Molecular Phylogenetics and Evolution
Vol. 12, No. 3, August, pp. 320–332, 1999
Article ID mpev.1999.0641, available online at http://www.idealibrary.com on
Vicariant Patterns of Fragmentation among Gekkonid Lizards of the
Genus Teratoscincus Produced by the Indian Collision: A Molecular
Phylogenetic Perspective and an Area Cladogram for Central Asia
J. Robert Macey,*,1 Yuezhao Wang,† Natalia B. Ananjeva,‡ Allan Larson,*
and Theodore J. Papenfuss§
*Department of Biology, Box 1137, Washington University, St. Louis, Missouri 63130; †Chengdu Institute of Biology, Chengdu,
Sichuan, China; ‡Zoological Institute, Russian Academy of Sciences, St. Petersburg, Russia; and §Museum of Vertebrate Zoology,
University of California, Berkeley, California 94720
Received October 21, 1998; revised December 29, 1998
A well-supported phylogenetic hypothesis is presented for gekkonid lizards of the genus Teratoscincus.
Phylogenetic relationships of four of the five species are
investigated using 1733 aligned bases of mitochondrial
DNA sequence from the genes encoding ND1 (subunit one
of NADH dehydrogenase), tRNAIle, tRNAGln, tRNAMet, ND2,
tRNATrp, tRNAAla, tRNAAsn, tRNACys, tRNATyr, and COI
(subunit I of cytochrome c oxidase). A single most parsimonious tree depicts T. przewalskii and T. roborowskii as
a monophyletic group, with T. scincus as their sister taxon
and T. microlepis as the sister taxon to the clade containing the first three species. The aligned sequences contain
341 phylogenetically informative characters. Each node is
supported by a bootstrap value of 100% and the shortest
suboptimal tree requires 29 additional steps. Allozymic
variation is presented for proteins encoded by 19 loci but
these data are largely uninformative phylogenetically.
Teratoscincus species occur on tectonic plates of Gondwanan origin that were compressed by the impinging
Indian Subcontinent, resulting in massive montane uplifting along plate boundaries. Taxa occurring in China
(Tarim Block) form a monophyletic group showing vicariant separation from taxa in former Soviet Central Asia
and northern Afghanistan (Farah Block); alternative biogeographic hypotheses are statistically rejected. This vicariant event involved the rise of the Tien Shan-Pamir
and is well dated to 10 million years before present. Using
this date for separation of taxa occurring on opposite
sides of the Tien Shan-Pamir, an evolutionary rate of
0.57% divergence per lineage per million years is calculated. This rate is similar to estimates derived from fish,
bufonid frogs, and agamid lizards for the same region of
the mitochondrial genome (D0.65% divergence per lineage per million years). Evolutionary divergence of the
mitochondrial genome has a surprisingly stable rate
across vertebrates.
r 1999 Academic Press
1
To whom correspondence should be addressed. Fax: (314) 935-4432.
1055-7903/99 $30.00
Copyright r 1999 by Academic Press
All rights of reproduction in any form reserved.
Key Words: Reptilia; Sauria; Gekkota; Gekkonidae; biogeography; Indian collision; mitochondrial DNA; replication; phylogenetics; Asia; China; Kazakhstan; Pakistan.
The gekkonid lizard genus Teratoscincus is endemic
to the desert regions of central and southwest Asia.
Five species of Teratoscincus are currently recognized
(Macey et al., 1997a). Two species, T. bedriagai and T.
microlepis, are restricted to desert regions south of the
Hindu Kush in Iran, Afghanistan, and Pakistan (Fig.
1). Teratoscincus scincus occurs both in the region of
southwest Asia where T. bedriagai and T. microlepis
are found (Anderson, 1993), as well as to the north in
the Central Asian republics of the former USSR (Kazakhstan, Kyrgyzistan, Tadjikistan, Turkmenistan, and
Uzbekistan; Szczerbak and Golubev, 1996). The two
remaining species occur in China and Mongolia. Teratoscincus przewalskii is found in desert regions of the
Taklimakan, Hami Depression, and low-elevation Gobi.
Teratoscincus roborowskii is restricted to the Turpan Depression in China, which is the second lowest depression in
the world. These two species do not overlap in distribution
and are geographically separated from the remaining
three species by the Tien Shan and Pamir (Macey et al.,
1997a). Prior to the taxonomic revision of Macey et al.
(1997a), T. roborowskii was considered conspecific with T.
scincus, suggesting that populations occurring east of the
Tien Shan-Pamir do not form a monophyletic group (see,
Szczerbak and Golubev, 1996; Zhao and Adler, 1993).
Phylogenetic patterns of fragmentation are investigated
in taxa occurring on different tectonic plates of Gondwanan
origin now separated by mountain uplifting that resulted
from the Indian collision. We predict that phylogenetic
relationships in Teratoscincus reflect the historical formation of faunal barriers by the uplift of the Karakorum,
Pamir, Tien Shan, and Hindu Kush mountains (Fig. 1).
Data are reported for nuclear-encoded allozymic
variation representing 19 loci and mitochondrial DNA
320
VICARIANT PATTERNS IN ASIAN Teratoscincus LIZARDS
321
FIG. 1. Map of central and southwest Asia illustrating the approximate distribution of Teratoscincus species relative to major mountain
belts and basins. Montane regions that separate Teratoscincus species are depicted with stippling. Note that the mountain chain consisting of
the Hindu Kush, Karakoram, Himalaya, Pamir, and Tien Shan all connect, causing a dramatic barrier to the distribution of Teratoscincus
species. In addition, the Kopet-Dagh is connected to the Hindu Kush by the Badkyz Plateau (not illustrated) causing a barrier between the
Central Asian republics of the former USSR (Kazakhstan, Kyrgyzistan, Tadjikistan, Turkmenistan, and Uzbekistan) and Southwest Asia. The
Pamir-Tien Shan is particularly noteworthy because it separates T. przewalskii and T. roborowskii from T. scincus.
sequences from the genes encoding ND1 (subunit one of
NADH dehydrogenase), tRNAIle, tRNAGln, tRNAMet,
ND2, tRNATrp, tRNAAla, tRNAAsn, tRNACys, tRNATyr,
and COI (subunit I of cytochrome c oxidase). Both data
sets include taxa that occur on each side of the Tien
Shan and Pamir (T. scincus to the west, and T. przewalskii and T. roborowskii to the east); the phylogenetic
relationships of these taxa are a major focus of this
study. Unfortunately, T. bedriagai, known only from
extreme eastern Iran and adjacent Afghanistan, is not
available for either data set. The sequence data include
representatives of the other four Teratoscincus species
(T. microlepis, T. przewalskii, T. roborowskii, and
T. scincus). Allozymic data were not collected for
T. microlepis, which occurs exclusively in southwest
Asia south of the Hindu Kush. Gekko gecko serves as an
outgroup for both data sets. In the allozymic data, an
additional gekkonine outgroup, Cyrtodactylus tibetanus, is used. An eublepharine species, Eublepharus
turkmenicus, serves as an additional outgroup for the
mitochondrial DNA sequence data.
