Cryopreservation of Persimmon Shoot Tips from Dormant Buds Using the D Cryo-plate Technique
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- Matsumoto Toshikazu
- Faculty of Life and Environmental Science, Shimane University
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- Yamamoto Shin-ichi
- National Institute of Agrobiological Sciences (NIAS)
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- Fukui Kuniaki
- National Institute of Agrobiological Sciences (NIAS)
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- Rafique Tariq
- National Institute of Agrobiological Sciences (NIAS)
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- Engelmann Florent
- Institut de Recherche pour le Développement (IRD)
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- Niino Takao
- Gene Research Center, University of Tsukuba
Bibliographic Information
- Other Title
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- D クライオプレート法によるカキ冬芽茎頂の超低温保存
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Abstract
Cryopreservation has become a very important tool for the long-term storage of plant germplasm. A new cryopreservation protocol based on air dehydration of explants placed on aluminum cryo-plates, termed the D cryo-plate technique, was developed. In this study, the most suitable conditions of cryopreservation for dormant shoot tips of Japanese persimmon (Diospyros kaki Thunb. ‘Saijo’) using the D cryo-plate technique were investigated. Dormant one-year-old shoots of persimmon were collected from the experimental farm of Shimane University in January 2013 and stored at 2°C until use. After surface sterilization, shoot tips of about 1 mm in size were dissected from the dormant buds and precultured overnight at 25°C on solidified 1/2MS medium containing 0.3 M sucrose. Precultured shoot tips were placed on aluminum cryo-plates and embedded in calcium alginate gel. Osmoprotection of shoot tips was performed by immersing the cryo-plates for 30 min at 25°C in an LS solution containing 2 M glycerol + 1.0 M sucrose in 1/2MS solution. For the D cryo-plate technique, encapsulated shoot tips were dehydrated by placing the cryo-plates in the air current of a laminar flow cabinet for 30–90 min. Cooling was performed by placing the cryo-plates in uncapped cryotubes, which were immersed in liquid nitrogen. For rewarming, the cryo-plates were immersed in 1/2MS medium containing 1.0 M sucrose for 20 min at 25°C. In this study, the preculture did not improve the regrowth after cryopreservation; however, we consider that it should be performed in the D cryo-plate procedure for application of other cultivars and utilization in genebanks. A high regrowth rate of cryopreserved shoot tips (84%) was achieved after dehydration for 30 min. This optimized procedure was applied to 10 additional persimmon cultivars, resulting in regrowth rates ranging between 67 and 97%, with an average of 87%. As shoot tips derived from dormant buds proved to be highly tolerant to liquid nitrogen exposure, the D cryo-plate technique may facilitate long-term conservation of persimmon germplasm.
Journal
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- The Horticulture Journal
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The Horticulture Journal 84 (2), 106-110, 2015
The Japanese Society for Horticultural Science
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Keywords
Details 詳細情報について
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- CRID
- 1390282680736902912
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- NII Article ID
- 120006867455
- 130005066784
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- NII Book ID
- AA12708073
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- ISSN
- 21890110
- 21890102
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- NDL BIB ID
- 026322078
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- Text Lang
- en
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- Data Source
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- JaLC
- IRDB
- NDL
- Crossref
- CiNii Articles
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- Abstract License Flag
- Disallowed