoxide (DMSO), alone or in combination with 5% glucose, sucrose or sorbitol was most effective as a cryoprotectant for carnation shoot apices. The shoot apices survived slow freezing at about -70℃ in the presence of 10% DMSO. About 80% of the shoot apices survived freezing at the temperature of liquid nitrogen after prefreezing at -50℃ or below, regardless of the rewarming rates. Shoot apices in the presence of 10% DMSO were cooled at different rates then rewarmed rapidly. The survival rate gradually decreased to zero as the cooling rate increased from about 0.5℃/min to 50℃/min. At cooling rates higher than 50℃/min, no survival was observed even at 5 × 104℃/min. However, in apices prefrozen at -15℃ or below then cooled ultrarapidly at 104℃/min, all remained alive with subsequent rapid rewarming. These apices developed normal young plants. This ultrarapid cooling method combined with prefreezing seems to be useful for the cryopreservation of shoot apices from various plants.
日本植物生理学会の論文