MATERIALS AND METHODS
Specimen Information
Museum numbers and localities for voucher specimens from which DNA was extracted and GenBank
accession numbers are presented below. Acronyms are
CAS for California Academy of Sciences, San Francisco
322
MACEY ET AL.
and MVZ for Museum of Vertebrate Zoology, University
of California at Berkeley. The acronym followed by a
dash RM represents a field number of the first author
for an uncatalogued specimen being deposited at the
Museum of Vertebrate Zoology. The previously reported
sequence for T. przewalskii (Macey et al., 1997b) has
been extended by 19 bases to include 6 additional
amino acid positions and a third codon position of the
ND1 gene; the GenBank accession has been updated
accordingly.
Eublepharus turkmenicus: sequence CAS 184771,
AF114248, Elev. 300 m, 39° 068 N 55° 088 E, vicinity of
Temen Spring, 2.5 km west of Danata (39° 078 N 55° 088
E) on paved Rd. from Danata to the Ashgabad (Ashkabad) to Krasnovodsk Rd., then 5.4 km south on Dirt
Rd., Krasnovodsk Region, Turkmenistan. Cyrtodactylus tibetanus: allozymes CAS 171751–171760, Elev.
3700 m, at base of mountains, approx 3 km WNW
(airline) of the Potala Palace, Lhasa (29° 398 N 91° 068
E), Lhasa Municipality, Xizang (Tibet) Autonomous
Region, China. Gekko gecko: allozymes MVZ 215269,
215312, 215314, 215356, 215358–215360, sequence
MVZ 215314, AF114249, Patong Beach, Kathu District,
Phuket Island, Phuket Province, Thailand. Teratoscincus microlepis: sequence MVZ-RM10464, AF114250,
captive bred from wild-caught specimens probably originating from Pakistan. Teratoscincus scincus: allozymes
MVZ 216056–216065, sequence MVZ 216056,
AF114251, near Alma-Ata (43° 158 N 76° 578 E), AlmaAta Region, Kazakhstan. Teratoscincus przewalskii:
allozymes CAS 171010–171019, sequence CAS 171010,
U71326 (Macey et al., 1997b,c), Elev. 1000 m, 19.5 km
east of the Uygur girl-Hami Mellon monument in the
center of Hami (42° 488 N 93° 278 E), also at km 177.9
from the Gansu Province line on the Lanzhou-Urumqi
Rd., then 9.0 km NE on dirt road to Mirowlu, Hami
(Kumul) Prefecture, Xinjiang Uygur Autonomous Region, China. Teratoscincus roborowskii: allozymes CAS
171203–171212, sequence CAS 171203, AF114252, Elev.
470 m, 1.7 km south of the Lanzhou-Urumqi Rd. on
Shanshan main street, then 13.3 km east of Shanshan
main street on Ma Chang Dadu Rd., Ma Chang Dadu
District, Shanshan (42° 528 N 90° 108 E), Turpan
Prefecture, Xinjiang Uygur Autonomous Region, China.
Laboratory Protocols
Tissues were taken in the field and immediately
frozen in liquid nitrogen and later transferred to an
ultracold freezer and maintained at ⫺80°C. For analysis of allozymic variation, liver tissue was homogenized
separately from other tissue. Horizontal starch-gel
electrophoresis was employed to differentiate variation
in 19 presumptive loci. The 19 loci and five buffer
conditions utilized to resolve them are displayed in
Table 1. Allozymes were stained using standard methods (Harris and Hopkinson, 1976; Murphy et al., 1990;
Richardson et al., 1986; Selander et al., 1971). Carbox-
TABLE 1
The 19 Allozymic Systems Scored and the
Five Electrophoretic Conditions within Which
They Were Resolved
Enzyme
1. Aconitase hydratase
2. Aconitase hydratase
3. Aspartate aminotransferase
4. Carboxylic ester
hydrolase
5. Fructose-bisphosphate aldolase
6. Glucose-6-phosphate
isomerase
7. Glycerol-3-phosphate
dehydrogenase
8. L-Iditol dehydrogenase
9. Isocitrate dehydrogenase
10. Isocitrate dehydrogenase
11. L-Lactate dehydrogenase
12. L-Lactate dehydrogenase
13. Malate dehydrogenase
14. Malate dehydrogenase
15. Peptidase B
16. Peptidase D
17. Phosphogluconate
dehydrogenase
18. Purine-nucleoside
phosphorylase
19. Superoxide dismutase
Abbreviation E. C. No.
Electrophoretic
conditions a
ACOH-1
ACOH-2
4.2.1.3
4.2.1.3
1
1
AAT
2.6.1.1
3
EST-D b
3.1.1.–
3
FBA
4.1.2.13
4
GPI
5.3.1.9
2
G3PDH
1.1.1.8
1
IDDH
1.1.1.14
4
IDH-1
1.1.1.42
1
IDH-2
1.1.1.42
1
LDH-1
1.1.1.27
3
LDH-2
1.1.1.27
3
MDH-1
1.1.1.37
1
MDH-2
PEP-B
PEP-D
1.1.1.37
3.4.11.4
3.4.13.9
1
4
5
PGDH
1.1.1.44
1
PNP
2.4.2.1
5
SOD
1.15.1.1
2
a Electrophoretic conditions: (1) Amine–citrate (morpholine) pH
6.0, 250 v for 6 h or 300 v for 5 h (Clayton and Tretiak, 1972); (2)
Histidine–citrate pH 7.8, 150 v for 8 h (Harris and Hopkinson, 1976);
(3) Lithium–borate/Tris–citrate pH 8.2, 250 v for 6 h or 300 v for 5 h;
(4) Phosphate–citrate pH 7.0, 120 v for 7 h; (5) Tris–HCL pH 8.5, 250
v for 4 1/2 h (all, Selander et al., 1971).
b EST-D, dimeric esterase.
ylic ester hydrolase (Dimeric Esterase) was resolved
using 4-methylumbelliferyl acetate as the substrate;
Peptidase B (PEP-B) was resolved using L-leucyl-Lalanine as the substrate, and Peptidase D (PEP-D)
with the use of L-phenylalanyl-L-proline as the substrate. The isozymes, and loci if more than one, were
labeled according to their migration from anode to
cathode.
