Harry L. Taylor

Natural Hybridization Between the Teiid Lizards Cnemidophorus tesselatus (Parthenogenetic) and C. tigris marmoratus (Bisexual): Assessment of Evolutionary Alternatives

Abstract Annual hybridization is taking place between representatives of the parthenogenetic lizard Cnemidophorus tesselatus (2n = 46, 47) and males of the bisexual species C. tigris marmoratus (2n = 46) in desert grassland habitats at Arroyo del Macho, Chaves County, New Mexico. This raises the question of whether a new triploid parthenogenetic species may be originating as a consequence of this activity. Hybrids were collected in each of four years (1996–1999), and 20 of 21 hybrids collected (12 males and 8 females) were available for study. Although a triploid parthenogenetic species (Cnemidophorus exsanguis 3n = 69) and a diploid bisexual species (C. inornatus 2n = 46) were also found at the hybridization site, the genealogy of the hybrids was determined unequivocally with karyotypic and electrophoretic evidence (34 loci tested). The specimens examined electrophoretically included an adult female and one of her laboratory-reared daughters, which demonstrated for the first time clonal inheritance in C. tesselatus pattern class E. The population of C. tesselatus at Arroyo del Macho is characterized by two karyotypic cytotypes. The ancestral one (2n = 46) occurs at about half the frequency of the derived cytotype (2n = 47), which apparently was produced by centric fission of the ancestral X-chromosome from C. tigris In contrast, the occurrence of the two cytotypes was reversed and strongly asymmetrical in the hybrids; only one of nine hybrids possessed the fissioned X-chromosome. This individual was significantly different in 12 meristic characters from the sample of hybrids with intact X-chromosomes. Predictably, principal components scores for this individual fell outside the 95% confidence ellipse of scores of the other eight hybrids that were karyotyped. The skewed ratio and multiple phenotypic differences suggest that hybrids inheriting a fissioned X-chromosome might be at a selective disadvantage compared to hybrids with intact X-chromosomes. All 20 hybrids closely resemble C. tesselatus in most color pattern features. However, these hybrids, like C. tigris marmoratus lack lateral stripes. Because the population of C. tesselatus at Arroyo del Macho has lateral stripes (or their remnants), hybrids can be readily distinguished from C. tesselatus by this color pattern feature. Compared to the two parental species, hybrids had a significantly lower mean number of scales around midbody, but hybrids resembled either C. tesselatus or C. tigris marmoratus in other univariate meristic characters. This mosaic pattern of resemblance was simplified to a three-dimensional depiction of variation using principal components analysis. Each of two principal components expressed the resemblance of hybrids to one of the two parental species. A third component reflected the difference between hybrids and both parental species. A canonical variate analysis of meristic characters demonstrated the multivariate distinctiveness of each group—hybrids, C. tesselatus and C. tigris marmoratus However, based on Mahalanobis D2 distances, the closest morphological resemblance among hybrids and parental species was between hybrids and the maternal species, C. tesselatus Nine additional museum specimens, suspected of being C. tesselatus × C. tigris marmoratus hybrids, were identified, as such, by a canonical variate analysis using our samples of C. tesselatus, C. tigris marmoratus and hybrids from Arroyo del Macho as a priori groups. These nine individuals document hybridizations between C. tesselatus and C. tigris marmoratus at two additional localities in Chaves County, New Mexico, two localities in Sierra County, New Mexico, and a cluster of sites near Presidio, Presidio County, Texas. Previously, several of these hybrids had been misidentified as male C. tesselatus The reproductive systems of female and male hybrids were compared histologically to those of C. tesselatus and C. tigris marmoratus respectively. Sexually mature and reproductive adults of C. tesselatus usually have oocytes in the ovary, complete and well-organized ovarian follicle walls, inconspicuous connective tissue and fewer vacuoles in the well-vascularized ovary, the distal oviduct with a thin mucosa, well-developed alveolar glands restricted to the middle oviduct, a proximal oviduct with a thick mucosa and well-developed folds, and small mesonephric tubules. Female hybrids have a poorly defined follicular epithelium with little vascularization in small ovaries, empty or fluid-filled follicles without oocytes, few or no cilia in the middle oviduct, and numerous abnormally large mesonephric tubules. There is no evidence that Cnemidophorus tesselatus × C. tigris marmoratus females can produce viable and fertile eggs. Although hybrid males are capable of producing sperm that appear normal and were present in the epididymides, the allotriploid chromosome complement reduces the chance that sperm would carry genetically balanced sets of information. Although the annual production of hybrids could affect the long-term success of this local population of C. tesselatus two lines of evidence indicate that hybridization is unlikely to result in its extirpation. First, the population of C. tigris marmoratus at Arroyo del Macho is tightly associated with a microhabitat dominated by creosote bush. Because creosote bush is distributed there in small, widely scattered patches, the density of C. tigris marmoratus is relatively low, and many individuals of C. tesselatus escape insemination. This was evident from an absence of sperm in the reproductive tracts of 11 individuals of C. tesselatus collected during the peak reproductive season (May and June) of three different years. Second, reproductively mature individuals of C. tesselatus are significantly larger than comparable females of C. tigris marmoratus This translates into larger clutches, with the mean clutch size of C. tesselatus being twice as large as that of C. tigris marmoratus The disparity in mean clutch size in conjunction with habitat constraints on C. tigris marmoratus probably explains why C. tesselatus outnumbers both C. tigris marmoratus and hybrids by a ratio of approximately 2:1 at the hybridization site. Although hybridization between C. tesselatus and C. tigris marmoratus appears to be an annual event at Arroyo del Macho, there is no evidence that a new triploid parthenogenetic species is resulting from this hybridization activity—all female hybrids examined were sterile. Nevertheless, the hybridization taking place at Arroyo del Macho is a remarkable natural experiment in progress, with either evolutionary alternative—speciation vs. destabilizing hybridization—adding to an understanding of the dynamics between parthenogenetic and bisexual species in sympatric associations.

