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

Published online by Cambridge University Press:  28 November 2011

Shin-ichi Yamamoto ,
Kuniaki Fukui ,
Tariq Rafique ,
Nayyar Iqbal Khan ,
Carlos Roman Castillo Martinez ,
Kentaro Sekizawa ,
Toshikazu Matsumoto  and
Takao Niino
Shin-ichi Yamamoto
Affiliation:
Genebank, National Institute of Agrobiological Sciences, Tsukuba 305-8602, Japan
Kuniaki Fukui
Affiliation:
Genebank, National Institute of Agrobiological Sciences, Tsukuba 305-8602, Japan
Tariq Rafique
Affiliation:
Plant Genetic Resources Programme, National Agricultural Research Center, Islamabad, Pakistan
Nayyar Iqbal Khan
Affiliation:
NWFP Agricultural Research System, Abottabad, Pakistan
Carlos Roman Castillo Martinez
Affiliation:
National Genetic Resources Center, INIFAP, Tepatitlan, Mexico
Kentaro Sekizawa
Affiliation:
National Center for Seeds and Seedlings, Tsukuba 305-0852, Japan
Toshikazu Matsumoto
Affiliation:
Faculty of Life and Environmental Science, Shimane University, Matsue 690-1102, Japan
Takao Niino*
Affiliation:
Genebank, National Institute of Agrobiological Sciences, Tsukuba 305-8602, Japan
*
*Corresponding author. E-mail: [email protected]
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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.

Keywords

aluminium platecryo-platestrawberryV-Cryo-plate methodvitrification
Type
Research Article
Information
Plant Genetic Resources , Volume 10 , Issue 1 , April 2012 , pp. 14 - 19
Copyright
Copyright © NIAB 2011

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