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Significance Organisms frequently exchange costly resources with other species. Theory suggests that this paradoxical cooperation between species might have its origins in waste consumption. When a species benefits from the waste of another, the recipient can evolve to aid the waste producer. The waste producer could then be selected to provide costly resources in return. We previously demonstrated the first step of this theorized process: Salmonella enterica evolved to secrete a costly amino acid to increase access to a byproduct generated by Escherichia coli . Here, we provide demonstration of a waste producer switching to costly cooperation. E. coli repeatedly evolved novel secretion of sugar to feed S. enterica . The results validate long-standing theory about the evolutionary origins of costly mutualism.
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