Cryopreservation of garlic bulbil primordia by the droplet-vitrification procedure.
The droplet-vitrification protocol, a combination of droplet-freezing and solution-based vitrification was applied for cryopreserving garlic bulbil primordia. The highest survival and regeneration percentages of cryopreserved primordia (90.1 to 95.0 percent and 82.7 to 85.0 percent, respectively) were achieved after preculture for 2-4 days at 10 degree C on solid medium with 0.1 - 0.3 M sucrose, loading for 50 minutes in liquid medium with 2 M glycerol + 0.5 M sucrose, dehydration with PVS3 vitrification solution for 90-150 min, cooling primordia in 5 microl droplets of PVS3 vitrification solution placed on aluminum foil strips by dipping these strips in liquid nitrogen, warming them by plunging the foil strips into pre-heated (40 degree C) 0.8 M sucrose solution for 30 s and further incubation in the same solution for 30 minutes. The optimized droplet-vitrification protocol was successfully applied to bulbil primordia of five garlic varieties originating from various countries and to immature bulbils of two vegetatively propagated Allium species, with regeneration percentages ranging between 77.4 - 95.4 percent.
Kim
Haeng-Hoon
HH
National Institute of Agricultural Biotechnology, RDA, Suwon 441-707, Korea. hkim@rda.go.kr
Lee
Joung-Kwan
JK
Yoon
Ju-Won
JW
Ji
Jae-Jun
JJ
Nam
Sang-Sik
SS
Hwang
Hae-Sung
HS
Cho
Eun-Gi
EG
Engelmann
Florent
F
D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D046508:Culture Techniques; D005737:Garlic / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D018520:Plant Shoots / Q000187:drug effects / Q000502:physiology*
D003451:Cryoprotective Agents