Defining intraspecific conservation units in the endemic Cuban Rock Iguanas (Cyclura nubila nubila).

Defining conservation units is an important step in species management and requires interpretation of the genetic diversity and ecological function of the taxon being considered. We used the endemic Cuban Rock Iguanas (Cyclura nubila nubila) as a model to highlight this challenge and examined patterns of its intraspecific genetic diversity across Cuba. We evaluated nuclear (microsatellite loci) and mitochondrial diversity across eight populations from the island and its off-shore cays, and applied the population genetics results for assignment of Management Unit (MU) status and Evolutionary Significant Units (ESUs) based on phylogeographic and time of divergence information. We identified at least six distinct Cuban Rock Iguana MUs, encompassing demographically isolated and genetically differentiated populations across Cuba, most with low effective population size, declining populations, and with high risk of inbreeding and genetic drift. Hence, each MU should be considered of urgent conservation priority. Given the key ecological seed dispersal role of C. n. nubila, the disappearance of any MU could trigger the loss of local ecological functional diversity and major negative impacts on their ecosystems. Two divergent ESUs were also identified, exhibiting an historical east-west geographic separation on Cuba. Based on a Caribbean phylogeographic assessment, our findings strengthen the conclusion that all geographically and evolutionarily differentiated Cyclura species and subspecies across the archipelago warrant ESU distinction.

Effects of phylogenetic uncertainty on fossil identification illustrated by a new and enigmatic Eocene iguanian.

Fossil identifications made in a phylogenetic framework are beholden to specific tree hypotheses. Without phylogenetic consensus, the systematic provenance of any given fossil can be volatile. Paleobiogeographic and divergence time hypotheses are contingent on the accurate systematic placement of fossils. Thus, fossil diagnoses should consider multiple topologies when phylogenetic resolution or clear apomorphies are lacking. However, such analyses are infrequently performed. Pleurodonta (Squamata: Iguania) is an ancient and frequently-studied lizard clade for which phylogenetic resolution is notoriously elusive. I describe a skull fossil of a new pleurodontan lizard taxon from the Eocene deposits of the Willwood Formation, Wyoming, and use the new taxon as a case-study to explore the effects of phylogenetic uncertainty on fossil identification. The relationships of the new taxon differ considerably among analyses, and resulting interpretations are correspondingly disparate. These results illustrate generalizable and severe issues with fossil interpretations made without consideration of alternative phylogenetic hypotheses.

Never judge an iguana by its spines: Systematics of the Yucatan spiny tailed iguana, Ctenosaura defensor (Cope, 1866).

Spiny tailed iguanas are highly diverse clade of lizards in Mesoamerica, ranging from northern Mexico through Panama. Utilizing 2 regions of mitochondrial DNA (1948bp) and 4 nuclear loci (2232bp) we explored the relationships between these species and the phylogeographic history of the major clades. We discovered that the lineage endemic to the Yucatan Peninsula renders the genus Ctenosaura paraphyletic. To resolve this non-monophyly, we resurrect the taxon Cachryx Cope, 1866, and provide a new diagnosis for the genus. We also find that small body-size and highly spinose tails in the species previously referred to the subgenus Enyaliosaurus, have evolved independently 3 times. Cachryx were recovered as sister to the lineage of iguanines endemic to the Galapagos Islands, and we discuss biogeographic scenarios to explain this relationship as well as those among the primary clades of Ctenosaura in Mesoamerica.

A new species of iguanid lizard, genus Stenocercus (Squamata, Iguania), from the Central Andes in Peru.

We describe a new species of Stenocercus from the montane forest of the right margin of the Marañón river in the northern portion of the Central Andes in northern Peru (Amazonas and La Libertad departments), at elevations ranging from 2300 to 3035 m. Stenocercus omari sp. nov. differs from other Stenocercus species, with the exception of S. amydrorhytus, S. chrysopygus, S. cupreus, S. johaberfellneri, S. latebrosus, S. melanopygus, S. modestus, S. ornatissimus, S. orientalis, and S. stigmosus, by having granular scales on the posterior surfaces of thighs, a conspicuous antehumeral fold and by lacking a vertebral crest. However, Stenocercus omari sp. nov. is easily distinguished from the aforementioned species, except S. orientalis, by the presence of prominently keeled dorsal head scales. The new species differs from S. orientalis by lacking a prominent oblique neck fold and by having a distinct deep postfemoral mite pocket.

