Establishment of plant regeneration and cryopreservation system from zygotic embryo-derived embryogenic cell suspension cultures of Ranunculus kazusensis

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Title
Establishment of plant regeneration and cryopreservation system from zygotic embryo-derived embryogenic cell suspension cultures of Ranunculus kazusensis
Author(s)
Suk Weon Kim; Myung Jin Oh
Bibliographic Citation
Methods in Molecular Biology, vol. 547, pp. 107-115
Publication Year
2009
Abstract
This chapter describes culture conditions for high-frequency plant regeneration via somatic embryogenesis and cryopreservation from cell suspension cultures of Ranunculus kazusensis. Zygotic embryos form white nodular structures and pale-yellow calli at a frequency of 84.9% on half-strength Schenk and Hildebrandt (SH) medium supplemented with 0.1 mg/L 2,4-dichlorophenoxyacetic acid (2,4- D). However, the frequency of white nodular structure and off-white callus formation decreases to 25% with an increasing concentration of 2,4- D up to 10 mg/L cell suspension cultures are established from zygotic embryo-derived pale-yellow calli using half-strength SH medium supplemented with 0.1 mg/L 2,4- D. Upon plating onto half-strength SH basal medium, over 90% cell aggregates give rise to numerous somatic embryos and develop into plantlets. Regenerated plantlets are transplanted to pots filled with soil and grown to maturity at 90% survival rate in a growth chamber. Furthermore, we have developed the cryopreservation system using embryogenic cell suspension cultures of Ranunculus kazusensis. The re-growth rate of cryopreserved cells in 20% glycerol and 10% dimethylsulfoxide (DMSO) is 10% and 28.3%, respectively. These results show that DMSO is more effective cryoprotectant than glycerol in long-term preservation of embryogenic cell suspension cultures. The plant regeneration and cryopreservation system established in this study could be applied for mass propagation and ex situ conservation of this plant species.
Keyword
cryopreservationplant regenerationranunculus kazusensissomatic embryogenesissuspension culture
ISSN
1064-3745
Publisher
Humana Press
DOI
http://dx.doi.org/10.1007/978-1-60327-287-2_9
Type
Article
Appears in Collections:
Jeonbuk Branch Institute > Biological Resource Center > 1. Journal Articles
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