Finding arboreal snakes in an evolutionary tree: phylogenetic placement and systematic revision of the Neotropical birdsnakes

Abstract

The genus Pseustes Fitzinger, 1843 is composed of three recognized species, Pseustes poecilonotus, P. shropshirei and P. sulphureus, which may be the largest sized colubrid snake in the New World. The group has a complex systematic history that has yet to be untangled using modern molecular phylogenetic approaches. The systematic position, within-group diversity and distribution are therefore uncertain. We obtained samples of four species from multiple specimens across their distribution and analysed one nuclear and two mitochondrial genes to determine the phylogenetic placement of the genus and infer relationships among Pseustes lineages. We find strong support for the paraphyly of Pseustes with respect to the monotypic genus Spilotes, both of which are nested within a clade of at least 23 other New World Colubrinae genera. Based on our results, we formally revise the taxonomy of P. poecilonotus and P. sulphureus, resurrecting the taxon P. polylepis for populations of P. poecilonotus from South America and allocating P. sulphureus to the genus Spilotes which renders both genera monophyletic. Additionally, we identify two lineages that are putatively new and currently unrecognized species. Finally, the placement of P. sulphureus, the type species of Pseustes, in the genus Spilotes, requires the allocation of the senior synonym Phrynonax be considered for the remaining Pseustes taxa.

Full text

Short Communication
1
Department of Ecology and Evolution, University of Colorado at Boulder, Boulder CO, USA;
2
Department of Biology, Northeastern
Illinois University, Chicago IL, USA;
3
Department of Biology, The Graduate School and University Center, The City University of
New York, New York NY, USA;
4
Department of Biology, The College of Staten Island, CUNY, Staten Island NY, USA;
5
Museo de
Biolog

ıa, Facultad Experimental de Ciencias, La Universidad del Zulia, Maracaibo Venezuela;
6
Sam Noble Museum and Department
of Zoology, University of Oklahoma, Norman OK, USA;
7
Instituto de Biodiversidad Tropical, Uvita Puntarenas, Costa Rica
Finding arboreal snakes in an evolutionary tree: phylogenetic placement and
systematic revision of the Neotropical birdsnakes
ROBERT C. JADIN
1,2
,FRANK T. BURBRINK
3,4
,GILSON A. RIVAS
5
,LAURIE J. VITT
6
,C