Genomic DNA was extracted from liver using the
Qiagen QIAamp tissue kit (Qiagen Inc., Hilden, Germany). Amplification of genomic DNA was conducted
using a denaturation at 94°C for 35 s, annealing at
50°C for 35 s, and extension at 70°C for 150 s with 4 s
VICARIANT PATTERNS IN ASIAN Teratoscincus LIZARDS
added to the extension per cycle, for 30 cycles. Negative
controls were run for all amplifications. Amplified
products were purified on 2.5% Nusieve GTG agarose
gels (FMC Bioproducts, Rockland, ME) and reamplified
under similar conditions. Reamplified double-stranded
products were purified on 2.5% acrylamide gels (Maniatis et al., 1982). Template DNA was eluted from
acrylamide passively over 3 days with Maniatis elution
buffer (Maniatis et al., 1982). Cycle-sequencing reactions were run using the Promega fmol DNA-sequencing system (Fisher Scientific, Pittsburgh, PA) with a
denaturation at 95°C for 35 s, annealing at 45–60°C for
35 s, and extension at 70°C for 1 min for 30 cycles.
Sequencing reactions were run on Long Ranger sequencing gels (FMC Bioproducts, Rockland, ME) for 5–12 h at
38–40°C.
Amplifications were done using either primers L3881,
L3887, L4160, L4178a, or L4178b in combination with
primer H5934. In addition, some taxa were amplified
with primers L3002 and H4419. Both strands were
sequenced using the primers in Table 2. Primer numbers refer to the 38 end on the human mitochondrial
genome (Anderson et al., 1981), where L and H correspond to light and heavy strands, respectively.
Sequence Alignment
Reported sequences are presented in Fig. 2 and
correspond to positions 4161 to 5936 on the human
mitochondrial genome (Anderson et al., 1981). This
sequence contains the genes encoding ND1 (subunit
TABLE 2
Primers Used in This Study a
Position b
L3002
L3887
L4160
L4178a
L4178b
L4221
H4419
L4437
H4645
L5002
H5540
L5556
L5638
H5692
H5934
H5937
Gene
Primer sequence c
16S rRNA
ND1
ND1
ND1
ND1
tRNAIle
tRNAMet
tRNAMet
ND2
ND2
tRNATrp
tRNATrp
tRNAAla
tRNAAsn
COI
COI
58-TACGACCTCGATGTTGGATCAGG-38
58-GACCTAACAGAAGGAGAATCAGA-38
58-CGATTCCGATATGACCARCT-38
58-CARCTWATACACYTACTATGAAA-38
58-CAACTAATACACCTACTATGAAA-38
58-AAGGATTACTTTGATAGAGT-38
58-GGTATGAGCCCAATTGCTT-38
58-AAGCTTTCGGGCCCATACC-38
58-ACAGAAGCCGCAACAAAATA-38
58-AACCAAACCCAACTACGAAAAAT-38
58-TTTAGGGCTTTGAAGGC-38
58-GCCTTCAAAGCCCTAAA-38
58-CTGAATGCAACTCAGACACTTT-38
58-TTGGGTGTTTAGCTGTTAA-38
58-AGRGTGCCAATGTCTTTGTGRTT-38
58-GTGCCAATGTCTTTGTG-38
a All primers are from Macey et al. (1997b) except L4160 which is
from Kumazawa and Nishida (1993).
b Primers are designated by their 38 ends which correspond to the
position in the human mitochondrial genome (Anderson et al., 1981)
by convention. H and L designate heavy-strand and light-strand
primers, respectively.
c Positions with mixed bases are labeled with the standard oneletter code: R ⫽ G or A, W ⫽ A or T, Y ⫽ C or T.
323
one of NADH dehydrogenase), tRNAIle, tRNAGln,
tRNAMet, ND2, tRNATrp, tRNAAla, tRNAAsn, tRNACys,
tRNATyr, and COI (subunit I of cytochrome c oxidase).
Protein-coding sequences were translated to amino
acids using MacClade (Maddison and Maddison, 1992)
for alignment. Transfer-RNA secondary structure was
determined manually using the criteria of Kumazawa
and Nishida (1993) to ensure proper alignment (Macey
and Verma, 1997).
Phylogenetic Analysis
Phylogenetic trees were estimated using PAUP (Swofford, 1993) with exhaustive searches. Bootstrap resampling was applied to assess support for individual nodes
with 1000 bootstrap replicates using branch-and-bound
searches. Decay indices (⫽‘‘branch support’’ of Bremer,
1994) were calculated for all internal branches of the
tree. Branch-and-bound searches retained suboptimal
trees. The decay index for a particular node was tabulated
as the difference in length between the overall shortest tree
and the shortest tree lacking that node.
Wilcoxon signed-ranks tests (Felsenstein, 1985;
Templeton, 1983) were applied to examine statistical
significance of the shortest tree relative to alternative
hypotheses. This test asks whether the most parsimonious tree is significantly shorter than an alternative or
whether their differences in length can be attributed to
chance alone (Larson, 1998). Wilcoxon signed-ranks
tests were conducted as two-tailed tests. Felsenstein
(1985) showed that the two-tailed test is conservative.
The test statistic Ts was compared with critical values
for the Wilcoxon rank sum in table B.11 of Zar (1984).
Alternative phylogenetic hypotheses were tested using the most parsimonious phylogenetic topologies compatible with them. Each branch in the overall shortest
tree was tested statistically by comparing the overall
shortest tree to the shortest tree(s) that did not contain
the node being examined. Alternative tree(s) were
found with branch-and-bound searches that retained
suboptimal trees using PAUP (Swofford, 1993). Statistical tests were conducted using the ‘‘compare trees’’
option of MacClade (Maddison and Maddison, 1992).
Cladistic Analyses of Allozymic Data
Allozymic data were coded in two ways for cladistic
phylogenetic analysis. While presence–absence coding
of alleles has received considerable criticism for a lack
of independence of alleles and the possibility of no allele
being reconstructed for an ancestral node (Swofford
and Olsen, 1990), it remains the method that provides
the greatest amount of resolution. Alternatively, combinations of alleles for a particular locus may be coded as
discrete character states (Buth, 1984). If step matrices
are used to connect character states, a greater amount
of information can be retained (Mabee and Humphries,
1993). In our analysis, step matrices were constructed
on the basis of gains and losses of alleles. For example, a
324
MACEY ET AL.
VICARIANT PATTERNS IN ASIAN Teratoscincus LIZARDS
325
FIG. 3. Overlap between the tRNACys gene and a presumptive nonfunctional OL. Pseudo-OL is depicted as template heavy-strand sequence
and the tRNACys gene, which is encoded on the light strand, is depicted as the presumptive transcribed tRNA. Boldface bases are used in the
function of light-strand replication and also tRNACys. Three bases of the tRNACys gene, two of which encode the AA-stem, also would be used in
the OL stem. Another three bases of the tRNACys gene encoding the AA-stem also would be used in initiation of light-strand replication
(required for in vitro replication in humans; Hixson et al., 1986). Hence, five out of seven bases encoding one side of the tRNACys AA-stem also
would be used in initiation of light-strand replication. The heavy-strand template sequence identified as the point of light-strand elongation in
mouse 38-GCC-58 (Brennicke and Clayton, 1981) is not present. The heavy-strand sequence conserved in lizards as 38-GBCCB-58 (Macey et al.,
1997b) related to the 38-GGCCG-58 sequence found to be required for in vitro replication in humans (Hixson et al., 1986) also is missing
(underlined with arrows). This missing sequence has been hypothesized to be the result of an insertion to the D-loop/stem region of the
tRNACys gene during replication. This insertion resulted in a realigned AA-stem on the opposite side, causing compensatory mutation to the OL
functional regions and disrupting the initiation of light-strand replication (Macey et al., 1997c,d).
fixed difference between two alleles was counted as two
steps, one allele lost and another gained. In the case of a
two-allele polymorphism in one population with one allele
shared with another monomorphic population, a single
gain or loss was counted as a single step. Additional
polymorphisms were counted in the same manner.