Congruent Patterns of Genetic and Morphological Variation in the Parthenogenetic Lizard Aspidoscelis tesselata (Squamata: Teiidae) and the Origins of Color Pattern Classes and Genotypic Clones in Eastern New Mexico

Abstract Aspidoscelis tesselata exhibits significant clonal diversity despite its recent origin (from hybridization between A. tigris marmorata and A. gularis septemvittata) and its parthenogenetic mode of reproduction. Two hypotheses have been advanced to explain the derivation of its genetic and morphological variation: (1) separate parthenogenetic lineages derived from several different F1 hybrid zygotes, and (2) postformational mutations occurring in a parthenogenetic lineage derived from a single F1 hybrid zygote. We evaluated these competing hypotheses with evidence from skin transplant studies, protein electrophoresis, multivariate analyses of morphological characters, and geographic distributions of pertinent groups. Starting with the clonal diversity at Conchas Lake State Park, San Miguel County, New Mexico, we expanded the study to include populations at Sumner Lake State Park and Fort Sumner (De Baca County), Puerto de Luna (Guadalupe County), and Arroyo del Macho and Roswell (Chaves County). This enabled us to resolve origins of color pattern classes and genotypic clones in eastern New Mexico. We used pattern class designations C-E and E-C to signify that elements of both pattern classes were expressed in populations at Conchas Lake and Arroyo del Macho. The two pattern classes at Conchas Lake (C-E and D) had the same F1 hybrid karyotype (2n = 46), with haploid sets of 23 chromosomes characteristic of each progenitor species of A. tesselata. Clonal variation was found at 4 of the 35 gene loci examined electrophoretically: GPI (glucose-6-phosphate isomerase), EST2 (a muscle esterase), sACOH (aconitase hydratase), and MPI (mannose-6-phosphate isomerase). The strong congruence between genotype and morphological variation facilitated the characterization of three morphological subgroups of C-E. Although these subgroups lacked individually distinctive color patterns, they were discriminated effectively in canonical variate analyses based on scalation characters and a priori groups of known genotype. Nine individuals of Conchas C-E and four individuals of Conchas D have histocompatibility data from a recent skin transplant study (Cordes and Walker, 2003). The subgroup identities of the C-E specimens document histocompatibility among the three morphological subgroups of C-E and between each subgroup and representatives of pattern class D. This evidence, together with Maslins (1967) report of histocompatibility between pattern classes C and E, suggests that all color pattern classes, morphological subgroups, and genotypic clones of A. tesselata can be traced back to a single ancestral F1 hybrid zygote. A pair of pale broken lines in the middorsal region distinguishes pattern class D from the other pattern classes. However, Conchas ID shared the GPI −100/−96, EST2 100/96 genotype with Conchas IC-E, and individuals of these pattern classes were very similar in multivariate meristic characters. Sumner D expressed the same type of relationship, resembling the syntopic population of Sumner C rather than the other population of D. In addition, certain individuals of Sumner C had partially divided (D-like) vertebral lines—additional evidence that Sumner C was ancestral to Sumner D. We conclude that pattern class New Mexico D is polyphyletic, having originated twice from different individuals of C-E and C in the vicinities of Conchas and Sumner Lakes. The northern position of pattern classes C and C-E in the range of A. tesselata is consistent with recent colonizations by individuals from more southerly populations. A candidate source population, based on its extensive color pattern and meristic variation, is E-C at Arroyo del Macho. The strong morphological resemblance of several northern populations to Macho E-C rather than to either syntopic clones or geographically proximate populations of other pattern classes supports this possibility. Evidence from geographic distributions, patterns of genotypic and meristic variation, and histocompatibility identifies postformational mutations as the likely basis for the genetic and morphological variation found in A. tesselata. This variation also includes different life-history characteristics between pattern classes C and E at Sumner Lake State Park. The name tesselata is presently associated indirectly with pattern class C through the neotype of A. tesselata. The neotype is a specimen of Colorado D, a derivative of pattern class C. With respect to pattern classes E-C, E, and other southern variants, taxonomic restructuring would confront mosaic patterns of genotypic, phenotypic, and geographic variation—patterns expected from random mutations in clonally reproducing species. Aspidoscelis tesselata has exploited a variety of ecological opportunities despite the constraints of clonal inheritance. Postformational mutations in the generalized genotype acquired from its progenitor species may have contributed to its ecological success.

