Caryologia. International Journal of Cytology, Cytosystematics and Cytogenetics 73(2): 15-25, 2020 Firenze University Press www.fupress.com/caryologiaCaryologia International Journal of Cytology, Cytosystematics and Cytogenetics ISSN 0008-7114 (print) | ISSN 2165-5391 (online) | DOI: 10.13128/caryologia-672 Citation: F. Ito, D.J. Gama-Maia, D.M.A. Brito, R.A. Torres (2020) Title. Caryologia 73(2): 15-25. doi: 10.13128/ caryologia-672 Received: October 23, 2019 Accepted: March 27, 2020 Published: July 31, 2020 Copyright: © 2020 F. Ito, D.J. Gama- Maia, D.M.A. Brito, R.A. Torres. This is an open access, peer-reviewed article published by Firenze University Press (http://www.fupress.com/caryologia) and distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distri- bution, and reproduction in any medi- um, provided the original author and source are credited. Data Availability Statement: All rel- evant data are within the paper and its Supporting Information files. Competing Interests: The Author(s) declare(s) no conflict of interest. Gene flow patterns reinforce the ecological plasticity of Tropidurus hispidus (Squamata: Tropiduridae) Fernanda Ito, Danielle J. Gama-Maia, Diego M. A. Brito, Rodrigo A. Torres* LAGEA – Laboratório de Genômica Evolutiva & Ambiental, Departamento de Zoologia, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Brazil *Corresponding author: rodrigotorres@ufpe.br Abstract. The analysis of gene flow patterns can provide important insights into pop- ulation dynamics in the context of landscape ecology. In lizards, this approach has been used to evaluate patterns related to climate change, habitat fragmentation, and taxonomic uncertainties. Tropidurus hispidus is an ecologically plastic species, which presents some evidence of population structuring. In the present study, we investi- gated the potential structuring of T. hispidus populations across a gradient of tropical biomes, including the Amazon and Atlantic rainforests, the Caatinga dry forest, the Caatinga-Atlantic Forest transition zone (Agreste), coastal Restinga, and urban envi- ronments. Nuclear ISSR markers were obtained by PCR/electrophoresis, and a num- ber of population parameters were estimated and analyzed. Despite the extreme envi- ronmental discontinuities found across the vast study area, the results revealed a high degree of genetic connectivity among the different demes. This pattern indicates that the species can be considered to be a single evolutionary taxon with gene flow among all populations, despite the marked environmental discontinuities. Tropidurus hispidus clearly has a marked capacity for dispersal, which may be favored by its intrinsic genet- ic diversity. Keywords: Tropidurus hispidus, ISSRs, gene flow, dispersal capacity, population con- nectivity. INTRODUCTION Gene flow is one of the most important components in population struc- ture because it can determine how much populations have evolved indepen- dently (Slatkin 2018). Therefore gene flow patterns can also provide impor- tant insights for studies on population ecology and also on population genet- ics based on a landscape ecology approach. The genetic admixture resulting from gene flow may contribute to a short-term increase in population fitness (Facon et al. 2005) and adaptive potential (Verhoeven et al. 2011). However, the approach usually focuses on micro-evolutionary phenomena and process- es that lead to intraspecific discontinuities (Holderegger and Wagner 2006). 