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Chromosome localization of sequences homologous to λ20p1.4 of the Drosophila melanogaster nuclear lamina DNA (nlDNA) was established by in situ hybridization in species of the melanogastersubgroup. DNA of the λ20p1.4 clone was shown to be located in the chromocenter in all the species examined. Laboratory strains of D. simulans, D. mauritiana, andD. sechellia exhibited interspecific differences in localization of λ20p1.4 nlDNA on chromosome arms. In eight natural populations, intraspecific polymorphism of λ20p1.4 nlDNA chromosome localization was shown to be present in D. simulans but absent in D. melanogaster. The possible participation of transposable elements in λ20p1.4 nlDNA relocation is discussed.