Ectothermic species face problems of water balance under colder and drier climatic conditions in montane localities. We investigated five ecophysiological traits (body melanization, desiccation resistance, rate of water loss or gain and body size) in eight populations of Drosophila immigrans from an altitudinal gradient (600–2226 m) in the Western Himalayas. The traits showed bell-shaped variability patterns characteristic of quantitative traits. For body melanization, we observed high heritability (0.65) on the basis of parent–offspring regression. A comparison of highland versus foothill populations showed significant divergence for all the traits except body size. Darker flies from the highlands exhibited higher desiccation resistance but reduced rate of water loss or gain as compared with lighter flies from the foothills, which showed lower desiccation resistance and higher rates of water loss as well as gain. Lack of differences in the amount of epicuticular lipids cannot account for differential reduction in cuticular water loss in altitudinal populations. However, within- as well as between-population differences in body melanization can account for changes in desiccation resistance and reduction in cuticular water loss. Analyses of highland versus lowland populations as well as in assorted darker and lighter flies from a highland population have shown differences in haemolymph and dehydration tolerance. For the mechanistic basis of desiccation resistance, our results on wild populations of Drosophila immigrans are not in agreement with those reported for laboratory-selected desiccation-resistant strains in Drosophila melanogaster. Thus, ecophysiological mechanisms could be different under laboratory versus field selection.