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Accepted Manuscript

Molecular phylogenetics of the mud and musk turtle family Kinosternidae

John B. Iverson, Minh Le, Colleen Ingram

PII: S1055-7903(13)00251-0
DOI: http://dx.doi.org/10.1016/j.ympev.2013.06.011
Reference: YMPEV 4635

To appear in: Molecular Phylogenetics and Evolution

Received Date: 25 November 2012


Revised Date: 15 May 2013
Accepted Date: 18 June 2013

Please cite this article as: Iverson, J.B., Le, M., Ingram, C., Molecular phylogenetics of the mud and musk turtle
family Kinosternidae, Molecular Phylogenetics and Evolution (2013), doi: http://dx.doi.org/10.1016/j.ympev.
2013.06.011

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Molecular phylogenetics of the mud and musk turtle family Kinosternidae

a* b,c,d d
John B. Iverson , Minh Le , Colleen Ingram
a
Department of Biology, Earlham College, Richmond, Indiana 47374 United States
b
Faculty of Environmental Sciences, Hanoi University of Science, 334 Nguyen Trai Road,
Hanoi, Vietnam
c
Centre for Natural Resources and Environmental Studies, Vietnam National University, 19 Le
Thanh Tong Street, Hanoi, Vietnam
d
Department of Herpetology, American Museum of Natural History, Central Park West at 79th
Street, New York 10024 United States [Present address: Department of Biology,
University of Virginia, Charlottesville, Virginia 22904 United States]

ABSTRACT

The turtle family Kinosternidae comprises 25 living species of mud and musk turtles confined to
the New World. Previous attempts to reconstruct a phylogenetic history of the group have
employed morphological, isozyme, and limited mitochondrial DNA sequence data, but have not
been successful in producing a well-resolved phylogeny. With tissues from every recognized
species and most subspecies, we sequenced three mitochondrial (cyt b, 12S, 16S) and three
nuclear markers (C-mos, RAG1, RAG2). Our analyses revealed the existence of three well-
resolved clades within the Kinosterninae (aged > 22 mya), only two of which have been named:
Sternotherus and Kinosternon. We here describe the third clade as a new genus. The
evolutionary relationships among most species were well resolved, although those belonging to
the K. scorpioides species group will require more extensive geographic and genetic sampling.
Divergence time estimates and ancestral area reconstructions permitted the development of the
first rigorous hypothesis of the zoogeographic history of the group, including support for three
separate dispersals into South America, at least two of which preceded the closure of the
Panamanian portal.

Key words: Claudius, DNA, evolution, Kinosternon, Staurotypus, Sternotherus

*Corresponding author. Phone 765-983-1045; FAX +1 765 983 1497.


E-mail address: johni@earlham.edu (J. B. Iverson)
Iverson, Le, and Ingram 2

1. Introduction

The turtle family Kinosternidae includes 25 recognized extant aquatic to semiaquatic species
(38 taxa including subspecies) distributed in the New World from Canada to Argentina (Iverson,
1992a; TTWG, 2012). Although the greatest living diversity is in Mesoamerica (Iverson, 1992a,
1992b), fossil taxa are most diverse in the Eocene and Oligocene of Wyoming and South Dakota
(Hutchison, 1991). Two subfamilies, the Staurotypinae (including the genera Staurotypus and
Claudius) and the Kinosterninae (including Kinosternon and Sternotherus), have been generally
recognized as distinct, with some authors elevating them to the family level (Bickham and Carr,
1983; Vetter, 2005).
Referred to as “mud” or “musk” turtles, members of this family are small, secretive,
malodorous, and generally non-descript (Bonin et al., 2006; Schilde, 2001; Vetter, 2005). As a
result of this combination of traits, recognition of the living species diversity in the family has
been delayed compared to most other turtle families. For example, ten species (40% of total) and
8 subspecies (of 13 total) have been described since 1922 (TTWG, 2012), with one or two
species described each decade since then. Furthermore, because of a paucity of meristic
characters and the existence of significant (though drab) color variation within and among
populations, species boundaries have often been difficult to establish (e.g., Bourque, 2012c;
Iverson, 2010; Lamb and Lovich, 1990; Serb et. al., 2001). Nonetheless, undescribed taxonomic
diversity is suspected to exist (Webb, 1984; Iverson, unpublished).
The family Kinosternidae also exhibits a stunning diversity of life history traits when
compared to other turtle families. It ranges from north temperate to tropical habitats, and from
rain forest to grasslands to desert (Bonin et al., 2006). It includes totally aquatic to semi-
terrestrial species, with adult carapace lengths of 10 to 38 cm (Bonin et al., 2006), and female-
dominated to male-dominated sexual size dimorphism (Ceballos et al. 2103). At least one species
exhibits close to the maximum skeletal mass relative to body mass among all vertebrates
(Iverson, 1984). Some species have a greatly reduced plastron, whereas others have a plastron so
extensive as to completely close the shell (Hutchison, 1991). The group includes members
capable of submerged, fully aquatic respiration (Belkin, 1968), and others capable of estivating
underground for up to two years (Rose, 1980). Some species produce a single clutch in the
spring, others nest multiple times in the summer, and others nest nearly year-round (Iverson,
2010), with clutches ranging from one or two relatively huge eggs to ten or more relatively tiny
eggs (Iverson, 1999). Embryonic development is direct in some species, whereas others exhibit
early embryonic diapause and/or late embryonic estivation, with incubation times from 56 to
over 366 days (Ewert, 1991). Finally, sex determination in the family ranges from genetic (with
sex chromosomes) to temperature-dependent (Ewert et al., 2004). Unfortunately, understanding
the evolution of these diverse traits has been impeded by the lack of a well-resolved phylogeny
for the group.
Published phylogenetic hypotheses to date have been based on morphology (Iverson, 1991),
protein electromorph data (Iverson, 1991; Seidel et al., 1986), and small segments of the
mitochondrial genome, using limited taxonomic sampling (Iverson, 1998; Serb et al., 2001).
None employed complete taxonomic sampling or nuclear markers, nor applied modern
phylogenetic methods. Hence, the phylogenetic structure for the family is not well-resolved (e.g.,
see Iverson, 1998). To solve this deficiency, we sequenced 3 mitochondrial (cyt b, 12S, and 16S)
and 3 nuclear (C-mos, RAG1, and RAG2) markers from representatives of every recognized
species and most subspecies in this family (a total of 34 samples). Contemporary phylogenetic
methods were used to test previous hypotheses regarding the relationships among the known
Iverson, Le, and Ingram 3

kinosternid turtles and to direct future research in clarifying cryptic diversity in the family. In
addition, the recovered phylogeny was calibrated using known fossils to permit a reconstruction
of the zoogeographic history of the family, particularly the timing of its multiple dispersals into
South America.

2. Materials and methods

2.1 Sampling and laboratory methods

All 25 species of the family Kinosternidae were included in the study (Table S1). Outgroup
polarity was provided by the sister family Dermatemydidae (Barley et al., 2010; Krenz et al.,
2005; Near et al., 2005). We sequenced three regions of the mitochondrial genome, the complete
cytochrome b (cyt b) sequence, and the partial 12S and 16S rRNA genes (1958 total aligned bp).
We also sequenced three nuclear fragments of the C-mos, RAG1, and RAG2 genes (2553 total
aligned bp). Primers used for this study are listed in Table S2.
DNA was extracted from tissues and blood samples using the DNeasy kit (QIAGEN,
Valencia, CA, USA) following manufacturer’s instructions for animal tissues. We also extracted
DNA from bone samples using the procedures described in Le et al. (2007). PCR volume
consisted of 30µl (9µl of water, 2µl of each primers, 15µl of HotStar Taq Master Mix (QIAGEN,
Valencia, CA, USA), and 2µl of diluted DNA). PCR conditions for the mitochondrial genes
were: 95°C for 15 min to activate the Taq; with 42 cycles at 95°C for 30 s, 45°C for 45 s, 72°C
for 60 s; and a final extension of 72°C for 6 min. Nuclear DNA was amplified using the same
PCR conditions, while the annealing temperatures were 52°C for RAG1 and the second fragment
of RAG2, 56°C for the first fragment of RAG2, and 58°C for C-mos.
PCR products were visualized using electrophoresis through a 2% low melting-point agarose
gel (NuSieve GTG, FMC Biopolymers) stained with ethidium bromide and/or Safe DNA
(SYBR®). PCR products were cleaned using PerfectPrep® PCR Cleanup 96 plate (Eppendorf
Scientific Inc., Hamburg, Germany) and cycle sequenced using ABI prism big-dye terminator
(Applied Biosystems, Foster City, CA, USA) according to manufacturer recommendation.
Sequences were generated in both directions on an ABI 3130xl Genetic Analyzer (Applied
Biosystems, Foster City, CA, USA).

