BUSCADOR RECOLECTA

Supplementary Material 1, Biotechnology and Biological Sciences Research Council; Horizon 2020 Framework Programme; Wellcome Trust, Peer reviewed

Pleistocene Pongo teeth show substantial variation in size and morphology, fueling taxonomic debates about the paleodiversity of the genus. We investigated prominent features of the enamel-dentine-junction junction (EDJ)–phylogenetically informative internal structures–of 71 fossil Pongo lower molars from various sites by applying geometric morphometrics and conducted paleoproteomic analyses from enamel proteins to attempt to identify extinct orangutan species. Forty-three orangutan lower molars representing Pongo pygmaeus and Pongo abelii were included for comparison. The shape of the EDJ was analyzed by placing five landmarks on the tip of the main dentine horns, and 142 semilandmarks along the marginal ridges connecting the dentine horns. Paleoproteomic analyses were conducted on 15 teeth of Late Pleistocene Pongo using high-resolution tandem mass spectrometry. The geometric morphometric results show variations in EDJ shape regarding aspects of the height and position of the dentine horns and connecting ridges. Despite the issue of molar position and sample size, modern molars are distinguished from fossil counterparts by their elongated tooth outline and narrowly positioned dentine horns. Proteomic results show that neither a distinction of P. pygmaeus and P. abelii, nor a consistent allocation of fossil specimens to extant species is feasible. Based on the EDJ shape, the (late) Middle to Late Pleistocene Pongo samples from Vietnam share the same morphospace, supporting the previous allocation to P. devosi, although substantial overlap with Chinese fossils could also indicate close affinities with P. weidenreichi. The hypothesis that both species represent one chronospecies cannot be ruled out. Two fossil specimens, one from Tam Hay Marklot (Laos, Late Pleistocene), and another from Sangiran (Java, Early to Middle Pleistocene), along with some specimens within the Punung sample (Java), exhibit affinities with Pongo abelii. The Punung fossils might represent a mix of early Late Pleistocene and later specimens (terminal Pleistocene to Holocene) related to modern Pongo. The taxonomy and phylogeny of the complete Punung sample needs to be further investigated., Peer reviewed

Cross-validated classification results in frequencies based on M1–M2., Peer reviewed

Cross-validated classification results in percentages based on M1–M2., Peer reviewed

Cross-validated classification results in frequencies based on M3., Peer reviewed

Cross-validated classification results in percentages based on M3., Peer reviewed

Pleistocene Pongo teeth show substantial variation in size and morphology, fueling taxonomic debates about the paleodiversity of the genus. We investigated prominent features of the enamel-dentine-junction junction (EDJ)–phylogenetically informative internal structures–of 71 fossil Pongo lower molars from various sites by applying geometric morphometrics and conducted paleoproteomic analyses from enamel proteins to attempt to identify extinct orangutan species. Forty-three orangutan lower molars representing Pongo pygmaeus and Pongo abelii were included for comparison. The shape of the EDJ was analyzed by placing five landmarks on the tip of the main dentine horns, and 142 semilandmarks along the marginal ridges connecting the dentine horns. Paleoproteomic analyses were conducted on 15 teeth of Late Pleistocene Pongo using high-resolution tandem mass spectrometry. The geometric morphometric results show variations in EDJ shape regarding aspects of the height and position of the dentine horns and connecting ridges. Despite the issue of molar position and sample size, modern molars are distinguished from fossil counterparts by their elongated tooth outline and narrowly positioned dentine horns. Proteomic results show that neither a distinction of P. pygmaeus and P. abelii, nor a consistent allocation of fossil specimens to extant species is feasible. Based on the EDJ shape, the (late) Middle to Late Pleistocene Pongo samples from Vietnam share the same morphospace, supporting the previous allocation to P. devosi, although substantial overlap with Chinese fossils could also indicate close affinities with P. weidenreichi. The hypothesis that both species represent one chronospecies cannot be ruled out. Two fossil specimens, one from Tam Hay Marklot (Laos, Late Pleistocene), and another from Sangiran (Java, Early to Middle Pleistocene), along with some specimens within the Punung sample (Java), exhibit affinities with Pongo abelii. The Punung fossils might represent a mix of early Late Pleistocene and later specimens (terminal Pleistocene to Holocene) related to modern Pongo. The taxonomy and phylogeny of the complete Punung sample needs to be further investigated., Peer reviewed

Semilandmark coordinates for each specimen used in this study., Peer reviewed

a Cross-validated CVA of the EDJbased on M1–M2. Jackknife cross-validation was performed using the same groups as in the CVA (Fig 1A) to validate group discrimination in Fig 1A. b Cross-validated CVA of the EDJ based on M3s. Jackknife cross-validation was performed using the same groups as in the CVA (Fig 1B) to validate group discrimination in Fig 1B. (ZIP), Peer reviewed

Maximum Likelihood phylogenetic tree created by using PhyML based on protein sequences of several great ape species (Gorilla gorilla, Gorilla beringei graueri, Pan paniscus, Pan troglodytes schweinfurthii, Pan troglodytes ellioti, Pongo pygmaeus, Pongo abelii, Pongo tapanuliensis and Homo sapiens) and fossil Pongo samples included. Nomascus leucogenys was added as an outgroup. In this tree both modern and fossil Pongo are placed within the same clade but with moderate support. Some substructure can be observed on the tree but without any concrete support. (TIF), Peer reviewed