16 Fernanda Ito et al. Genetic studies, especially in the Neotropical region, and in particular for reptile species, have been more frequent in the last years focusing on questions related to climate change, habitat fragmentation, and taxonomic uncer- tainties (e.g. Ricketts 2001; Stow et al. 2001; Berry et al. 2005; Driscoll and Hardy 2005; Sumner 2005; Hoehn et al. 2007; O’Neill et al. 2008; Tolley et al. 2009; Freedman et al. 2010; Levy et al. 2010; Werneck et al. 2015; Men- ezes et al. 2016; Fazolato et al. 2017; Cacciali and Köhler 2018; Oliveira et al. 2018). However studies focused on landscape genetics and on the gene flow patterns are still scarce in Neotropical region. Tropidurus hispidus is one of the largest species of the genus, reaching a rostrum-caudal length (RCL) of 114 mm (Kolodiuk et al. 2010). It is found on a variety of substrates such as sand, tree trunks, and rocky outcrops, but it is primarily saxicolous, given that rocks provide space for foraging, shelter, nesting, and thermoregula- tion (Pelegrin et al. 2017). Also, this species is commonly found in urban areas, foraging and thermoregulating on walls and fences (Rodrigues 1987; Abreu et al. 2002; Pel- egrin et al. 2017). The ecological tolerance of T. hispidus allows this species to occupy a number of distinct mor- phoclimatic domains, such as the Brazilian Atlantic For- est, coastal shrubby vegetation (Restinga), transition are- as between the Caatinga scrub (in Portuguese Agreste), the Atlantic Forest, Cerrado savanna, and rocky out- crops in the Amazon basin (Vanzolini et al. 1980; Vitt 1995; Abreu et al. 2002; Carvalho 2013). The species is a habitat generalist, able to colonize a wide range of microhabitats (Rodrigues 1987; Vitt 1995; Vitt and Car- valho 1995; Vitt et al. 1997; Pelegrin et al. 2017). This species is also an opportunistic sit-and-wait predator with a diverse trophic niche, feeding mainly on arthropods, in particular ants, but in some areas they may include plant material in their diet, especially flow- ers (Van Sluys et al. 2004; Ribeiro and Freire 2011; Pel- egrin et al. 2017). Differences in the composition of the diet among biomes reinforce the ecological plasticity of the species (Pelegrin et al. 2017), but may also reflect distinct selective pressures on different populations. In addition to these dietary differences, there is some evidence of genetic structuring among popula- tions. Three distinct karyotypes have been found in six populations from different ecosystems in eastern Brazil (Kasahara et al. 1987; Kasahara et al. 1996). All karyo- types had 2n = 36 and XX/XY sex chromosomes, but three variants (prominent, mild or absent) were found in a secondary constriction of the second chromosome pair, which appeared to be typical of specific sites, sug- gesting genetic variation on an inter-population level. However, specimens from the six populations are mor- phologically indistinguishable (Kasahara et al. 1987; Kasahara et al. 1996). Also there is a clear evidence for cryptic diversity in T. hispidus as revealed by karyo- type and DNA barcode sequences analyses (Matos et al. 2016). Tropidurus hispidus is abundant across an extremely diverse ecological landscape (Carvalho 2013). From the coast of Pernambuco (north-eastern Brazil) to the Ama- zon basin there is a major shift in the geographical and ecological landscape, in which environmental variation may be reflected into distinct selective regimes, as previ- ously suggested by the chromosomal and molecular evi- dences. Then, given previous ecological, distributional, karyotypical, and molecular evidence, we tested for the hypothesis of the existence of population-level divisions in Tropidurus hispidus along a highly diverse adaptive landscape in Brazil, using nuclear DNA markers adopt- ing a gene flow approach. MATERIALS AND METHODS A total of 155 specimens of Tropidurus hispidus were captured at sites representing the distinct phytophysi- ognomic domains found across the landscape between the Pernambuco and Paraiba coasts in eastern Bra- zil, and the Amazon basin, in the north of the country (Table 1; Figure 1). The specimens were identified using the taxonomic key of Rodrigues (1987). Liver and mus- cle samples for DNA analyses were collected from each specimen. These samples were immersed in 96% etha- nol and stored in a freezer at -20°C. Tissue was also obtained from three specimens of Tropidurus torquatus from