2.2. Phylogenetic analyses

DNA sequences were edited, checked for ambiguities, and aligned using Geneious v5.4
(Drummond et al., 2011). For coding regions, alignments were refined by eye to translated
sequences to confirm reading frame conservation and checked for premature stop codons.
The loci were analyzed individually, mtDNA only (1958 bp), nuDNA only (2553 bp) and as
a single concatenated dataset (4511 bp) under maximum likelihood (GARLI 2.0: Zwickl, 2006),
and Bayesian inference (MrBayes v3.1.2). Coding regions were partitioned into first, second, and
third position. jModelTest 2.1.1 (Darriba et al., 2012) was used to determine the appropriate
model of sequence evolution for each partition for all model-based analyses. The best models
were GTR + G + I for the mtDNA regions 16S rRNA, 12S rRNA, and the first and second
positions of cyt b; GTR + G for the third position of cyt b; HKY+ I for the first and second
positions of C-mos; GTR for the third position of C-mos and first and second positions of RAG1;
GTR + I for the third position of RAG1; HKY+G for the first and second positions of RAG2; and
HKY for the third position of RAG2. For the ML analyses, four independent searches were
performed on the concatenated mtDNA, nuDNA, and combined datasets with partitions modeled
Iverson, Le, and Ingram 4

separately. All other parameters of GARLI were left at the default settings. The four independent
searches were compared to confirm that the heuristic searches were converging on the same
likelihood and topology, and the topology with the highest likelihood value was considered the
best tree. 100 non-parametric ML bootstrap replicates were examined to determine support for
each node.
Bayesian posterior probabilities were calculated using the Metropolis-coupled Markov chain
Monte Carlo (MC3) sampling approach in MrBayes v3.01 (Ronquist et al., 2012). Four
independent searches were performed for each dataset; each search consisted of a cold chain and
6 heated chains. All searches started with random trees and uniform prior probabilities for all
possible trees. For all datasets, the original Markov chains were run for 4 X107 generations and
trees were sampled every 10000 generations. To determine that stationarity had been reached, we
compared both the fluctuating values of the likelihood from the four independent searches using
TRACER v1.4 (Rambaut and Drummond, 2007) and convergence rates of posterior split
probabilities and branch lengths using AWTY (Nylander et al., 2008). The “burn-in” value was
conservatively set at 1,000; the first 1,000 (10,000,000 generations) trees were eliminated from
the approximation of posterior probabilities. The trees retained from each run were combined
and a 50% majority rule consensus tree was produced. The ML trees from each independent
GARLI and MrBayes search were compared. The likelihood scores from the “best tree”
recovered using GARLI and the ML tree from each MrBayes search were optimized using
PAUP* for comparison of ML and topology testing. Uncorrected p distances between each
sample pair were calculated for the cyt b sequence data using PAUP* (Swofford, 2002).

2.3 Divergence time estimates

Divergence times were estimated using an uncorrelated log-normal relaxed clock model
(Drummond et al., 2006) as implemented in the program BEAST v.1.7.5 (Drummond and
Rambaut, 2007), using the subroutine BEAUti v1.7.5 to set the analysis parameters. The model
of evolution for partitions that fit GTR were conservatively set to HKY. This was only after
initial runs using GTR had low ESS values for prior and posterior, with one of the relative rates
in each of the GTR modeled partition going to zero. Reducing the model to HKY fixed this issue
without any change in results. Two calibration points were used. The minimum age for the
divergence between the Dermatemydidae and Kinosternidae was constrained to 74.8 mya, based
on stem kinosternid fossils from the lower third of the Kaiparowits Formation (Brinkman and
Rodriguez De La Rosa, 2006; Brinkman et al. in press; Hutchison et al. in press; Knauss et al.,
2011). Hutchison (1991) described the oldest known kinosternine fossils from the early Eocene
(Lysitian), and suggested the divergence of the subfamilies Kinosterninae and Staurotypinae at
between 56 and 65 mya. For our analysis we constrained the minimum age of the
Kinosterninae/Staurotypinae split to 53 mya based on Koch and Morrill (2000). In each analysis,
the Markov chain was run for 100 million generations and sampled every 10,000. Convergence
to stationarity was checked using TRACER v. 1.5 (Rambaut and Drummond, 2007) for each
search and compared across all runs. The search results were summarized using TreeAnnotator
v. 1.7.5 and visualized using Figtree v. 1.3.1 (http://tree.bio.ed.ac.uk/software/figtree/).

2.4 Biogeography

Ancestral distributions of all extant kinosternid turtles were reconstructed using both the
Statistical Dispersal-Vicariance Analysis (S-DIVA) and the Bayesian Binary method (BBM)
implemented in RASP (Reconstruct Ancestral State in Phylogenies: Yu et al., 2011). We used a
Iverson, Le, and Ingram 5

time-calibrated phylogeny that included representatives from all 25 species of Kinosternidae and
a single outgroup (Dermatemys). We coded eleven geographical areas: A -South America, B -
Central America (south of Mesoamerica, Honduras to Panama), C - Mesoamerica (Isthmus of
Tehuantepec to NW Honduras), D - Atlantic Mexico (Tampico embayment, Tamaulipas to
northern Veracruz), E - Mexican Plateau, F - Pacific coastal Mexico (Sinaloa to Oaxaca), G -
Northwest Mexico (Sonora), H - Southwest USA (Arizona), I - Central USA, J - Southeast USA,
and K - Northeast USA. Although more fine scale coding is possible, we find this simplistic
matrix more appropriate for the questions at hand, rather than making data overly complex and
over-parameterized, exceeding their explanatory value. Because most sampled taxa occupied
only one or two areas, and only one occupied three (leucostomum), we set the maximum number
of areas for reconstruction in both S-DIVA and BBM at three.

3. Results

3.1. Phylogenetic analyses

The final data matrix contained 4511 aligned characters, and among the six markers, cyt b
and 16S were the most phylogenetially informative (Table 1). Phylogenies estimated for each
locus were variable in the amount of resolution; mitochondrial gene trees were resolved with
support while the nuclear gene trees showed variable and poor resolution distributed across the
tree (not shown). Phylogenies based on the combined nuclear data also demonstrated minimal
resolution (e.g., Fig. S1), whereas those based on the combined mitochondrial sequences were
almost fully resolved (e.g., Fig. S2) and nearly identical to those based on the combined nuclear
and mitochondrial data set. While these datasets varied in the amount of phylogenetic
information, comparisons do not reveal any major conflict; areas that were not completely
congruent among trees were restricted to nodes involving short branches and/or weak bootstrap
support, and therefore we focused on the results from the total concatenated dataset.
The four different analyses (ML and BI, codon and non-codon-based) recovered almost
identical topologies, differing only in node support values (Fig. 1). The two previously
recognized subfamilies, Staurotypinae and Kinosterninae (TTWG, 2012), were recovered as
monophyletic in all analyses (ML bootstrap [BP] = 100%, BI posterior probability [PP] = 100%;
only codon-based support values reported in the text, but see Fig. 1).
Within the Kinosterninae, three major clades were resolved with high support (ML BP = 89-
97%, BI PP = 100%), although the relationship among the three clades was not well resolved
(ML BP <50%, BI PP = 51%). A monophyletic genus Sternotherus was strongly supported (ML
BP = 96%, BI PP = 100%); however, the genus Kinosternon as previously recognized was
recovered consistently as paraphyletic with respect to Sternotherus. Most former members of
Kinosternon (sensu lato) belong to the clade including K. scorpioides (type species of the genus
Kinosternon; Iverson, 1992a), with high support (ML BP = 89%, BI PP = 100%. However, the
leucostomum group (also including acutum, angustipons, creaseri, dunni, and herrerai) was
recovered as monophyletic with respect to Sternotherus and the restricted genus Kinosternon
with strong support (ML = 97%, BI = 100%). Mean uncorrected p distances for cyt b among
species within each clade (0.058 – 0.076) were substantially lower than average distances
between clades (0.10 – 0.12: Table 2). No unique genus name has ever been proposed for any
member of the leucostomum group (Fritz and Havas, 2007).
Within Sternotherus, odoratus was weakly supported as sister to the carinatus complex
(including minor and depressus; see also Tinkle, 1958; ML = 56%, BI = 82%), suggesting that
Iverson, Le, and Ingram 6

the divergence of odoratus and the carinatus group was nearly coincident in time. The mtDNA
only dataset strongly supported S. odoratus as sister to S. carinatus (ML = 96%, BI = 100%; Fig.
S2), while the nuDNA only analysis does not recover odoratus within the Sternotherus clade.
This may be explained by the limited amount of nuDNA signal and the lack of RAG1 sequence
for S. odoratus.
Within the unnamed clade including leucostomum, two radiations were well supported. The
“northern” group (ML BP = 100%, BI PP = 100%) included the three parapatric taxa found on
the Atlantic versant of Mexico, Guatemala, and Belize, whereas the “southern” group (ML =
73%, BI = 94%) included the two disjunct, southern, small-plastroned species angustipons
(Nicaragua to Panama) and dunni (Colombia), and the wide-ranging leucostomum (Veracruz to
Peru; Iverson, 1992a), which is sympatric with those two species as well as the southern
members of the “northern” clade. The remaining kinosternine species belong to the restricted,
well-supported (see above), monophyletic genus Kinosternon, with a distribution across the USA
and Mexico (where its diversity is greatest), but also into Central and South America (Iverson,
1992a).
Within Kinosternon (sensu stricto) those taxa found in the central and eastern USA represent
a well-separated monophyletic clade divergent from the other included taxa (ML = 88%, BI =
100%). The latter includes the previously recognized “scorpioides group” (as defined by Berry et
al., 1997 and Iverson, 1991, but excluding acutum, creaseri, chimalhuaca, and alamosae), which
is resolved as monophyletic (ML = 98% BI = 100%). However, the relationships among the
members of this derived group are not resolved with high support values (Fig. 1). On the
contrary, the relationships among nearly all of the other members of the restricted genus
Kinosternon were consistently resolved.

