The effect of light on the biosynthesis and the assembly of rod photoreceptor outer segment membranes was analyzed in vitro using retinas from Xenopus laevis. The number of open discs at the base of the outer segment was used as a morphologic index to evaluate relative differences in rates of membrane assembly. Assembly was stimulated in vitro by light, as evidenced by a greater number of open discs that accumulated in light-exposed retinas than that in retinas maintained under the same conditions but in darkness. Quantitative autoradiography indicated that unlike membrane assembly, the incorporation of 3H-leucine or 3H-mannose into proteins by rods was nearly identical in light-stimulated and dark-maintained retinas. The synthesis of opsin, a protein destined for the rod outer segment, was not affected by light. Apparently, the enhanced rate of outer segment membrane assembly that occurs during the early light phase of the diurnal cycle takes place in the absence of an increase in the rate of biosynthesis of opsin, the major outer segment membrane protein.