Heterochronic truncation of odontogenesis in theropod

dinosaurs provides insight into the macroevolution

of avian beaks
Shuo Wanga,b,1, Josef Stieglerc, Ping Wud, Cheng-Ming Chuongd, Dongyu Hue, Amy Balanofff, Yachun Zhoug,h,
and Xing Xug
a
Laboratory of Vertebrate Evolution, College of Life Science, Capital Normal University, Beijing 100048, China; bState Key Laboratory of Palaeobiology and
Stratigraphy, Nanjing Institute of Geology and Palaeontology, Chinese Academy of Sciences, Nanjing, 210008, China; cDepartment of Biological Sciences,
The George Washington University, Washington, DC 20052; dDepartment of Pathology, Keck School of Medicine, University of Southern California, Los
Angeles, CA 90033; ePaleontological Institute of Shenyang Normal University, Paleontological Museum of Liaoning, Key Laboratory for Evolution of Past
Life in Northeast Asia, Ministry of Land and Resources, Shenyang 110034, China; fCenter for Functional Anatomy and Evolution, Johns Hopkins University,
Baltimore, MD 21205; gKey Laboratory of Vertebrate Evolution and Human Origin of Chinese Academy of Sciences, Institute of Vertebrate Paleontology
and Paleoanthropology, Chinese Academy of Sciences, Beijing 100044, China; and hUniversity of Chinese Academy of Sciences, Beijing 100049, China

Edited by Neil H. Shubin, The University of Chicago, Chicago, IL, and approved August 18, 2017 (received for review May 15, 2017)

Beaks are innovative structures characterizing numerous tetrapod related to dietary specialization appear salient for at least initial
lineages, including birds, but little is known about how develop- rostral beak formation in theropods.
mental processes influenced the macroevolution of these important Here we provide fossil evidence consistent with postnatal
structures. Here we provide evidence of ontogenetic vestigialization truncation of odontogenesis in several lineages of theropod di-
of alveoli in two lineages of theropod dinosaurs and show that nosaurs that eventually reach complete edentulism. We hy-

EVOLUTION
these are transitional phenotypes in the evolution of beaks. One of pothesize that BMP4 (bone morphogenetic protein 4) mediated
the smallest known caenagnathid oviraptorosaurs and a small peramorphic expansions of keratinized epithelia underlying
specimen of the Early Cretaceous bird Sapeornis both possess shal- caruncles (egg teeth) are linked with progenetic truncation of
low, empty vestiges of dentary alveoli. In both individuals, the sys- odontogenesis in beaked theropod lineages, and these hetero-
tem of vestiges connects via foramina with a dorsally closed canal chronic processes combined to generate the repeated evolution
homologous to alveoli. Similar morphologies are present in Limu- of nonavian and avian beaks when induced by selection for spe-
saurus, a beaked theropod that becomes edentulous during ontog- cialized diets.
eny; and an analysis of neontological and paleontological evidence
shows that ontogenetic reduction of the dentition is a relatively Results
common phenomenon in vertebrate evolution. Based on these lines Postnatal Dental Reduction. Among oviraptorosaurian theropods,
of evidence, we propose that progressively earlier postnatal and teeth are only present in some Early Cretaceous taxa (1317),
embryonic truncation of odontogenesis corresponds with expansion whereas all other oviraptorosaurs are edentulous (18). Cae-
of rostral keratin associated with the caruncle, and these progenesis nagnathidae is a group of edentulous oviraptorosaurs that is
and peramorphosis heterochronies combine to drive the evolution characterized by a series of lateral grooves separate from each
of edentulous beaks in nonavian theropods and birds. Following other by lateral ridges (sensu ref. 19) present on the occlusal
initial apomorphic expansion of rostral keratinized epithelia in peri- surface of the dentary (Fig. 1 AC) (19), which together consti-
natal toothed theropods, beaks appear to inhibit odontogenesis as tute the lingual groove. Our computed tomography (CT) results
they grow postnatally, resulting in a sequence of common morphol- reveal that in addition to the dentarys mandibular canal (inferior
ogies. This sequence is shifted earlier in development through phy-
logeny until dentition is absent at hatching, and odontogenesis is
Significance
inhibited by beak formation in ovo.

