Cryopreservation of potato cultivated varieties and wild species: critical factors in droplet vitrification.
The applicability of cryopreservation protocols to a broad range of genotypes is a key issue for genebanks. We tried to identify the critical factors causing differences in survival of cryopreserved shoot tips using potato varieties coming from cultivated and wild species. The droplet-vitrification method, a combination of droplet-freezing and solution-based vitrification, was selected from several protocols. High survival after freezing was observed after dehydration with PVS2 for 20 min, cooling shoot tips placed in a droplet of PVS2 solution on aluminum foil strips by immersing the foil strips in liquid nitrogen, warming them by plunging the foil strips into a 0.8 M sucrose solution (at 40 degrees C) for 30 s and unloading in 0.8 M sucrose for 30 min. This optimized protocol was successfully applied to 12 accessions with survival ranging between 64.0 and 94.4%.
Kim
H H
HH
National Institute of Agricultural Biotechnology, RDA, Suwon 441-707, Korea. hkim@rda.go.kr
Yoon
J W
JW
P
Y E
YE
Cho
E G
EG
Sohn
J K
JK
Kim
T K
TK
Engelmann
F
F
D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D003247:Conservation of Natural Resources; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D003470:Culture Media / Q000494:pharmacology; D046508:Culture Techniques / Q000379:methods; D004305:Dose-Response Relationship, Drug; D005838:Genotype; D005990:Glycerol / Q000494:pharmacology; D018520:Plant Shoots / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D032322:Solanum / Q000166:cytology / Q000235:genetics* / Q000502:physiology*; D013395:Sucrose / Q000494:pharmacology; D013696:Temperature; D013997:Time Factors
D003451:Cryoprotective Agents; D003470:Culture Media; D013395:Sucrose; D005990:Glycerol