Maricá, Rio de Janeiro, south-eastern Brazil, and one Eurolophosaurus divaricatus from Alagoado, Bahia, north-eastern Brazil, for inclusion in the study as out- groups. DNA extraction and ISSR amplification The extraction of DNA was conducted using the Sambrook and Russell (2001) procedure. The integrity of the DNA was checked by electrophoresis in agarose gel and the concentration was estimated by visual com- parison with the intensity of the DNA of the Lambda phage. The DNA was then diluted to a standard concen- tration of 5 ng/ul for the PCR-ISSR reactions. Inter sim- ple sequence repeats (ISSRs) are PCR-amplified nuclear genomic regions using primers anchored at microsatel- lite regions (SSRs) (Gupta et al. 1994; Zietkiewicz et al. 1994). These markers have been considered of low cost and highly reproducible (Sarwat 2012), and very effective 17Gene flow patterns reinforce the ecological plasticity of Tropidurus hispidus (Squamata: Tropiduridae) in terms of studying the genetic variation and popula- tion cohesiveness in several biological groups (Gama- Maia and Torres 2016; Al Salameen et al. 2018; Hassani- em & Al Rashada 2019). The PCRs were carried out in a final volume of 20 µL in which consisted of 0.2 units of Taq DNA polymerase (New England/Biolabs), 1x buffer, 50 mM MgCl2, 50 mM of primer, 0.2 mM dNTP and 20 ng of genomic DNA. The PCR reactions were run in a Biocycler thermocycler and comprised a cycle of 4 min at 94°C, 39 cycles of 40 s at 94°C, 40 s at the specific temperature of each primer (Table 2), and 120 s at 72°C, with a final annealing cycle of 7 minutes. All reactions were run with a negative control. Horizontal electrophoresis was conducted in 1.8% agarose gel containing 0.5X TBE buffer diluted from an original 10X solution (0.89 M Tris, 0.89 M boric acid and EDTA, 0.01M, pH = 8.3) for 4 hours at 60 volts. In each well of the gel we placed a solution containing 10 µL of the PCR product in 1.5 mL of gel loading dye blue (6x) and 1.5 mL of gel green (0.5 ml 10,000x in H20). To support the analysis of bands, we inserted 2 µL of 1 Kb DNA ladder marker with 1.5 mL of gel loading dye blue (6x) in one well. After the run, all gels were photo- graphed using a transilluminator under an ultraviolet light source. Data analyses Initially, 17 different random ISSR primers were tested for their reproducibility and their degree of pol- ymorphism. They were tested in five specimens from four sites using different PCR reagents from Fermentas (Thermo Fisher Scientific) and New England Biolabs Inc. (Table 2). The 10 most polymorphic primers were then selected for the amplification of the DNA of all the specimens (Table 2), with the objective of generating at least 60 polymorphic loci, as recommended by Telles et al. (2001) and Nelson and Anderson (2013). After pho- tographic documentation, the gels were transformed into a binary matrix of presence and absence (0 = absent and 1= presence) of the DNA bands. In order to avoid the misinterpretation of valid markers, only clear and well- defined bands were assigned as markers. It is important to note that to increase sample size, the animals sampled in the localities of Caraguetama and Tamandaré were treated as a single sample in all analyzes, since both are- as represent the same adaptive landscape (named Restin- Table 1. Number (N) of Tropidurus hispidus, T. torquatus, and Eurolophosaurus divaricatus specimens captured in each site along the study area. Species Municipality* or state** Geographic coordinates Acronyms Biome N Camaragibe* 8º02’31”S 35º06’17”W AF Atlantic Forest 24 Canguaretama* 6º22’58”S 35º07’29”W Rest Restinga 2 Gravatá* 8º16’02”S 35º27’35” W TZ Transition zone 25 Tropidurus hispidus Manaus* 3º09’34”S 59º36’10”W AM Amazon Forest 19 Petrolândia* 9º05’37”S 38º15’05”W Ca1 Caatinga 26 Recife* 8º10’43”S 34º42’46”W UZ Urban zone 25 Serra Talhada* 7°59’7”S 38°17’34”W Ca2 Caatinga 30 Tamandaré* 8º45’28”S 35º06’18”W Rest Restinga 4 Total 155 