3.2 Divergence estimates and biogeography

Program BEAST estimated the divergence of the three extant kinosternine clades at 22-25
mya (Fig. 2; Table S4), but most of the diversification of extant kinosternine species took place
between 5 and 10 mya. At the species level, only the divergence of triporcatus from salvinii,
acutum from creaseri and depressus from minor were aged at less than 4 mya. The topology of
the BEAST tree at the species level was identical to that in the ML and BI trees, except for the
placement of S. odoratus.
Bayesian (BBM) and event based (S-DIVA) method reconstruction of ancestral areas based
on extant taxa produced conflicting results for the basal nodes. The BBM method in RASP (Fig.
3) supported a Mesoamerican origin (Node 69, marginal probability = 99%) for the family
Kinosternidae during the late Cretaceous. In this reconstruction, the two subfamilies diverged
within Mesoamerica (Node 68, marginal probability = 97%) during the Paleocene, with the
Staurotypinae confined to Mesoamerica (Nodes 66 and 67, marginal probability > 99%).
Mesoamerica is hypothesized as the ancestral area for the extant Kinosterninae (Node 65, 84%)
followed by the divergence of the Mesoamerican clade (the leucostomum group; Node 42, 65%)
from the ancestor of the remaining Kinosternon plus Sternotherus which is hypothesized to have
dispersed into the Southeastern USA (Node 64, marginal probability = 87%). S-DIVA (Fig. S3)
reconstructed the ancestral areas for these nodes with much larger distributions, with support
distributed across a number of combinations including large areas of the current distribution of
the Kinosternidae. This is not unexpected due to the nature of the different methods; S-DIVA
maximizes vicariance and minimizes dispersal/extinction leading to a preference for larger
ancestral areas (Yu et al., 2011). At almost every node, BBM showed strong support for a much
Iverson, Le, and Ingram 7

smaller geographic area, typically one to at most three areas, while S-DIVA increased the
number of areas from the tips to the base of the tree, with the basal node reconstructed as
potentially including nearly the entire geographic range the Kinosternidae. Predicted ancestral
areas for the BBM analyses were not impacted by the maximum number of areas chosen (i.e.,
three, six, or twelve; latter not shown), whereas ancestral areas were strongly influenced by the
number of areas for S-DIVA, increasing to the maximal setting (whether three, six or twelve;
latter not shown).

4. Discussion

4.1 Taxonomic implications

Our analysis confirmed the monophyly of the two previously recognized clades within of the
family Kinosternidae, one including the genera Staurotypus and Claudius, and the other
including Sternotherus and Kinosternon. The fossil record dates the divergence of these two
clades at >54 mya (see Section 2.3). Given 1) that the other North American turtle subfamilies
date from only 34 mya (emydids) to 52 mya (geoemydids; Spinks and Shaffer, 2009), 2) that
other speciose cryptodiran turtle families date from ca. 52 to 70 mya (Testudinidae and
Geoemydidae), 60 to 90 mya (Emydidae), and 100-129 mya (Trionychidae)(Lourenço et al.,
2012; Spinks and Shaffer, 2009; Wang et al., 2012), and 3) that the two subfamilies
Staurotypinae and Kinosterninae are unambiguously distinct morphologically (Hutchison, 1991)
and in their sex-determining mechanisms (Fig. 1; Ewert et al., 2004), we follow Bickham and
Carr (1983; among others) in recognizing these two clades as separate families (Staurotypidae
and Kinosternidae; Table 3).
Our analysis also resolved three relatively old (22-25 my), distinctly monophyletic clades
within the restricted Kinosternidae (Fig. 2). The age of these clades exceeds the estimated ages
of most of the recognized genera in the other primarily North American radiation (the Emydidae)
for which data are available. For example, Martin et al. (2013) provided the following estimates:
Actinemys and Emys (ca. 7.5 mya), Emydoidea (ca. 12.5 mya), Glyptemys (20 mya), and
Terrapene and Clemmys (ca. 21 mya); and Spinks and Shaffer (2009) dated the genera
Trachemys and Graptemys at only ca. 15 mya. Furthermore, mean cyt b uncorrected p distances
between members of these three kinosternid clades range from 10.3 to 12.3% (Table 1),
generally exceeding distances found among species in the same genus (reviewed by Vargas-
Ramirez et al., 2010). The three clades are also differentiated by at least their carination and life
style patterns (Fig. 1; see also below).
These data argue that the three kinosternid clades merit recognition as genera, but only two
of the clades have previously been so named (Sternotherus and Kinosternon). Because members
of the third radiation have been known since at least 1831, but the distinction of that clade has
been unrecognized and undiagnosed until now, we here describe this “hidden”, early, tropical
radiation of the turtle subfamily Kinosterninae as a distinct genus (Table 3).

Family Kinosternidae Agassiz 1857


Tribe Kinosternini Hutchison 1991
Cryptochelys gen. nov.

Synonymy: Kinosternon Duméril and Bibron (in Duméril and Duméril 1851:17 (in part)[and
nearly all subsequent authors].
Iverson, Le, and Ingram 8

Etymology: From the Greek, kruptos (cryptic, hidden) and chelus (tortoise, turtle). The
genus is feminine, requiring a feminine suffix for adjectival species names.
Type species: Kinosternon leucostomum Duméril and Bibron (in Duméril and Duméril
1851)[= Cryptochelys leucostoma]
Content: Cryptochelys acuta (Gray 1831), C. angustipons (Legler, 1965), C. creaseri
(Hartweg, 1934), C. dunni (Schmidt, 1947), C. herrerai (Stejneger, 1925), and C. leucostoma
(Duméril and Bibron in Duméril and Duméril 1851).
Diagnosis: Kinosternid (sensu stricto) turtles lacking an entoplastron (present in Baltemys
and Xenochelys), with reduced carination (basically unicarinate; usually tricarinate in Baltemys,
Xenochelys, Sternotherus, and Kinosternon, though nearly acarinate in some in the latter genus),
a reduced neural series (typically five bones, all posteriorly symmetric; six in C. creaseri) not in
contact with the nuchal bone (usually six with neural contact in other kinosternids; Iverson,
1988b), the presence of clasping organs on the posterior crus and thigh (except absent in C. acuta
and C. creaseri; also present in Sternotherus, but absent in many Kinosternon), the anterior end
of the anterior musk duct groove reaching only to the anterior half of the third peripheral
(unknown for dunni; reaching to the second peripheral in Sternotherus and most Kinosternon,
and to the first peripheral in Baltemys and Xenochelys), a gular scute of intermediate width
(much narrower in Sternotherus and usually broader in Kinosternon; see Appendix in Iverson,
1991), and distinctive mitochondrial DNA.
Phylogenetic definition: All members of the Kinosternini more closely related to
Cryptochelys leucostoma than to Sternotherus odoratus or Kinosternon flavescens.
Distribution: Atlantic versant of Mexico, Central, and extreme northwestern South America,
and Pacific versant of South America from Panama to northern Peru.
Fossil history: Langebartel (1953) reported post-Pleistocene remains on the Yucatan that may
represent C. creaseri, and Cadena et al. (2007) described kinosternid fragments from the late
Pleistocene of Colombia that likely represent C. leucostoma (see 4.3 below).

4.2 Phylogenetics

The relationships among the species of Cryptochelys have previously been obscure, primarily
because of the unavailability of tissues from rare taxa (especially angustipons, dunni, and
creaseri). Iverson (1988a) first proposed that acuta and creaseri were sister taxa, based on their
parapatry and similar morphology and ecology. On morphological grounds, Legler (1965)
suggested that angustipons and dunni were sister taxa, and using combined morphology and
preliminary mtDNA sequence data, Iverson (1998) first noted the close relationship of
leucostoma and dunni, and of acuta and herrerai. Our analysis is the first to clarify the
relationships among the included taxa and the monophyly of this new genus. However, more
thorough geographic sampling is needed for the wide-ranging species leucostoma, since
preliminary morphological data indicate the existence of undescribed variation (Berry, 1978;
Iverson, unpublished).
Within the genus Sternotherus the most primitive species has been hypothesized to be S.
carinatus (Zug, 1966) or S. odoratus (Iverson, 1998; Tinkle, 1958). Our ML and BI analyses
supported the latter, though with relatively low support indices (Fig. 1), and our BEAST analysis
placed odoratus as sister to carinatus (Fig. 2). These data suggest that the divergence of
odoratus, carinatus, and minor (including depressus) may have been nearly simultaneous. S.
depressus was consistently (ML BP = 100%, BI PP = 100%) found to be sister to S. m. peltifer,
from which it has long been assumed to have been derived (e.g., Iverson, 1977; Tinkle, 1958;
Iverson, Le, and Ingram 9