RESULTS
Sequences ranging in size from 1763 to 1777 bases of
mitochondrial DNA for six taxa of gekkonid lizards are
presented as 1815 aligned positions in Fig. 2.
Authentic Mitochondrial DNA
Several observations suggest that the DNA sequences analyzed here are from the mitochondrial
genome and not nuclear-integrated copies of mitochondrial genes (see Zhang and Hewitt, 1996). Protein-
coding genes have no premature stop codons, suggesting that these sequences represent functional copies
that encode a protein. Transfer-RNA genes appear to
encode tRNAs with stable secondary structures, indicating functional genes. Strand bias further supports our
conclusion that the six DNA sequences analyzed here
are from the mitochondrial genome. The strong bias
against guanine on the light strand (G ⫽ 11.0–13.8%,
A ⫽ 29.8–34.3%, T ⫽ 22.8–27.3%, and C ⫽ 27.4–33.0%)
is characteristic of the mitochondrial genome but not
the nuclear genome. See Macey et al. (1997b,d) for
similar strand bias across most squamate-reptile families for the same region of the mitochondrial genome.
Origin for Light-Strand Replication and tRNACys
Two structural features of the mitochondrial genome
(origin for light-strand replication and secondary structure of tRNACys ) show derived states in Gekko gecko
FIG. 2. Length-variable regions among the 1815 aligned mitochondrial DNA sequences as used in the phylogenetic analysis. Six regions
totaling 82 positions are excluded from the analysis and are denoted by an ‘‘x’’ above the sequence. Sequences in the tRNACys gene of Gekko
gecko treated as missing are doubly underlined. The peudo-start codon of ACG for COI in Gekko gecko also is doubly underlined. The codon
directly following these three nucleotides is ATG, the typical start codon for most mitochondrial proteins. Positions 401–1300 from the ND2
gene are not shown because this region has no length variation. Sequences are presented as light-strand sequence and tRNA secondary structure is
designated above the sequence. Stems are indicated by arrows in the direction encoded: AA, amino acid-acceptor stem; D, dihydrouridine stem; AC,
anticodon stem; T, T⌿C stem. The tRNA anticodons are designated COD. Asterisks indicate the unpaired 38 tRNA position 73. Periods represent bases
located outside stem regions; 1 depicts the first codon position of protein-coding sequences. STP represents stop codons.
326
MACEY ET AL.
(Fig. 3). Most vertebrates have a mitochondrial gene
order of ND1, tRNAIle, tRNAGln, tRNAMet, ND2, tRNATrp,
tRNAAla, tRNAAsn, OL (origin for light-strand replication), tRNACys, tRNATyr, and COI. In Gekko gecko, a
stem-and-loop structure is present in the typical vertebrate position for OL between the tRNAAsn and tRNACys
genes. This sequence does not have the functional
characteristics of OL identified in studies of mammalian mitochondrial replication (Brennicke and Clayton,
1981; Hixson et al., 1986). We therefore interpret these
sequences as nonfunctional. In addition, the tRNACys
gene of Gekko gecko encodes a tRNA that lacks a
D-stem and instead contains a D-arm replacement loop
(Fig. 3). The sequences reported here provide further
evidence that loss of a recognizable origin for light-strand
replication between the tRNAAsn and tRNACys genes and
changes in secondary structure of tRNACys may be evolutionarily coupled (Macey et al., 1997b,c,d, 1998c).
Assessment of Homology and Sequence Alignment
Among protein-coding genes, a length-variable region encoding the C-terminus of ND1 extending beyond
that of Eublepharus is excluded from analyses (positions 101–118). As aligned in Fig. 2, this region contains
no phylogenetically informative sites. Teratoscincus
scincus has two extra amino acids in the ND2 gene and
gaps are placed in the other taxa at positions 1330–
1335. Gekko gecko appears to have a pseudo-start codon
of ACG for COI in the position homologous with the start
codon of other gekkonid lizards. The codon directly following these three nucleotides is ATG, the typical start codon
for most mitochondrial proteins (Fig. 2).
In two tRNA genes, both the dihydrouridine (D) and
the T⌿C (T) loops are excluded from analyses because
of questionable alignment. In the tRNATrp gene, the
positions excluded are 1398–1409 (D-loop) and 1441–
1448 (T-loop); in the tRNACys gene, the positions excluded are 1658–1665 (T-loop) and 1697–1699 (D-loop).
In addition, Gekko gecko has a tRNACys gene encoding a
tRNA that lacks a D-stem and instead contains a D-arm
replacement loop (Macey et al., 1997c). Transfer-RNA
gene sequences that encode tRNAs with D-arm replacement loops are subject to stem realignments and shifts
in bases from one stem to another, making homology of
bases difficult to assess (Macey et al., 1997c). Therefore,
sequences in the AA-stem and adjacent bases of the
tRNACys gene from Gekko gecko are coded as missing in
phylogenetic analyses (positions 1645–1657, 1704–
1712). In addition, the origin for light-strand replication (OL ) is unalignable between Gekko gecko and the
other taxa and therefore excluded from phylogenetic
analyses (positions 1612–1644).
Of the 1815 aligned positions, 1733 are used in the
phylogenetic analysis. The excluded positions are less
than 5% of the aligned positions.
Genic Variation
Different levels of variation are observed among the 3
protein-coding genes, 8 tRNA-coding genes, and 2
noncoding regions (Table 4). All 11 genes sequenced
contain phylogenetically informative characters. The 8
tRNA genes each have phylogenetically informative
sites in stem and nonstem regions except nonstem
regions of the tRNATrp gene. Each of the three proteincoding genes contains phylogenetic information in first,
second, and third codon positions. Most of the variation
and phylogenetically informative sites are from proteincoding regions. Only 23% (194 sites) of the variable and
19% (65 sites) of the phylogenetically informative sites
are from tRNA genes and noncoding regions. Of the 276
phylogenetically informative characters from proteincoding regions, 159 are from third positions of codons.
Third-position sites account for nearly half of the
phylogenetically informative sites in the total data set.
Only 12% (40 sites) of the phylogenetically informative
sites occur in regions encoding stems of tRNAs, suggesting that compensatory substitutions do not compromise
the phylogenetic analysis.