ESAR L. BARRIO-AMOR

OS
7
and
ROBERT P. GURALNICK
1
Abstract
The genus Pseustes Fitzinger, 1843 is composed of three recognized species, Pseustes poecilonotus,P. shropshirei and P. sulphureus, which may be
the largest sized colubrid snake in the New World. The group has a complex systematic history that has yet to be untangled using modern molecular
phylogenetic approaches. The systematic position, within-group diversity and distribution are therefore uncertain. We obtained samples of four species
from multiple specimens across their distribution and analysed one nuclear and two mitochondrial genes to determine the phylogenetic placement of
the genus and infer relationships among Pseustes lineages. We find strong support for the paraphyly of Pseustes with respect to the monotypic genus
Spilotes, both of which are nested within a clade of at least 23 other New World Colubrinae genera. Based on our results, we formally revise the tax-
onomy of P. poecilonotus and P. sulphureus, resurrecting the taxon P. polylepis for populations of P. poecilonotus from South America and allocating
P. sulphureus to the genus Spilotes which renders both genera monophyletic. Additionally, we identify two lineages that are putatively new and
currently unrecognized species. Finally, the placement of P. sulphureus, the type species of Pseustes, in the genus Spilotes, requires the allocation of
the senior synonym Phrynonax be considered for the remaining Pseustes taxa.
Key words: Colubridae –phylogeny –Phrynonax –Pseustes –Serpentes –Spilotes
Introduction
Origin and evolution of the Neotropical fauna remains of great
interest, especially given the complex geological history of the
area, habitat heterogeneity and high levels of endemicity of the
region (Cracraft and Prum 1988; Duellman 1999; Daza et al.
2009). However, well-resolved phylogenies are still lacking for
many organisms, and thus, the evolutionary histories as well as
spatial and ecological distributions remain poorly known for a
majority of taxa (Daza et al. 2009; Schargel et al. 2010). Despite
recent advancements in our understanding of evolutionary rela-
tionships of Neotropical colubrid snakes, which represent a
majority of snake taxa in the New World (e.g. Pyron et al.
2011), most taxa, including several genera, have yet to be exam-
ined using molecular phylogenetic approaches. Here, we begin to
assemble one piece of that puzzle by focusing on the Neotropical
snake genus Pseustes, commonly referred to as ‘birdsnakes’or
‘puffing snakes’. Three species are currently recognized in the
genus (Rivas et al. 2012): Pseustes poecilonotus G€
unther 1858;
P. shropshirei Barbour and Amaral 1924; and P. sulphureus
Wagler 1824; which is among the largest sized colubrid snake in
the New World (P
erez-Santos and Moreno 1988). Other species
of Pseustes mentioned in Uetz (2013), such as P. cinnamomeus
Wagler 1824 and P. sexcarinatus Wagler 1824; are considered
in the synonymy of Chironius scurrulus Wagler, 1924 and
C. quadricarinatus Boie 1827; respectively (Dixon et al. 1993;
Giraudo and Scrocchi 2002).
To date, evolutionary relationships within this genus have not
been assessed, rendering their evolutionary relationships and spe-
cies boundaries poorly understood. Additionally, accurate taxon-
omy of Pseustes remains questionable as species in the genus
have been classified as belonging to numerous other genera,
including Ahaetulla,Chironius,Coluber,Dipsas,Herpetodryas,
Natrix,Phrynonax,Spilotes,Synchalinus,Thamnobius and
Tropidodipsas (Uetz, 2013). Recently, Pyron et al. (2013) found
support that Pseustes sulphureus is the sister taxon to Spilotes
pullatus, both included within the Colubrinae. However, the esti-
mated phylogenetic position of this species was based on a sin-
gle 12S gene fragment as part of a large, multigene concatenated
phylogenetic analysis of 4161 species of squamate reptiles.
Therefore, increased individual, taxonomic and gene sampling is
warranted to confirm this finding.
We utilize samples collected from multiple individuals of
Pseustes across Central and South America in order to conduct a
molecular phylogenetic analysis of this genus to (i) infer the phy-
logenetic position of Pseustes within the subfamily Colubrinae,
(ii) assess the relationship between Spilotes and Pseustes, (iii)
determine whether species of the genus Pseustes form a mono-
phyletic group, (iv) infer phylogenetic relationships within Pseus-
tes and (v) assess species-level diversity to resolve historical
taxonomic debates. We provide a solid foundation for under-
standing the evolutionary history of the genus Pseustes, which
will aid future work on this and other understudied groups of
Neotropical snakes.
Methods
Molecular sampling
Genomic DNA was isolated from muscle tissue of 17 specimens of
Pseustes and Spilotes using a Qiagen DNeasy extraction kit and protocol.
Corresponding author: Robert C. Jadin (rcjadin@gmail.com)
Contributing authors: Frank T. Burbrink (burbrink@mail.csi.cuny.edu),
Gilson A. Rivas (anolis30@hotmail.com), Laurie J. Vitt (vitt@ou.edu),
C
esar L. Barrio-Amor
o (cesarlba@yahoo.com), Robert P. Guralnick
(robert.guralnick@colorado.edu)
J Zoolog Syst Evol Res (2014) 52(3), 257--264
Accepted on 1 November 2013
© 2013 Blackwell Verlag GmbH J Zoolog Syst Evol Res doi: 10.1111/jzs.12055

