Cryopreservation of in vitro-grown apical meristems of wasabi (Wasabia japonica) by vitrification and subsequent high plant regeneration |
| |
Authors: | T Matsumoto A Sakai K Yamada |
| |
Institution: | (1) Shimane Agricultural Experiment Station, 693 Izumo, Japan;(2) Asabucho 1-5-23, Kitaku, 001 Sapporo, Japan |
| |
Abstract: | Summary
In vitro-grown apical meristems of wasabi (Wasabia japonica Matsumura) were successfully cryopreserved by vitrification. Excised apical meristems precultured on solidified M S medium containing 0.3M sucrose at 20°C for 1 day were loaded with a mixture of 2M glycerol and 0.4M sucrose for 20 min at 25°C. Cryoprotected meristems were then sufficiently dehydrated with a highly concentrated vitrification solution (designated PVS2) for 10 min at 25°C prior to a plunge into liquid nitrogen. After rapid warming, the meristems were expelled into 2 ml of 1.2M sucrose for 20 min and then plated on solidified culture medium. Successfully vitrified and warmed meristems remained green after plating, resumed growth within 3 days, and directly developed shoots within two weeks. The average rate of normal shoot formation amounted to about 80 to 90% in the cryopreserved meristems. This method was successfully applied to three other cultivars of wasabi. This vitrification procedure promises to become a routine method for cryopreserving meristems of wasabi.Abbreviations BA
6-benzylaminopurine
- DMSO
dimethylsulfoxide
- EG
ethylene glycol
- LN
liquid nitrogen
- MS medium
Murashige and Skoog medium (1962)
- PVS2
vitrification solution |
| |
Keywords: | cryopreservation apical meristems vitrification wasabi (Wasabia japonica) |
本文献已被 SpringerLink 等数据库收录! |