Droplet vitrification cryopreservation of Rosa canina and Rosa rubiginosa using shoot tips from in situ plants
2014
Pawłowska, Bożena | Szewczyk-Taranek, Bożena
In this study two wild Rosa species in vivo shoot tips were successfully cryopreserved using the droplet vitrification technique. The growth recovery of cryopreserved shoot tips was satisfactory when the explants were isolated from the lateral buds in situ plants at the end of the winter dormancy period (February) and following the disinfection of the buds surface with 70% ethanol. The optimal condition included treatment for 20min in the loading solution (2M glycerol and 0.4M sucrose), treatment with plant vitrification solution (PVS2) containing 30% glycerol, 15% ethylene glycol, 15% DMSO and 0.4M sucrose for 20min (Rosa canina) or 30min (Rosa rubiginosa). A rapid freezing was conducted on strips of aluminium and, after cryopreservation, followed by a rapid, immersed re-warming for 20min in the recovery solution, which contained 1.2M sucrose and, additionally, 0.2% of sodium hypochlorite. Post-freezing regeneration was performed by the in vitro culture on MS media with 5μM BA, 1.5μM and 0.087M sucrose. Under these conditions, 93.5–96% of the shoot tips survived, and over 83.3–86.7% of these regenerated axillary shoots. This procedure facilitates the routine use of cryopreservation of rose in vivo shoot tips and eliminates the maintenance of in vitro cultures as a source of explants for cryopreservation.
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