Two mitochondrial [NADH dehydrogenase subunit 4 (ND4) and cyto-
chrome b(cyt b)] and one nuclear oocyte maturation factor Mos (c-mos)
gene fragments were independently PCR-amplified using GoTaq
â
Green
master mix by Promega, Madison, WI, USA. Protocols for amplification
were carried out as described in Ar
evalo et al. (1994), Burbrink et al.
(2000), de Queiroz et al. (2002) and Lawson et al. (2005) incorporating
the primer pairs ND4 +LEU, L14910 +H16064 and S77 +S78 and
annealing temperatures 48, 46 and 55°C, respectively. Sequencing was
performed in both forward and reverse directions using the PCR primers
on a Beckman Coulter automated capillary sequencer, and sequence chro-
matographs were edited using Sequencher 4.2, Gene Codes Corporation,
Ann Arbor, MI, USA. Sequences for each gene fragment were aligned
separately, first automatically using the program MUSCLE (Edgar 2004),
and then manually rechecked using Se-Al v2.0a11 (Rambaut 2002). No
internal stop codons were found in these protein-coding gene fragments.
Previously published sequences of snakes within the family Colubridae
were downloaded from GenBank (see Appendix 1) and were combined
with new sequence data generated in this study (GenBank accession
numbers KF669664–KF669710, Table 1).
Our preliminary analyses based on taxa throughout the family Colubri-
dae found Pseustes to have a close affinity to members the subfamily Co-
lubrinae, and we therefore incorporated taxa in that subfamily in our final
analyses. Grayia tholloni,Pseudorabdion oxycephalum,Scaphiodontophis
annulatus and Storeria dekayi were used as outgroup taxa to root our Co-
lubrinae phylogenetic tree of 95 species.
Phylogenetic analyses
We conducted mixed-model analyses on a concatenated data set (2340 bp
in total), partitioned by gene as well as by codon, resulting in a total of
nine partitions. We used Akaike information criterion (AIC) to identify the
best-fit models of nucleotide substitution for both Bayesian inference (BI)
and maximum-likelihood (ML) analyses. For this, we implemented the
program MrModeltest v2.2 (Nylander 2004), run in PAUP*v4.0b10
(Swofford 2002), which recovered the GTR +I+Γmodel for all three
codon positions of ND4 and cyt bwhile finding models HKY +Γfor the
first and second position of c-mos and GTR +Γfor the third position.
We inferred phylogenetic relationships using Bayesian inference crite-
rion implementing MrBayes v3.0b4 (Ronquist and Huelsenbeck 2003).
Two simultaneous runs were conducted (with the default Markov chain
Monte Carlo [MCMC] settings), for a total of 10.0 910
6
generations
per run, sampling trees and parameters every 100 generations. We used
potential scale reduction factor values (output by MrBayes), together with
plots of cold-chain likelihood values and parameter estimates visualized
in Tracer v1.5.4 (Rambaut and Drummond 2009) to confirm stationarity
and convergence of MCMC runs. Based on this evaluation, the first
2.5 910
6
generations from each run were discarded as burn-in.
Using the same partitioning scheme described above, we inferred the
ML tree using RAxML 7.2.8 under the GTRCAT model and assessed
tree support with the rapid-bootstrapping algorithm using 1000 nonpara-
metric bootstraps (Stamatakis 2006; Stamatakis et al. 2008). Additionally,
