Matsumoto T. auteur aut IRD Yamamoto S. auteur aut IRD Fukui K. auteur aut IRD Rafique T. auteur aut IRD Engelmann Florent auteur aut IRD Niino T. auteur aut IRD text journalArticle eng text/pdf born digital access 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 degrees 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 degrees 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 degrees 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 degrees 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 cryoplate technique may facilitate long-term conservation of persimmon germplasm. specialized dormant shoot tips genebank kaki osmoprotection 076 084 84 2 106-110 2015 2189-0102 https://www.documentation.ird.fr/hor/fdi:010064162 10.2503/hortj.MI-043 2189-0102 [F B010064162] https://www.documentation.ird.fr/hor/fdi:010064162 https://www.documentation.ird.fr/intranet/publi/2015/05/010064162.pdf Accès réservé (Intranet de l'IRD) IRD - Base Horizon / Pleins textes 2015-06-04 2017-08-23 fdi:010064162 fre