A photomap of the polythenic chromosomes of D. malerkotliana was constructed to facilitate the identification of the chromosomal arms and their sections, in order to allow the identification of the breaking points of inversions and the location of the bands marked by in situ hybridization. The photomap of the polythenic chromosomes of D. malerkotliana was constructed with the six chromosome arms corresponding to pairs I (chromosome X), II and III, which are submetacentric and pair IV and Y chromosome were not included because they were not visualized in the material examined, probably due to their small size. The in-situ hybridization was performed on polythenic chromosome slides of 3 individuals. Fourteen marks were observed in Sect. 98 of the chromosomal arm IIIR, representing 70% of the markings. Other marks were found in Sect. 91 (2 marks, 10%), 87 (2 marks, 10%) and 85 (1 mark, 5%) of the same arm and in Sect. 25 (1 mark, 5%) of the IIL chromosomal arm. In this work, a photomap was made and mapped by in situ hybridization a physiologically important gene, the first being mapped in D. malerkotliana, representing the beginning of a complete study of the chromosomal evolution of the species, which will also include the induction of poufs by stress or hormones such as ecdysone.