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Cryopreservation of Thymus moroderi shoot tips by vitrification and encapsulation-dehydration
Research Areas
  • Botany
Thymus moroderi Pau ex Martínez (Labiatae) is an endemism of the Southeast of Spain, classified as "vulnerable" according to the IUCN categories. In order to preserve this species, vitrification and encapsulation-dehydration methods were tested. For vitrification, shoot tips (1mm in length) were loaded for 20 minutes at room temperature with the loading solution (2M glycerol + 0.4M sucrose) and then dehydrated with the PVS2 solution for 15-105 min at 0ºC. Shoot tips were rapidly immersed in liquid nitrogen (LN). Rewarming was carried out by rapid immersion of cryovials in a water bath at 40°C for 2 min. Subsequently shoot tips were kept on unloading solution (MS + 1.2M sucrose) for 15 min at room temperature. After that, shoot tips were placed onto one sterile filter paper on top of MS solid medium containing 0.3M sucrose. After 24h in the dark, they were transferred onto regeneration medium .The first week they were incubated in darkness at 25°C. Survival rates of cryopreserved meristems increased as PVS2 exposure time increased up to 60 min (67% survival). In the case of the encapsulation-dehydration method, the influence of sucrose preculture, time of bead desiccation on silica gel and cold hardening was tested. The best results were obtained with shoot tips excised from in vitro plantlets that had been cold-hardened (10ºC under a 16h light/8h dark photoperiod, and a 10 umol/m2 s irradiance) for 5 weeks; then precultured for 48h in MS with 0.08M sucrose and dehydrated to 22% moisture before plunging in LN. Comparing both cryopreservation methods for T. moroderi, the higher survival and regeneration rates were obtained with the vitrification protocol.
Annual Meeting of The Society for Low Temperature Biology
Copenague, Dinamarca
Start Date
End Date
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Cryoletters 30(2)
  • Autor: J.L. CASAS
  • Autor: M. Elena Gonzalez Benito (UPM)
  • Autor: A. MARCO-MEDINA
Research Group, Departaments and Institutes related
  • Creador: Grupo de Investigación: Biodiversidad y conservación de recursos fitogenéticos
  • Departamento: Biología Vegetal
S2i 2020 Observatorio de investigación @ UPM con la colaboración del Consejo Social UPM
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