Evidence for Specific Recognition of the San Esteban Whiptail Lizard (Cnemidophorus estebanensis)

The divergent whiptail lizard of Isla de San Esteban, Gulf of California, differs from Cnemidophorus tigris in more morphological characters than any other form of the complex. C. estebanensis, characterized by impressive differences in size, color pattern, and scutellation, is regarded herein as specifically distinct from C. tigris. The whiptail lizard of Isla Tiburon, with which C. estebanensis has sometimes been confused, is allocated to C. tigris gracilis instead of C. tigris aethiops, which is restricted to southern Sonora, Mexico. i i t e San Esteban hiptail sis)

Laboratory Hybridization Among North American Whiptail Lizards, Including Aspidoscelis Inornata Arizonae × A. tigris marmorata (Squamata: Teiidae), Ancestors of Unisexual Clones in Nature

ABSTRACT The natural origin of diploid parthenogenesis in whiptail lizards has been through interspecific hybridization. Genomes of the parthenogens indicate that they originated in one generation, as the lizards clone the F1 hybrid state. In addition, hybridization between diploid parthenogens and males of bisexual species has resulted in triploid parthenogenetic clones in nature. Consequently, the genus Aspidoscelis contains numerous gonochoristic (= bisexual) species and numerous unisexual species whose closest relatives are bisexual, and from whom they originated through instantaneous sympatric speciation and an abrupt and dramatic switch in reproductive biology.

Application of the Evolutionary Species Concept to Parthenogenetic Entities: Comparison of Postformational Divergence in Two Clones of Aspidoscelis tesselata and between Aspidoscelis cozumela and Aspidoscelis maslini (Squamata: Teiidae)

Abstract Sumner Lake State Park, De Baca County, New Mexico, is the only known locality where three pattern classes of diploid, parthenogenetic Aspidoscelis tesselata (Sumner C, Sumner D, and Sumner E) coexist in syntopy. Reciprocal skin transplants confirmed that the pronounced phenotypic differences between Sumner C and Sumner E represent postformational genetic changes rather than separate hybridization origins. Sumner D is meristically indistinguishable from Sumner C and is considered to be a recent mutational derivative of the latter. In contrast, Sumner E is distinctly different from Sumner C in multivariate meristic characters and several important life-history characteristics. Discordant patterns of phenotypic variation characterize many geographically disjunct groups of A. tesselata classified as pattern class E, thus defying a cohesive diagnosis. Therefore, based on the evolutionary species concept (ESC), we consider Sumner C and Sumner E to be divergent clonal groups in the same species. We contrast this example with a parthenogenetic complex on the Yucatán Peninsula in which formal recognition of Aspidoscelis maslini and Aspidoscelis cozumela can be accommodated under the ESC.

