Donkey domestication represents a pivotal milestone in human history, driving the expansion of trade networks and adaptation to arid environments. This chapter synthesizes archaeological, genomic, and ecological evidence to reconstruct the origins, global dispersal, and breed diversification of donkeys (Equus asinus). Comprehensive analyses of mitochondrial haplogroups, nuclear genome variation, and Y-chromosome patterns reveal a single domestication event in Northeast Africa within a genetically diverse wild ass population, giving rise to Haplogroups A and B. Subsequent dispersals across Africa, Asia, and Europe were shaped by environmental constraints, hybridization events, and selective breeding. Genome sequencing highlights functional adaptations in energy metabolism, immunity, and coat color genetics, while molecular differentiation studies illuminate breed-specific traits and phenotypic diversity, such as long-hair phenotypes and color variants linked to TBX3, ASIP, and KIT genes. Conservation concerns emerge from ongoing population declines, driven by mechanization, market pressures (e.g., Ejiao industry), and neglect of traditional breeds. Integrating molecular insights with historical context underscores the importance of genomic tools for safeguarding donkey genetic diversity, informing conservation strategies, and unraveling the evolutionary dynamics of one of humanity’s oldest domesticated species.