SignificanceWhile tobacco is one of the most heavily consumed (and abused) plant substances of the modern era, with profound global health consequences, its early use remains poorly understood. Here we report a substantial direct biomolecular record of ancient tobacco smoking by hunter-gatherers of interior northwestern North America. Nicotine-positive samples demonstrate deep time continuity of indigenous tobacco smoking in a place where tobacco has been depicted as being introduced by early Euro-American traders and explorers. The spread of domesticated trade tobacco seems to have overtaken and obscured ancient indigenous tobacco practices. The information—represented here by the longest continuous biomolecular record of tobacco use from a single region—informs programs designed to combat persistent commercial tobacco use rates among modern Tribal communities.
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