Tropidurus torquatus (og) Rio de Janeiro** 22º22’28”S 42º57’01”W 3 Eurolophorus divaricatus (og) Bahia** 13º07’07”S 38º28’50”W 1 Figure 1. South America/Brazil map depicting capture sites of T. hispidus. In evidence are the different capture sites in Pernam- buco (PE) Brazilian state. The biomes accessed are written here within parentheses as follows: 1. Manaus (Amazon Forest-AM), 2. Canguaretama-state of Paraíba (Restinga-Rest), 3. Serra Talhada (Caatinga-Ca2), 4. Petrolândia (Caatinga-Ca1), 5. Gravatá (Transi- tion zone between Caatinga and Atlantic Forest-TZ), 6. Camara- gibe (Atlantic Forest-AF), 7. Recife (Urban zone-UZ), 8. Tamandaré (Restinga-Rest). 18 Fernanda Ito et al. ga) (Table 1). The overall genetic variation was measured in percentage by the proportion of the polymorphic loci having the total number of observed loci as 100%. To evaluate the existence of potential genetic and/ or evolutionary groupings among biomes, multi-dimen- sional scaling (MDS) with neighbour-joining (NJ) genetic distances was applied on local and regional scales through the simple matching technique (Primer software) (Clarke and Gorley 2006). An additional NJ topology was also obtained by using PAUP* v.4.0b10 (Swofford 2000) in order to observe alternative group- ings among sampled specimens. A Maximum parsimony (MP) method was also used in order to test for hidden evolutionary diversity in T. hispidus across those differ- ent adaptive landscapes (biomes) having Eurolophosau- rus divaricatus and Tropidurus torquatus as outgroups given their phylogenetic proximity to the study spe- cies (Frost et al. 2001; Passoni et al. 2008). These analy- ses were run in PAUP* v.4.0b10 (Swofford 2000), in its graphic interface PaupUp v.1.0.3.1 (Calendini and Mar- tin 2005). A maximum number of 100,000 random trees with 5000 replications were computed. The robustness of the branches was tested by the bootstrap method with 1000 random replicates. Population structuring was tested by the Bayesian approach using the Structure 2.3.3 software (Pritchard et al. 2000; Falush et al. 2003, 2007; Hubisz et al. 2009). In order to determine the number of populations (K) within the complete data set, ten independent runs for K= 1-10 and 100,000 MCMC (Markov Chains Monte Carlo) inter- actions after burn-in period were computed. The analysis was performed by using both the admixture model of pop- ulation structure and allele frequencies correlated among populations. The number of populations (K) was estimated using the protocol described by Evanno et al. (2005). In addition, we conducted an analysis of molecular variance (AMOVA) to check for patterns of genetic iso- lation within and among local populations (Excoffier et al. 1992) with Arlequin v.3.5.1.2 (Excoffier and Lischer 2010). This method also permits the calculation of the global fixation index (ΦST). Parameters of genetic dif- ferentiation among populations (GST) and the number of migrants per generation (Nm – gene flow) were calculat- ed with PopGene 1.3.2 (Yeh et al. 1999). RESULTS Based on the 10 ISSR primers selected, a total of 283 loci were observed. Overall, 99.2% of the observed loci were polymorphic. Mean genetic neighbor-joinning dis- tances among local populations varied from 0.12720 to 0.48763. Table 2. ISSR primers tested in this study with sequence and annealing temperature. Selected primers are marked with (*). Primer Sequence (5’ – 3’) Annealing temperature (ºC) ISSR 1* (AG)8T 50,4 ISSR 2* (AG)8C 52,8 ISSR 3* (GA)8T 50,4 ISSR 4 (GA)8C 52,8 ISSR 5* (CT)8G 52,0 ISSR 6* (AG)8YC 52,8 ISSR 7 (AG)8YA 54,0 ISSR 8* (GA)8YT 52,8 ISSR 9* (GA)8YC 52,8 ISSR 10 (GA)8YG 54,0 ISSR 11 (CT)8RA 50,0 ISSR 12 (AC)8YG 54,0 ISSR 13* (GGAC)3A 51,0 ISSR 14 (GGAC)3C 51,0 ISSR 15 (GGAC)3T 51,0 ISSR 16* (AACC)4 51,0 ISSR 17* (GGAC)4 51,0 Figure 2. Multi-dimensional scaling plots of the genetic similarities (simple matching index) among the Tropidurus hispidus popula- tions sampled in the present study. See Table 1 and text for acro- nyms. The graphs a/b show the different domains and localities respectively. 