|
ontogenetic edentulism beak evolution | tooth reduction | We identified truncation of tooth development during post-
|
Caenagnathidae Sapeornis
natal ontogeny in two theropod dinosaurs, a caenagnathid
oviraptorosaur and the Early Cretaceous bird Sapeornis. De-
velopmental and paleontological evidence each suggests den-

A t least seven transitions to edentulism occurred indepen-

dently in theropod dinosaurs (13), all presumably ac-
companied by the appearance of a horny beak (1). Although
tal reduction and beak evolution are coupled, and a sequence
of common morphologies is identified that characterizes the
multiple transitions to toothless beaks in theropod dinosaurs
the structure and morphogenetic events of beak formation have and birds. Shifts toward earlier cessation of postnatal tooth
been well studied in extant birds (46), evolutionary de- development can be identified in fish, amphibians, and mam-
velopmental mechanisms linking beak formation and tooth loss mals that are edentulous as adults; therefore the identification
have proven difficult to test, given extant models. Previous au- of similar transitions in multiple Mesozoic theropod dinosaur
thors hypothesized that avian tooth loss was due to inactivation lineages strongly implies that heterochronic truncation of
of odontogenic signaling pathways (7), but acknowledged that odontogenesis played an important role in the macroevolution
regional tooth loss initially accompanied various acquisition of of beaks in modern birds.
beaks in Cretaceous birds (8, 9). Therefore, degradation of the
odontogenic program alone cannot provide a developmental Author contributions: S.W. and J.S. designed research; S.W., J.S., P.W., C.-M.C., D.H., A.B.,
explanation for the coupling of tooth loss and beak formation Y.Z., and X.X. performed research; S.W., J.S., P.W., and C.-M.C. analyzed data; and S.W.,
(9). Macroevolutionary hypotheses for these phenomena have J.S., P.W., C.-M.C., and X.X. wrote the paper.
included weight-saving in response to the evolution of flight (1, The authors declare no conflict of interest.
10) and efficient processing of herbivorous diets (3). Weight- This article is a PNAS Direct Submission.
saving hypotheses have been rejected by recent studies (11, 12) 1
To whom correspondence should be addressed. Email: [email protected].
and fail to explain the tradeoff between tooth loss and beak This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.
development in nonvolant theropod lineages, while hypotheses 1073/pnas.1708023114/-/DCSupplemental.

www.pnas.org/cgi/doi/10.1073/pnas.1708023114 PNAS Early Edition | 1 of 6

Fig. 1. Dinosaurian and crocodilian dentaries illustrated by CT data, showing the presence of the alveolar canal and other alveolar vestiges. (A) Dorsal view
and (B) coronal section of Caenagnathasia sp. (IVPP V20377). (C) Dorsal view of rostral dentary of cf. Chirostenotes TMP 2012.12.12, and (Inset) entire
mandible. Photographs in Fig. 1C are reproduced with permission from ref. 24. Copyright 2008 Canadian Science Publishing or its licensors. (D) Mirror
symmetry of right dentary of Sapeornis chaoyangensis (LPM B00015) in lingual view showing vestigial alveoli and foramina present on the dorsolingual aspect
of the dentary. (EK) Coronal dentary sections of (E) S. chaoyangensis (LPM B00015), (F) subadult Limusaurus inextricabilis (IVPP V15923), (G) juvenile
L. inextricabilis (IVPPV15301), (H) extant Alligator sinensis (IVPP 1361), (I) extant Pavo sp. (IVPP 1032), (J) Confuciusornis sp. (IVPP V23275), and (K) Khaan
mckennai (IGM 100/973). av, alveolar vestige; mc, mandibular canal (V3); mf, Meckelian foramen; mg, Meckelian groove; sd, supradentary; t, tooth. White
stars, swollen ridge present on the lingual aspect of the dentary; red arrows, lingual groove; white arrows, foramina piercing the lingual groove on the medial
aspect of the dentary; dashed lines in A and D mark the positions of the slices shown in B and E, respectively; light yellow dashed lines in F and G mark the
dentary; orange dashed line in G marks a replacement tooth present inside an enclosed dentary alveolus. (Not to scale.)

