Cryopreservation of hairy roots of Rubia akane (Nakai) using a droplet-vitrification procedure.
Cryo Letters
; 31(6): 473-84, 2010.
Article
en En
| MEDLINE
| ID: mdl-21410016
ABSTRACT
An efficient protocol for the cryopreservation of madder (Rubia akane Nakai) hairy root cultures was developed using droplet-vitrification and alternative loading and vitrification solutions formulated previously in our laboratory. Among eight preculture treatments tested, the highest post-cryopreservation regeneration was obtained for explants incubated in liquid half-strength MS medium with progressively increased sucrose concentration (0.3 M for 54 h, then 0.5 M for 16 h). Loading of precultured explants improved their post-cryopreservation regeneration by 50-75% compared with non-loaded control. Combining loading solution C4 (35% PVS3) and vitrification solution B5 (80% PVS3) was the most effective, while applying six PVS2-based solutions at room temperature resulted in low post-cryo regeneration. Treatment duration was optimized to 30 min for loading and to 10-20 min for vitrification solution. Apices of primary and secondary hairy roots showed similar post-cryo regeneration (88 and 95%, respectively), which was significantly higher than regeneration of root sections without apices (65%). Droplet-vitrification produced higher post-cryo regeneration than 'classical' vitrification in cryovials. Our results suggest that droplet-vitrification using alternative loading and vitrification solutions is an efficient method for cryopreservation of R. akane hairy root cultures.
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Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Criopreservación
/
Crioprotectores
/
Rubia
Tipo de estudio:
Guideline
Idioma:
En
Revista:
Cryo Letters
Asunto de la revista:
BIOLOGIA
/
QUIMICA
Año:
2010
Tipo del documento:
Article