The phenotypic effects of genetic and environmental manipulations have been rarely investigated simultaneously. In addition to phenotypic plasticity, their effect on the amount and directions of genetic and phenotypic variation is of particular evolutionary importance because these constitute the material for natural selection. Here, we used heterozygous insertional mutations of 16 genes involved in the formation of the Drosophila wing. The flies were raised at two developmental temperatures (18°C and 28°C). Landmark-based geometric morphometries was used to analyze the variation of the wing size and shape at different hierarchical levels: among genotypes and temperatures; among individuals within group; and fluctuating asymmetry (FA). Our results show that (1) the phenotypic effects of the mutations depend on temperature; (2) reciprocally, most mutations affect wing plasticity; (3) both temperature and mutations modify the levels of FA and of among individuals variation within lines. Remarkably, the patterns of shape FA seem unaffected by temperature whereas those associated with individual variation are systematically altered. By modifying the direction of available phenotypic variation, temperature might thus directly affect the potential for further evolution. It suggests as well that the developmental processes responsible for developmental stability and environmental canalization might be partially distinct.