1. The frequency of the chromosomal types of several western Mediterranean populations of D. subobscura, distributed in a zone running north–south, is analysed and compared with that from an Edinburgh (Knight, 1961) site at a similar meridian as the Mediterranean populations.2. North–south clines are found in the frequencies of several chromosomal types. Some types are more frequent in the north, decreasing gradually southwards; others show the reverse trend of variation.3. The comparison with a similar array of populations from Central Europe and the Central Mediterranean area, indicates that the chromosomal types more frequent in the northern populations are mostly the same, i.e. the standard orders. But chromosomal types with complex inversion orders are the most frequent in southern populations: in some chromosomes, different orders are predominant in Israel, southern Italy and southern Spain.4. The Pyrenees, acting as an ecological barrier, strongly influence the diversity of the populations north and south of the range. This result and the latitudinal clines support the adaptive significance of the chromosomal polymorphism in D. subobscura.5. The index of free recombination of the population from Malaga (in the south of Spain) is higher than in the populations from the northern Mediterranean area. D. subobscura apparently supports the claims of da Cunha & Dobzhansky (1954) and Carson (1955) that a higher level of chromosomal polymorphism occurs in the central areas of the distribution of a species.