Walker et al., 1998b). The controversy concerning the recognition of depressus as a species or
subspecies (Walker et al., 1998b) is still an open question.
The restricted genus Kinosternon comprises two well-supported clades, the “subrubrum
group” (previously identified by Iverson, 1998; including, baurii, flavescens, and subrubrum
[including steindachneri], for which the name Thyrosternum Agassiz 1857 could be applied as a
subgenus), and the remaining species. Within the former, the paraphyly of K. subrubrum (as
previously recognized; e.g., Walker et al., 1998a) was fully supported in all analyses. This
suggests that steindachneri may represent a distinct species (see also Bourque, 2012a). Our
analysis also supports the conclusion that K. durangoense and K. arizonense, once considered
subspecies of K. flavescens because of obvious morphological similarity (Iverson, 1979a, 1979b,
1989b), are independent radiations and hence, separate species (Serb et al., 2001).
With two exceptions, the relationships among the remaining members of the genus
Kinosternon (sensu stricto) are not well resolved. First, the “scorpioides group” (as defined by
Berry et al., 1997, but excluding K. alamosae and K. chimalhuaca; Fig. 1 Node 24) is supported
with high confidence (ML BP = 98%; BI PP = 100%), although the four currently recognized
(parapatric) subspecies of K. scorpioides were recovered as paraphyletic with respect to K.
integrum and K. oaxacae in each analysis, with generally weak support (e.g., Fig. 1). This
suggests that K. scorpioides likely represents a multispecies complex (see also Iverson, 2010),
but much more complete geographic sampling will be necessary to clarify species boundaries in
this clade. Future work should also reconsider the validity of the South American taxa currently
synonymized under K. s. scorpioides (TTWG, 2012): carajascensis da Cunha 1970 in central
Brazil and (especially) the disjunct seriei Freiberg 1936 in Paraguay, Argentina, and Bolivia.
Second, samples of K. integrum from Colima, Puebla, and Oaxaca were resolved (together)
as monophyletic with complete support in all analyses. Webb (1984) suggested that integrum is
polytypic, but confirmation of that will require much more thorough geographic sampling,
including comparisons with Pleistocene fossils from the Mexican Plateau (Cruz et al., 2009;
Mooser, 1980).
Iverson (1981, 1998) hypothesized that the morphologically similar, precisely parapatric K.
sonoriense and K. hirtipes were sister taxa. That relationship was resolved with reasonably high
support in this study (ML BP = 75%; BI PP = 100%), although additional sampling across both
of these wide-ranging taxa is needed, particularly given the subspecific variation in morphology
that has been described in both (Iverson, 1981).
The resolution of K. alamosae and K. chimalhuaca outside the K. scorpioides group is an
enigma. All previous authors examining the group have concurred with their inclusion therein,
and their close relationship with K. integrum (Berry and Legler, 1980; Berry et al., 1997;
Iverson, 1991, 1998), albeit on morphological grounds (e.g., each share the assumed
synapomorphy of the lack of clasping organs; Iverson, 1991). Since its description (Berry and
Legler, 1980), K. alamosae has been assumed to be most closely related to K. integrum, despite
its general external similarity to K. arizonense (Iverson, 1989a).
The identification of K. chimalhuaca as sister to K. hirtipes with very high support (87-98%)
in all analyses is even more surprising. The range of K. chimalhuaca is completely within that of
K. integrum (Berry et al., 1997), with which it is parapatric, and the nearest population of K.
hirtipes lies at least 100 km to the northeast, across at least two mountain ranges (with only
integrum inhabiting the valley between them). Genetic sampling across the range of integrum
may help explain the puzzling resolution of alamosae and chimalhuaca in our analyses.
Although it is clear that species (or genus) boundaries should not be based on such variable
measures as uncorrected p distances, those values can still be useful as relative measures of
Iverson, Le, and Ingram 10

taxonomic divergence. For example, published values of p distances for the cyt b gene between
species of Asian pond turtles, Palearctic tortoises, and Asian and North American softshells
range from about 2.8 to 13.9% (review in Vargas-Ramirez et al., 2010).
With only a few exceptions, p distances among kinosternid species fall in the middle of this
range or higher (Table S3), and minimum distances between samples of the three proposed
genera were approximately 9% and averaged 10-12% (Table 1). Interestingly, we found K.
subrubrum steindachneri to be less divergent from the broadly sympatric K. baurii (4.2%) than it
was from purportedly conspecific K. s. hippocrepis (6.8%); however, we did not sequence the
more geographically proximate K. s. subrubrum. Nevertheless, these data do support Bourque’s
(2012a) recommendation that steindachneri be elevated to species status (Table 3). In addition,
K. scorpioides abaxillare, isolated in the Central Valley of Chiapas, is >4% divergent from all
other samples except the distinctly different K. oaxacae (3.7%), including the supposedly
conspecific K. s. cruentatum, K. s. albogulare, and K. s. scorpioides. Given its morphological
distinction (Berry, 1978), its allopatry with other Kinosternon, and its genetic divergence, we
recommend the elevation of abaxillare to species status. Unexpectedly, we found a distance of
only 1.2% between the parapatric sister taxa C. creaseri and C. acuta, but because of their
morphometric distinction (Iverson, 1988a), we continue to support their retention as separate
species.

4.3 Historical biogeography

Our analysis confirmed the reciprocal monophyly of the previously recognized Staurotypinae
and Kinosterninae (here recognized as separate families). The former has a meager fossil record
in Central America (Cadena et al., 2012), but is currently distributed only in Mesoamerica, where
it may initially have been isolated in sympatry with Cryptochelys (see below). When only
considering extant taxa, the BBM analyses reconstructed a clean history for the group,
originating in Mesoamerica, then expanding its range into North America and South America
(Fig. 3). Given the evidence provided by the fossil record, the early biogeographic history for
this group must be more complex and may be more closely aligned with the prediction of S-
DIVA (Fig. S3). For example, aquatic fossil kinosternids (Baltemys and Xenochelys) are
hypothesized to have occurred all over North America in the late Paleocene, since Eocene
kinosternine fossils are known from at least 23 sites from Florida to Arkansas to Texas to New
Mexico to Wyoming to northern Canada (33 to 55 mya; Bourque, 2012c; Eberle and Greenwood,
2012; Hutchison, 1991). Indeed, maximum known Cenozoic turtle diversity in at least western
North America was in the Wasatchian of the early Eocene (50-55 mya; Hutchison, 1982; Zachos
et al., 2001). Those early kinosternids (with an entoplastron and a small non-hinged plastron)
likely diversified across the mid-continent during the warm Eocene (Zachos et al., 2001) as noted
above. However, during the cooling period in the late Eocene and early Oligocene, the group
likely survived only in the more tropical regions in Meso- and Central America (and Florida; see
Bourque, 2012c), as hypothesized for other reptiles and amphibians (Savage, 2002; Le and
McCord 2008). The well-documented decline in aquatic turtle diversity in at least western North
America in the early Oligocene, between 33 and 28 mya (Corsini et al., 2011; Hutchison, 1992,
2005), was presumably due to these cooling and drying conditions (Zachos et al., 2001). Hence,
the hypothesized ancestral area for the restricted family Kinosternidae in Mesoamerica (Fig. 3) is
actually a relict distribution compared to its former wide distribution and high diversity in the
Paleocene and Eocene.
Iverson, Le, and Ingram 11

The Oligocene Mesoamerican relict kinosternids apparently retained the tricarinate shell and
small plastron (but lost the entoplastron) of Baltemys and Xenochelys, and were presumably
aquatic. But when the climate warmed in the mid to late Oligocene (ca. 28 mya), the group
apparently dispersed both northward and southward. The common ancestor of Sternotherus and
the restricted Kinosternon (node 10; Fig. 2) is hypothesized to have dispersed north and east
along the Gulf Coast, where one subclade, primarily aquatic, with a reduced plastron, diverged as
the genus Sternotherus. The second subclade (Kinosternon sensu stricto), with a more extensive
plastron, dispersed westward and apparently adapted to the increased seasonal aridity in North
America at that time (Hay, 1908:39; Zachos et al., 2001; as evidenced by the significant
transition in general turtle diversity from primarily aquatic to primarily terrestrial; Hutchison,
1982, 1992, 2005). The third group (Cryptochelys) remained tropical and dispersed into Central
America (i.e., to the new Chortis Block; Savage, 2002).
During the warming and drying of the early Miocene (20-25 mya; Woodburne, 2004), the
Mesoamerican Cryptochelys clade is hypothesized to have dispersed northward and southward
and the ancestral range divided (node 5; ca. 18 mya) by an unknown vicariant event into a
northern element (to become herrerai, creaseri, and acuta) and a southern element
(leucostoma/dunni/angustipons). This was likely the same event that accounted for the
divergence of the 17 mya fossil Staurotypus moschus in Panama from its Mesoamerican
congeners (Cadena et al., 2012). From Central America, Cryptochelys twice dispersed into South
America, once by C. leucostoma and once by ancestral C. dunni (Fig. 2). Unfortunately, timing
of the dispersal of the first clade will require future extensive geographic sampling across the
range of C. leucostoma. However, based on our chronogram (Fig. 2), the second dispersal must
have occurred by about 8 mya (Node 7), well before the closure of the Panamanian portal (ca.
2.5 to 4 mya; Coates and Obando, 1996; Iturralde-Vinent, 2006; Iturralde-Vinent, and MacPhee,
1999; Kirby et al., 2008; Woodburne, 2004). It is noteworthy that an increasing list of reptile and
amphibian taxa (among others; Cody et al., 2010) show evidence of dispersal to South America
in the mid- to late Miocene (before portal closure): Rhinoclemmys, 20-22 mya (Le and McCord,
2008); Bolitiglossa, 18 mya (Hanken and Wake, 1982); Lachesis, 6-18 mya (Zamudio and
Greene, 1997); Apalone, >5 mya (Head et al., 2006); and others might also have done so (e.g.,
the turtles Chelydra, Trachemys, and Cryptochelys leucostoma).
Within Sternotherus (node 11; Fig 2), one species (odoratus) evolved as an aquatic habitat
generalist (perhaps primarily northern in distribution), whereas the others (minor/carinatus)
adapted to the permanent rivers and streams of the Gulf Coast of southeastern North America.
Based primarily on the mammalian fauna, Webb et al. (1981) reported that the “Gulf Coast
seems to have been a separate biotic province during most of the Miocene”, and the
carinatus/minor complex apparently diverged there during that time. One member of that group
(carinatus, in the western Gulf) became the most specialized with its high, tent-like carapace.
The eastern forms retained lower, basically tricarinate shells (node 13), with the population in the
upper Black Warrior River in Alabama (depressus) only recently evolving its very low shell.
Curiously, the oldest fossil record for Sternotherus is only ca. 7 mya (Bourque, 2011).
The ancestral restricted genus Kinosternon is hypothesized to have originated in eastern
North America between 17 and 22 mya, but must have dispersed westward as far as
southwestern North America (Sonora, according to the BBM) by 14-17 mya, because fossil
fragments of Kinosternon ranging in age from ca. 14 to 18 mya are known from Delaware,
extreme northern Florida, Nebraska, and New Mexico (Bourque, 2012a, 2012b; Holman, 1998;
Hutchison, 1991). This ancestor must have been flavescens-like in morphology and ecology
(e.g., reduced carination and semiaquatic), given the retention of those traits in both descendent
Iverson, Le, and Ingram 12

