Tissue homogenates, etioplasts, and developing chloroplasts were prepared from cucumber (Cumucis sativus L.) cotyledons in tris-sucrose. They were incubated aerobically in the dark or in the light at pH 7.7 in the presence or absence of a cofactor mixture containing coenzyme A, glutathione, potassium phosphate, methyl alcohol, magnesium, nicotinamide adenine dinucleotide, and adenosine triphosphate. These (1,6,8,9). These organelles appear to possess certain amounts of RNA and DNA as well as the machinery for the biosynthesis of lipid, protein, and nucleic acids (9). They seem capable of dividing in vitro (17) and of forming their own protochlorophyll and chlorophyll from simple substrates (14,15,22). The differentiation of etioplasts into chloroplasts during greening is accompanied by substantial synthesis and accumulation of chlorophyll (11), carotenoids (12, 13), insoluble proteins, and colorless lipids (10). Likewise, dividing chloroplasts must grow before reaching full size (17). An assessment of cellular control during chloroplast differentiation, growth, maintenance, and division is therefore indispensable in evaluating the degree of autonomy of this organelle. (14,15). Cotyledons to be preirradiated were harvested with full hypocotyl hook as previously described (7). They were placed in beakers with enough distilled H20 to keep them moist and were illuminated with 240 ft-c of white fluorescent light at 28 C for 2.5 or 4.5 hr (15).Chemicals. The commercial sources of chemicals were reported elsewhere (14, 15). Preparation of Unfortified and Fortified Crude Homogenates. "Unfortified" crude homogenates were prepared from 4.5-day-old cotyledons as follows. Four grams of etiolated or greening cotyledons were gently ground with mortar and pestle without sand in 6.0 ml of 0.5 M sucrose and 0.2 M tris-HCl, pH 8.0, at 0 to 5 C. The brei was filtered through four layers of cheesecloth. About 5 ml of unfortified crude homogenates were obtained. "Fortified" crude homogenates were prepared by grinding 4 g of etiolated or greening cotyledons in 6.0 ml of fortified tris-HCl and 0.