Gillott M A, Erdös G, Buetow D E
Center for Electron Microscopy, University of Illinois, Urbana, Illinois 61801.
Plant Physiol. 1991 Jul;96(3):962-70. doi: 10.1104/pp.96.3.962.
Suspension cultures of SB-P cells of soybean (Glycine max) provide a novel, reproducible, and readily manipulable greening system useful for inducing chloroplast differentiation. The cells are subcultured and grown heterotrophically (3% sucrose) in the dark for at least three successive 14-day periods, subcultured and grown in the dark for 7 days more, and finally placed under white light and grown photoautotrophically. Chlorophyll begins to accumulate by 1 hour of light and continues up to 12 days. The chlorophyll a:chlorophyll b ratio is 3:1. Dark-grown cells contain a small amount of total carotenoids which increase 10-fold during greening. Chloroplast differentiation is strictly light dependent, with photosynthetic pigments accumulating in the light and being lost from cells returned to the dark. In the dark, the chloroplasts dedifferentiate to amyloplasts as the organized thylakoid network is lost and starch accumulates. Under continuous light, the amyloplasts differentiate into mature chloroplasts as the organelle elongates, becomes spanned by several bands of thylakoids, and undergoes grana formation. Chloroplast differentiation in SB-P cells is similar to that in intact angiosperms developing under normal light-dark cycles.
大豆（Glycine max）SB-P细胞的悬浮培养提供了一种新颖、可重复且易于操作的绿化系统，可用于诱导叶绿体分化。细胞在黑暗中以异养方式（3%蔗糖）继代培养并生长至少三个连续的14天周期，再继代培养并在黑暗中生长7天，最后置于白光下进行光自养生长。光照1小时后叶绿素开始积累，并持续至12天。叶绿素a与叶绿素b的比例为3:1。黑暗生长的细胞含有少量总类胡萝卜素，在绿化过程中增加10倍。叶绿体分化严格依赖光照，光合色素在光照下积累，而当细胞回到黑暗中时会丢失。在黑暗中，随着有组织的类囊体网络消失且淀粉积累，叶绿体去分化为造粉体。在连续光照下，随着细胞器伸长、被几条类囊体带跨越并形成基粒，造粉体分化为成熟叶绿体。SB-P细胞中的叶绿体分化与在正常光暗周期下发育的完整被子植物中的叶绿体分化相似。