clades within Kinosternon (baurii, subrubrum, flavescens, arizonense, and durangoense; Iverson,
1991; Serb et al. 2001), and the presence of that general morphology in the two New Mexico
Miocene species, K. pojoaque (ca. 14 mya; Bourque, 2012a) and K. skullridgescens (ca. 15 mya;
Bourque, 2012b).
Within the restricted genus Kinosternon, because of a vicariant event ca. 17 mya (node 14;
perhaps the continued faulting of the Basin and Range province [Woodburne, 2004] coupled
with warm, dry conditions in the early Miocene), the clade in the central and eastern USA (the
“subrubrum group”, also including flavescens, baurii, and steindachneri) diverged from the
clade of other Kinosternon in the southwestern USA and northern and central Mexico. Although
the zoogeographic history of the latter clade was poorly resolved in both the RASP analyses, the
scorpioides group apparently dispersed via the Pacific coast into southeastern Mexico, through
Central America, into sympatry with Cryptochelys, and eventually into South America.
Amazingly, this scorpioides group is currently distributed from Sonora and Tamaulipas in
Mexico southward to Argentina. And although our sampling of the group is incomplete
geographically, our results suggest that this group dispersed from coastal Mexico to at least
northern South America (based on our Venezuelan scorpioides sample) in only about 2 million
years (compare nodes 21 and 24), by ca. 7 mya, and well before the closure of the Panamanian
portal (references above). K. scorpioides was thus the third kinosternid clade to disperse into
northern South America, where it dispersed to the Atlantic versant (as did Trachemys; e.g.
Iverson, 1992a), while the genus Cryptochelys dispersed primarily to the Pacific versant (as did
Chelydra; e.g., Iverson 1992a). The absence of Cryptochelys in Atlantic drainages beyond
Colombia, and the generalized morphology and ecology of K. scorpioides, likely facilitated the
latter’s very rapid dispersal southward across the Amazon basin to the more seasonally dry
conditions of Bolivia, Paraguay, and Argentina. Soon after this extensive dispersal (i.e., beyond
node 24), and despite the evident vagility of the ancestral form, tectonic and/or climatic events
apparently isolated allopatric populations along coastal Oaxaca (oaxacae), possibly coastal
Colima/Jalisco (chimalhuaca; see above), the Central Valley of Chiapas (abaxillare), South
America (scorpioides), and the remainder of Mesoamerica and Central America
(cruentatum/albogulare), each of which diverged at least morphologically. Clearly, the
scorpioides group represents the most successful and rapid radiation in this family, and deserves
much more complete genetic sampling to clarify its phylogeny and taxonomy.

4.4 Morphological implications

Within the Kinosternidae, the plastron is much reduced (with reduced kinesis) in several
taxa, including all Sternotherus, Kinosternon herrerai, K. angustipons, and some populations of
K. hirtipes (Iverson, 1991). Bramble et al. (1984) hypothesized that Sternotherus and K. herrerai
evolved first among the kinosternids, and that anterior plastral lobe kinesis was ancestral, with
hindlobe kinesis evolving later as an adaptation to terrestriality. That scenario would require the
independent evolution of dual kinesis at least twice in the family (in Cryptochelys and in
Kinosternon).
Second, although the lack of clasping organs has previously been considered a
synapomorphy defining the scorpioides group (Berry and Legler, 1980; Iverson, 1991), it seems
more likely that clasping organs were lost independently at least twice (in the C. acuta/creaseri
clade and the K. scorpioides clade). In addition, their absence in alamosae and chimalhuaca,
seemingly outside the latter group, may be additional independent evolutionary events. Thorough
Iverson, Le, and Ingram 13

character reconstruction studies will be necessary to evaluate the evolutionary history of these
and other key kinosternid traits.

4.5 Concluding thoughts

Despite the resolution of many of the relationships among the kinosternids, and the
establishment of a reasonable chronology of diversification, several key evolutionary questions
remain. Because most of the diversification within the genus Kinosternon (s.s.) occurred within
the last 10 my (especially within the scorpioides group), we were not able to resolve their
relationships with high confidence. Doing so will require sampling additional genetic markers, as
well as much more thorough sampling across the distributions of such wide-ranging taxa as K.
scorpioides, K. integrum, and Cryptochelys leucostoma. Such work is certain to uncover
additional unappreciated variation in these groups.
Second, although the fossil record for Kinosternon (s.s.) dates from the Miocene (16-18 mya;
see above), older fossil crown group kinosternids (to perhaps 22 mya) must exist. But even more
surprising is the lack of fossils older than 7 my for the genus Sternotherus (Bourque, 2011) and
only 0.5 my for the genus Cryptochelys (Cadena et al. 2007). Significant fossils likely remain to
be discovered for this group, particularly in the eastern USA and Mesoamerica.

Acknowledgments

Tissue samples were generously provided by T. S. Akre, R. J. Burke, J. Campbell, J. C. Carr,


O. Victoria Castano, C. R. Etchberger, M. Ewert, M. J. Forstner, C. J. Franklin, M. Gaston, D.
Gicca, D. Greene, S. Guzman, D. R. Jackson, M. Klemens, K. L. Krysko, B. Lamar, J. E.
Lovich, R. E. Lovich, K. Marion, C. May, W. P. McCord, F. Medem, P. Meylan, P. Moler, S.
Pasachnik, C. Phillips, S. Platt, S. Poulin, J. Reyes Velasco, E. Rickart, P. Rosen, J. Serb, E.
Smith, N. Soule, P. Stone, P. Vander Schouw, T. Tuberville, W. Van Devender, T. R. Van
Devender, and G. Weatherman. Critical literature was provided by D. Brinkman and J. Bourque.
P. Meylan accompanied Iverson during field work in Mexico and American Southwest, and his
expert field assistance was critical to early work. Financial support for this project was provided
by the American Philosophical Society, the American Museum of Natural History, the Joseph
Moore Museum of Natural History at Earlham College, and Grant 106.15-2010.30 from
Vietnam’s National Foundation for Science and Technology Development (NAFOSTED) to ML.
Comments on early drafts of the manuscript by R. Bour, J. Bourque, S. Pasachnik, M. E. Seidel,
and two anonymous reviewers were greatly appreciated.

Appendix A. Supplemental material

Supplemental data can be found associated with this article can be found in the on-line version,
at doi: xxxx.

Table S1. Origin of samples sequenced in this study with GenBank numbers.

Table S2. Primers used in this study.


Iverson, Le, and Ingram 14

Table S3. Compilation of uncorrected p distances between samples of kinosternid turtles based
on cyt b sequence data.

Table S4. Estimates and variance of ages for each node as numbered in text Figure 2.

Fig. S1. GARLI partitioned maximum likelihood tree for the Kinosternidae based on
concatenated nuclear DNA data only. Numbers at nodes are bootstrap support values.

Fig. S2. GARLI partitioned maximum likelihood tree for the Kinosternidae based on
concatenated mitochondrial DNA data only. Numbers at nodes are bootstrap support
values.

Fig. S3. Ancestral distributions of extant kinosternid turtles reconstructed using the Statistical
Dispersal-Vicariance Analysis (S-DIVA) method implemented in RASP (Reconstruct
Ancestral State in Phylogenies) and color key to ancestral areas. [COLOR- ON WEB
ONLY]

References

Agassiz, L., 1857. Contributions to the Natural History of the United States of America. Vol 1-2.
Little, Brown and Co., Boston.
Barley, A.J., Spinks, P.Q., Thomson, R.C., Shaffer, H.B., 2010. Fourteen nuclear genes provide
phylogenetic resolution for difficult nodes in the turtle tree of life. Mol. Phylo. Evol. 55,
1189-1194.
Belkin, D.A., 1968. Aquatic respiration and underwater survival of two freshwater turtle species.
Respir. Physiol. 4, 1-14.
Berry, J.F., 1978. Variation and systematics in the Kinosternon scorpioides and K. leucostomum
complexes (Reptilia: Testudines: Kinosternidae) of Mexico and Central America.
Unpublished doctoral dissertation, University of Utah, Salt Lake City, 325 pp.
Berry, J.F., Legler, J.M., 1980. A new turtle (genus Kinosternon) from Sonora, Mexico.
Contrib. Sci. Natur. Hist. Mus. Los Angeles Co. 325, 1-12.
Berry, J. F., Seidel, M.E., Iverson. J.B., 1997. A new species of mud turtle (genus Kinosternon)
from Jalisco and Colima, Mexico, with notes on its natural history. Chelonian Conserv.
Biol. 2, 329-337.
Bickham, J.W., Carr, J.L., 1983. Taxonomy and phylogeny of the higher categories of
cryptodiran turtles based on a cladistic analysis of chromosomal data. Copeia 1983, 918-
932.
Bonin, F., Devaux, B., Dupre, A., 2006. Turtles of the World. Johns Hopkins Univ. Press,
Baltimore, Maryland.
Bourque, J.R., 2011. Reassessment of a putative fossil stinkpot (Kinosterndae: Sternotherus)
from the late Miocene (Clarendonian) of Kansas. J. Herpetol. 45, 234-237.
Bourque, J.R., 2012a. An extinct mud turtle of the Kinosternon flavescens group (Testudines,
Kinosternidae) from the Middle Miocene (Late Barstovian) of New Mexico. J. Vert.
Paleo. 32, 68-81.
Bourque, J.R., 2012b. A fossil mud turtle (Testudines, Kinosternidae) from the early Middle
Miocene (Early Barstovian) of New Mexico. J. Vert. Paleo. 32, 836-853.
Iverson, Le, and Ingram 15