we performed the SHL test (Shimodaira and Hasegawa 1999; Anisimova
and Gascuel 2006) in RAxML 7.2.8 to provide another maximum-likeli-
hood measure of support. Support for the SHL test is measured as 1 - p,
where p is equivalent to the probability of obtaining a particular test sta-
tistic under the null hypothesis that the maximum-likelihood estimate of
the branch is not significantly more likely than any nearest-neighbour
rearrangements of that branch.
Results
Our phylogenetic analyses are very similar to recent molecular
phylogenies of the Colubrinae (e.g. Pyron et al. 2011, 2013).
Our analyses recovered the genus Pseustes nested within a clade
of 23 other New World Colubrinae genera (Fig. 1), and paraphy-
letic with respect to the closely related genus Spilotes (Fig. 2).
Unfortunately, our phylogenetic analyses do not resolve the sister
Table 1. GenBank numbers for DNA sequences generated in this study (KF669664–KF669710)
Species Voucher Locality ND4 cyt bc-mos
Pseustes poecilonotus USNM 564157 Gracias a Dios, Tapalw
as, Honduras
(14°51′N, 84°32′W)
KF669690 KF669674 KF669708
Pseustes poecilonotus LSUMZ H-14673 Rio San Juan, Nicaragua: Ca.
15 km S. El Castillo on north bank
Rio San Juan at Isla el Diamante
KF669689 KF669673 KF669707
Pseustes poecilonotus JMR 725 Near La Mica Biological Station, El
Cop
e, Cocl
e, Panama (8°37′12″N,
80°36′0″W)
KF669687 KF669671 KF669705
Pseustes poecilonotus JMR 744 Parque Nacional G. D. Omar Torrijos
Herrera, Cocl
e, Panama (8°40′N, 80°37′W)
KF669688 KF669672 KF669706
Pseustes polylepis LSUMZ H-17739 Rond^
onia, Brazil KF669693 KF669676 KF669710
Pseustes polylepis UTA R-55965 Morona-Santiago, Ecuador: Road to
Mendez (2.65590°S; 78.20707°W)
KF669692 KF669675 KF669709
Pseustes polylepis LSUMZ 42718 “Suriname”KF669691 ––
Pseustes sp. UTA R-46140 Alta Verapaz, Guatemala: Cob
an, Parque
Nacional. Laguna Lachu
a. ca. 175 MSNM
–KF669670 KF669704
Pseustes sp. LSUMZ 36746 “Honduras”KF669686 KF669669 KF669703
Pseustes sp. LSUMZ 39592 North coast of Honduras KF669685 KF669668 KF669702
Pseustes sp. LSUMZ H-7806 North coast of Honduras KF669684 KF669667 KF669701
Spilotes sp. LSUMZ H-14026 Amazonas, Brazil: Rio Ituxi at the Madeirera
Scheffer (8°20′47″N, 65°42′57.9″W)
KF669679 –KF669696
Spilotes pullatus UTA R-52006 Pet
en, Guatemala: La Libertad, Parque
Nacional Sierra Lacand
on, Distrito Guayac
an
KF669677 –KF669694
Spilotes pullatus LSUMZ 36738 “Honduras”KF669678 –KF669695
Pseustes sulphureus LSUMZ H-14023 Amazonas, Brazil KF669681 KF669665 KF669698
Pseustes sulphureus LSUMZ 43274 Pasco Department, Peru; 41 km Villa Rica
Puerto. Bermudez Hwy, 750 m elev.
KF669680 KF669664 KF669697
Pseustes sulphureus LSUMZ 42645 “Suriname”KF669682 KF669666 KF669699
Pseustes sulphureus EBRG 5107 Bolivar, Venezuela: 25 km W Santa Elena
de Uairen via El Paují
KF669683 –KF669700
Abbreviations of institutions and individuals for voucher specimens are as follows: EBRG (Museo de la Estacion Biologica Rancho Grande, Maracay,
Venezuela), JMR (Julie M. Ray field series), LSUMZ (Louisiana State Museum of Natural Science), USNM (Smithsonian Institution, National
Museum of Natural History), UTA (Amphibian and Reptile Diversity Research Center, University of Texas, Arlington).
J Zoolog Syst Evol Res (2014) 52(3), 257--264
©2013 Blackwell Verlag GmbH
258 JADIN,BURBRINK,RIVAS,VITT,BARRIO-AMOR

