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KMID : 0903619940350050499
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Journal of the Korean Society for Horticultural Science
1994 Volume.35 No. 5 p.499 ~ p.506
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Survival by Vitrification of Dendranthema grandiflorum L . Shoot Tips for Cryopreservation
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Abstract
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Effect of cooling and rewarming rates on the survival of Dendranthema grandiflorum L.shoot tips frozen to the temperature of liquid nitrogen were investigated. Excised shoot tips from plants were precultured on plant hormon free agar medium containing 0.4M sucrose at 27℃ for 12 hours. Shoot tips following preculture were transferred to a 1.8㎖ plastic cryotube and then a highly concentrated cryoprotective solution (designated PVS2) was added at 25℃ or 0℃. The shoot tips treated with PVS2 at 25℃ for 20 min or at 0℃ for 50 min were directly plunged into liquid nitrogen. After rapid thawing at 40℃, the shoot tips were expelled into 1.8㎖ of 1.2M sucrose solution and then plated on agar hyponex medium. The average rate of survival, as evaluated by fluorescein diacetate (FDA) and phenosafranine double staining, was about 80%. The survival rates did not decrease when the shoot tips were stored for 30 days in LN. This procedure for cryopreservation of Dendranthema grandiflorum L. is very simple and appears promising as a routine method for cryopreserving shoot tip tissues from various plants.
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