1. Cryopreservation of Limonium serotinum apical meristems from in vitro plantlets using droplet-vitrification
- Author
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Giuseppe Barraco, Isabelle Sylvestre, Giovanni Iapichino, Florent Engelmann, Barraco, G, Sylvestre, I, Iapichino, G, and Engelmann, F
- Subjects
Cryopreservation ,Sucrose ,Chromatography ,Callus formation ,Limonium ,Settore AGR/04 - Orticoltura E Floricoltura ,Horticulture ,Biology ,Limonium serotinum ,Vitrification solution ,biology.organism_classification ,chemistry.chemical_compound ,Cryopreservation Droplet-vitrification Limonium serotinum Statice Sucrose pretreatment Vitrification solution ,chemistry ,Droplet-vitrification ,Shoot ,Botany ,Glycerol ,Statice ,Sucrose pretreatment ,Vitrification ,Ethylene glycol - 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.
- Published
- 2011