Rediscovery of the 220-year-old holotype of the Banded Iguana, Brachylophus fasciatus (Brongniart, 1800) in the Paris Natural History Museum.

The Paris Natural History Museum herpetological collection (MNHN-RA) has seven historical specimens of Brachylophus spp. collected late in the 18th and early in the 19th centuries. Brachylophus fasciatus was described in 1800 by Brongniart but its type was subsequently considered as lost and never present in MNHN-RA collections. We found that 220 year old holotype among existing collections, registered without any data, and we show that it was donated to MNHN-RA from Brongniart's private collection after his death in 1847. It was registered in the catalogue of 1851 but without any data or reference to its type status. According to the coloration (uncommon midbody saddle-like dorsal banding pattern) and morphometric data given in its original description and in the subsequent examination of the type in 1802 by Daudin and in 1805 by Brongniart we found that lost holotype in the collections. Another MNHN-RA specimen has Horn Islands (Wallis and Futuna) as the collection location but we show that most of the collections given to MNHN-RA by its collector, Louis Arnoux, have mixed localities in the MNHN-RA catalogues. We thus conclude that the locality is wrong and that the species never inhabited those islands located west of Western Samoa and north-east of Fiji.

The complete mitochondrial genomes of the Galápagos iguanas, Amblyrhynchus cristatus and Conolophus subcristatus.

The Galápagos iguanas are among the oldest vertebrate lineages on the Galápagos archipelago, and the evolutionary history of this clade is of great interest to biologists. We describe here the complete mitochondrial genomes of the marine iguana, Amblyrhynchus cristatus (Genbank accession number: KT277937) and the land iguana Conolophus subcristatus (Genbank accession number: KT277936). The genomes contain 13 protein-coding genes, 22 transfer RNAs, and two ribosomal RNAs genes, as well as a control region (CR). Both species have an identical gene order, which matches that of Iguana iguana. The CR of both Galápagos iguanas features similar tandem repeats units, which are absent in I. iguana. We present a phylogeny of the Iguanidae based on complete mitochondrial genomes, which confirms the sister-group relationship of Galápagos iguanas. These new mitochondrial genomes constitute an important data source for future exploration of the phylogenetic relationships and evolutionary history of the Galápagos iguanas.

MIG-seq: an effective PCR-based method for genome-wide single-nucleotide polymorphism genotyping using the next-generation sequencing platform.

Restriction-enzyme (RE)-based next-generation sequencing methods have revolutionized marker-assisted genetic studies; however, the use of REs has limited their widespread adoption, especially in field samples with low-quality DNA and/or small quantities of DNA. Here, we developed a PCR-based procedure to construct reduced representation libraries without RE digestion steps, representing de novo single-nucleotide polymorphism discovery, and its genotyping using next-generation sequencing. Using multiplexed inter-simple sequence repeat (ISSR) primers, thousands of genome-wide regions were amplified effectively from a wide variety of genomes, without prior genetic information. We demonstrated: 1) Mendelian gametic segregation of the discovered variants; 2) reproducibility of genotyping by checking its applicability for individual identification; and 3) applicability in a wide variety of species by checking standard population genetic analysis. This approach, called multiplexed ISSR genotyping by sequencing, should be applicable to many marker-assisted genetic studies with a wide range of DNA qualities and quantities.

A likely microendemic new species of terrestrial iguana, genus Chalarodon, from Madagascar.

A new species of the hitherto monotypic genus Chalarodon is described from southern Madagascar and a lectotype (ZMB 4360) is designated for C. madagascariensis Peters, 1854. The new species of terrestrial iguana, Chalarodon steinkampi sp. nov., is defined by several morphological characters and by concordant differentiation in mitochondrial and nuclear DNA with >5% uncorrected pairwise genetic distance in the 16S rRNA gene. It can be most clearly recognized by the presence of smooth (vs. keeled) gular and ventral scales, a spotted pattern extending from flanks onto belly, and an unpigmented throat. The new species is known from only a small area between the villages of Amboasary Sud and Esomony, located west of the Andohahela Massif, while C. madagascariensis appears to be widespread over much of southern and western Madagascar. We highlight the need for further exploration of this unprotected region which might host several other microendemic species.

Inferred vs realized patterns of gene flow: an analysis of population structure in the Andros Island Rock Iguana.