Phylogenetic Relationships
The allozymic data are unable to resolve relationships among Teratoscincus species (Table 3). When the
data are coded as combinations of alleles for a particular locus and used as discrete character states, no
informative characters are found. When step matrices
are used to connect these character states, three equally
most parsimonious trees are produced with a length of
99 steps. A strict consensus tree of these three trees
does not contain a monophyletic Teratoscincus; instead,
the grouping of T. przewalskii and Cyrtodactylus is
found (results not shown). This grouping collapses in a
single step and therefore is not a significant result.
When these allozymic data are coded as the presence
or absence of alleles, 13 informative characters identify
two equally most parsimonious trees of 78 steps (consistency index 0.987). A strict consensus of these two trees
depicts a monophyletic Teratoscincus, which is supported by a bootstrap value of 100% and a decay index
of 11 (results not shown). One of the shortest trees
groups T. scincus and T. przewalskii as sister species
and the other tree shows T. scincus and T. roborowskii
as sister species. The third possibility of a sister-taxon
relationship between T. przewalskii and T. roborowskii
requires only a single extra step.
In contrast to the allozymic data, the DNA sequence
data provide exceptionally good phylogenetic resolution. The shortest estimate of phylogeny derived from
the 1733 aligned positions (341 informative) is stable
on all branches until trees with lengths of 29 steps
longer are retained in a parsimony search (Fig. 4). In
addition, the bootstrap analysis finds no other trees;
thus, all branches receive a 100% bootstrap value.
The sister-group relationship of the Chinese T. przew-
327
VICARIANT PATTERNS IN ASIAN Teratoscincus LIZARDS
TABLE 3
Distribution of Allelic Variation in Cyrtodactylus, Gekko, and Teratoscincus Species
1
ACOH1
N
2
ACOH2
3
AAT
4
EST-D
5
FBA
6
GPI
7
G3PDH
8
IDDH
9
IDH1
10
IDH2
b
c
e
d
c 0.90
d 0.10
a 0.05
c 0.05
f 0.90
a 0.05
c 0.90
e 0.05
c
c
Cyrtodactylus tibetanus
Gekko gecko
10
7
g
g
b
a
a
b
a
e
e
d
b
a
c
d
Teratoscincus scincus
10
c 0.20
d 0.70
e 0.10
b
b 0.10
c 0.90
b
a 0.20
c 0.80
a 0.20
b 0.80
Teratoscincus przewalskii
10
c 0.15
d 0.05
e 0.75
f 0.05
a
e
d 0.50
f 0.50
a 0.35
c 0.65
d
b
c
c
a
b
c
Teratoscincus roborowskii
10
b
f
b
c 0.60
d 0.40
a
a
b
b 0.05
c 0.95
b 0.10
c 0.90
N
11
LDH1
12
LDH2
13
MDH1
14
MDH2
15
PEP-B
16
PEP-D
17
PGDH
18
PNP
19
SOD
Cyrtodactylus tibetanus
Gekko gecko
10
7
b
a
a
b
b
a
c
a
c
a
c
a
c
a
c
b
Teratoscincus scincus
10
a
c
a
b
b
b
b
a
Teratoscincus przewalskii
Teratoscincus roborowskii
10
10
a
a
c
c
a
a
b
b
b
b
d
a 0.14
b 0.86
c 0.95
f 0.05
e
c
b
b
b
b
a
a
TABLE 4
Distribution of Phylogenetically Informative and Variable Positions
ND1
Codon positions
Informative sites
Variable sites
tRNAIle
2nd
3rd
Stem
Nonstem
Stem
Nonstem
Stem
Nonstem
7
19
4
8
13
30
6
10
3
12
8
22
3
15
1
8
1
9
tRNATrp
a
tRNAAla
tRNAAsn
1st
2nd
3rd
Stem
Nonstem
Stem
Nonstem
Noncoding b
region 1
61
185
40
112
143
286
6
17
—
2
5
17
3
7
—
1
tRNACys
Informative sites
Variable sites
tRNAMet
1st
ND2
Codon positions
Informative sites
Variable sites
tRNAGln
a
Stem
Nonstem
6
19
6
8
COI
Codon positions
tRNATyr
Stem
Nonstem
Stem
Nonstem
Noncoding b
region 2
1st
2nd
3rd
6
11
2
5
2
18
6
12
1
1
4
5
1
3
3
9
Protein coding
codon positions
tRNA
Total
1st
2nd
3rd
Stem
Nonstem
Noncoding
regions
All aligned
sequence
Informative sites
Variable sites
72
209
45
123
159
325
40
122
24
70
1
2
341
851
a
b
Not including D- and T-loops, which are excluded from the analyses.
Noncoding region 1 is between the tRNAAla and the tRNAAsn genes. Noncoding region 2 is between the tRNATyr and the COI genes.
328
MACEY ET AL.
alskii and T. roborowskii is the weakest branch
with a decay index of 29. This clade is the sister taxon
to T. scincus from the former USSR and southwest
Asia; monophyly of the group containing T. przewalskii, T. roborowskii, and T. scincus is supported
by a decay index of 30. Hence, T. microlepis from
the Helmand Basin and adjacent regions in southwest Asia is the basal taxon among Teratoscincus species sampled. The monophyly of Teratoscincus is extremely well supported with a decay index
of 62.
The phylogenetic results provide an area cladogram
for Central Asia. To confirm these results and to test
support for the origins of clades found in separate
historical regions, the Wilcoxon signed-ranks test
(Felsenstein, 1985; Templeton, 1983) is applied.
When the overall shortest tree (Fig. 4) is compared
independently to the shortest alternative trees (Appendix 1) not showing each of the nodes in the overall shortest tree, these alternative trees are rejected
in favor of the overall shortest tree (see Appendix 1;
A-1 and 2, n ⫽ 53, Ts ⫽ 324; B-1, n ⫽ 58, Ts ⫽ 413;
C-1, n ⫽ 128, Ts ⫽ 2128.5; all P ⬍ 0.001). All suboptimal trees are rejected in favor of the overall shortest
tree.
FIG. 4. Single most parsimonious tree produced from analysis of
the 1733 aligned (341 phylogenetically informative) positions from
mitochondrial DNA sequences. The tree has a length of 1268 steps
and a consistency index of 0.864. Bootstrap values are presented
above branches and decay indices below branches.
DISCUSSION
The DNA sequences recover phylogenetic relationships on all branches of the tree with remarkable
support (Fig. 4). Phylogenetic analysis is most conducive to reconstructing branching events that are well
spaced in time, and the geologic history of Asia suggests
that divergence events among Teratoscincus species are
separated by sizable intervals.