alveolar canal, V3), a neomorphic canal is present in caenagnathid or alveolar homologs at the anterior end of the dentary which
oviraptorosaurs situated above the Meckelian groove and below are shallowest anteriorly and deepest posteriorly (Fig. 1D).
the system of lateral occlusal grooves and ridges (Fig. 1 A and B) These features are shallower than the alveolar homologs in
(2). In small caenagnathid specimens [e.g., IVPP V20377 (20) and IVPP V20377 and instead resemble the condition in large
CCMGE 401/12457 (21); see SI Appendix for the institutional caenagnathid dentaries (22). The posteriormost alveolar ho-
abbreviations] the lateral grooves are relatively deep, the lateral molog is interpreted as the position of the rostral-most dentary
ridges extend across the whole lingual groove to contact the lin- tooth in subadult Sapeornis (e.g., STM 1618) based on the
gual ridge, and the neomorphic canal communicates dorsally with relative anterior extension of the Meckelian groove (23). Poste-
lateral grooves via several small foramina (Fig. 1A in ref. 20). In rior to this alveolus there is a lingual groove situated above a canal
larger caenagnathid dentaries (e.g., Caenagnathus collinsi CMN similar in morphology and position to the one present in the
8776) (22) the lateral grooves are bounded medially by a shallow caenagnathid IVPP V20377 (Fig. 1 B and E). Several concentrated
lingual ridge, the lateral ridges fail to reach the lingual ridge, and foramina pierce the lingual groove to connect with the neo-
the grooves resemble the fossa-like alveolar vestiges present in the morphic canal where the second dentary tooth is present in other
anterior dentary of young juvenile Limusaurus (e.g., IVPP Sapeornis specimens (Fig. 1 D and E), suggesting the alveolus for
V15301) (2) and the Early Cretaceous bird Sapeornis (Fig. 1D and the second dentary tooth was remodeled in life. In LPM
SI Appendix, Table S1) (23). Rather than exiting vertically from B00015 and other Sapeornis specimens with dentary teeth (e.g.,
the floor of lateral grooves as in IVPP 20377, foramina commu- STM 1618, figure 6A in ref. 23), several slit-like foramina enter
nicating with the interior dentary in larger specimens (Fig. 1C) the lingual groove posteriorly to connect with the neomorphic
(22, 24) are minute and extend onto the medial face of the den- canal, each of them slightly shorter than the anteroposterior length
tarys buccal edge as anastomosing vascular grooves, an osteo- of the anterior alveolar homologs. The length and regular spacing of
logical correlate of a keratinous rhamphotheca (25). these foramina suggest they are also alveolar vestiges.
Although extant birds lack dentition, most known Mesozoic An unusual form of edentulism known as ontogenetic edentulism
birds possess teeth and exhibit various tooth reduction patterns occurs in some extant vertebrates (26), and we reported it
(1). Some specimens of Sapeornis (STM 1618 and STM 157) recently in the Jurassic ceratosaurian theropod Limusaurus
(23) possess a combination of two dentary teeth and three fossa- inextricabilis, whereby toothed jaws in the juvenile individuals
like alveolar homologs anterior to those teeth, whereas larger transitioned to a completely toothless beaked jaw in more
specimens have a toothless dentary (e.g., BMNH C-PH1067; SI mature individuals during ontogeny (2). In mature Limusau-
Appendix, Table S2). The specimen LPM B00015 preserves no rus, vestigial dentary alveoli remain after tooth loss but are
dentary teeth (SI Appendix, Fig. S1), but there are four alveoli enclosed dorsally and modified into an alveolar canal dorsal to

