Changes in the plastid ultrastructure during greening in cultured soybean cells
Mesophyll cells isolated from soybeans (Glycine max, L. Merr. var. Corsoy) can be grown photoautotrophically in suspension culture. The SB-P cell line can be bleached by maintaining them in total darkness in sucrose supplemented media for several weeks, and will regain photosynthetic competency when returned to the light. This system is ideally suited for the study of gene regulation and the biochemical and ultrastructural changes which occur during the greening process.Cells were fixed for electron microscopy after 8 weeks of growth in total darkness and at intervals of 1 h to 12 d after transfer to the light. Chlorophyll measurements were determined for each sample. For transmission electron microscopy, the cells were fixed for 1-2 h in 4% glutaraldehyde in 0.1M Pipes buffer, pH 7.4, washed in the same buffer, then postfixed for 1 h in 1% OsO4 in Pipes, pH 6.8. Following a graded ethanol dehydration series the cells were transferred into propylene oxide and embedded in Epon. Sections were stained with uranyl acetate and observed on a JEOL 100C TEM operated at 80 kV.