Ecological data, the primary source of information on patterns and rates of migration, can be integrated with genetic data to more accurately describe the realized connectivity between geographically isolated demes. In this paper we implement this approach and discuss its implications for managing populations of the endangered Andros Island Rock Iguana, Cyclura cychlura cychlura. This iguana is endemic to Andros, a highly fragmented landmass of large islands and smaller cays. Field observations suggest that geographically isolated demes were panmictic due to high, inferred rates of gene flow. We expand on these observations using 16 polymorphic microsatellites to investigate the genetic structure and rates of gene flow from 188 Andros Iguanas collected across 23 island sites. Bayesian clustering of specimens assigned individuals to three distinct genotypic clusters. An analysis of molecular variance (AMOVA) indicates that allele frequency differences are responsible for a significant portion of the genetic variance across the three defined clusters (Fst =  0.117, p<<0.01). These clusters are associated with larger islands and satellite cays isolated by broad water channels with strong currents. These findings imply that broad water channels present greater obstacles to gene flow than was inferred from field observation alone. Additionally, rates of gene flow were indirectly estimated using BAYESASS 3.0. The proportion of individuals originating from within each identified cluster varied from 94.5 to 98.7%, providing further support for local isolation. Our assessment reveals a major disparity between inferred and realized gene flow. We discuss our results in a conservation perspective for species inhabiting highly fragmented landscapes.

A new species of spiny-tailed iguanid lizard (Iguania: Stenocercus) from northwestern Peru.

We describe a new species of Stenocercus from the interandean valley of Río Chotano on the Amazonian slope of the northern portion of the Cordillera Occidental of Peru (Cajamarca Region), at elevations of between 1997 and 2318 m. Stenocercus arndti sp. nov. differs from other Stenocercus, except from S. bolivarensis, S. carrioni, S. chlorostictus, S. crassicaudatus, S. empetrus, S. eunetopsis, S. simonsii, and S. torquatus, in having granular scales on the posterior surface of the thighs, two caudal whorls per autotomic segment, mucronate caudal scales, and distinct longitudinal row of enlarged vertebral scales. However, Stenocercus arndti sp. nov. is easily distinguished from these species in having a bold black transversal band at midbody that extends ventrolaterally in adult males.

Species delimitation using Bayes factors: simulations and application to the Sceloporus scalaris species group (Squamata: Phrynosomatidae).

Current molecular methods of species delimitation are limited by the types of species delimitation models and scenarios that can be tested. Bayes factors allow for more flexibility in testing non-nested species delimitation models and hypotheses of individual assignment to alternative lineages. Here, we examined the efficacy of Bayes factors in delimiting species through simulations and empirical data from the Sceloporus scalaris species group. Marginal-likelihood scores of competing species delimitation models, from which Bayes factor values were compared, were estimated with four different methods: harmonic mean estimation (HME), smoothed harmonic mean estimation (sHME), path-sampling/thermodynamic integration (PS), and stepping-stone (SS) analysis. We also performed model selection using a posterior simulation-based analog of the Akaike information criterion through Markov chain Monte Carlo analysis (AICM). Bayes factor species delimitation results from the empirical data were then compared with results from the reversible-jump MCMC (rjMCMC) coalescent-based species delimitation method Bayesian Phylogenetics and Phylogeography (BP&P). Simulation results show that HME and sHME perform poorly compared with PS and SS marginal-likelihood estimators when identifying the true species delimitation model. Furthermore, Bayes factor delimitation (BFD) of species showed improved performance when species limits are tested by reassigning individuals between species, as opposed to either lumping or splitting lineages. In the empirical data, BFD through PS and SS analyses, as well as the rjMCMC method, each provide support for the recognition of all scalaris group taxa as independent evolutionary lineages. Bayes factor species delimitation and BP&P also support the recognition of three previously undescribed lineages. In both simulated and empirical data sets, harmonic and smoothed harmonic mean marginal-likelihood estimators provided much higher marginal-likelihood estimates than PS and SS estimators. The AICM displayed poor repeatability in both simulated and empirical data sets, and produced inconsistent model rankings across replicate runs with the empirical data. Our results suggest that species delimitation through the use of Bayes factors with marginal-likelihood estimates via PS or SS analyses provide a useful and complementary alternative to existing species delimitation methods.

A new species of arboreal iguanid lizard, genus Stenocercus (Squamata: Iguania), from central Peru.

