Callus formation from protoplasts of culturedSpinacia oleracea cells

Cell suspensions were initiated from plumule derived calli ofSpinacia oleracea. Some of these cell lines could be maintained in culture for at least three years without a reduced growth rate. A high yield of protoplasts was obtained from the cell suspensions. When protoplasts were cultured in Murashige and Skoog medium with naphthaleneacetic acid and 6-benzyladenine, cell wall formation was observed after three days. The cultured protoplasts produced numerous cell-clusters within two weeks. However only protoplasts isolated from suspensions which were in a rapidly dividing phase were able to divide with a high frequency and give rise to callus colonies.