Abstract
Efficient plant regeneration was obtained from a cryopreserved embryogenic cell suspension of sugarcane established from leaf derived callus. Pregrowing the cells for three days in MS basal medium supplemented with 0.33 M sorbitol was essential to the process. The cells were cooled at a rate of 0.5°C/min to −40°C and then stored in liquid nitrogen. Thawing was carried out rapidly in water at +40°C, and the cells were then plated without washing onto filter paper discs placed on a semi-solid regeneration medium (MS basal + 3% sucrose + 0.13 mg/1 2,4-D +0.25 mg/1 BAP + 0.25 mg/1 kinetin + 0.25 mg/1 zeatin). The filter paper discs, along with the cells, were transferred to the same, fresh medium after five hours. After 24 hours the cells were scraped off, placed on fresh semi-solid medium and incubated at 28°C in the dark for two weeks before transfer to light. A regeneration efficiency of 92% was obtained (regenerated plants, expressed as a percent of unfrozen control). Plants regenerated from cryopreserved cells, and grown to maturity in the greenhouse, were morphologically identical to regenerated control plants.
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Abbreviations
- DMSO:
-
dimethyl sulfoxide
- PEG:
-
polyethylene glycol
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- BAP:
-
benzyl aminopurine
- TTC:
-
2,3,5-triphenyl tetrazolium chloride
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Gnanapragasam, S., Vasil, I.K. Plant regeneration from a cryopreserved embryogenic cell suspension of a commercial sugarcane hybrid (Saccharum sp.). Plant Cell Reports 9, 419–423 (1990). https://doi.org/10.1007/BF00232263
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DOI: https://doi.org/10.1007/BF00232263