19Gene flow patterns reinforce the ecological plasticity of Tropidurus hispidus (Squamata: Tropiduridae) The simple matching MDS analysis revealed a single grouping comprising all sampled populations on both regional and local scales (Figure 2a-b). The NJ topology showed also no particular genetic groupings among T. hispidus sampled from different biomes (Figure 3). The maximum parsimony (MP) analysis revealed 2 constant and 281 informative characters. The majority-rule con- sensus topology (Supplemental material) had a length (L) of 6236, a consistency index (Ci) of 0.045, and a reten- tion index (R) of 0.310. This topology also failed to iden- tify any evolutionary differentiation among the popula- tions analysed. The analysis identified a total of 283 loci and more than 90% were variable in terms of the proportion of polymorphic loci. This amount of molecular informa- tion satisfies the recommendation of Nelson and Ander- Figure 3. Neighbor-Joinning topology from Tropidurus hispidus specimens for ISSR markers. See Table 1 and text for acronyms. 20 Fernanda Ito et al. son (2013) for the application of AMOVA and Bayesian structuring analyses. The AMOVA indicated that 90.99% of the total genetic variance was found within popula- tions and only 9.01% among populations (Table 3). The Bayesian structuring analysis revealed the existence of two genetic populations (K= 2; Figure 4), and these genetic profiles were clearly distributed in all specimens throughout the geographic areas sampled. The global GST value was 0.07, while the Nm was 6.59. The pairwise analyses showed values ranging from 0.03 to 0.06 for GST and from 7.75 to 15.03 for Nm (Table 4). DISCUSSION The genetic evidence of this study indicates strong connectivity among local T. hispidus groups, despite the intense ecological distinctiveness of the landscapes seen in the study area. The MDS (Figure 2), the NJ topology (Figure 3), and the MP topology (Supplemental material) evidenced a lack of any genetic or evolutionary differen- tiation among T. hispidus groups, pointing to a high dis- persive behaviour in this species. Tropidurus hispidus is widely distributed in the Caatinga and can also be found along the Brazilian coast and in the Amazon Forest (Carvalho 2013). The extent of its distribution range would suggest a high probability of differentiation due to strong and diverse evolutionary pressures imposed to the populations (Kisel and Barra- clough 2010). However, the clustering produced by the Bayesian analyses showed also no genetic structuring (Figure 4). Although there are two genetic populations, the analysis indicated a clear admixture of these two T. hispidus gene pools among the demes studied. The lack of genetic structuring resulted from an intense gene flow among populations as indicated by the degree of migrants per generation (Table 4). The observed global Nm value (6.59), as well as the pairwise ones (7.75 – 16.03), support the hypothesis of strong evo- lutionary cohesion, since Nm ≥ 1 indicates a minimum amount of genetic migration capable of homogenizing demes within species (Mills and Allendorf 1996), includ- ing in lacertids (Levy et al. 2010). This feature of a highly cohesive species could be also explained by a recent irra- diation phenomenon. However this hypothesis requires a robust phylogeographic study offering coalescence-dat- ing analyses. Considering all the results, it is possible to argument in favour to the hypothesis of panmixia in T. hispidus, despite the discontinuity and historical changes seen in the biomes studied. This is surprising since T. hispidus individuals are sit-and-wait predators, territorialists, and oviparous that would suggest a tendency for struc- turings (Prieto et al. 1976; Van Sluys et al. 2010; Ribeiro and Freire 2011). Besides not dispersing through long distances (Pontes et