To obtain a suitable explant and efficient culture medium for plant regeneration in spinach, two cultivars of Melody and Karaj local seedlings were chosen. A hypocotyl and cotyledon segments as well as shoot tip explants were dissected from seedlings. The explants were then cultured on MS medium supplemented with IAA, GA₃, NAA, and BAP and their response to this media was studied. A completely randomized design with different replicates was used to conduct the experiments. Callus was formed at the base of the hypocotyl explants on the medium containing 15 mg.l^-1 IAA, and 3.4 mg.l^-1 GA₃. Calli capable of regeneration was obtained after subculturing on the medium containing 2 mg.l^-1 IAA, and 3.4 mg.l^-1 GA₃ at the rate of 38 percent. The vitrified plantlets were abnormally glassy, and translucent which might have high water content. It was a physiological disorder which was overcome when an improved agar medium raising to 9 gl^-1 was used. Callus has been obtained only from the hypocotyl explants, while regeneration has been obtained from shoot tip cultured on the medium containing 0.02 mgl^-1 BAP at the rate of 80 percent.

Keywords: Spinacia oleracea L., Organogenesis, Hypocotyl, Indoleacetic acid (IAA ), Gibberellic acid (GA3), 6- Benzylaminopurin ( BAP )

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