This paper reports the demonstration, using fluorescence microscopy, of nucleolar DNA in two species of Drosophila. In Drosophila fulvimaculoides, the nucleolar DNA presents a variable morphology, suggestive of puffing activity. This material, which sometimes shows a banded structure like that of the polytene chromosomes, is shown not to be coextensive with the Y chromosome. Nucleolar DNA is demonstrated in Drosophila tumiditarsus also, and previous reports of an association of the dot chromosome with the nucleolus in this species are confirmed. The special usefulness of these two species for various sorts of investigation is pointed out.