ISHS Acta Horticulturae 271: VII International Asparagus Symposium
CRYOPRESERVATION BY VITRIFICATION OF CULTURED CELLS AND SOMATIC EMBRYOS FROM MESOPHYLL TISSUE OF ASPARAGUS |
| Authors: | A. Uragami, A. Sakai, M. Nagai, T. Takahashi |
| Abstract:
Mechanically isolated single cells of asparagus (Asparagus officinalis L.) regenerated plantlets through somatic embryogenesis. Cultured cells and embryos originated from single cells were cryopreserved by vitrification. The vitrification solution (PVS1) contains (w/v) 22% glycerol, 15% ethylene glycol, 15% propylene glycol and 7% DMSO in 0.5M sorbitol Murashige-Skoog medium (MS). After cryoprotection with 0.5M-sorbitol MS medium containing 12% ethylene glycol, cells or embryos were exposed stepwise to 85% PVS1 at 0°C. They were loaded in 0.25 ml transparent straw, and then plunged directly into liquid nitrogen. After rapid warming, PVS1 was removed and diluted stepwise. The highest survival of vitrified cells and embryos were about 65% and 50%, respectively. Survived embryos developed into plantlets. | |
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