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the mandibular canal (Fig. 1F) (2), as in the theropods described fails to reject this hypothesis and shows that development of the
above. A swollen ridge medial to the lingual grooves in both superficially initiated null-generation dentition is inhibited in all
caenagnathids and Sapeornis (Fig. 1 AE) resembles a similar extant tetrapods with rhamphothecae or occlusal keratinization
structure that accommodates the teeth in other archosaurs, of embryonic mouthparts (SI Appendix and SI Appendix, Fig. S2).
including juvenile Limusaurus (Fig. 1G), but is reduced in Among Archosauromorpha, the cessation of null-generation
subadult Limusaurus (Fig. 1F). Unlike these taxa, most eden- tooth development occurs penecontemporaneously with first
tulous vertebrates lack an alveolar canal bounded medially by keratinization of the caruncle in birds (7) and turtles (39),
this projecting ridge and instead possess only the mandibular whereas keratinization of the Alligator caruncle begins simulta-
canal (Fig. 1 IK). neous with initiation of the final null-generation premaxillary
These lines of evidence confirm the vestigial alveoli seen in teeth (40). All extant amniotes that possess nonvestigial true egg
Sapeornis and suggest that the lateral grooves and ridges present teeth [a hypothesized serial homolog of null-generation teeth (41,
in caenagnathid oviraptorosaurs (Fig. 1 A and C) are vestigial 42)] either lack caruncles and rhamphothecae altogether, or kera-
alveoli and interdental septa, respectively. The isolated occur- tinize the rhamphotheca postnatally [e.g., monotremes (43)], sug-
rence of most known caenagnathid mandibles and absence of gesting that transitions between aquatic and terrestrial oviparity,
ontogenetic series from a limited stratigraphic horizon in both and perhaps hard-shelled and soft-shelled oviparity or viviparity,
Caenagnathidae and Sapeornis complicates ontogenetic com- involve trade-offs between egg-pipping mechanisms that are con-
parisons, though differences in dentary tooth count in Sapeornis sequential for the phylogenetic distribution of rhamphothecae.
(2, 1, or 0) correspond roughly with body size and osteohis- We conducted tests for character correlation (44) between
tological indicators (see also SI Appendix, Table S2). Given terrestrial oviparity, mineralized columnar shell units, the pres-
the recent description of postnatal ontogenetic edentulism in ence of caruncles, and the presence of rhamphothecae on
Limusaurus (2), the identification of alveolar homologs in a species-level time-calibrated supertree of extant Tetrapoda
Caenagnathidae and Sapeornis consistent with morphologies (45). The independent evolution of caruncles in amniotes and
occurring in Limusaurus suggests that ontogenetic truncation multiple lineages of direct-developing frogs was significantly
of odontogenesis produced the observed morphologies in these correlated with terrestrial oviparity; and the evolution of rham-

