ISSN:
1573-5044
Keywords:
freeze-preservation
;
genetic conservation
;
nucellar callus
;
suspension culture
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract Suspension cultured cells of nucellar callus of navel orange (Citrus sinensis Osb. var. brasiliensis Tanaka) were successfully cryopreserved. The nucellar cells were cryoprotected in Murashige-Tucker basal medium supplemented with 5% DMSO+1.2 M sucrose in an ice bath for 1 h, and then were frozen in this solution at a cooling rate of 0.5°C/min to −40°C prior to immersion in LN2. After rapid thawing in a +40°C water bath, regrowth was achieved by transferring the treated cells, without washing, onto filter paper discs over nutrient media solidified with agar. The viability after thawing, as evaluated by FDA and phenosafranine double staining, was about 70% of controls. The revived cells resumed growth within 3 days and produced cotyledonary embryos that developed into plants within 2 to 6 months of culture. Plants regenerated from cryopreserved cells were morphologically uniform and had the characteristics typical of navel orange.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00116084
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