Journal of the Japanese Society for Horticultural Science
Online ISSN : 1880-358X
Print ISSN : 0013-7626
ISSN-L : 0013-7626
Stable Maintenance of an Integrated Gene in Nucellar Cells of Navel Orange (Citrus sinensis Osb.) Under Storage in LN2
Shozo KobayashiAkira SakaiToshifumi OhgawaraYuri Nakamura
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1994 Volume 63 Issue 3 Pages 553-558


Transformed nucellar cells of navel orange obtained by electroporation, were successfully cryopreserved in liquid nitrogen (LN2) by a vitrification method. The cells treated with a concentrated cryoprotective solution (PVS2) at 25 °C for 3 min were transferred to a 1.8 nil plastic cryotube and then directly plunged into LN2. After storage for 1 year in LN2, cell viability was about 90%, as evaluated by fluorescein diacetate (FDA) and phenosafranine double staining. Southern blot analysis showed that the integrated neomycinphosphotransferase II (NTPII) gene was stably maintained in the cryopreserved-regrown cells without DNA alteration. No morphological differences were observed between plants regenerated from cryopreserved cells and those from non-cryopreserved cells.

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