Cryopreservation of nucellar cells of navel orange (Citrus sinensis Osb.) by a simple freezing method
1991; Elsevier BV; Volume: 74; Issue: 2 Linguagem: Inglês
10.1016/0168-9452(91)90052-a
ISSN1873-2259
AutoresAkira Sakai, Shozo Kobayashi, I. Oiyama,
Tópico(s)Transgenic Plants and Applications
ResumoSuspension cultured cells of nucellar cells of navel orange (Citrus sinensis Osb. var. brasiliensis Tanaka) was cryopreserved without using a programmable freezer and dimethyl sulfoxide (DMSO) as a cryoprotectant. The nucellar cells were cryoprotected with a mixture of 2 or 3 M glycerol and 0.4 M sucrose dissolved in Murashige-Tucker basal medium (MT) for 10 min at 25°C. A cell suspension of about 0.2 ml was loaded into a 0.5-ml trasparent straw and frozen spontaneously by placing straws in a freezer at −30°C for 20–30 min prior to direct immersion into LN. The average rate of survival after thawing, as evaluated by fluorescein diacetate and phenosafranin double staining, was about 90% of non-frozen controls. We also obtained a similar survival rate using a 1.8-ml cryotube when the cell suspension was held at −30° for 40–50 min. The revived cells resumed growth within 3 days and developed into plantlets via embryogenesis. This simple procedure for cryopreservation seems promising for cultured plant cells and meristems.
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