Cryopreservation and post freeze molecular and biosynthetic stability in transformed roots of Beta vulgaris and Nicotiana rustica |
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| Authors: | Erica E. Benson John D. Hamill |
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| Affiliation: | (1) Department of Agriculture and Horticulture, Nottingham University School of Agriculture, Sutton Bonington, LE12 5RD Loughborough, Leics, UK;(2) Plant Cell Culture Group, AFRC Food Research Council Institute, Norwich Colney Lane, NR4 7UA Norwich, UK;(3) Present address: Department of Biology, University of Crete, P.O. Box 1470, 71110 Heraklion, Crete, Greece;(4) Present address: Department of Genetics and Developmental Biology, Monash University, Clayton 3168, Melbourne, Victoria, Australia |
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| 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.Abbreviations DMSO dimethyl sulphoxide - 2,4-D dichlorophenoxyacetic acid - FDA fluoroscein diacetate - IAA indole-3-acetic acid - NAA  -napthalene acetic acid |
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| Keywords: | Agrobacterium rhizogenes cryopreservation hairy roots molecular stability secondary metabolites transformation |
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