We describe a new species of Stenocercus from an interandean valley of the upper Río Huallaga on the Amazonian slope of central Peru (Región Huánuco), at an elevation of 1700-1900 m. The new species differs from other Stenocercus, except S. boettgeri, S. haenschi, S. humeralis, and S. varius, by the combination of the following characters: presence of granular scales on the posterior surface of the thighs, enlarged vertebrals, three caudal whorls per autotomic segment, a medially complete antegular fold, non-spinose caudals, and by males lacking a black transverse band on the ventral surface of the neck. However, the new Stenocercus differs from these, with the exception of S. humeralis, by having more scales around the midbody (104-107, =105.66) than S. boettgeri (79-104, Mean= 88.61), S. haenschi (57-64, Mean=60.50), and S. varius (74-88, Mean=82.35); and from S. humeralis by having the scales in the frontonasal region nearly equal in size to the scales in the occipitoparietal region, while in S. humeralis the scales on the frontonasal region are twice or three times longer than the scales on the occipitoparietal region.

Basic ecology of the Oaxacan Spiny-tailed Iguana Ctenosaura oaxacana (Squamata: Iguanidae), in Oaxaca, Mexico

The Oaxacan Spiny-tailed Iguana Ctenosaura oaxacana is a restricted species to the Isthmus of Tehuantepec in Southern Oaxaca, Mexico. This reptile is one of the less known iguanid species. We censustracked a population in the South of Niltepec, Oaxaca, Mexico from May 2010 to April 2011. Throughout one year, a total of 10 line transects were situated and recorded in the study area to determine relative abundance and density, and habitat type use (dry forest, Nanchal, grassland, riparian vegetation, and mangrove) by the species. This study reports a new C. oaxacana population on the Southeastern limit of species range. Although this species has a very restricted distribution and is in danger of extinction, C. oaxacana has a high population density when compared to other Ctenosaura species. A total of 108 individuals were recorded throughout the study. Dry forest (33.75ind/ha) and Nanchal (18.75ind/ha) were the habitats with higher densities. Comparisons between habitat types showed no significant differences between dry forest and Nanchal (W=15, p=0.0808). Results between seasons were similar. The Oaxacan Spiny tailed Iguana preferred first the dry forest, and then Nanchal, while avoided grassland, riparian vegetation, and mangroves. There was no difference in habitat use between males and females. Mean perch heights were 1.23±0.32 (n=30) in Nanchal, 2.11±0.30 (n=9) in grassland, 1.90±0.56 (n=54) in dry forest, 1.91±0.28 (n=9) in mangrove and 2.30±0.37 (n=6) in riparian vegetation. Species observed as refuge and perch were B. crassifolia (Nanchal); C. alata (grassland); Tabebuia sp., Genipa americana, G. sepium, Acacia sp., Ficus sp. and Haematoxylon sp. (dry forest); G. sepium, Acacia sp. and Guazuma ulmifolia (riparian vegetation); and C. erecta (mangrove). Live trees hollows and branches were used by species. Main threats to the species are excessive hunting and habitat loss. Furthermore, grassland fires are still common in the study ...

La Iguana de Cola Espinosa Ctenosaura oaxacana es una especie restringida al Istmo de Tehuantepec en el Sureste de Oaxaca, México. Este reptil es uno de los iguánidos menos conocidos. Entre Mayo 2010 y Abril 2011 hicimos el seguimiento de una población al Sur de Niltepec, Oaxaca, México. A lo largo de un año, se colocaron y monitorearon un total de diez transectos en el área de estudio, para determinar la abundancia relativa, densidad y uso de hábitat (selva seca, Nanchal, pastizal, vegetación riparia y manglar) de la especie. Esta investigación reporta una nueva población de C. oaxacana en el límite sureste de la distribución conocida para la especie. Aunque esta especie posee una distribución muy restringida y se encuentra en peligro de extinción, C. oaxacana presenta una alta densidad poblacional comparada con otras especies de Ctenosaura. Se registró un total de 108 individuos a lo largo del estudio. La selva seca (33.75ind/ha) y el Nanchal (18.75ind/ha) fueron los hábitats con densidad más alta. No se encontró diferencia significativa entre la selva seca y el Nanchal (W=15, p=0.0808). Al comparar entre estaciones el resultado fue similar. La Iguana de Cola Espinosa prefirió la selva seca y después al Nanchal, mientras que evitó el pastizal, la vegetación riparia y el manglar. No se registró diferencia en el uso de hábitat entre machos y hembras. La altura media de las perchas fue 1.23±0.32 (n=30) en Nanchal, 2.11±0.30 (n=9) en pastizal, 1.90±0.56 (n=54) en selva baja, 1.91±0.28 (n=9) en manglar y 2.30±0.37 (n=6) en vegetación riparia. Las especies observadas como refugio y percha fueron B. crassifolia (Nanchal); C. alata (pastizal); Tabebuia sp., Genipa americana, G. sepium, Acacia sp., Ficus sp. y Haematoxylon sp. (selva seca); G. sepium, Acacia sp. y Guazuma ulmifolia (vegetación riparia); y C. erecta (manglar). La especie utilizó agujeros y ramas de árboles vivos. Las principales amenazas para la especie son la cacería excesiva y la pérdida de hábitat. Además, la quema de pastizales sigue siendo común en el área de estudio durante la temporada seca, lo que puede resultar en la pérdida de hábitat y el desplazamiento territorial de los individuos.

