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Cryopreservation of in vitro-grown shoot tips of strawberry by the vitrification method using aluminium cryo-plates

  • Shin-ichi Yamamoto (a1), Kuniaki Fukui (a1), Tariq Rafique (a2), Nayyar Iqbal Khan (a3), Carlos Roman Castillo Martinez (a4), Kentaro Sekizawa (a5), Toshikazu Matsumoto (a6) and Takao Niino (a1)...

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.

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A Sakai , D Hirai and T Niino (2008) Development of PVS-based vitrification and encapsulation–vitrification protocols. In: B Reed (ed.) Plant Cryopreservation: A Practical Guide. New York: Springer LLC, pp. 3358.

K Sekizawa , S Yamamoto , T Rafique , K Fukui and T Niino (2011) Cryopreservation of in vitro-grown shoot tips of carnation (Dianthus caryophyllus L.) by vitrification method using aluminium cryo-plates. Plant Biotechnology 28: 401405.

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Plant Genetic Resources
  • ISSN: 1479-2621
  • EISSN: 1479-263X
  • URL: /core/journals/plant-genetic-resources
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