Recent studies by various authors suggest that variation in gene regulation may be common in nature, and might be of great evolutionary consequence; but the ascertainment of variation in gene regulation has proven to be a difficult problem. In this study, we explore this problem by measuring alcohol dehydrogenase (ADH) activity in Drosophila melanogaster strains homozygous for various combinations of given second and third chromosomes sampled from a natural population. The structural locus (Adh) coding for ADH is on the second chromosome. The results show that: (1) there are genes, other than Adh, that affect the levels of ADH activity; (2) at least some of these "regulatory" genes are located on the third chromosome, and thus are not adjacent to the Adh locus; (3) variation exists in natural populations for such regulatory genes; (4) the effect of these regulatory genes varies as they interact with different second chromosomes; (5) third chromosomes with high-activity genes are either partially or completely dominant over chromosomes with low-activity genes; (6) the effects of the regulatory genes are pervasive throughout development; and (7) the third chromosome genes regulate the levels of ADH activity by affecting the number of ADH molecules in the flies. The results are consistent with the view that the evolution of regulatory genes may play an important role in adaptation.

The activity levels of alcohol dehydrogenase and alpha-glycerophosphate dehydrogenase were compared among nine species of Drosophila representing three phylogenetic groups. For any given life stage , interspecific variability in activity level was much greater for ADH than for d-GPDH. Patterns of ontogenetic expression of enzyme activity were also much more variable among species for ADH than for alpha-GPDH. These results are consistent with the interpretation that alpha-GPDH is involved with a relatively uniform adaptive function among species, whereas ADH levels may reflect variable adaptive capabilities. There is a significant correlation between ADH activities and survivorship on alcohol-treated media for these nine species.

Previous workers (McKenzie and Parsons, 1972, 1974; McKenzie, 1974; Briscoe et al., 1975) have found anomalous distributions of species of Drosophila, of sexes of D. melanogaster, and of Adh alleles in and around wineries in Australia and Spain. Field studies in California's Sonoma Valley provide evidence that the explanations advanced for these distributions may incorrect. The anomalous distribution of species was attributed to alcohol, either as a selective agent or as a behavioral stimulus. We find a virtually identical species distribution in the absence of environmental alcohol. The anomalous sex ratio was attributedd to differential survivall of the sexes when raised on alcohol. We present crude evidence thatehe difference may simply be a behavioral response to some product of fermentation, which need not be alcohol. Finally, the allele frequency difference reported from Spain was attributed to differential adult mortality on alcohol. We do not find an allele frequency difference even when alcohol is exposed, and therefore suggest that selection is occurring in pre-adult stages.

Starch and polyacrylamide gel electrophoresis were used to ascertain the substrate specificities of alcohol-oxidizing enzymes in 13 Drosophila species. The substrates used were a variety of long- and short-chain aliphatic alcohols, one aromatic alcohol, and benzaldehyde. Only one enzyme (product of a single-gene locus) showed significant NAD+-dependent alcohol dehydrogenase activity with short-chain aliphatic alcohols. The 13 species, belonging to four different Drosophila groups, all showed a similar complement of alcohol-oxidizing enzymes, although differences in electrophoretic mobility and in levels of activity existed from species to species. These findings are relevant to the adaptation of Drosophila to alcohol environments.