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Free Content Long-term Preconditioning of Plantlets: a Practical Method for Enhancing Survival of Pineapple (Ananas Comosus (l.) Merr.) Shoot Tips Cryopreserved Using Vitrification

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BACKGROUND: The purpose of this study was to develop an efficient cryopreservation protocol for pineapple (Ananas comosus (L.) Merr.) shoot tips. OBJECTIVE: The optimal state of pineapple plantlets was investigated by using sucrose preconditioning to enhance survival after cryostorage. METHODS: To achieve a suitable state of plantlets before cryopreservation, 0.2 M to 0.4 M sucrose concentrations combined with short- (0–7 days), medium- (15–30 days), and long-term (75–150 days) preconditioning periods were compared. RESULTS AND CONCLUSION: The highest survival (100%) was achieved using the following procedure: intact plantlets underwent long-term preconditioning with 0.2 M sucrose for 135 days, dissected shoot tips were treated with a loading solution containing 2.0 M glycerol + 0.4 M sucrose for 60 min at 25°C and the shoot tips were dehydrated in PVS2 for 2 h at 0°C before being plunged in liquid nitrogen. Rewarming was conducted in a water-bath for 30 s at 40°C and PVS2 was replaced with a 1.2 M sucrose solution for 30 min at 25°C. The shoot tips were transferred on semisolid medium and left in the dark for 1 week, then in dim light for 3 weeks.

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Document Type: Research Article

Publication date: July 1, 2015

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  • CryoLetters is a bimonthly international journal for low temperature sciences, including cryobiology, cryopreservation or vitrification of cells and tissues, chemical and physical aspects of freezing and drying, and studies involving ecology of cold environments, and cold adaptation

    The journal publishes original research reports, authoritative reviews, technical developments and commissioned book reviews of studies of the effects produced by low temperatures on a wide variety of scientific and technical processes, or those involving low temperature techniques in the investigation of physical, chemical, biological and ecological problems.

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