Bourque, J.R., 2012c. Fossil Kinosternidae from the Oligocene and Miocene of Florida, USA. In:
Brinkman, D.B., Holroyd, P.A., Gardner, J.D. (Eds.), Morphology and Evolution of
Turtles: Origin and Early Diversification. Springer, Dordecht, The Netherlands, pp. 459-
475.
Bramble, D.M., Hutchison, J.H., Legler, J.M., 1984. Kinosternid shell kinesis: structure,
function, and evolution. Copeia 1984, 456-475.
Brinkman, D.B., Newbrey, M.G., Neuman, A.G., Eaton, J.G., In press. Freshwater Osteichthyes
from the Cenomanian to Late Campanian of Grand Staircase-Escalente National
Monument, Utah. In: Titus, A.L., Loewn, M.A. (Eds.), At the Top of the Staircase: The
Late Cretaceous of Southern Utah. Indiana Univ. Press, Bloomington, pp. xx-xx (publ.
date 9/20/13).
Brinkman, D.B., Rodriguez de la Rosa, R., 2006. Nonmarine turtles from the Cerro del Pueblo
Formation (Campanian), Coahuila State, Mexico. New Mexico Mus. Nat. Hist. Sci. Bull.
35, 229-233
Cadena, E.R., Jaramillo, C.M., Paramo, M., 2007. The first late Pleistocene record of
Kinosternon (Cryptodira: Kinosternidae) turtles for northern South America, Pubenza
Locality, Colombia. S. Amer. J. Herpetol. 2, 201-205.
Cadena, E.R., Bourque, J.R., Rincon, A.F., Bloch, J.I., Jaramilliop, C.A., McFadden, B.J., 2012.
New turtles (Chelonia) from the Late Eocene through Late Miocene of the Panama Canal
Basin. J. Paleo. 86, 539-557.
Ceballos, C.P., Adams, D.C., Iverson, J.B., and Valenzuela, N., 2013. Evolution of male and
female body size in turtles: Implications for the evolution of sexual size dimorphism and
Rensch’s rule in long-lived vertebrates. Evol. Biol. 40, in press-available on-line.
Coates, A.G., Obando, J.A., 1996. The geologic evolution of the Central American Isthmus. In:
Jackson, J.B.C., Budd, A.F., Coates, A.G. (Eds.), Evolution and Environment in Tropical
America. University of Chicago, Chicago, pp. 21-56.
Cody, S., Richardson, J.E., Rull, V., Ellis, C., Pennington, R.T., 2010. The Great American
Biotic Interchange revisited. Ecography 33, 326-332.
Corsini, J.A., Smith, T.R., Leite, M.B., 2011. Turtle decline in the early Oligocene of western
Nebraska. J. Herpetol. 45, 238-243.
Cruz, J. A., Arroyo-Cabrales, J., Viñas-Vallverdu, R., 2009. Tortugas fósiles del Pleistoceno
tradío de Santiago Chazumba, Oaxaca. Bol. Soc. Geol. Mexicana 61, 225-232.
da Cunha, O.R., 1970. Uma nova subespécie de quelônio, Kinosternon scorpioides carajasensis
da Serra dos Carajás, Pará. Bol. Mus. Paraense Emilio Goeldi 73, 1-11.
Darriba, D, Taboada, G.L., Doallo, R., and Posada D., 2012. jModelTest 2: more models, new
heuristics and parallel computing. Nature Methods 9, 772.
Drummond, A.J, Ashton, B., Buxton, S., Cheung, M., Cooper, A., Duran, C., Field, M., Heled,
J., Kearse, M., Markowitz, S., Moir, R., Stones-Havas, S., Sturrock, S., Thierer, T.,
Wilson, A., 2011. Geneious v5.4, Available from http://www.geneious.com/
Drummond, A.J., Ho, S.Y.W., Phillips, M.J., Rambaut, A., 2006. Relaxed phylogenetics and
dating with confidence. PloS Biol. 4, 699-710.
Drummond, A.J., Rambaut, A., 2007. BEAST: Bayesian evolutionary analysis by sampling trees.
BMC Evol. Bio. 7, 214. Available from: http://beast.bio.ed.ac.uk/.
Duméril, A.M.C., Duméril, A.H.A., 1851. Catalogue Methodique de la Collection des Reptiles
du Museum d'Histoire Naturelle. Gide and Boudry, Paris.
Eberle, J.J., Greenwood, D.R., 2012. Life at the top of the greenhouse Eocene world- A review
of the Eocene flora and vertebrate fauna from Canada’s High Arctic. GSA Bull. 124, 3-23.
Iverson, Le, and Ingram 16

Ewert, M.A., 1991. Cold torpor, diapause, delayed hatching and aestivation in reptiles and birds.
In: Deeming, D.C., Ferguson, M.W.J. (Eds.), Egg Incubation: Its Effect on Embryonic
Development in Birds and Reptiles. Cambridge Univ. Press, Cambridge, England, pp.
173-191.
Ewert, M.A., Etchberger, C.R., Nelson, C.E., 2004. Turtle sex-determining modes and TSD
patterns, and some TSD pattern correlates. In: Valenzuela, N., Lance. V. (Eds.),
Temperature-dependent Sex Determination in Vertebrates. Smithsonian Inst.,
Washington, D.C., pp. 21-32.
Freiberg, M., 1936. Una nueva tortuga del norte Argentina. Physis 12(43), 169-173.
Fritz, U., Havas, P., 2007. Checklist of chelonians of the world. Vertebrate Zoology 57, 149-368.
Gray, J.E., 1831. Synopsis Reptilium or Short Descriptions of the Species of Reptiles. Part I.
Cataphracta, Tortoises, Crocodiles, and Enaliosaurians. Treuttel, Wurz Co., London.
Hanken, J., Wake, D.B., 1982. Genetic differentiation among plethodontid salamanders (genus
Bolitoglossa) in Central and South America: implications for the South American
invasion. Herpetologica 38, 272-287.
Hartweg, N., 1934. Description of a new kinosternid from Yucatan. Occas. Pap. Mus. Zool.,
Univ. Michigan 277, 1-2.
Hay, O.P., 1908. The fossil turtles of North America. Carnegie Inst. Washington. Publ. 75, 1-
568.
Head, J.J., Aguilera, O.A., Sanchez-Villagra, M.R., 2006. Past colonization of South America by
trionychid turtles: fossil evidence from the Neogene of Margarita Island, Venezuela. J.
Herpetol. 40, 378-381.
Holman, J.A., 1998. Reptiles of the lower Miocene (Hemingfordian) Pollack Farm Fossil Site,
Delaware. In: Benson, R.N. (Ed.), Geology and Paleontology of the Lower Miocene
Pollack Farm Fossil Site, Delaware. Delaware Geological Survey, Special Publication 21,
pp. 141-147.
Hutchison, J.H., 1982. Turtle, crocodilian and champsosaur diversity changes in the Cenozoic of
the north-central region of the western United States. Palaeogeog. Palaeoclim. Palaeoecol.
37, 149-164.
Hutchison, J.H., 1991. Early kinosterninae (Reptilia: Testudines) and their phylogenetic
significance. J. Vert. Paleont. 11, 145-167.
Hutchison, J.H., 1992. Western North American reptile and amphibian record across the
Eocene/Oligocene boundary and its climatic implications. In: Prothero, D.R., Berggren,
W.A. (Eds.), Eocene-Oligocene Climate and Biotic Evolution. Princeton Univ, Press,
Princeton, New Jersey, pp. 451-463.
Hutchison, J. H., 2005. Testudines. In: Prothero, D.R., Emry, R.J. (Eds.), The Terrestrial Eocene-
Oligocene Transition in North America. Cambridge Univ. Press, Cambridge,
Massachusetts, pp. 337-353.
Hutchison, J.H., Brinkman, D.B., Knell, M J., In press. Turtles from the Kaiparowits Formation,
Utah. In: Titus, A.L., Loewn, M.A. (Eds.), At the Top of the Staircase: The Late
Cretaceous of Southern Utah. Indiana Univ. Press, Bloomington, pp. xx-xx. (publ. date
9/20/13)
Iturralde-Vinent, M.A., 2006. Meso-Cenozoic Caribbean paleogeography: implications for the
historical biogeography of the region. Internat. Geo. Rev. 48, 791-827.
Iturralde-Vinent, M.A., MacPhee, R.D.E., 1999. Paleogeography of the Caribbean region:
implications for Cenozoic biogeography. Bull. Amer. Mus. Nat. Hist. 238, 1-95.
Iverson, J.B., 1977. Geographic variation in the musk turtle, Sternotherus minor. Copeia 1977,
502-517.
Iverson, Le, and Ingram 17

