Polyribosome Formation in Relation to Cytokinin-induced Cell Division in Suspension Cultures of Glycine max [L.] Merr.

Abstract

We have investigated the relationship between cell proliferation and protein synthetic capacity in a cytokinin-requiring strain of cultured soybean cells (Glycine max [L.] Merr. cv. Sodifuri, of cotyledonary origin) in suspension culture. When transferred to a defined medium lacking cytokinin, very little cell division or cell enlargement took place over the course of a 6-day culture period. Cells transferred to medium of the same composition, but containing 0.5 μm zeatin, exhibited rapid initial growth, with maximum mitotic activity occurring after 24 hours in culture, and a doubling of the cell population within the first 36 hours of the culture period. The polyribosomal RNA content of the cells decreased over the course of the first 24 hours of the growth cycle while the polyribosome to monoribosome (P/M) ratio increased. The increase in the P/M ratio was greater in the cytokinin-treated cells. This apparent relationship between cytokinin-induced cell proliferation and polyribosome formation was examined further. Polyribosome formation was stimulated when zeatin was added directly to cell populations which had been cultured for 24 hours in medium lacking a cytokinin. Transfer to fresh medium alone also stimulated polyribosome formation, whether this medium contained a cytokinin or not. The magnitude of transfer-induced polyribosome formation depended upon the initial cell density (number of cells/ml of medium). Regardless of the initial cell density and independent of the P/M ratios attained, the cytokinin-treated cell populations divided while the cytokinin-deprived cell populations did not. In vivo labeling with [³⁵S]methionine and slab gel electrophoretic separation of sodium dodecyl sulfate derivatives of the labeled polypeptides demonstrated qualitative changes in the spectrum of proteins synthesized by the cytokinin-treated cells. These qualitative changes were independent of the cell density (and hence, independent of the P/M ratio) but they preceded cytokinin-induced cell division.