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The history and origins of the donkey are particularly interesting because, together with the horse, the donkey (Equus asinus) has been used for the transportation of people, possessions, and produce in many cultures. This domestication marks a major cultural shift away from sedentary, agrarian life-styles toward more extensive movement and trade. Sparse archaeological evidence from Egypt suggests that donkeys, like horses, were domesticated 5000 years ago. Exactly where this occurred is still unclear (1, 2). We assessed domestic donkey origins by sampling donkeys from 52 countries across the Old World (table S1) and sequencing 479 base pairs (bp) of the mitochondrial DNA (mtDNA) control region (3). Phylogenetic analyses identified two highly divergent phylogenetic groups (Fig. 1) with an average corrected sequence divergence of 15.73 0.61 substitutions. Sequences from the same mtDNA control …