A number of experimental populations of Drosophila melanogaster either monochromosomal (e.g. isogenic) or polychromosomal (e.g. random heterozygous) with respect to their second chromosome were obtained by use of the L Cy/Pm marker strain. All populations were initially lethal free. Every other generation was, however, subsequently treated with the mutagen EMS. Between the EMS treatments samples were taken and analyzed with respect to second chromosome lethal frequency. In spite of a great fluctuation in lethal frequencies, the general tendency of the accumulation of recessive lethals was higher in monothan in polychromosomal populations. Allelism tests showed a very high rate of identical lethals in monochromosomal as well as in polychromosomal populations. Hence, some of the induced lethals seem to be heterotie. Viability tests were performed in some of the initial populations and again after EMS treatment in generation 19. The mean viabilities of homozygotes for second chromosomes were rather constant in a polyehromosomal population but were highly reduced in three out of four monochromosomal populations. This, again, may be due to a higher tendency of load accumulation in monochromosomal populations.