Abstract
Crypopreservation methods were firstly developed for root-tips from hairy root cultures of Beta vulgaris, established after transformation by Agrobacterium rhizogenes. The effects of culture age, pre-growth, cryoprotection, freezing rate and post-freeze culture conditions were determined. The resulting freezing protocol was then used to cryopreserve transformed root cultures of Nicotiana rustica. Both species were viable after freezing (ca. 80%), according to fluorescein diacetate vital staining. However, on average the regeneration of proliferating roots from surviving root-tips was low (<20%). Growth rates, secondary metabolite production and T-DNA structure of a number of hairy root lines were examined and found to be unchanged after cryopreservation.
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Abbreviations
- DMSO:
-
dimethyl sulphoxide
- 2,4-D:
-
dichlorophenoxyacetic acid
- FDA:
-
fluoroscein diacetate
- IAA:
-
indole-3-acetic acid
- NAA:
-
α-napthalene acetic acid
References
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Benson, E.E., Hamill, J.D. Cryopreservation and post freeze molecular and biosynthetic stability in transformed roots of Beta vulgaris and Nicotiana rustica . Plant Cell Tiss Organ Cult 24, 163–172 (1991). https://doi.org/10.1007/BF00033472
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DOI: https://doi.org/10.1007/BF00033472