The phylogenetic estimate recovered is consistent
with geological predictions. A sequential uplifting of
mountains resulted from compression of Gondwanan
plates along the southern margin of Eurasia as the
impinging Indian subcontinent moved northward. Teratoscincus species may be assigned to tectonic plates of
Gondwanan origin that are now in Asia. While the
evolutionary history of Teratoscincus postdates the
Indian collision, which occurred in the middle Eocene
[Dewey et al., 1989 (45 MYBP); Windley, 1988 (50
MYBP)], the pre-Eocene events produced the geological
conditions necessary for the Eocene and later events
that are directly related to the phylogenetic history
under investigation (Fig. 5).
Approximately 300 MYBP, the Tarim Plate (Taklimakan Desert) collided with the Siberian and Kazakhstan
blocks of Laurasia (Feng et al., 1989; Kwon et al., 1989)
forming the paleo-Tien Shan (mountains) in the vicinity of what is now the western Chinese–former Soviet
border (Fig. 5). A complex series of events followed with
the successive accretion of blocks to the southern
margin of Eurasia. The blocks termed the Cimmerian
Continent broke from Gondwanaland in the late Permian (250 MYBP), migrated northward across the
Tethys Sea, and subsequently broke apart along the
way. The individual blocks collided with Eurasia, and
most completed suturing from late Triassic to middle
Jurassic (225–175 MYBP; for details see Sengör, 1984;
Sengör et al., 1988). These blocks now are situated from
Turkey through Iran, Afghanistan, and Tibet. They
include the North Tibet Block (Qiangtang and SongbanGanzi Terranes), South Tibet Block (Lhasa Terrane),
Farah Block (northern Afghanistan), and Helmand
Block (southern Afghanistan). Figure 5 shows the
current positions of these blocks.
Paleo-sutures of these blocks were reactivated as
strike-slip faults by the Indian collision, and dramatic
mountain building occurred along these faults, separating Teratoscincus species. A sequential uplifting of
mountain ranges is observed, with the ordering largely
dependent on geographic distance from the contact of
the collision. The Indian collision first caused the
trans-Himalayan uplift (Hindu Kush, Karakorum, and
Himalaya), which was followed by the Pamir-Tien Shan
uplift.
Teratoscincus microlepis can be assigned unambiguously to the Helmand Block south of the Hindu Kush.
Teratoscincus przewalskii and T. roborowskii can be
VICARIANT PATTERNS IN ASIAN Teratoscincus LIZARDS
329
FIG. 5. Movements of tectonic plates. During the middle Eocene, 50–45 MYBP, India first contacted Eurasia. Since that time, India and
Laurasian plates have converged 2365 km in the west, 2475 in the center, and 2750 km in the east (Dewey et al., 1989; Molnar et al., 1987). The
high altitudes now present in the Hindu Kush, Karakoram, Tien Shan, and Pamir are attributed to the Indian collision (Dewey et al., 1988,
1989). The Hindu Kush between the Helmand and Farah blocks is associated with the uplift of the trans-Himalaya, which includes the
Karakoram and is one of the earlier uplifting events. The Tien Shan and Pamir, which now separate the Taklimakan Desert (Tarim Plate) from
the former Soviet deserts and the Farah Block, were formed approximately 10 MYBP (Abdrakhmatov et al., 1996; Tapponier et al., 1981).
Crustal shortening and deformation rates are from Dewey et al. (1989). The map is modified from Tapponier et al. (1981).
assigned unambiguously to the Tarim Block east of the
Tien Shan and Pamir. Teratoscincus scincus can be
assigned either to the Farah or the Helmand blocks,
north or south, respectively, of the Hindu Kush. Note
that Laurasian plates north of the Farah Block underwent periods of flooding by the Paratethys Sea until 3.5
MYBP (Dercourt et al., 1986; Steininger and Rogl,
1984) and therefore are unlikely to be a region of
historical endemism for T. scincus. The Tien ShanPamir is well dated at 10 MYBP (Abdrakhmatov et al.,
1996; Tapponier et al., 1981). The separation between
T. scincus from the ancestor of T. przewalskii and
T. roborowskii is expected to be coincident with the
uplift of the Tien Shan-Pamir (10 MYBP). If this
hypothesis is correct, T. scincus would be assigned to
the Farah Block north of the Hindu Kush. Hence, the
formation of the Hindu Kush, which predates the Tien
Shan-Pamir uplift, would have separated T. microlepis
on the Helmand Block from T. scincus on the Farah
Block.
Using the date of 10 MYBP for the formation of the
Tien Shan-Pamir uplift, which separates T. scincus on
the Farah Block from T. przewalskii and T. roborowskii
on the Tarim Block, a rate of evolutionary divergence of
0.57% per lineage per million years is estimated. This
estimate approximates the rate of 0.65% divergence per
lineage per million years suggested for the same segment of the mitochondrial genome from fish, bufonid
frogs, and agamid lizards (Bermingham et al., 1997;
Macey et al., 1998a,b). If our interpretation of T. scincus
being from the Farah Block is correct, then the formation of the Hindu Kush would separate T. scincus on the
330
MACEY ET AL.
TABLE 5
Pairwise Comparisons of DNA Sequences Between
Eublepharus, Gekko, and Teratoscincus Species a
1
1.
2.
3.
4.
5.
6.
Eublepharus
Gekko
T. microlepis
T. scincus
T. przewalskii
T. roborowskii
—
591
527
525
516
521
2
3
4
5
6
35.5%
31.0%
30.8% 30.3% 30.6%
—
30.7%
32.1% 32.0% 32.5%
510
—
16.0% 15.9% 16.6%
534
272
—
11.1% 11.6%
533
271
190
—
6.5%
541
282
198
112
—
a Percentage sequence divergence is shown above the diagonal and
the number of base substitutions between sequences is shown below
the diagonal. Taxa are abbreviated with T. representing Teratoscincus.
second, the formation of the Tien Shan-Pamir occurred
between the Farah and the Tarim blocks at 10 MYBP.
ACKNOWLEDGMENTS
This work was supported by grants from the National Science
Foundation (predoctoral fellowship to J.R.M.; DEB-9726064 to A.L.,
J.R.M. and T.J.P.), National Geographic Society (4110-89 and 4872-93
to T.J.P. and J.R.M.), Russian Foundation of Basic Research (N
97-04-50093 to N.B.A.), and the California Academy of Sciences. We
thank Zhili Fang, Tatjana Dujsebayeza, and Ermi Zhao for field
assistance, and Alok Verma for assistance in the laboratory. The first
author thanks David B. Wake and Margaret F. Smith for the
opportunity to collect allozymic data at the Museum of Vertebrate
Zoology.
APPENDIX 1
Farah Block from T. microlepis on the Helmand Block.
This event predates the formation of the Tien ShanPamir uplift and these taxa should show more than 10
million years of sequence divergence. The average
pairwise sequence divergence across the Tien ShanPamir is 11.4% and that between T. microlepis and
T. scincus, presumably across the Hindu Kush, is 15.9%
(Table 5). Because saturation of mitochondrial sequences occurs past 10 MYBP (Moritz et al., 1987), this
latter split is probably much older than 10 MYBP.
