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

Effects of cooling and rewarming rates on the survival of carnationshoot apices frozen to the temperature of liquid nitrogen wereinvestigated. Ten percent dimethyl sulfoxide (DMSO), alone orin combination with 5% glucose, sucrose or sorbitol was mosteffective as a cryoprotectant for carnation shoot apices. Theshoot apices survived slow freezing at about –70?C inthe presence of 10% DMSO. About 80% of the shoot apices survivedfreezing at the temperature of liquid nitrogen after prefreezingat –50?C or below, regardless of the rewarming rates.Shoot apices in the presence of 10% DMSO were cooled at differentrates then rewarmed rapidly. The survival rate gradually decreasedto zero as the cooling rate increased from about 0.5?C/min to50?C/min. At cooling rates higher than 50?C/min, no survivalwas observed even at 5?10⁴?C/min. However, in apices prefrozenat –15?C or below then cooled ultrarapidly at 10⁴?C/min,all remained alive with subsequent rapid rewarming. These apicesdeveloped normal young plants. This ultrarapid cooling methodcombined with prefreezing seems to be useful for the cryopreservationof shoot apices from various plants. ¹Contribution No. 2207 from the Institute of Low TemperatureScience, Hokkaido University. This work was supported in partby a Grant-in-Aid (No. 434035) for Scientific Research fromthe Ministry of Education, Science and Culture. (Received November 13, 1979; )