ISHS Acta Horticulturae 538: Eucarpia symposium on Fruit Breeding and Genetics
CRYOPRESERVATION OF IN VITRO-CULTURED AXILLARY SHOOT TIPS OF VITIS BY VITRIFICATION |
| Authors: | T. Matsumoto, A. Sakai |
| Keywords: | cryopreservation, shoot tips, Vitis, vitrification, PVS2, osmoprotection |
| Abstract:
In vitro-grown axillary shoot tips of Vitis were successfully cryopreserved by vitrification. Excised shoot tips were precultured on solidified 1/2 MS medium containing 0.3 M sucrose for 3 days at 25oC, then osmoprotected a mixture of 2 M glycerol plus 0.4 M sucrose for 20 min at 25oC before dehydration with PVS2 solution for 80 min at 0oC (a two-step vitrification). The post liquid nitrogen (LN) recovery rate by this procedure was approximately 60%. To improve the recovery rate, precultured shoot tips were previously dehydrated with a 1/2PVS2 solution (half strength of glycerol, ethylene glycol and dimethylsulphoxide DMSO) for 30 min, followed by PVS2 solution for 50 min at 0oC prior to a plunge into LN (a three-step vitrification). This three-step vitrification procedure gave a considerably higher rate of recovery growth (approx. 85%) over the two-step vitrification. This three-step procedure was successfully applied for other 7 Vitis plants. Thus, this three-step vitrification procedure appears promising for cryopreservation of Vitis germplasm. | |
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