Cryopreservation of Limonium serotinum apical meristems from in vitro plantlets using droplet-vitrification |
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| Authors: | Giuseppe Barraco Isabelle Sylvestre Giovanni Iapichino Florent Engelmann |
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| Institution: | 1. IRD, UMR DIADE, 911 avenue Agropolis, BP 64501, 34394 Montpellier cedex 5, France;2. Università degli Studi di Palermo, Facoltà di Agraria, Viale delle Scienze ed. 4, 90128 Palermo, Italy;3. Bioversity International, Via dei Tre Denari 472/a, 00057 Maccarese (Fiumicino), Rome, Italy |
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| Abstract: | In this study in vitro shoot tips of a Sicilian genotype of Limonium serotinum were successfully cryopreserved using the droplet-vitrification technique. Growth recovery of cryopreserved shoot tips was possible only when samples were pretreated for 16 h in liquid medium with 0.3 M sucrose, then for 5 h in liquid medium with 0.7 M sucrose before performing the cryopreservation protocol. Optimal conditions included treatment for 20 min in a loading solution containing 1.9 M glycerol + 0.5 M sucrose, treatment with vitrification solution B5 (glycerol 40.0%, sucrose 40.0%, w/v) for 60 and 90 min or vitrification solution A9 (glycerol 30.0%, dimethylsulfoxide 20.0%, ethylene glycol 20.0%, sucrose 15.0%) for 20 min, rapid cooling in minute droplets of vitrification solution, rapid rewarming by immersion for 20 min in unloading solution containing 1.2 M sucrose. Under these conditions, 37% recovery of cryopreserved shoot tips was achieved. Regrowth of cryopreserved samples was slow but always direct, without callus formation. |
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| Keywords: | Cryopreservation Droplet-vitrification Limonium serotinum Statice Sucrose pretreatment Vitrification solution |
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