Basic ecology of the Oaxacan Spiny-tailed Iguana Ctenosaura oaxacana (Squamata: Iguanidae), in Oaxaca, Mexico.

The Oaxacan Spiny-tailed Iguana Ctenosaura oaxacana is a restricted species to the Isthmus of Tehuantepec in Southern Oaxaca, Mexico. This reptile is one of the less known iguanid species. We census-tracked a population in the South ofNiltepec, Oaxaca, Mexico from May 2010 to April 2011. Throughout one year, a total of 10 line transects were situated and recorded in the study area to determine relative abundance and density, and habitat type use (dry forest, Nanchal, grassland, riparian vegetation, and mangrove) by the species. This study reports a new C. oaxacana population on the Southeastern limit of species range. Although this species has a very restricted distribution and is in danger of extinction, C. oaxacana has a high population density when compared to other Ctenosaura species. A total of 108 individuals were recorded throughout the study. Dry forest (33.75ind/ha) and Nanchal (18.75ind/ha) were the habitats with higher densities. Comparisons between habitat types showed no significant differences between dry forest and Nanchal (W=15, p=0.0808). Results between seasons were similar. The Oaxacan Spiny tailed Iguana preferred first the dry forest, and then Nanchal, while avoided grassland, riparian vegetation, and mangroves. There was no difference in habitat use between males and females. Mean perch heights were 1.23 +/- 0.32 (n=30) in Nanchal, 2.11 +/- 0.30 (n=9) in grassland, 1.90 +/- 0.56 (n=54) in dry forest, 1.91 +/- 0.28 (n=9) in mangrove and 2.30 +/- 0.37 (n=6) in riparian vegetation. Species observed as refuge and perch were B. crassifolia (Nanchal); C. alata (grassland); Tabebuia sp., Genipa americana, G. sepium, Acacia sp., Ficus sp. and Haematoxylon sp. (dry forest); G. sepium, Acacia sp. and Guazuma ulmifolia (riparian vegetation); and C. erecta (mangrove). Live trees hollows and branches were used by species. Main threats to the species are excessive hunting and habitat loss. Furthermore, grassland fires are still common in the study area during the dry season, which can result in habitat loss and territorial displacement of individuals.

Diversification in the Mexican horned lizard Phrynosoma orbiculare across a dynamic landscape.

The widespread montane Mexican horned lizard Phrynosoma orbiculare (Squamata: Phrynosomatidae) represents an ideal species to investigate the relative impacts of Neogene vicariance and Quaternary climate change on lineage diversification across the Mexican highlands. We used mitochondrial DNA to examine the maternal history of P. orbiculare and estimate the timing and tempo of lineage diversification. Based on our results, we inferred 11 geographically structured, well supported mitochondrial lineages within this species, suggesting P. orbiculare represents a species complex. Six divergences between lineages likely occurred during the Late Miocene and Pliocene, and four splits probably happened during the Pleistocene. Diversification rate appeared relatively constant through time. Spatial and temporal divergences between lineages of P. orbiculare and co-distributed taxa suggest that a distinct period of uplifting of the Transvolcanic Belt around 7.5-3 million years ago broadly impacted diversification in taxa associated with this mountain range. To the north, several river drainages acting as filter barriers differentially subdivided co-distributed highland taxa through time. Diversification patterns observed in P. orbiculare provide additional insight into the mechanisms that impacted differentiation of highland taxa across the complex Mexican highlands.

An overlooked pink species of land iguana in the Galapagos.

