Southeast Asia is home to rich human genetic and linguistic diversity, but the details of past population movements in the region are not well known. Here, we report genome-wide ancient DNA data from 18 Southeast Asian individuals spanning from the Neolithic period through the Iron Age (4100 to 1700 years ago). Early farmers from Man Bac in Vietnam exhibit a mixture of East Asian (southern Chinese agriculturalist) and deeply diverged eastern Eurasian (hunter-gatherer) ancestry characteristic of Austroasiatic speakers, with similar ancestry as far south as Indonesia providing evidence for an expansive initial spread of Austroasiatic languages. By the Bronze Age, in a parallel pattern to Europe, sites in Vietnam and Myanmar show close connections to present-day majority groups, reflecting substantial additional influxes of migrants.

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Thalassemias are inherited blood disorders that are found in high prevalences in the Mediterranean, Southeast Asia and the Pacific. These diseases provide varying levels of resistance to malaria and are proposed to have emerged as an adaptive response to malaria in these regions. The transition to agriculture in the Holocene has been suggested to have influenced the selection for thalassemia in the Mediterranean as land clearance for farming encouraged interaction between Anopheles mosquitos, the vectors for malaria, and human groups. Here we document macroscopic and microscopic skeletal evidence for the presence of thalassemia in both hunter-gatherer (Con Co Ngua) and early agricultural (Man Bac) populations in northern Vietnam. Firstly, our findings demonstrate that thalassemia emerged prior to the transition to agriculture in Mainland Southeast Asia, from at least the early seventh millennium BP, contradicting a long-held assumption that agriculture was the main driver for an increase in malaria in Southeast Asia. Secondly, we describe evidence for significant malarial burden in the region during early agriculture. We argue that the introduction of farming into the region was not the initial driver of the selection for thalassemia, as it may have been in other regions of the world.

Between 4500 and 3500 years ago, partially intrusive Neolithic populations in the riverine basins of mainland Southeast Asia began to form mounded settlements and to develop economies based on rice cultivation, fishing, hunting, and the domestication of animals, especially pigs and dogs. A number of these sites have been excavated in recent years and they are often large mounds that can attain several meters in depth, comprising successive layers of alluvial soil brought in periodically to serve as living floors. The site of An Son is of this type and lies in a small valley immediately north of the Mekong Delta in southern Vietnam. Excavated on five occasions since 1978, and most recently in 2009, it was occupied from the late third into the late second millennium b.c . An Son has produced evidence that attests the domestication of pigs and dogs in all layers apart (perhaps) from the most basal one, which was not investigated in 2009, together with the growing of rice of the subspecies Oryza sativa japonica , of Chinese Neolithic origin. The oldest pottery has simple incised and punctate zoned decoration with parallels in central Thailand, especially in the basal phases at Nong Nor and Khok Phanom Di. From its middle and later occupation phases (1800–1200 b.c .), An Son has produced a number of supine extended burials with finely decorated pottery grave goods that carry some unique forms, especially vessels with wavy or serrated rims. The An Son burials represent a Neolithic population that expressed a mixture of both indigenous Hoabinhian and more northerly (probably Neolithic southern Chinese) cranial and dental phenotypes, perhaps representing a likely ancestral population for some of the modern Austroasiatic-speaking populations of mainland Southeast Asia.