OS and GURALNICK

clade to Pseustes +Spilotes; multiple other lineages of New
World Colubrinae snakes form a basal polytomy at this node.
Within the Pseustes +Spilotes clade, we identify six diver-
gent lineages, three within each of two larger clades (Fig. 2).
Our results strongly support Pseustes sulphureus being more
closely related to Spilotes pullatus lineages than other species
of Pseustes.Pseustes poecilonotus as currently recognized is
composed of two distinct, geographically separated lineages
based on current sampling, one of which is found in Central
America and the other in South America. Sister to this P. poe-
cilonotus clade is a lineage that occurs in northern Central
America and may be sympatric with Honduran populations of
P. poecilonotus. Finally, our findings show a deep split
between Spilotes pullatus from Brazil and other S. pullatus
populations in Central America, potentially being indicative of
cryptic diversity.
(a)
Fig. 1. Phylogenetic estimate of relationships between genera and species within the Colubrinae. The tree was estimated from a Bayesian 50% major-
ity-rule consensus composed from a concatenated multigene data set (ND4, cyt band c-mos; total of 2340 bp). Numbers at nodes represent values of
Bayesian posterior probabilities (PP, above) and maximum-likelihood bootstraps and SHL tests (BS/SHL, below). Values are provided for nodes
supported by ≥95% PP and ≥70 BS/SHL
J Zoolog Syst Evol Res (2014) 52(3), 257--264
©2013 Blackwell Verlag GmbH
Systematic Revision of the Birdsnakes 259