Hence, the DNA sequence data are consistent with
our interpretation of two vicariant events across paleosutures of accreted Gondwanan plates (Fig. 6). First,
the formation of the Hindu Kush between the Helmand
and the Farah blocks occurred before 10 MYBP and,
Alternative hypotheses used in Wilcoxon signedranks tests (Felsenstein, 1985; Templeton, 1983).
Lengths of trees and consistency indices (CI) (Swofford,
1993) are given in parentheses.
The most parsimonious trees that do not contain a
monophyletic grouping of T. przewalskii and T. roborowskii (length of 1297 steps and a CI of 0.845): A-1 ⫽
(Eublepharus, (Gekko, (T. microlepis, ((T. scincus,
T. roborowskii), T. przewalskii)))). A-2 ⫽ (Eublepharus,
(Gekko, (T. microlepis, ((T. scincus, T. przewalskii),
T. roborowskii)))).
The most parsimonious tree that does not contain a
monophyletic grouping of T. scincus, T. przewalskii, and
T. roborowskii (length of 1298 steps and a CI of 0.844):
B-1 ⫽ (Eublepharus, (Gekko, ((T. microlepis, (T. przewalskii, T. roborowskii)), T. scincus))).
The most parsimonious tree that does not contain a
monophyletic grouping of all Teratoscincus species
(length of 1330 steps and a CI of 0.824): C-1 ⫽
(Eublepharus, ((Gekko, T. microlepis), (T. scincus,
(T. przewalskii, T. roborowskii)))).
REFERENCES
FIG. 6. Area cladogram for the tectonic plates derived from
Gondwanaland based on fragmentation patterns among Teratoscincus species. Approximate dates for branching events are shown. As
the Indian subcontinent impinged into Eurasia following collision,
paleo-sutures were reactivated as strike-slip faults that formed
mountain chains (Fig. 5). Two events are suggested to have caused
vicariant speciation among Teratoscincus species. First, the formation of the Hindu Kush between the Helmand and the Farah blocks
occurred prior to 10 MYBP and, second, the formation of the Tien
Shan-Pamir occurred between the Farah and the Tarim blocks at 10
MYBP. The biological and geological data are in complete agreement.
Abdrakhmatov, K. Ye., Aldazhanov, S. A., Hager, B. H., Hamburger,
M. W., Herring, T. A., Kalabaev, K. B., Makarov, V. I., Molnar, P.,
Panasyuk, S. V., Prilepin, M. T., Reilinger, R. E., Sadybakasov, I. S.,
Souter, B. J., Trapeznikov, Yu. A., Tsurkov, V. Ye., and Zubovich,
A. V. (1996). Relatively recent construction of the Tien Shan
inferred from GPS measurements of present-day crustal deformation rates. Nature 384: 450–453.
Anderson, S., Bankier, A. T., Barrell, B. G., de Bruijn, M. H. L.,
Coulson, A. R., Drouin, J., Eperon, I. C., Nierlich, D. P., Roe, B. A.,
Sanger, F., Schreier, P. H., Smith, A. J. H., Staden, R., and Young,
I. G. (1981). Sequence and organization of the human mitochondrial genome. Nature 290: 457–465.
Anderson, S. C. (1993). A note on the syntopic occurrence of three
species of Teratoscincus in eastern Iran. Dactylus 1(4): 8–10.
Bermingham, E., McCafferty, S. S., and Martin, A. P. (1997). Fish
biogeography and molecular clocks: Perspectives from the Panamanian Isthmus. In ‘‘Molecular Systematics of Fishes’’ (T. D. Kocher
and C. A. Stepien, Eds.), pp. 113–128. Academic Press, San Diego.
VICARIANT PATTERNS IN ASIAN Teratoscincus LIZARDS
Bremer, K. (1994). Branch support and tree stability. Cladistics 10:
295–304.
Brennicke, A., and Clayton, D. A. (1981). Nucleotide assignment of
alkali-sensitive sites in mouse mitochondrial DNA. J. Biol. Chem.
256: 10613–10617.
Buth, D. G. (1984). The application of electrophoretic data in systematic studies. Annu. Rev. Ecol. Syst. 15: 501–522.
Clayton, J. W., and Tretiak, D. N. (1972). Amine-citrate buffers for pH
control in starch gel electrophoresis. J. Fish. Res. Board Canada
29: 1169–1172.
Dercourt, J., Zonenshain, L. P., Ricou, L.-E., Kazmin, V. G., Le
Pichon, X., Knipper, A. L., Grandjacquet, C., Sbortshikov, I. M.,
Geyssant, J., Lepvrier, C., Pechersky, D. H., Boulin, J., Sibuet,
J.-C., Savostin, L. A., Sorokhtin, O., Westphal, M., Bazhenov, M. L.,
Lauer, J. P., and Biju-Duval, B. (1986). Geological evolution of the
tethys belt from the Atlantic to the Pamirs since the Lias. Tectonophysics 123: 241–315 and maps (plates I–X).
Dewey, J. F., Shackleton, R. M., Chang, C., and Sun, Y. (1988). The
tectonic evolution of the Tibetan Plateau. Phil. Trans. R. Soc. Lond.
A 327: 3–16.
Dewey, J. F., Cande, S., and Pitman, W. C., III. (1989). Tectonic
evolution of the India/Eurasia collision zone. Eclogae Geol. Helv.
82: 717–734.
Felsenstein, J. (1985). Confidence limits on phylogenies with a
molecular clock. Syst. Zool. 34: 152–161.
Feng, Y., Coleman, R. G., Tilton, G., and Xiao, X. (1989). Tectonic
evolution of the west Junggar region, Xinjiang, China. Tectonics 8:
729–752.
Harris, H., and Hopkinson, D. A. (1976 et seq.). ‘‘Handbook of Enzyme
Electrophoresis in Human Genetics,’’ Oxford, North-Holland, Amsterdam. [Loose leaf with supplements in 1977 and 1978]
Hixson, J. E., Wong, T. W., and Clayton, D. A. (1986). Both the
conserved and divergent 58-flanking sequences are required for
initiation at the human mitochondrial origin of light strand
replication. J. Biol. Chem. 261: 2384–2390.
Kumazawa, Y., and Nishida, M. (1993). Sequence evolution of mitochondrial tRNA genes and deep-branch animal phylogenetics. J.
Mol. Evol. 37: 380–398.
Kwon, S.-T., Tilton, G. R., Coleman, R. G., and Feng, Y. (1989).
Isotopic studies bearing on the tectonics of the west Junggar
region, Xinjiang, China. Tectonics 8: 719–727.
Larson, A. (1998). The comparison of morphological and molecular
data in phylogenetic systematics. In ‘‘Molecular Approaches to
Ecology and Evolution’’ (R. DeSalle and B. Schierwater, Eds.), pp
275–296. Birkhäuser, Basel.
