Natural alleles at the <i>Doa</i> locus underpin evolutionary changes in <i>Drosophila</i> lifespan and fecundity.

'Evolve and resequence' (E&amp;R) studies in <i>Drosophila melanogaster</i> have identified many candidate loci underlying the evolution of ageing and life history, but experiments that validate the effects of such candidates remain rare. In a recent E&amp;R study we have identified several alleles of the LAMMER kinase <i>Darkener of apricot</i> (<i>Doa</i>) as candidates for evolutionary changes in lifespan and fecundity. Here, we use two complementary approaches to confirm a functional role of <i>Doa</i> in life-history evolution. First, we used transgenic RNAi to study the effects of <i>Doa</i> at the whole-gene level. Ubiquitous silencing of expression in adult flies reduced both lifespan and fecundity, indicating pleiotropic effects. Second, to characterize segregating variation at <i>Doa</i>, we examined four candidate single nucleotide polymorphisms (SNPs; <i>Doa-1</i>, -<i>2</i>, -<i>3</i>, <i>-4</i>) using a genetic association approach. Three candidate SNPs had effects that were qualitatively consistent with expectations based on our E&amp;R study: <i>Doa-2</i> pleiotropically affected both lifespan and late-life fecundity; <i>Doa-1</i> affected lifespan (but not fecundity); and <i>Doa-4</i> affected late-life fecundity (but not lifespan). Finally, the last candidate allele (<i>Doa-3</i>) also affected lifespan, but in the opposite direction from predicted.