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- Title
Cryopreservation of in vitro-grown shoot tips of strawberry by the vitrification method using aluminium cryo-plates.
- Authors
Yamamoto, Shin-ichi; Fukui, Kuniaki; Rafique, Tariq; Khan, Nayyar Iqbal; Castillo Martinez, Carlos Roman; Sekizawa, Kentaro; Matsumoto, Toshikazu; Niino, Takao
- Abstract
Cryopreservation using an aluminium cryo-plate was successfully applied to in vitro-grown strawberry (Fragaria × ananassa Duch.) shoot tips. The shoots were cold-hardened at 5°C for 3 weeks with an 8-h photoperiod. The shoot tips (1.5–2.0 mm × 0.5–1.0 mm) were dissected from the shoot and pre-cultured at 5°C for 2 d on Murashige and Skoog medium containing 2 M glycerol and 0.3 M sucrose. The pre-cultured shoot tips were placed on the aluminium cryo-plate containing ten wells embedded in alginate gel. Osmoprotection was performed by immersing the cryo-plates in a loading solution (2 M glycerol and 0.8 M sucrose) for 30 min at 25°C. Dehydration was performed by immersing the cryo-plates in plant vitrification solution 2 for 50 min at 25°C. Then, the cryo-plate with shoot tips was transferred into an uncapped cryotube that was held on a cryo-cane and directly immersed into liquid nitrogen (LN). After storage in LN, shoot tips attached to the cryo-plate were directly immersed into 2 ml of a 1 M sucrose solution for regeneration. Using this procedure, the average regrowth level of vitrified shoot tips of 15 strawberry cultivars reached 81%. This new method has many advantages and will facilitate the cryostorage of strawberry germplasm.
- Subjects
CRYOPRESERVATION of plant cells &; tissues; PLANT growth; PLANT shoots; STRAWBERRIES; ALUMINUM plates; ALGINATES; PLANT germplasm; LIQUID nitrogen
- Publication
Plant Genetic Resources: Characterisation & Utilisation, 2012, Vol 10, Issue 1, p14
- ISSN
1479-2621
- Publication type
Article
- DOI
10.1017/S1479262111000906