Characteristics of Cold-Preserved Embryogenic Suspension Cells in Fennel, Foeniculum vulgare Miller

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An embryogenic suspension cell line of fennel, Foeniculum vulgare Miller, directly induced from hypocotyl, was developed. The suspension cells were cold-preserved at 4 °C for up to 12 weeks and then cultured at 25 °C for 2 weeks. The packed cell volume of the cultured cells decreased as the cold-preservation period became longer. The cells, with a size of 32-82μm and having embryogenic potential, diminished suddenly with the passage of the cold-preservation period. The cells cold-preserved for 2 to 6 weeks were capable of forming normal somatic embryos that were identical to those obtained from the cells without cold-preservation. The resulting somatic embryos regenerated to develop into normal plantlets when cultured in hormone-free Murashige and Skoog medium under illumination. Esterase isozyme analysis on polyacrylamide gel electrophoresis revealed the appearance of different bands in the embryos treated with 4-week and 6-week cold-preservation, in contrast to the embryos from the cells without cold-preservation. Anethole was detected in the methanol extracts of embryos induced from the cold-preserved cells at the same level as that of embryos induced from the cells without cold-preservation.