al. 2008), sit-and-wait predators are usually opportunists and can feed on a variety of food items according to the local availability (Rodrigues 1987, 1988; Vitt 1991; Bergallo and Rocha 1993; Vitt 1995; Vitt et al. 1997; Pontes et al. 2008) suggesting ecologi- cal plasticity. Ecological plasticity predicts high genetic diversity. Higher genetic diversity tends to favour a bet- ter adaptation at the population, community and eco- system levels (Hughes et al. 2008). The feeding plasticity observed in T. hispidus (Pelegrin et al. 2017) is related to its high success in attempts to colonize new sites (Teixei- Table 3. Results of the AMOVA for the Tropidurus hispidus popula- tions in the study area. (p < 0.01). Source of variation Degrees of freedom Sum of squares Components of variance % of variance Among populations 6 729.782 3818.75 va 9.01 Within populations 148 5709.625 38,578.55 vb 90.09 Total 154 6439.407 42,397.30 100 ΦST 0.09007 Figure 4. Bayesian structuring analysis. The Y axis indicates the probability-based assignments for the genetic composition of each specimen analyzed (vertical bars). Note (1) Ca2, (2) Ca1, (3) AF, (4) TZ, (5) UZ, (6) AM, (7) Rest (including specimens from Canguare- tama and Tamandaré) . For a description of the sites, see Table 1. Table 4. Pairwise GST (above diagonal) and Nm (below diagonal) values recorded between Tropidurus hispidus populations. For acro- nyms, please refer to Table 1. Ca2 Ca1 TZ AF UZ AM Rest Ca2 - 0.0420 0.0363 0.0363 0.0322 0.0360 0.0527 Ca1 11.4029 - 0.0457 0.0379 0.0368 0.0435 0.0511 TZ 13.2673 10.4325 - 0.0383 0.0388 0.0458 0.0605 AF 13.2673 12.7004 12.5522 - 0.0302 0.0341 0.0455 UZ 15.0321 13.0902 12.3747 16.0316 - 0.0341 0.0505 AM 13.3976 10.9953 10.4261 14.1587 14.1595 - 0.0492 Rest 8.9877 9.2855 7.7588 10.4786 9.4000 9.6556 - 21Gene flow patterns reinforce the ecological plasticity of Tropidurus hispidus (Squamata: Tropiduridae) ra and Giovanelli 1999) and to its capacity of expanding towards new habitats (Levy et al. 2010; Breininger et al. 2012). This hypothesis is supported by the high degree of genetic diversity observed herein in T. hispidus and this feature might be favouring historically the species to a better adaptation to different biomes. These combined evidences suggest the stepping-stone model of range expansion as a probable explanation for wide distribu- tion of T. hispidus. Molecular studies with species of Tropidurus have revealed different patterns of evolutionary cohesion among populations, depending on the species studied. For instance, in Tropidurus semitaeniatus and T. hygomi, populations tend to be highly structured, but due to dif- ferent processes. In T. semitaeniatus the process of pop- ulation structuring was mediated by the course of the River São Francisco (Northeastern Brazil; Werneck et al. 2015). In T. hygomi, the population structuring was associated to different marine transgression/regression events, which isolated or connected regions along the Brazilian coastal plains (Fazolato et al. 2017). Tropidurus hispidus was expected to show the same pattern of genetically structured populations due to geographic isolation by different ecological pressures of morphoclimatic domains of humid forests and the Caat- inga (González et al. 2011; Matos et al. 2016). The data supporting these conclusions were the karyotypic struc- ture and COI gene sequences, respectively (Matos et al. 2016). However, our data failed to reinforce this idea and the analyses of the hypervariable regions of ISSR nuclear markers strongly pointed to panmixia. This occurred despite the geographical distances and the different selective pressures among studied biomes. A likely explanation for these contrasting evidences could be an intense dispersive behavior showed by T. hispidus males. Indeed, the use of bi-parental genetic markers has been recommended as a strategy to understand patterns of gene flow and demography (Goudet et al. 2002). The