EVOLUTION
theropod taxa. phothecae was significantly correlated with the presence of a
Alveolar remodeling occurs primarily on the occlusal and caruncle rather than a true egg-tooth (Table 1). These results
dorsolingual margins of the alveoli in Limusaurus, Sapeornis, and should be interpreted in light of the susceptibility of character
Caenagnathidae. In the latter two there are no preserved rem- correlation tests to Type I error (46), especially given the in-
nants of replacement teeth (Fig. 1 B and E), so a replacement frequent origins of rhamphothecae among extant taxa; however,
tooth present inside an enclosed dentary alveolus in young juv- independent acquisitions of rhamphothecae in the fossil record
enile Limusaurus deserves special note (Fig. 1G). Since the al- are heavily concentrated within Archosauromorpha, nearly all
veolus is completely enclosed on the occlusal surface, inhibiting extant members of which possess caruncles (SI Appendix and SI
normal tooth replacement, a signal extrinsic to the odontogenic
program likely initiated remodeling of the alveolus in this case.
Midtooth row alveolar closure is unusual in reptiles but is known
to result from traumatic pathology and has been observed in
extant (27) and fossil Archosauromorpha (28), including a
nonavian theropod (29). These examples confirm that epigenetic
factors such as wound healing or other tissue interactions can
inhibit odontogenesis and result in permanent closure of alveoli
with normally continuous tooth replacement. Though the precise
mechanism by which keratinized oral epithelia would affect the
aborally located dental lamina is unclear, acceleration of post-
natal growth of the keratinized beak could overgrow alveoli and
would likely induce alveolar remodeling. Keratinous epithelial
appendages are associated with delayed tooth eruption and
truncation of odontogenesis in a range of tetrapods at various
embryonic and postnatal ontogenetic stages (26, 3032), and
cytokeratin expression is up-regulated during degradation of the
successional dental lamina in monophyodont (33, 34) and di-
phyodont (35) amniotes. Given that a functional dental lamina
and appropriate expression of odontogenic pathways are essen-
tial prerequisites for vertebrate tooth formation (5, 7, 36), dental
lamina damage, loss of epithelialmesenchymal interaction, or Fig. 2. Development of chicken beak keratin. (AC) Sagittal sections of the
even misexpression of odontogenic signals caused by jaw bone snout showing expression patterns of beak keratin in wild type chicken: (A)
remodeling and/or rhamphotheca growth could have led to the E7, (B) E12, and (C) E20. Inset in A shows whole-mount in situ hybridization
tooth reduction in theropods. of beak keratin, and red arrows in AC mark beak keratin expression in
E7 egg tooth, and E12 and E20 upper and lower beaks. (DF) Sagittal sec-
Embryonic Dental Reduction. During the evolution of birds, tions of the snout showing expression patterns of beak keratin in developing
odontogenic truncation could be achieved progressively early chicken with molecular perturbation: (D) control; (E) BMP4 overexpression;
through heterochronic development. Just as postnatal growth of and (F) noggin overexpression. (G and H) Stereophotos showing ectopic
beaks and other keratinous oral structures serves to inhibit tooth expression of beak keratin caused by BMP4 overexpression: (G) control (left
lateral view); (H) ectopic expression of beak keratin on face (left lateral
replacement, the embryonic development of these structures
view); (I) coronal section of the snout in H showing in situ hybridization of
appears to play a role in the loss of early tooth generations (null beak keratin. Dashed line in H marks the position of the slice shown in I. Red
and/or first-generation teeth) as well. Previous authors noticed arrows mark positions where normal keratin is present, green arrows mark
the broad coincidence of rhamphothecae and perinatal caruncles the anteroposterior length of the keratin expression domain in the upper
in amniotes and hypothesized that the two structures are itera- beak, and blue arrows mark ectopic expression of beak keratin on the face.
tive homologs (37, 38). A survey of embryonic/larval dentitions et, egg tooth; lb, lower beak; ub, upper beak. (Scale bars, 1 mm.)

Wang et al. PNAS Early Edition | 3 of 6

Table 1. Tests of phylogenetic character correlations keratin growth (Fig. 2 GI). Though BMP4 is a necessary
Characters I (LnL) D (LnL) p odontogenic agonist, the absence of BMP4 antagonists is known
to cause incisor agenesis or malformation in Nog / (47) and
Terrestrial oviparitycaruncle 1139.8 1047.7 9.34e39 Grem2/ (48) mice, suggesting that rhamphotheca hypertrophy
Carunclebeak 147.9 137.2 0.0003 induced by BMP4 overexpression could cause an imbalance in
D, dependent model; I, independent model; LnL, mean log-likelihood.
agonist/antagonist relationships necessary for normal tooth de-
velopment. In addition, the spatiotemporal pattern of BMP4 ex-
pression in mouse oral epithelium matches apoptosis associated
Appendix, Fig. S3). Beak development in birds is initiated by with regression of vestigial tooth germs in murine diastemata (49),
cornification of epithelium underlying the premaxillary caruncle and BMPs are known to regulate apoptosis in the chicken man-
on the premaxilla and a corresponding proliferative zone on the dible (50). Therefore, both alteration of epithelialmesenchymal
interactions due to keratinization of oral epithelium and early
rostral end of the mandible (Fig. 2 AC) and proceeds posteri-
disruption of odontogenic developmental pathways are potential
orly to form the premaxillary and mandibular nails (38). Growth mechanisms for inhibition of embryonic tooth development by
of rhamphothecae can be modulated in vivo by overexpression of rhamphothecae.
BMP4 or its antagonist noggin (NOG), producing peramorphic
or paedomorphic beak phenotypes at comparable ontogenetic Macroevolutionary Model for Edentulism. Evidence for postnatal
stages (Fig. 2 DF), respectively, and/or producing ectopic and embryonic ontogenetic reduction of dental tissues can be

