Synchronization of protoplasts from Glycine max (L.) Merr. and Brassica napus (L.)

Abstract

Cells of Glycine max originating in a suspension culture and cells of Brassica napus prepared from hypocotyls were synchronized. Synchronization was achieved by preparing protoplasts in the usual way and subsequently letting the protoplasts regenerate into cells by removing the cell-wall-digesting enzymes. More than 70% of the cells had divided synchronously at the end of the first cycle as determined by the mitotic index. The high frequency of mitosis critically depended on the osmolality of the medium. The duration of the S-phase was estimated by measuring the activity of thymidylate kinase as well as incorporation of [(3)H]deoxythymidine into acid-insoluble material. The data indicate that synchronization is induced by resetting the cell cycle.