ISSRs markers analysed in this study agree with this rec- ommendation and allow inferences about the gene flow among T. hispidus demes. Cases of male-mediated dispersion in lizards have been documented in the literature in the last years (e.g. Johansson et al. 2008; Mouret et al. 2011; Ferchaud et al. 2015). When considering mitochondrial markers, of female inheritance, populations seem structured, (Matos et al. 2016) but when the male genetic pools is also ana- lysed such structuring disappears, as seen here using the ISSRs markers. This supports the hypothesis that the expansion of T. hispidus distribution range, and there- fore, new colonisations, would depend on a higher eco- logical ability of males to disperse farther than females. Mark-recapture studies of males and females could con- firm the explanations given herein. The occurrence of T. hispidus in urban areas, and its use of anthropogenic structures, (Carvalho 2013; Pel- egrin et al. 2017) could lead to facilitated dispersion and extend its distribution range. Human-facilitated disper- sion occurs in other lizard species, including exotic and invasive species (Vanzolini 1978; Mausfeld et al. 2002; Anjos and Rocha 2008). The T. hispidus population of Manaus (Amazon), which was recently invaded by indi- viduals from Roraima (Northern Brazil), is an example of this phenomenon (Ávila-Pires 1995; Carvalho 2013). However, this is speculative since we lack genetic data from Roraima. On the other hand, the individuals from Manaus had the same genetic profiles as the populations from Pernambuco, and did not show any type of genetic structuring, corroborating the hypothesis of panmixia along our study area. Our results revealed also that Tropidurus hispidus has a genetic variation above 90%. This points to an excellent conservation status along the studied area, con- sidering that low genetic variation would decrease this species’ ability to adapt to current and stochastic selec- tive pressures (Frankham and Ralls 1998; Frankham et al. 2002; Allendorf and Lundquist 2003). Indeed, T. his- pidus seems to have a high tolerance to habitat modifi- cations (Rodrigues 1987; Ávilla-Pires 1995), and it is a generalist regarding its microenvironmental require- ments (Vitt 1995; Mendonça and Moura 2011; Pelegrin et al. 2017). Therefore, our data reinforces this biological attribute (evolutionary potential), due to the high genetic variation observed. To conclude, according to our results, the sharing of a high genetic variation among the several T. hispi- dus population demes from different morphoclimatic domains seems to explain its ecological plasticity/evo- lutionary potential. According to Vitt et al. (1997) and Ivkovich et al. (2010) this is common in species with wide distribution ranges. This is a testable hypothesis that could be further tested in other Neotropical lizard species that have distribution patterns similar to T. his- pidus. ACKNOWLEDGMENTS This study was financed in part by the Coorde- nação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) – Brazil - Finance Code 001. Funds supporting this study were also provided by Facepe. F Ito and DMA Brito are grateful to CNPq for the Master fellowships provided (Graduate Program in Animal Biology-UFPE 22 Fernanda Ito et al. and grant number 552364/2010-0). RA Torres is espe- cially grateful to CNPq for the research fellowships pro- vided (grant numbers 301208/2012-3 and 306290/2015- 4) and to Drs. Miguel Trefaut Rodrigues and Marco Aurélio de Sena for providing tissue samples of E. divari- catus and T. torquatus. REFERENCES Abreu MLS, Frota JG, Yuki RN. 2002. Geographic distri- bution of Tropidurus hispidus. Herpetological Review. 33:66. Al Salameen F, Habibi N, Kumar V, Al Amad S, Dashti J, Talebi L, Al Doaij B. 2018 Genetic diversity and pop- ulation structure of Haloxylon salicornicum moq. in Kuwait by ISSR markers. PLoS One 13(11): e0207369. 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