Iverson, J.B., 1979a. On the validity of Kinosternon arizonense Gilmore. Copeia 1979, 175-177.
Iverson, J.B., 1979b. A taxonomic reappraisal of the yellow mud turtle, Kinosternon flavescens
(Testudines: Kinosternidae). Copeia 1979, 212-225.
Iverson, J.B., 1981. Biosystematics of the Kinosternon hirtipes species group (Testudines:
Kinosternidae). Tulane Stud. Zool. Bot. 23, 1-74.
Iverson, J.B., 1984. Proportional skeletal mass in turtles. Florida Sci. 47, 1-11.
Iverson, J.B., 1988a. Distribution and status of Creaser's mud turtle (Kinosternon creaseri).
Herpetol. J. 1, 285-291.
Iverson, J.B., 1988b. Neural bone patterns and the phylogeny of the turtles of the subfamily
Kinosterninae. Milwaukee Public Mus. Contrib. Biol. Geol. 75, 1-12.
Iverson, J.B., 1989a. Natural history of the Alamos mud turtle, Kinosternon alamosae
(Kinosternidae). Southwest. Nat. 34, 134-142.
Iverson, J.B., 1989b. The Arizona mud turtle Kinosternon flavescens arizonense (Kinosternidae)
in Arizona and Sonora. Southwest. Nat. 34, 356-368.
Iverson, J.B., 1991. Preliminary phylogenetic hypotheses for the evolution of modern
kinosternine turtles. Herpetol. Monogr. 5, 1-27.
Iverson, J.B., 1992a. A Revised Checklist with Distribution Maps of the Turtles of the World.
Privately printed, Richmond, Indiana.
Iverson, J.B., 1992b. Species richness maps of the freshwater and terrestrial turtles of the world.
Smithsonian Herpet. Inform. Serv. 88, 1-18.
Iverson, J.B., 1998. Molecules, morphology, and mud turtle phylogenetics. Chelon. Conserv.
Biol. 3, 113-117.
Iverson, J.B., 1999. Reproduction in the Mexican mud turtle Kinosternon integrum. J. Herpetol.
33,145-149.
Iverson, J.B., 2010. Reproduction in the Red-Cheeked Mud Turtle (Kinosternon scorpioides
cruentatum) in southeastern Mexico and Belize, with comparisons across the species
range. Chelon. Conserv. Biol. 9, 250-261.
Kirby, M.X., Jones, D.S., McFadden, B.J., 2008. Lower Miocene stratigraphy along the Panama
Canal and its bearing on the Central American peninsula. PloS One 3, 2791 (1-14).
Knauss, G.E., Joyce, W.G., Lyson, T.R., Pearson, D., 2011. A new kinosternoid from the late
Cretaceous Hell Creek Formation of North Dakota and Montana and the origin of the
Dermatemys mawii lineage. Paläontol. Z. 85, 125-142.
Koch, P.L., Morrill, C. 2000. Paleocene-Eocene climate change in southwestern Wyoming:
isotopic records from paleosol carbonates and freshwater bivalves. GFF 122, 89-91.
Krenz, J.G., Naylor, G.J.P., Shaffer, H.B., Janzen, F.J., 2005. Molecular phylogenetics and
evolution of turtles. Molecular Phylogenetics and Evolution 37, 178-191.
Lamb, T., Lovich, J.E., 1990. Morphometric validation of the striped mud turtle (Kinosternon
baurii) in the Carolinas and Virginia. Copeia 1990, 613-618.
Langebartel, D.A., 1953. Faunal and archeological researches in Yucatan caves. Part 4. The
reptiles and amphibians. Cranbrook Inst. Sci. Bull. 33, 97-108.
Le, M., McCord, W.P., 2008. Phylogenetic relationships and biogeographical history of the
genus Rhinoclemmys Fitzinger, 1835 and the monophyly of the turtle family Geoemydidae
(Testudines: Testudinoidea). Zool. J. Linn. Soc. 153, 751-767.
Le, M., McCord, W.P., Iverson, J.B., 2007. On the paraphyly of the genus Kachuga
(Testudines: Geoemydidae). Mol. Phylo. Evol. 45, 398-404.
Legler, J.M., 1965. A new species of turtle, genus Kinosternon from Central America. Univ.
Kansas Publ. Mus. Natur. Hist. 15, 615-625.
Iverson, Le, and Ingram 18

Lourenço, J.M., Claude, J., Galtier, N., Chiari, Y., 2012. Dating crytodiran nodes: origin and
diversification of the turtle superfamily Testudinoidea. Mol. Phylo. Evol. 62, 496-507.
Martin, B.T., Bernstein, N.P., Birkhead, R.D., Koukl, J.F., Mussmann, S.M., Placyk Jr., J.S.,
2013. Sequence-based molecular phylogenetics and phylogeogaphy of the American box
turtles (Terrapene spp.) with support from DNA barcoding. Mol. Phylo. Gen., in press.
[http://dx.doi.org/10.1016/j.ympev.2013.03.006]
Mooser, O., 1980. Pleistocene fossil turtles from Aguascalientes, State of Aguascalientes. Univ.
Nac. Autón. Méx., Inst. Geol., Rev. 4(1), 63-66.
Near, T.J., Meylan, P.A., Shaffer, H.B., 2005. Assessing concordance of fossil calibration points
in molecular clock studies: an example using turtles. Amer. Nat. 165, 137-146.
Nylander, J.A. A., Wilgenbusch, J.C., Warren, D.L., Swofford, D.L., 2008. AWTY (are we there
yet?): a system for graphical exploration of MCMC convergence in Bayesian
phylogenetics. Bioinformatics 24, 581-584.
Rambaut, A., Drummond, A.J., 2007. Tracer v1.4, Available from
http://beast.bio.ed.ac.uk/Tracer
Ronquist, F.; M. Teslenko, P. van der Mark, D.L. Ayres, A. Darling, S. Höhna, B. Larget, L. Liu,
M.A. Suchard & J.P. Huelsenbeck. 2012. MrBayes 3.2: Efficient Bayesian Phylogenetic
Inference and Model Choice Across a Large Model Space. Syst. Bio. 61, 539-542.
Rose, F.L., 1980. Turtles in arid and semiarid regions. Bull. Ecol. Soc. Amer. 61, 89.
Savage, J.M., 2002. The Amphibians and Reptiles of Costa Rica. University of Chicago,
Chicago, Illinois.
Schilde, M., 2001. Schlammschildkröten: Kinosternon, Sternotherus, Claudius, und Staurotypus.
Natur und Tier-Verlag, Münster, Germany.
Schmidt, K.P., 1947. A new kinosternid turtle from Colombia. Fieldiana Zool. 31(13), 109-112.
Seidel, M.E., Iverson, J.B., Adkins, M.D., 1986. Biochemical comparisons and phylogenetic
relationships in the family Kinosternidae (Testudines). Copeia 1986, 285-294.
Serb, J., Phillips, C., Iverson, J.B., 2001. Molecular phylogeny and biogeography of Kinosternon
flavescens based on complete mitochondrial control region sequences. Mol. Phylo. Evol.
18, 149-162.
Spinks, P.Q., Shaffer, H.B., 2009. Conflicting mitochondrial and nuclear phylogenies for the
widely disjunct Emys (Testudines: Emydidae) species complex, and what they tell us
about biogeography and hybridization. Syst. Bio. 58, 1-20.
Stejneger, L., 1925. New species and subspecies of American turtles. J. Washington Acad. Sci.
15(20), 462-463.
Swofford, D.L., 2002. PAUP*: Phylogenetic Analysis Using Parismony Version 4.0b10. Sinauer
Associates, Sunderland, MA.
Tinkle, D.W., 1958. The systematics and ecology of the Sternothaerus carinatus complex
(Testudinata, Chelydridae). Tulane Stud. Zool. 6, 3-56.
Turtle Taxonomy Working Group (van Dijk, P.P., Iverson, J.B., Shaffer, H.B., Bour, R.,
Rhodin, A.G.J.), 2012. Turtles of the World: 2012 update. Annotated checklist of
taxonomy, synonymy, distribution, and conservation status. In: Rhodin, A,G J.,
Pritchard, P.C.H., van Dijk, P.P., Saumure, R.A., Buhlmann, K.A., Iverson, J.B.,
Mittermeier, R.A. (Eds.), Conservation Biology of Freshwater Turtles and
Tortoises. Chelon. Res. Monog. 5, pp. 0.243-0.328.
Vargas-Ramirez, M., Vences, M., Branch, W.R., Daniels, S.R., Glaw, F., Hofmeyr, M.D.,
Kuchling, G., Maran, J., Papenfuss, T.J., Siroky, P., Vieites, D.R., Fritz, U., 2010.
Deep genealogical lineages in the widely distributed African helmeted terrapin:
Iverson, Le, and Ingram 19

evidence from mitochondrial and nuclear DNA (Testudines: Pelomedusidae:


