%0 Journal Article
%9 ACL : Articles dans des revues avec comité de lecture répertoriées par l'AERES
%A Kim, H. H.
%A Popova, E. V.
%A Yi, J. Y.
%A Cho, G. T.
%A Park, S. U.
%A Lee, S. C.
%A Engelmann, Florent
%T Cryopreservation of hairy roots of Rubia akane (Nakai) using a droplet-vitrification procedure
%D 2010
%L fdi:010053113
%G ENG
%J Cryoletters
%@ 0143-2044
%K droplet-vitrification ; hairy roots ; loading ; preculture ; Rubia akane ; vitrification solution
%M ISI:000286697900005
%N 6
%P 473-484
%U https://www.documentation.ird.fr/hor/fdi:010053113
%> https://www.documentation.ird.fr/intranet/publi/2011/03/010053113.pdf
%V 31
%W Horizon (IRD)
%X 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.
%$ 076