Discussion
Phylogeny, species boundaries and taxonomic recommenda-
tions within Pseustes
Our study reveals that the genus Pseustes can formally be placed
inside a New World Colubrinae clade (Fig. 1) and that this
genus is paraphyletic with respect to Spilotes (Fig. 2). In particu-
lar, we find the species Pseustes sulphureus to be the sister taxon
of Spilotes pullatus, which together are sister to the other Pseus-
tes species. Given that the genus Spilotes Linneaus, 1758 is older
than Pseustes Fitzinger 1843; we recommend changing Pseustes
sulphureus to Spilotes sulphureus. This taxonomic change will
resolve the paraphyly of Pseustes and render both genera mono-
phyletic. Additionally, this relationship is supported by a unique
(b)
Fig. 1. (continued)
*/59
**/**
**/**
**/**
**/**
**/**
**/**
**/**
**/**
**/**
**/**
**/**
**/**
unknown, genbank
Guatemala, UTA R-52006
Guatemala, UTA R-46140
Panama, JMR 725
Panama, JMR 744
Nicaragua, H14673
Honduras, LSUMZ 36738
Honduras, H7806
Honduras, LSUMZ 39592
Honduras, USNM 564157
Peru, LSUMZ 43274
Brazil, H14023
Brazil, H14026
Brazil, H17739
Suriname, LSUMZ 42645
Suriname, LSUMZ 42718
Venezuela, EBRG 5107
Ecuador, UTA R-55965
Spilotes pullatus
Spilotes sulphureus
Spilotes sp.
Pseustes polylepis
Pseustes sp.
Pseustes poecilinotus
unknown, LSUMZ 36746
Fig. 2. Phylogenetic estimate of relationships within the Pseustes and Spilotes clade, resulting from a Bayesian 50% majority-rule consensus phylo-
gram. ** = 100 posterior probability and 95–100 bootstrap/SHL support, respectively
J Zoolog Syst Evol Res (2014) 52(3), 257--264
©2013 Blackwell Verlag GmbH
260 JADIN,BURBRINK,RIVAS,VITT,BARRIO-AMOR

