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Embryos were obtained from unfertilized and undeveloped seeds of satsuma  (Citrus unshiu  Marc.) cultured on a modified Murashige and Tucker (MT) medium. Embryogenic callus was induced from the hypocotyl region of the embryos. The callus was successfully maintained through subculturing on MT medium with 185 μ m  ade-nine, 2.8 μ m  GA 3  and 400 mg malt extract/liter, solidified with Gelrite. Somatic embryogenesis occurred from callus subculture on medium containing 50 g lactose/liter and in the absence of plant growth regulators. Somatic embryos developed into plants on medium with sucrose and GA 3 . Protoplasts isolated from this callus produced somatic embryos through colony formation: subsequently, normal, entire plants were regenerated.

Plant Regeneration from Protoplasts Isolated from Embryogenic Callus of Satsuma