Abstract ‘Evolve and resequence’ (E&amp;R) studies in Drosophila melanogaster 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 Darkener of apricot ( Doa ) as candidates for evolutionary changes in lifespan and fecundity. Here, we use two complementary approaches to confirm a functional role of Doa in life-history evolution. First, we used transgenic RNAi to study the effects of Doa 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 Doa , we examined four candidate single nucleotide polymorphisms (SNPs; Doa-1 , − 2 , − 3 , − 4 ) using a genetic association approach. Three candidate SNPs had effects that were qualitatively consistent with expectations based on our E&amp;R study: Doa-2 pleiotropically affected both lifespan and late-life fecundity; Doa-1 affected lifespan (but not fecundity); and Doa-4 affected late-life fecundity (but not lifespan). Finally, the last candidate allele ( Doa-3 ) also affected lifespan, but in the opposite direction than predicted.

Natural alleles at theDoalocus underpin evolutionary changes inDrosophilalifespan and fecundity