OS and GURALNICK

defensive behaviour, which includes an exaggerated neck posture
display and inflating of the gular region, not found in the remain-
ing species of Pseustes. Together, they also share a well-devel-
oped tracheal left lung with respect to their snout-vent length
(Rossman and Williams 1966). Although the other species of
Pseustes possesses a similar ‘puffing’behaviour (e.g. P. poecil-
onotus), this trait is more similar to typical colubrid snakes that
inflate their neck as a defensive behaviour when compared to the
more dramatic displays by Spilotes pullatus and S. sulphureus
(Rand and Ortleb 1969). Finally, S. pullatus and S. sulphureus
are notable for being among the largest known New World colu-
brid snakes (along with Clelia clelia and Drymarchon spp.),
reaching lengths upwards of three metres (P
erez-Santos and Mo-
reno 1988).
G€
unther (1858) described Pseustes poecilonotus from Hondu-
ras and Mexico, but Boulenger (1894) later restricted the species
to Honduras. Peters (1867) described P. polylepis (as Ahaetulla
polylepis) from Suriname, now considered one of four recog-
nized subspecies of P. poecilonotus. In this study, our sampling
included specimens from several localities throughout Central
and South America, including both Honduras and Suriname, and
we find that two lineages of P. poecilonotus are separated geo-
graphically somewhere between western Panama and northern
South America. Therefore, the name P. poecilonotus must be
applied only to the Mesoamerican populations, while South
American populations should be treated as the separate lineage
P. polylepis (Amazon and the Guianas).
Taxonomic recommendations for the genus Pseustes
Spilotes sulphureus is the type species of Pseustes Fitzinger
1843; and therefore, the remaining species in the genus Pseustes
must be allocated to Phrynonax Cope 1862 as a replacement.
We take this action based on the principle of priority (Interna-
tional Code of Zoological Nomenclature, article 23.9.9.1). In this
case, Amaral (1929) was the last author who used Phrynonax to
refer to a species of Pseustes, postdating the 1899 cut-off for
such a usage.
Cryptic diversity within Phrynonax and Spilotes
Our phylogenetic analyses provide a greater understanding of the
species composition within the Phrynonax and Spilotes clade. In
particular, our analyses detect two putative species-level lineages
that will likely need to be resurrected from previously described
species or elevated from subspecies status based on literature and
type material. The first undescribed lineage found within Phryno-
nax occurs in both Honduras and Guatemala, which may be
broadly sympatric with P. poecilonotus throughout much of its
range (Fig. 3, labelled Pseustes sp.). The second putative new
lineage we detected suggests that populations of Spilotes pullatus
from Central America are distinct from populations in South
America, which may correspond to subspecies that are occasion-
ally recognized for this taxon (Fig. 3, labelled Spilotes sp.). At
this time however, this inference is based on only one divergent
sample found in Brazil. A thorough sampling of specimens
throughout their range and an increased sampling of molecular
loci for lineages within these genera are needed for a more com-
plete systematic assessment of the diversity, distributions and
evolutionary history of these taxa.
Acknowledgements
We thank J.M. Ray (La MICA Biological Station, Panama), D.L. Ditt-
man, C. Duffie, J. Maley and R.T. Brumfield (LSU Museum of Natural
Science Collection of Genetic Resources), C.J. Franklin, E.N. Smith and
J.A. Campbell (Amphibian and Reptile Diversity Center, UTA) and J.R.
McCranie for loaning shed skins and/or tissues to this study. RCJ thanks
J.R. Dixon (TCWC), L.W. Porras and P. Uetz for sending some critical
literature references. Special thanks to T. Barros who always supports
GAR in his research. C
esar L. Barrio-Amor
os collected the Venezuelan
sample under permit number 2231 by the Ministerio del Ambiente de
Venezuela and deposited it in the Museo de Biologia de la Estacion Bio-
logica Rancho Grande, Maracay, Venezuela. Funding for the molecular
portion of this study was provided through grants to RCJ from the Joseph
B. Slowinski Memorial Research Fund (California Academy of Sciences),
Charles Edward Shaw Grant (East Texas Herpetological Society) and a
departmental grant from Ecology and Evolutionary Biology at CU Boul-
der. While conducting this study, RCJ received funding through an NSF
GK-12 Fellowship (DGE 0742544). Some specimens and tissues used in
this study were collected during field surveys funded by National Science
Foundation grants DEB-0415430, 9200779 and 9505518 to LJV and
Janalee P. Caldwell, DEB-0416160 to Eric N. Smith and DEB-0613802
and DEB-9705277 to Jonathan A. Campbell.
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Appendix 1. GenBank numbers for sequence data of New World Colubridae analysed in this study
(not including sequences listed in Table 1)
Species ND4 cyt bc-mos
Ahaetulla fronticincta (G€
unther, 1858) –AF471072 AF471161
Arizona elegans (Kennicott, 1859) DQ902279 DQ902101 DQ902058
Bogertophis rosaliae (Mocquard, 1899) DQ902280 DQ902102 DQ902059
Bogertophis subocularis (Brown, 1901) DQ902281 DQ902103 DQ902060
Boiga dendrophila (Boie, 1827) U49303 AF471089 AF471128
Cemophora coccinea (Blumenback, 1788) DQ902282 AF471091 AF471132
Chilomeniscus stramineus (Cope, 1860) U49305 GQ895856 GQ895800
Chionactis occipitalis (Hallowell, 1854) –GQ895857 GQ895801
Chironius carinatus (Linnaeus, 1758) –HQ529280 HQ529281
Chrysopelea paradisi (Boie, 1827) –GQ895858 GQ895802
Coelognathus flavolineatus (Schlegel, 1837) U49301 DQ902128 DQ902090
Coelognathus helena (Daudin, 1803) DQ902292 DQ902112 DQ902071
Coelognathus radiatus (Boie, 1827) DQ902317 DQ902121 DQ902079
Coluber constrictor (Linneaus, 1758) AY487041 AY486914 AY486938
Coluber dorri (Lataste, 1888) AY487042 AY188040 AY188001
Coluber zebrinus (Broadley and Sch€
atti, 2000) AY487058 AY188043 AY188004
Conopsis biserialis (Taylor and Smith, 1942) –GQ895860 GQ895804
Conopsis nasus G€
unther, 1858 –GQ895861 GQ895805
Coronella austriaca (Laurenti, 1768) AY487065 AY486930 AY486954
Coronella girondica (Daudin, 1803) AY487066 AF471088 AF471113
Crotaphopeltis tornieri (Werner, 1908) –AF471093 AF471112
Dasypeltis atra (Sternfeld, 1912) –AF471065 AF471136
Dendrelaphis caudolineatus (Gray, 1834) –GQ895864 GQ895808
Dendrophidion dendrophis (Schlegel, 1837) –GQ895865 GQ895809
Dinodon rufozonatum (Cantor, 1842) –AF471063 AF471163
J Zoolog Syst Evol Res (2014) 52(3), 257--264
©2013 Blackwell Verlag GmbH
262 JADIN,BURBRINK,RIVAS,VITT,BARRIO-AMOR

