Cryopreservation of Citrus madurensis embryonic axes by vitrification: importance of loading and treatment with vitrification solution.
This paper investigates the importance of loading and treatment with a vitrification solution on the survival of Citrus madurensis embryonic axes cryopreserved using a vitrification protocol. Among the seven different loading solutions tested, the solution containing 2 M glycerol + 0.4 M sucrose was the most efficient. Of the six vitrification solutions tested, the PVS2 vitrification solution, applied for 20 min at 25 degree C or for 60 min at 0 degree C, ensured the highest survival. A three-step vitrification protocol, involving the treatment of embryonic axes at 0 degree C with half strength PVS2 solution for 20 min then with full strength PVS2 for an additional 40 min was more efficient than a two-step protocol that involved treatment of axes directly with full strength PVS2 solution for 60 min. After rapid immersion in liquid nitrogen, rapid rewarming, unloading in a 1.2 M sucrose solution for 20 min, culture on solid medium with 0.3 M sucrose for 1 day and growth recovery for 4 weeks on standard medium, survival of C. madurensis embryonic axes reached 85 % following the three-step process, compared with 70 % for the two-step process.
Cho
Eun Gi
EG
International Plant Genetic Resources Institute (IPGRI), Regional Office for Asia, Pacific and Oceania, P.O. Box 236, UPM Post Office, 43400 Serdang, Selangor Darul Ehsan, Malaysia. e.cho@rda.go.kr.
Hor
Y L
YL
Kim
Haeng Hoon
HH
Rao
V Ramanatha
VR
Engelmann
Florent
F
D002957:Citrus / Q000196:embryology*; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D046508:Culture Techniques / Q000379:methods; D012639:Seeds / Q000187:drug effects* / Q000254:growth & development; D012996:Solutions; D015996:Survival Rate; D013696:Temperature; D013997:Time Factors
D003451:Cryoprotective Agents; D012996:Solutions