Cryopreservation of Ginkgo biloba cell culture: effect of pretreatment with sucrose and ABA

Cryo Letters. 2009 May-Jun;30(3):232-43.

Abstract

The report describes the impact of preculture with sucrose and sucrose + ABA on desiccation and cryopreservation tolerance of cell cultures of Ginkgo biloba L., an important landscape and medicinal tree. Callus clumps were incubated on MS medium supplemented with high sucrose concentrations (up to 24 percent, w/v), employed alone or with ABA (2-10 mg per L) for various durations followed by desiccation for 0-240 min and cryopreservation. The beneficial effect of preculture on regrowth after desiccation without cryopreservation was only observed for the cells with water content of 20 percent FW and was not influenced by presence of ABA. However, preculture of calli in presence of ABA resulted in a lower desiccation rate as compared with untreated controls and calli pretreated with sucrose alone. In calli precultured with sucrose alone, post-thaw regrowth was occasional regardless of the sugar concentration in the medium, while pretreatment of calli with ABA and sucrose ensured stable regrowth after cryopreservation. The highest post-thaw regrowth of 54 percent was achieved for calli precultured on medium supplemented with 10 percent (w/v) sucrose and 2 mg per L ABA for 21 days followed by desiccation for 150 min. The different effects of preculture treatments on post-thaw regrowth were associated with significant changes in content and in composition of endogenous soluble sugars in calli. Sucrose and glucose accumulated preferentially in ABA-precultured calli, while the fructose content was higher in calli precultured in absence of ABA. The possible role of preculture on desiccation and cryopreservation tolerance of G. biloba cell cultures is discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abscisic Acid / pharmacology*
  • Cell Survival / drug effects
  • Cell Survival / physiology
  • Cells, Cultured
  • Cryopreservation / methods*
  • Cryoprotective Agents / pharmacology
  • Dose-Response Relationship, Drug
  • Ginkgo biloba / cytology*
  • Ginkgo biloba / drug effects*
  • Ginkgo biloba / growth & development
  • Plant Growth Regulators / pharmacology
  • Sucrose / pharmacology*

Substances

  • Cryoprotective Agents
  • Plant Growth Regulators
  • Sucrose
  • Abscisic Acid