Pelomedusa subrufa). Mol. Phylo. Evol. 56, 428-440.
Vetter, H., 2005. Turtles of the World. Vol. 3. Central and South America. Chimaira, Frankfurt.
Walker, D., Moler, P.E., Buhlmann, K.A., Avise, J.C., 1998a. Phylogeographic patterns in
Kinosternon subrubrum and K. baurii based on mitochondrial DNA restriction analyses.
Herpetologica 54, 174-184.
Walker, D., Ortí, G., Avise, J.C., 1998b. Phylogenetic distinctiveness of a threatened aquatic
turtle (Sternotherus depressus). Conserv. Biol. 12, 639-645.
Wang, L., Zhou, X., Nie, L., Liu, L., Jiang, Y., Huang, Z., Jing, W., 2012. The complete
mitochondrial genome sequences of Chelodina rugosa and Chelus fimbriata (Pleurodira:
Chelidae): implications of a common absence of initiation sites (OL) in pleurodiran turtles.
Mol. Biol. Rep. 39, 2097-2107.
Webb, R.G., 1984. Herpetogeography in the Mazatlán-Durango region of the Sierra Madre
Occidental, Mexico. In Seigel, R.A., Hunt, L.E, Knight, J.L., Malaret, L., Zuschlog, N.L
(Eds.), Vertebrate Ecology and Systematics. A Tribute to Henry S. Fitch. Univ. Kansas
Mus. Nat. Hist. Spec. Publ. 10, pp. 217-241.
Webb, S.D., MacFadden, B.J., Baskin, J.A., 1981. Geology and paleontology of the Love Bone
Bed from the late Miocene of Florida. Amer. J. Sci. 281, 513-544.
Woodburne, M.O. (Ed.), 2004. Late Cretaceous and Cenozoic Mammals of North America:
Biostratigraphy and Geochronology. Columbia University Press, New York.
Yu, Y., Harris, A.J., He, X.J., 2011. RASP (reconstruct ancestral state in phylogenies).
http://mnh.scu.edu.cn/soft/blog/RASP.
Zachos, J., Pagani, M., Sloan, L., Thomas, E., Billups, K., 2001. Trends, rhythms, and
aberrations in global climate 65 ma to present. Science 292, 686-693.
Zamudio, K.R., Greene, H.W., 1997. Phylogeography of the bushmaster (Lachesis muta:
Viperidae): implications for neotropical biogeography, systematics, and conservation.
Biol. J. Linn. Soc. 62, 421-442.
Zug, G.R., 1966. The penial morphology and the relationships of cryptodiran turtles. Occas. Pap.
Mus. Zool., Univ. Michigan 647, 1-24.
Zwickl, D.J., 2006. GARLI-genetic algorithm for rapid likelihood inference.
http://www.bio.utexas.edu/faculty/antisense/garli.
Iverson, Le, and Ingram 20

Figure legends

Fig. 1. GARLI partitioned maximum likelihood tree for the Kinosternidae based on concatenated
sequences of three nuclear and three mitochondrial genes. Numbers at nodes are bootstrap
percentage support values (above: right, partitioned by codon position, and left, not so
partitioned) and Bayesian posterior probability (below: right, partitioned by codon
position, and left, not so partitioned). Asterisk indicates 100% value. Nodes lacking
support values have 100% support for all four partitions.

Fig. 2. Relaxed clock model of divergence times for kinosternid turtles from program BEAST
(see Methods). Shaded bars across nodes are 95% confidence intervals for age of node.
Nodes are numbered for text reference (Section 4.3). Numerical estimates of ages of
nodes are in Table S4. Calibration nodes are indicated by circled C. P+Q = Pliocene +
Quaternary.

Fig. 3. Graphical output from RASP (Reconstruct Ancestral State in Phylogenetics; see Section
2.4 for details). A) Reconstruction of ancestral distributions at each node based on the
Bayesian Binary Method (BBM). B. Color key to predicted ancestral ranges: A -South
America, B - Central America (south of Mesoamerica, Honduras to Panama), C -
Mesoamerica (defined as Isthmus of Tehuantepec to NW Honduras), D - Atlantic Mexico
(Tampico embayment, Tamaulipas to northern Veracruz), E - Mexican Plateau, F - Pacific
coastal Mexico (Sinaloa to Oaxaca), G - Northwest Mexico (Sonora), H - Southwest USA
(Arizona), I - Central USA, J - Southeast USA, and K - Northeast USA. [COLOR IN
PRINT AND ON WEB]
Iverson, Le, and Ingram 21

Table 1
Variation among markers used in our phylogenetic analysis of kinosternid turtles.

Marker Total # characters # variable # informative


---------------------------------------------------------------------------------------
cyt b 1085 471 355
12S 363 61 29
16S 510 141 108
C-mos 522 61 39
RAG1 872 56 23
RAG2 1159 90 35
---------------------------------------------------------------------------------------

Table 3
Explicit taxonomic changes recommended in this study.

Former name Proposed name


-------------------------------------------------------------------------
Staurotypinae Staurotypidae
Kinosternon (leucostomum group) Cryptochelys
K. leucostomum C. leucostoma
K. acutum C. acuta
K. subrubrum steindachneri K. steindachneri
K. scorpiodes abaxillare K. abaxillare
-------------------------------------------------------------------------
DERMATEMYDIDAE
Dermatemys mawii
STAUROTYPIDAE Claudius angustatus
*/98 Staurotypus salvinii
aquatic/tricarinate
q / */* Staurotypus triporcatus
entoplastron/GSD 73/73
semiaquatic 88/94 Kinosternon leucostomum leucostomum
unicarinate

Cryptochelys
Kinosternon angustipons
91/97 Kinosternon dunni
*/* Kinosternon herrerai
no hinge Kinosternon herrerai
entoplastron/small PL
entoplastron/small PL Ki
Kinosternon
t creaserii
aquatic Kinosternon acutum
tricarinate */99
KINOSTERNIDAE */* Kinosternon acutum
Sternotherus odoratus
99/96
aquatic/tricarinate Sternotherus carinatus
*/*
PL hinges/no entoplastron/TSD
PL hinges/no entoplastron/TSD Sternotherus minor minor Sternotherus
44/56 Sternotherus minor peltifer
53/82 Sternotherus depressus
67/‐
61/51 Kinosternon baurii
94/88 Kinosternon subrubrum steindachneri
*/* */99 Kinosternon flavescens
93/89 */* Kinosternon subrubrum hippocrepis
Kinosternon arizonense
semiaquatic */*
Kinosternon durangoense

Kinostern
reduced carination 91/91 Kinosternon alamosae
*/* 69/75
Kinosternon sonoriense
98/98 ‐/*
Kinosternon chimalhuaca
69/75 */* 98/87
*/98 Kinosternon
Ki t hi ti
hirtipes

non
‐/99
62/‐ Kinosternon scorpioides albogulare
60/‐ Kinosternon integrum
*/98 Kinosternon integrum
*/91 Kinosternon integrum
*/* */98
/
61/75 Kinosternon oaxacae
90/96 Kinosternon scorpioides abaxillare
44/‐ Kinosternon scorpioides cruentatum
58/54 Kinosternon scorpioides scorpioides
49/‐
0.03
0.03
66/62
Dermatemys mawii
3 g
Claudius angustatus
1 C Staurotypus triporcatus
Staurotypus salvinii
6 Kinosternon leucostomum leucostomum
Kinosternon angustipons
2 C 5 Kinosternon dunni
7 Kinosternon herrerai
Kinosternon herrerai
9 Kinosternon creaseri
8 Kinosternon acutum
4 11 Kinosternon acutum
Sternotherus carinatus
Sternotherus odoratus
13 Sternotherus minor minor
12 Sternotherus depressus
10 Sternotherus minor peltifer
16 Kinosternon
Ki t baurii
b ii
15 Kinosternon subrubrum steindachneri
17
Kinosternon flavescens
14 Kinosternon subrubrum hippocrepis
19
Kinosternon durangoense
18 Kinosternon arizonense
Kinosternon alamosae
20 22 Kinosternon sonoriense
Kinosternon chimalhuaca
21 Kinosternon hirtipes
Kinosternon scorpioides albogulare
Kinosternon integrum
23 Kinosternon integrum
Kinosternon integrum
Kinosternon scorpioides scorpioides
24 Kinosternon scorpioides
p cruentatum
Kinosternon scorpioides abaxillare
Kinosternon oaxacae
80.0 70.0 60.0 50.0 40.0 30.0 20.0 10.0 0.0

Cretaceous Paleocene Eocene Oligocene Miocene P + Q


Table 2
Summary of uncorrected p distances within and among proposed genera of kinosternid turtles
based on the cyt b sequence data. Mean distance of samples between genera appears below
diagonal; mean distance between samples within genera appears along diagonal. Ranges appear
in parentheses. Full compilation of distances is in Table S3.

Claudius Staurotypus Cryptochelys Sternotherus Kinosternon


-------------------------------------------------------------------------------------------------------------
Staurotypus 9.0 (8.7-9.3) 1.4
Cryptochelys 16.0 (15.1-17.3) 17.8 (17.0-18.3) 7.6 (1.2-10.6)
Sternotherus 16.3 (16.1-16.5) 16.8 (15.6-17.6) 11.3 (10.0-13.2) 5.8 (0.0-8.3)
Kinosternon 17.1 (15.7-18.2) 16.7 (14.9-17.7) 12.1 (9.0-14.3) 10.3 (8.7-12.4) 6.8 (2.0-9.8)
-------------------------------------------------------------------------------------------------------------
Iverson, Le, and Ingram 22

Highlights

We present a DNA-based phylogeny of the turtle Family Kinosternidae.


A fossil-calibrated chronogram is provided.
A previously unrecognized clade is described as a new genus.
A zoogeographic history of the family is hypothesized.