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Retention of the capacity to produce plants from protoplasts in cryopreserved cell lines of rice (Oryza sativa L.).

Authors
  • Meijer, E G
  • van Iren, F
  • Schrijnemakers, E
  • Hensgens, L A
  • van Zijderveld, M
  • Schilperoort, R A
Type
Published Article
Journal
Plant cell reports
Publication Date
Jul 01, 1991
Volume
10
Issue
4
Pages
171–174
Identifiers
DOI: 10.1007/BF00234288
PMID: 24221539
Source
Medline
License
Unknown

Abstract

A method is described for cryopreservation of cell suspension lines of rice (Oryza sativa L.) for use in protoplast research and as a way of retaining desirable characteristics of cell lines. The procedure involves pre-culture with mannitol, addition of a cryoprotectant solution of sucrose, dimethyl sulfoxide, glycerol and L-proline, two step freezing and storage in liquid nitrogen. Cells have been preserved for up to 14 months (the longest period tried in these experiments). Cryopreserved cells proliferated after plating on solid medium and new cell suspensions could be initiated within 15 days. Viable protoplasts, capable of divisions and callus formation, could be obtained 15-21 days after thawing. Variation between cell lines in terms of recovery rate after cryopreservation occurred. Differences between cell lines in plating efficiencies on solidified medium, however, contributed to this variation. Protoplasts from cryopreserved regenerable cell lines gave rise to embryogenic callus from which plants could be regenerated. These plants developed to maturity. A transformed cell line was also cryopreserved and it had retained the hygromycin resistance and regenerative capacity of the original cell line.

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