title: Cryopreservation of Citrus madurensis embryonic axes by vitrification: importance of loading and treatment with vitrification solution.
abstract: 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.
authors.affiliation: 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.
authors.forename: Eun Gi
authors.initials: EG
authors.lastname: Cho
authors.forename: Y L
authors.initials: YL
authors.lastname: Hor
authors.forename: Haeng Hoon
authors.initials: HH
authors.lastname: Kim
authors.forename: V Ramanatha
authors.initials: VR
authors.lastname: Rao
authors.forename: Florent
authors.initials: F
authors.lastname: Engelmann
mesh_terms: 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
chemical_list: D003451:Cryoprotective Agents; D012996:Solutions