Fig. 3. Summary of evidence for a macroevolutionary model of edentulism in vertebrates. (A) Evidence from extant nonavian vertebrate lineages with
complete or near edentulism, showing phylogenetic and trophic diversity of ontogenetic tooth loss. (B) Evidence from select theropod dinosaur lineages
focusing on Coelurosauria and showing independent evolution of adult edentulism at least seven times (17). ?, hypothesized state/partial evidence; B,
baleen; Bb, barbels; DR, sexually dimorphic reduction pattern; E, electroreception; F, filtration; G, gastric mill/ gizzard; H, expanded hypapophyses; R,
rhamphotheca; S, suction feeding; Sw, sword; T, projectile tongue. (Taxon silhouettes not to scale.)

4 of 6 | www.pnas.org/cgi/doi/10.1073/pnas.1708023114 Wang et al.

observed in numerous extant vertebrate lineages (Fig. 3A), in-
cluding the embryonic appearance of dental developmental ho-
mologs and their subsequent resorption in embryos of modern
birds (7). In most observable cases, a lineages initial phylogenetic
reduction in tooth number, size, tooth generations, and/or loca-
tion is followed by a stage of postnatal reduction and eventual
adult edentulism while juveniles retain teeth [e.g., Limusaurus
(2), Ornithorhynchus (51), and Acipenser (52, 53)].
This process is usually aided by a secondary morphological
and/or ethological tool for food acquisition or processing, the
appearance of which occurs phylogenetically before the loss of Fig. 4. Transformations involved in theropod tooth reduction. Right lateral
teeth in young juveniles (1, 26). The repeated evolution of view of the head (Left) and transverse view of the dentary (Right).
rhamphothecae and gastric mills in nonavian theropods and the (A) Normal tooth development and tooth replacement with apomorphic
addition of the crop in Cretaceous birds (Fig. 3B) provided keratinized rhamphotheca covering only the rostral-most portion of the
ample tools for food acquisition and mechanical digestion be- jaws. (B) Tooth replacement is impeded by external closure and/or con-
fore complete tooth loss in each lineage. Loss of the early tooth striction of alveoli and regional tooth reduction occurs. (C) As the kerati-
nized rhamphotheca enlarges, the remaining teeth are either functionally
generations present in young juveniles can be facilitated by
reduced or redundant. (D) Edentulous beak completely covered by rham-
parental behaviors that obviate juvenile foraging such as re- photheca. Green, mandibular canal; yellow, alveolus and alveolar canal;
gurgitation feeding in altricial birds, nursing in mammals, or orange, tooth. (Not to scale)
early competence of secondary foraging tools as in precocial
birds and anuran larvae.
Once juvenile tooth loss has occurred, relaxation of selection reptiles. BMP4-mediated expansion of keratinous caruncles in
leads to loss-of-function mutations in dental developmental ovo appears to have played an important role in initial rostral
genes and the successive vestigialization of the odontogenic beak formation and accompanying tooth loss, as well as the