OS and GURALNICK

Appendix 1. (continued)
Species ND4 cyt bc-mos
Dipsadoboa unicolor G€
unther, 1858 –AF471062 AF471139
Dispholidus typus (Smith, 1828) U49302 AY188012 AY187973
Dolichophis caspius (Gmelin, 1789) AY487039 AY376739 AY376797
Dolichophis jugularis (Linneaus, 1758) AY487046 AY486917 AY486941
Drymarchon corais (Boie, 1827) DQ902314 AF471064 AF471137
Drymobius rhombifer (G€
unther, 1860) –GQ927320 GQ927313
Drymoluber dichrous (Peters, 1863) –GQ895869 GQ895812
Eirenis eiselti (Schmidtler, 1978) AY487069 AY376747 AY376805
Eirenis levantinus (Schmidtler, 1993) AY487071 AY376765 AY376823
Eirenis modestus (Martin, 1838) AY487072 AY486933 AY486957
Eirenis punctatolineatus (Boettger, 1892) AY487073 AY376755 AY376813
Elaphe bimaculata (Schmidt, 1925) DQ902283 DQ902104 DQ902062
Elaphe climacophora (Boie, 1826) DQ902285 DQ902105 DQ902064
Elaphe quadrivirgata (Boie, 1826) DQ902300 DQ902120 DQ902078
Elaphe quatuorlineata (Bonnaterre, 1790) AY487067 AY486931 AY486955
Elaphe rufodorsata (Cantor, 1842) DQ902301 DQ902123 DQ902081
Euprepiophis conspicillata (Boie, 1826) DQ902286 DQ902106 DQ902065
Euprepiophis mandarinus (Cantor, 1842) DQ902294 DQ902115 DQ902073
Gonyosoma oxycephalum (Boie, 1827) DQ902309 AF471084 AF471105
Grayia tholloni DQ486326 DQ486351 DQ486175
Hemerophis socotrae (G€
unther, 1881) AY487055 AY188042 AY188003
Hemorrhois hippocrepis (Linneaus, 1758) AY487045 DQ451987 AY486940
Hemorrhois nummifer (Reuss, 1834) AY487049 AY376742 AY376800
Hierophis spinalis (Peters, 1866) AY487056 AY486924 AY486948
Hierophis viridiflavus (Lac
ep
ede, 1789) AY487057 AY486925 AY486949
Lampropeltis alterna (Brown, 1901) AY497307 AF337130 FJ627799
Lampropeltis Mexicana (Garman, 1884) AY497310 AF337146 FJ627800
Leptophis ahaetulla (Linnaeus, 1758) –GQ927321 GQ927316
Lycodon zawi (Slowinski et al., 2001) –AF471040 AF471111
Lytorhynchus diadema (Dum
eril et al., 1854) –AY188025 AY187986
Maculophis bella (Stanley, 1917) DQ902316 DQ902134 DQ902097
Masticophis flagellum (Shaw, 1802) AY487060 AY486928 AY234228
Mastigodryas boddaerti (Sentzen, 1796) –GQ895867 GQ895811
Mastigodryas melanolomus (Cope, 1868) –GQ895868 –
Oligodon cinereus (G€
unther, 1864) –AF471033 AF471101
Opheodrys aestivus (Linnaeus, 1766) –AF471057 AF471147
Oreocryptophis porphyracea (Cantor, 1839) DQ902298 DQ902118 DQ902076
Orthriophis hodgsoni (G€
unther, 1860) DQ902318 DQ902136 DQ902096
Orthriophis moellendorffi(Boettger, 1886) DQ902295 DQ902116 DQ902074
Oxybelis aeneus (Wagler, 1824) –AF471056 AF471148
Pantherophis guttatus (Linneaus, 1766) DQ902291 DQ902111 DQ902070
Philothamnus heterodermus (Hallowell, 1857) –AF471055 AF471149
Phyllorhynchus decurtatus (Cope, 1868) –AF471083 AF471098
Pituophis catenifer (Blainville, 1835) AF138764 AF337112 FJ627790
Pituophis deppei (Dum
eril, 1853) AF141096 FJ627818 FJ627801
Platyceps florulentus (Geoffroy, 1827) AY487043 AY486915 AY486939
Platyceps najadum (Eichwald, 1831) AY487038 AY486912 AY486936
Platyceps rhodorachis (Jan, 1865) AY487051 AY486921 AY486945
Pseudelaphe flavirufa (Cope, 1867) DQ902289 DQ902109 DQ902068
Pseudocyclophis persicus (Anderson, 1872) –AY376757 AY376815
Pseudoficimia frontalis (Cope, 1864) –GQ895886 GQ895827
Pseudorabdion oxycephalum (Boie, 1827) –AF471073 DQ112083
Pseustes sulphureus (Wagler, 1824)
Ptyas korros (Schlegel, 1837) AY487062 AY486929 AY486953
Rhadinophis frenatum (Gray, 1853) DQ902290 DQ902110 DQ902069
Rhadinophis prasina (Blyth, 1854) DQ902299 DQ902119 DQ902077
Rhinechis scalaris (Schinz, 1822) AY487068 AY486932 AY486956
Salvadora mexicana (Dum
eril et al., 1854) AY487075 AY486934 AY486958
Scaphiodontophis annulatus (Dum
eril et al., 1854) –GQ927323 GQ927318
Senticolis triaspis (Cope, 1866) AF138775 DQ902127 DQ902086
Sonora semiannulata (Baird and Girard, 1853) –AF471048 AF471164
Spalerosophis diadema (Schlegel, 1837) AY487059 AF471049 AF471155
Spilotes pullatus (Linnaeus, 1758) –AF471041 AF471110
Stenorrhina freminvillei (Dum
eril et al., 1854) –GQ895889 GQ895830
Storeria dekayi (Holbrook, 1839) EF417365 AF471050 AF471154
Sympholis lippiens (Cope, 1861) –GQ895890 GQ895831
Tantilla relicta (Telford, 1966) –AF471045 AF471107
Telescopus fallax (Fleischmann, 1831) –AF471043 AY188000
Thelotornis capensis (Smith, 1849) –AF471042 AF471109
J Zoolog Syst Evol Res (2014) 52(3), 257--264
©2013 Blackwell Verlag GmbH
Systematic Revision of the Birdsnakes 263

Appendix 1. (continued)
Species ND4 cyt bc-mos
Thrasops jacksonii (G€
unther, 1895) –AF471044 DQ112084
Trimorphodon biscutatus (Dum
eril et al., 1854) DQ497506 GQ927324 GQ927319
Zamenis hohenackeri (Strauch, 1873) DQ902320 DQ902137 DQ902098
Zamenis lineata (Camerano, 1891) DQ902319 AJ277674 DQ902099
J Zoolog Syst Evol Res (2014) 52(3), 257--264
©2013 Blackwell Verlag GmbH
264 JADIN,BURBRINK,RIVAS,VITT,BARRIO-AMOR

OS and GURALNICK