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.
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Abbreviations
- BA:
-
6-benzylaminopurine
- DMSO:
-
dimethylsulfoxide
- EG:
-
ethylene glycol
- LN:
-
liquid nitrogen
- MS medium:
-
Murashige and Skoog medium (1962)
- PVS2:
-
vitrification solution
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Matsumoto, T., Sakai, A. & Yamada, K. Cryopreservation of in vitro-grown apical meristems of wasabi (Wasabia japonica) by vitrification and subsequent high plant regeneration. Plant Cell Reports 13, 442–446 (1994). https://doi.org/10.1007/BF00231963
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DOI: https://doi.org/10.1007/BF00231963