EVOLUTION
program (9); and the final stages of embryonic reduction may final embryonic truncation of odontogenesis as lineages tran-
persist for tens of millions of years, as in birds and turtles (7, 39). sition to complete edentulism. Whereas birds (57) and turtles
Cases of complete embryonic or larval loss are only readily ap- (58) begin expanding the keratinized epithelium underlying the
parent in anurans, where larval keratinous structures overlie caruncle relatively early (shortly after the first elements ossify,
what will become dentigerous mouth parts (26), or where early Fig. 2 B, C, and G), crocodilians have formed multiple tooth
competence of secondary foraging or digestive features occurs, generations in ovo, and most of the skeleton has ossified by the
as in syngnathid and chanid fishes (54, 55). time rostral keratinization begins to expand beyond the caruncle
(59, 60).
Discussion Multiyear maturation and the large neonate-to-adult body size
The ubiquity of our models phenotypic succession in extant disparity of many dinosaurs and other archosauromorphs is
lineages with complete or near edentulism (Fig. 3A) and the predictive of ontogenetic niche partitioning and/or dietary
identification of these stages in various theropod dinosaur changes. Evidence for these phenomena have been observed
lineages (Fig. 3B) suggest that this heterochronic model of odon- directly in Limusaurus (2) and are probably widespread in di-
togenic truncation should be the null hypothesis for the macro- nosaurs given ontogenetic social segregation in many dinosaur
evolution of edentulism in crown birds. Heterochronic tooth clades (56) and examples of ontogenetic dietary change in many
reduction processes in nonavian and avian theropod lineages extant species within Archosauromorpha (6163). These life-
likely included the following transformations: (i) normal tooth history characteristics may have predisposed terrestrial arch-
development and tooth replacement with an apomorphic kera- osauromorphs transitioning from carnivory to omnivory or her-
tinized rhamphotheca covering only the rostral-most portion of bivory (3, 26) toward the evolution of rhamphothecae (Figs. 3B
the jaws; (ii) external closure and/or constriction of alveoli im- and SI Appendix, Fig. S3B). Tooth reduction is not as frequent or
peding tooth replacement and adjacent growth of the rham- complete in nonarchosauromorph amniotes, only rarely includes
photheca with regional tooth reduction; (iii) enlargement of the a rhamphotheca (e.g., Ornithorhynchus and Dicynodontia), and
keratinized rhamphotheca with either functional reduction or is mostly limited to aquatic, fossorial, oophagous, and myrme-
redundancy of the remaining teeth; and (iv), complete or nearly cophagous habits (Figs. 3A and SI Appendix, Fig. S3A).
complete remodeling of alveoli with complete coverage of the
edentulous beak by the rhamphotheca (Fig. 4). Morphologies Materials and Methods
consistent with this phenotypic sequence can be observed in both Details of the CT and synchrotron scanning and development experiment
ontogenetic and phylogenetic dimensions in theropods, including protocols are available in SI Appendix. For the character correlation analysis,
Limusaurus, Caenagnathidae, and Sapeornis, as well as other a species-level time-calibrated supertree was taken from Hedges et al.
Cretaceous birds (1), and it is likely only the rarity of strati- (45) (www.biodiversitycenter.org/ttol, TTOL_animals_unsmoothed) and
graphically controlled ontogenetic series and preservation biases pruned to include only Tetrapoda. The node ages of crown group Arch-
against young juveniles hindering more frequent documentation osauromorpha and Archosauria were modified to conform with minimum
of these phenomena in the fossil record. divergence dates based on the fossil record. Bayesian models of in-
This integrated developmental framework for ontogenetic dependent and dependent character evolution were estimated in Bayes-
Traits V3.0 and compared with likelihood ratio tests. See SI Appendix
tooth loss and beak formation is consistent with prevailing life-
for additional information on tree construction, sources of character in-
history characteristics of Mesozoic dinosaurs, including terres- formation, and character modeling.
trial oviparity, multiyear ontogenies with juvenile sociality, and
ontogenetic niche partitioning (2, 56). The results of the char- ACKNOWLEDGMENTS. We thank X. Ding for preparing the specimens;
acter correlation analysis show that the evolution of keratinous Y. Feng and Y. Hou for Mi-CT imaging and 3D reconstruction; and X. Ni,
rhamphothecae is correlated with the presence of a caruncle, J. Clark, and B. Zhu for discussion. S.W. was supported by the National
which is in turn correlated with terrestrial oviparity. These state- Natural Science Foundation of China (Grant 41602013), the Youth Innovative
Research Team Project of Capital Normal University, and the State Key Lab-
dependent correlations are supported by the absence of beaks in oratory of Palaeobiology and Stratigraphy, Nanjing Institute of Geology and
oviparous squamates (which have true egg teeth) and all vivip- Palaeontology, Chinese Academy of Sciences (Grant 173125); X.X. and S.W.
arous tetrapods, including mammals, squamates, and marine were supported by the National Natural Science Foundation of China (Grants

Wang et al. PNAS Early Edition | 5 of 6

41688103 and 41120124002); J.S. was supported by the U.S. National Science and C.-M.C. were supported by the National Institutes of Health (Grants
Foundation (Grants NSF EAR 0310217 and 0228559); D.H. was supported by AR47364 and AR60306); and A.B. was supported by the National Science
the National Natural Science Foundation of China (Grant 41172026); P.W. Foundation (Grant NSF DEB 1457181).

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