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  • 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
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 28 (1992), S. 261-266 
    ISSN: 1573-5044
    Keywords: apple ; cryopreservation ; fruit trees ; Malus ; pear ; Pyrus ; shoot tips ; vitrification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In vitro-grown shoot tips of apples (Malus domestica Borkh. cv. Fuji) were successfully cryopreserved by vitrification. Three-week-old in vitro apple plantlets were cold-hardened at 5°C for 3 weeks. Excised shoot tips from hardened plantlets were precultured on a solidified Murashige & Skoog agar medium (MS) supplemented with 0.7 M sucrose for 1 day at 5°C. Following preculture shoot tips were transferred to a 2 ml plastic cryotube and a highly concentrated cryoprotective solution (designated PVS2) was then added at 25°C. The PVS2 contains (W/V) 30% glycerol, 15% ethylene glycol and 15% dimethylsulfoxide in medium containing 0.4 M sucrose. After dehydration at 25°C for 80 min, the shoot tips were directly plunged into liquid nitrogen. After rapid warming, the shoot tips were expelled into 2 ml of MS medium containing 1.2 M sucrose and then plated on agar MS medium. Direct shoot elongation was observed in approximately 3 weeks. The average rate of shoot formation was about 80%. This vitrification method was successfully applied to five apple species or cultivars and eight pear cultivars. This method appears to be a promising technique for cryopreserving shoot tips from in vitro-grown plantlets of fruit trees.
    Type of Medium: Electronic Resource
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