The enhanced dietary flexibility of early hominins to include consumption of C 4 /crassulacean acid metabolism (CAM) foods (i.e., foods derived from grasses, sedges, and succulents common in tropical savannas and deserts) likely represents a significant ecological and behavioral distinction from both extant great apes and the last common ancestor that we shared with great apes. Here, we use stable carbon isotopic data from 20 samples of Australopithecus afarensis from Hadar and Dikika, Ethiopia (>3.4-2.9 Ma) to show that this species consumed a diet with significant C 4 / CAM foods, differing from its putative ancestor Au. anamensis. Furthermore, there is no temporal trend in the amount of C 4 / CAM food consumption over the age of the samples analyzed, and the amount of C 4 /CAM food intake was highly variable, even within a single narrow stratigraphic interval. As such, Au. afarensis was a key participant in the C 4 /CAM dietary expansion by early australopiths of the middle Pliocene. The middle Pliocene expansion of the eastern African australopith diet to include savannabased foods represents a shift to use of plant food resources that were already abundant in hominin environments for at least 1 million y and sets the stage for dietary differentiation and niche specialization by subsequent hominin taxa.O ne of the traits that distinguishes humans from our closest living relatives, Pan and Gorilla, is the inclusion of significant quantities of C 4 /crassulacean acid metabolism (CAM) foods in the diet. C 4 /CAM plants include grasses and sedges (C 4 ) common in tropical savannas and succulents (CAM) common in deserts. Therefore, the expansion of hominin diets to consume substantial amounts of C 4 /CAM foods signals a major ecological and adaptive divergence from the last common ancestor (LCA) that we shared with African great apes, which mostly occupy closed wooded habitats. C 4 /CAM food consumption is part of a general argument, based on several lines of evidence, that hominin diets diverged from the diets of the LCA during the shift to drier and more open environments in Africa during the Pliocene (1-4). This dietary transition occurred subsequent to the known fossil record of Ardipithecus ramidus from Ethiopia at 4.4 Ma (5), which shows little evidence of C 4 /CAM food consumption. The earliest previously published hominins to evince substantial C 4 /CAM food consumption are Au. africanus at ≤2.7 Ma (2, 6-8) as well as a small, poorly age-constrained (ca. 3.0-3.5 Ma) sample of Au. bahrelghazali from west-central Africa (9). Thus, current evidence places middle Pliocene Au. afarensis, a hominin with an extensive and well-defined stratigraphic range (3.7-3.0 Ma) in Ethiopia and elsewhere in eastern Africa at the crux of this hominin dietary change.In this paper, we use stable isotope analysis to directly investigate the diet of Au. afarensis from the Hadar Formation of Ethiopia. Stable isotope analysis of mammalian tooth enamel is a paleodietary tool that allows one to determine the proportions of dietary ca...