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  • 1990-1994  (2)
  • Cryopreservation  (2)
  • Azaindolidine derivative
  • 1
    ISSN: 1432-203X
    Keywords: Cryopreservation ; Vitrification ; Asparagus ; In vitro ; cultured bud clusters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A culture line of asparagus forming green bulbous structures consisting of numerous multiple bud clusters designated “bud clusters” was induced from a meristem culture of asparagus (Asparagus officinalis L.cv. Hiroshimagreen, 2n=30). Small cubic segments (2 mm3) cut from bud clusters were cryopreserved using three different cryogenic protocols. Only vitrification produced very high levels of shoot formation after cooling to −196°C. Segments were treated with a vitrification solution (PVS2) at 25°C for 45 min or at 0°C for 120 min prior to a direct plunge into liquid nitrogen. After rapid warming, the segments were expelled into Murashige and Skoog medium containing 1.2 M sucrose for 10 min and then plated on agar shoot outgrowth medium. The average rate of shoot formation of vitrified segments producing normal shoots was near 90% without any preculture and/or cold-acclimation treatment. Revived segments resumed growth within 3 days and developed about three shoots per segment. In vitro-cultured bud clusters appear promising as material for cryopreserving asparagus germplasm.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-203X
    Keywords: Cryopreservation ; Vitrification ; Nucellar cells ; Navel orange ; Citrus sinensis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nucellar cells of navel orange(Citrus sinensis Osb. var. brasiliensis Tanaka) were successfully cryopreserved by vitrification. In this method, cells were sufficiently dehydrated with highly concentrated cryoprotective solution(PVS2) prior to direct plunge in liquid nitrogen. The PVS2 contains(w/v) 30% glycerol, 15% ethylene glycol and 15% DMSO in Murashige-Tucker medium(MT) containing 0.15 M sucrose. Cells were treated with 60% PVS2 at 25°C for 5 min and then chilled PVS2 at 0°C for 3 min. The cell suspension of about 0.1 ml was loaded in a 0.5 ml transparent plastic straw and directly plunged in liquid nitrogen for 30 min. After rapid warming, the cell suspension was expelled in 2 ml of MT medium containing 1.2 M sucrose. The average rate of survival was about 80%. The vitrified cells regenerated plantlets. This method is very simple and the time required for cryopreservation is only about 10 min.
    Type of Medium: Electronic Resource
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