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[h1]An Ancient Harappan Genome Lacks Ancestry from Steppe Pastoralists or Iranian Farmers[/h1][h2]Summary[/h2]We report an ancient genome from the Indus Valley Civilization (IVC). The individual we sequenced fits as a mixture of people related to ancient Iranians (the largest component) and Southeast Asian hunter-gatherers, a unique profile that matches ancient DNA from 11 genetic outliers from sites in Iran and Turkmenistan in cultural communication with the IVC. These individuals had little if any Steppe pastoralist-derived ancestry, showing it was not ubiquitous in northwest South Asia during the IVC as it is today. The Iranian-related ancestry in the IVC derives from a lineage leading to early Iranian farmers, herders and hunter-gatherers before their ancestors separated, contradicting the hypothesis that the shared ancestry between early Iranians and South Asians reflects a large-scale spread of western Iranian farmers east. Instead, sampled ancient genomes from the Iranian plateau and IVC descend from different groups of hunter-gatherers who began farming without being connected by movement of people.

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Our results also have linguistic implications. One theory for the origins of the now-widespread Indo-European languages in South Asia is the "Anatolian hypothesis," which posits that the spread of these languages was propelled by movements of people from Anatolia across the Iranian plateau and into South Asia associated with the spread of farming. However, we have shown that the ancient South Asian farmers represented in the IVC Cline had negligible ancestry related to ancient Anatolian farmers, as well as an Iranian-related ancestry component distinct from sampled ancient farmers and herders in Iran. Since language spreads in pre-state societies are often accompanied by large-scale movements of people (Bellwood, 2013) these results argue against the model (Heggarty, 2019) of a trans-Iranian-plateau route for Indo-European language spread into South Asia. However, a natural route for Indo-European languages to have spread into South Asia is from Eastern Europe via Central Asia in the first half of the 2nd millennium BCE, a chain-of-transmission now documented in detail with ancient DNA. The fact that the Steppe pastoralist ancestry in South Asia matches that in Bronze Age Eastern Europe (but not Western Europe (de Barros Damgaard et al., 2018; Narasimhan et al., 2019)) provides additional evidence for this theory, as it elegantly explains the distinctive shared distinctive features of Balto-Slavic and Indo-Iranian languages (Ringe et al., 2002)

 
Skeletal DNA from a member of the ancient Indus Valley Civilization shows ancestry from ancient Iranians before their adoption of farming and from Southeast Asian hunter-gatherers, while completely lacking Steppe pastoralist ancestry.
 
However, a natural route for Indo-European languages to have spread into South Asia is from Eastern Europe via Central Asia in the first half of the 2nd millennium BCE, a chain-of-transmission now documented in detail with ancient DNA. The fact that the Steppe pastoralist ancestry in South Asia matches that in Bronze Age Eastern Europe (but not Western Europe (de Barros Damgaard et al., 2018; Narasimhan et al., 2019)) provides additional evidence for this theory, as it elegantly explains the distinctive shared distinctive features of Balto-Slavic and Indo-Iranian languages (Ringe et al., 2002)
 

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