Mabee, P. M., and Humphries, J. (1993). Coding polymorphic data:
Examples from allozymes and ontogeny. Syst. Biol. 42: 166–181.
Macey, J. R., Ananjeva, N. B., Wang, Y., and Papenfuss, T. J. (1997a).
A taxonomic reevaluation of the gekkonid lizard genus Teratoscincus in China. Russian J. Herpetol. 4: 8–16.
Macey, J. R., Larson, A., Ananjeva, N. B., Fang, Z., and Papenfuss,
T. J. (1997b). Two novel gene orders and the role of light-strand
replication in rearrangement of the vertebrate mitochondrial genome. Mol. Biol. Evol. 14: 91–104.
Macey, J. R., Larson, A., Ananjeva, N. B., and Papenfuss, T. J. (1997c).
Replication slippage may cause parallel evolution in the secondary
structures of mitochondrial transfer RNAs. Mol. Biol. Evol. 14:
31–39.
Macey, J. R., Larson, A., Ananjeva, N. B., and Papenfuss, T. J.
(1997d). Evolutionary shifts in three major structural features of
the mitochondrial genome among iguanian lizards. J. Mol. Evol.
44: 660–674.
Macey, J. R., Schulte, J. A., II, Ananjeva, N. B., Larson, A., RastegarPouyani, N., Shammakov, S. M., and Papenfuss, T. J. (1998a).
Phylogenetic relationships among agamid lizards of the Laudakia
331
caucasia species group: Testing hypotheses of biogeographic fragmentation and an area cladogram for the Iranian Plateau. Mol.
Phylogenet. Evol. 10: 118–131.
Macey, J. R., Schulte, J. A., II, Larson, A., Fang, Z., Wang, Y., Tuniyev,
B. S., and Papenfuss, T. J. (1998b). Phylogenetic relationships of
toads in the Bufo bufo species group from the eastern escarpment
of the Tibetan Plateau: A case of vicariance and dispersal. Mol.
Phylogenet. Evol. 9: 80–87.
Macey, J. R., Schulte, J. A., II, Larson, A., and Papenfuss, T. J.
(1998c). Tandem duplication via light-strand synthesis may provide a precursor for mitochondrial genomic rearrangement. Mol.
Biol. Evol. 15: 71–75.
Macey, J. R., and Verma, A. (1997). Homology in phylogenetic
analysis: Alignment of transfer RNA genes and the phylogenetic
position of snakes. Mol. Phylogenet. Evol. 7: 272–279.
Maddison, W. P., and Maddison, D. R. (1992). ‘‘MacClade, analysis of
phylogeny and character evolution, version 3.0,’’ Sinauer, Sunderland, MA.
Maniatis, T., Fritsch, E. F., and Sambrook, J. (1982). ‘‘Molecular
Cloning: A Laboratory Manual,’’ Cold Spring Harbor Laboratory
Press, Cold Spring Harbor, NY.
Molnar, P., Burchfiel, B. C., Liang, K., and Zhao, Z. (1987). Geomorphic evidence for active faulting in the Altyn Tagh and northern
Tibet and qualitative estimates of its contribution to the convergence of India and Eurasia. Geology 15: 249–253.
Moritz, C., Dowling, T. E., and Brown, W. M. (1987). Evolution of
animal mitochondrial DNA: Relevance for population biology and
systematics. Annu. Rev. Ecol. Syst. 18: 269–292.
Murphy, R. W., Sites, J. W., Jr., Buth, D. G., and Haufler, C. H. (1990).
Proteins I: Isozyme electrophoresis. In ‘‘Molecular Systematics’’
(D. M. Hillis and C. Moritz, Eds.), pp. 45–126. Sinauer, Sunderland, MA.
Richardson, B. J., Baverstock, P. R., and Adams, M. (1986). ‘‘Allozyme
Electrophoresis. A Handbook for Animal Systematics and Population Studies,’’ Academic Press, Sydney.
Selander, R. K., Smith, M. H., Yang, S. Y., Johnson, W. E., and Gentry,
J. R. (1971). Biochemical polymorphism and systematics in the
genus Peromyscus. I. Variation in the old-field mouse (Peromyscus
polionotus). Studies in Genetics VI. Univ. Texas Publ. 7103: 49–90.
Sengör, A. M. C. (1984). The Cimmeride orogenic system and the
tectonics of Eurasia. Geol. Soc. Am. Spec. Paper No. 195: 1–82.
Sengör, A. M. C., Altiner, D., Cin, A., Ustaomer, T., and Hsu, K. J.
(1988). Origin and assembly of the Tethyside orogenic collage at the
expense of Gondwana Land. In ‘‘Gondwana and the Tethys’’ (M. G.
Audley-Charles and A. Hallam, Eds.), pp. 119–181. Geol. Soc. Spec.
Publ. No. 37. Oxford Univ. Press, Oxford.
Steininger, F. F., and Rogl, F. (1984). Paleogeography and palinspastic reconstruction of the Neogene of the Mediterranean and Paratethys. In ‘‘The Geological Evolution of the Eastern Mediterranean’’
(J. E. Dixon and A. H. F. Robertson, Eds.), pp. 659–668. Blackwell
Sci., Oxford.
Swofford, D. L. (1993). ‘‘PAUP: Phylogenetic analysis using parsimony, version 3.1.1,’’ Illinois Natur. Hist. Survey., Urbana, IL.
Swofford, D. L., and Olsen, G. J. (1990). Phylogeny reconstruction. In
‘‘Molecular Systematics’’ (D. M. Hillis and C. Moritz, Eds.), pp.
411–501. Sinauer, Sunderland, MA.
Szczerbak, N. N., and Golubev, M. L. (1996). Gecko Fauna of the
USSR and Contiguous Regions. Soc. Study Amphib. Reptiles Contr.
Herpetol. 13: 1–233.
Tapponier, P., Mattauer, M., Proust, F., and Cassaigneau, C. (1981).
Mesozoic ophiolites, sutures, and large-scale tectonic movements
in Afghanistan. Earth Planet. Sci. Lett. 52: 355–371.
Templeton, A. R. (1983). Phylogenetic inference from restriction
endonuclease cleavage site maps with particular reference to the
evolution of humans and the apes. Evolution 37: 221–244.
332
MACEY ET AL.
Windley, B. F. (1988). Tectonic framework of the Himalaya, Karakorum and Tibet, and problems of their evolution. Phil. Trans. R. Soc.
Lond. A 326: 3–16.
Zar, J. H. (1984). ‘‘Biostatistical Analysis,’’ 2nd ed., Prentice Hall
International, Englewood Cliffs, NJ.
Zhang, D.-X., and Hewitt, G. M. (1996). Nuclear integrations: Challenges for mitochondrial DNA markers. Trends Ecol. Evol. 11:
247–251.
Zhao, E.-M., and Adler, K. (1993). Herpetology of China. Soc. Study
Amphib. Reptiles Contr. Herpetol. 10: 1–522.