Despite the attention given to them, the Galápagos have not yet finished offering evolutionary novelties. When Darwin visited the Galápagos, he observed both marine (Amblyrhynchus) and land (Conolophus) iguanas but did not encounter a rare pink black-striped land iguana (herein referred to as "rosada," meaning "pink" in Spanish), which, surprisingly, remained unseen until 1986. Here, we show that substantial genetic isolation exists between the rosada and syntopic yellow forms and that the rosada is basal to extant taxonomically recognized Galápagos land iguanas. The rosada, whose present distribution is a conundrum, is a relict lineage whose origin dates back to a period when at least some of the present-day islands had not yet formed. So far, this species is the only evidence of ancient diversification along the Galápagos land iguana lineage and documents one of the oldest events of divergence ever recorded in the Galápagos. Conservation efforts are needed to prevent this form, identified by us as a good species, from extinction.

Chromosome evolution and diversification in North American spiny lizards (genus Sceloporus).

The genus Sceloporus is a diverse clade of lizards that exhibits substantial variation in chromosome numbers and sex chromosome heteromorphisms, 2 features of the genome that are static among most other pleurodont iguanian lizards. Evolutionary changes to the fundamental number of chromosomes are hypothesized to be a primary factor responsible for driving the diversification of Sceloporus. We explore the patterns of chromosome evolution in Sceloporus using a combination of ancestral state estimations and species diversification tests. Phylogenetic relationships and divergence times within Sceloporus (53 species representing all 19 species groups) are estimated using 4 nuclear genes (>3.3 kb) and relaxed-clock analyses that incorporate a fossil calibration on the root of the tree. We test the hypothesis that chromosome evolution is correlated with shifts in species diversification using cross-validation predictive densities, a new Bayesian approach for modeling the number of species that are predicted to have evolved in the absence of a certain historical event (e.g., a change in chromosome numbers). Results of the cross-validation predictive densities approach indicate that chromosomal evolution is correlated with significantly higher species diversity than predicted under the background rate of diversification in Sceloporus. We conclude by discussing the future of comparative cytogenetic investigations in Sceloporus.

Chromosome variation, genomics, speciation and evolution in Sceloporus lizards.

The clade of the North American lizard genus Sceloporus and its relatives comprising the subfamily Phrynosomatinae (Iguanidae) includes perhaps 150 evolutionary lineages. The work reviewed here begins with the discovery of the concentration of Robertsonian chromosomal variability in Sceloporus more than 40 years ago and cytogenetic and genomic evidence of remarkable chromosomal variation within the S. grammicus complex associated with narrow zones of hybridization between different chromosomal races. These discoveries led to hypotheses about hybrid zones involving negative heterosis, possible modes of chromosomal speciation, and the potential roles of such speciation in phylogenesis. The radiation of Sceloporus has now been studied by many different workers extending and mapping the geographic distribution of cytogenetic and genomic variation to understand the biology of the chromosomal variation and to establish the phyletic relationships of the various lineages. The result is a robust phylogeny and a large and still growing database of genic, cytogenetic and other biological parameters. These materials provide a rich series of natural experiments to support both synthetic-comparative and analytical studies of the roles of chromosomal variation, hybrid zones and modes of speciation in phylogenesis and evolutionary success.

Diversification in the northern neotropics: mitochondrial and nuclear DNA phylogeography of the iguana Ctenosaura pectinata and related species.

While Quaternary climatic changes are considered by some to have been a major factor promoting speciation within the neotropics, others suggest that much of the neotropical species diversity originated before the Pleistocene. Using mitochondrial and nuclear sequence data, we evaluate the relative importance of Pleistocene and pre-Pleistocene events within the evolutionary history of the Mexican iguana Ctenosaura pectinata, and related species. Results support the existence of cryptic lineages with strong mitochondrial divergence (> 4%) among them. Some of these lineages form zones of secondary contact, with one of them hybridizing with C. hemilopha. Evolutionary network analyses reveal the oldest populations of C. pectinata to be those of the northern and southern Mexican coastal regions. Inland and mid-latitudinal coastal populations are younger in age as a consequence of a history of local extinction within these regions followed by re-colonization. Estimated divergence times suggest that C. pectinata originated during the Pliocene, whereas geographically distinct mitochondrial DNA lineages first started to diverge during the Pliocene, with subsequent divergence continuing through the Pleistocene. Our results highlight the influence of both Pliocene and Pleistocene events in shaping the geographical distribution of genetic variation within neotropical lowland organisms. Areas of high genetic diversity in southern Mexico were detected, this finding plus the high levels of genetic diversity within C. pectinata, have implications for the conservation of this threatened species.