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Cells of Euglena gracilis Klebs var. bacillaris Cori mutant W3BUL grown in darkness on Hutner's pH 3.5 medium without agitation accumulate wax ester. These cells have undeveloped proplastid remnants characteristic of this mutant. If these cells are transferred to an inorganic medium and bubbled with 2-3% CO2 in air, the wax disappears and the proplastid expands and develops in darkness to form prolamellar bodies and membrane vessicles within 96 h. No further development takes place in darkness, but if these cultures are illuminated at 96 h formation of prothylakoids is observed. Thus the wax ester accumulated during growth can be used subsequently to support proplastid development up to the prolamellar body stage, but the formation of prothylakoids is strictly light-dependent. Development in this system takes place at a slower rate than in cells grown with shaking and lacking wax which are transferred to resting medium. As previously shown, all of proplastid development requires light under these conditions. It is suggested that the oxygen-requiring utilization of wax in darkness can provide energy and metabolites for a part of proplastid development but the later steps in these cells, or the entire development in cells lacking wax is supported by paramylum degradation which is strictly light-dependent. However, a specific light reaction required for prothylakoid organization is not ruled out.

Light-independent and dependent phases of proplastid development in Euglena gracilis W3BUL.