Plant regeneration from protoplasts isolated from embryogenic callus of satsuma

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 µM ade- nine, 2.8 µM GA3 and 400 mg malt extract/liter, solidified with Gelrite. Somatic em- bryogenesis 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 GA3. Protoplasts isolated from this callus produced so- matic embryos through colony formation: subsequently, normal, entire plants were regenerated. Systems for somatic embryogenesis and