Males of three normally parthenogenetic species of teiid lizards (Genus Cnemidophorus)

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Hybridization Between the Endangered Unisexual Gray-Checkered Whiptail Lizard (Aspidoscelis dixoni) and the Bisexual Western Whiptail Lizard (Aspidoscelis tigris) in Southwestern New Mexico

Abstract Hybridization between the unisexual Aspidoscelis dixoni and the bisexual Aspidoscelis tigris punctilinealis in southwestern New Mexico is documented by observations and analyses of external morphology (coloration, size, scalation), chromosomes (karyotypes), nuclear gene products (allozymes), and mitochondrial DNA. The locality (Hidalgo County, Antelope Pass of the Peloncillo Mountains, centered at 10.5 km west of Animas), consisting of only a few square kilometers, is the only place where this particular unisexual clone of A. dixoni exists. Because of its extreme rarity in recent years, A. dixoni has been listed as an Endangered Species in New Mexico, and the status of its populations has received intense study. Today, the cause(s) of endangerment remains unknown, although we hypothesize that interspecific competition may be the problem. Aspidoscelis dixoni is a diploid unisexual species that normally reproduces by parthenogenetic cloning, as demonstrated here with genetic data from laboratory-reared lizards. However, fertilization of its eggs in Antelope Pass is possible if mating occurs with a male of the syntopic bisexual species A. tigris punctilinealis. The resulting hybrids closely resemble their maternal parent morphologically, but they are triploid and the females observed to date have been sterile. Aspidoscelis t. punctilinealis is a recent invader of southwestern New Mexico. It is the dominant species of whiptail lizard today in the low-elevation, semiarid habitat of creosote desertscrub in Antelope Pass. The present rarity of A. dixoni in Antelope Pass, in contrast to its abundance a few decades ago, may result from negative interactions with this dominant species, including asymmetrical destabilizing hybridization. Only a few other populations of A. dixoni are known to exist, each in a limited area in southwestern Texas, so there is a hiatus of nearly 500 km between the small and restricted populations in New Mexico and Texas. Comparative genetic data presented here indicate that although these populations are similar, the population in New Mexico represents a unique clone. It has three alleles at 3 nuclear gene loci (among 31 examined) that distinguish it from the Texan populations, and it lacks a microchromosome that occurs in Texan populations. In addition, in this paper we present new comparative genetic data confirming that the origin of A. dixoni itself was from a hybrid between an A. tigris marmorata ♀ × A. gularis septemvittata ♂, consistent with earlier studies.

Hybrid Cnemidophorus (Sauria: Teiidae) in Ninemile Valley of the Purgatoire River, Colorado

Canonical variate analysis (CVA) of color pattern and scutellation characters in lizards collected in Ninemile Valley of the Purgatoire River at Higbee, Otero Co., Colorado graphically depicted two forms of parthenogenetic Cnemidophorus tesselatus (diploid color pattern class C and triploid color pattern class B), gonochoristic C. sexlineatus, and hybrid C. tesselatus x C. sexlineatus as four morphologically distinctive groups. A newly obtained male specimen with a C. tesselatus-like color pattern entered as an unknown was classified to the hybrid group in the CVA. It represented the fourth tetraploid hybrid C. tesselatus B(3n) x C. sexlineatus collected in the valley. A female specimen was also classified to the hybrid group by the CVA. A triploid karyotype (3n = 69) and color pattern identified this female as a putative C. tesselatus C(2n) x C. sexlineatus hybrid. Whereas male hybrids are presumably sterile and biological novelties at most, the possibility remains that a female hybrid could be the founder of a new parthenogenetic lineage. Zweifel (1965) informally described six color pattern classes of parthenogenetic lizards (A, B, C, D, E, F) in the Cnemidophorus tesselatus complex (family Teiidae) of which four (A-D) were reported from Colorado. Wright and Lowe (1967a) found that pattern classes A and B were triploid hybrid derivatives and pattern classes CF were diploid hybrid derivatives. Analyses of morphology (Walker et al., 1990) and karyology (Cordes, 1991) identified three of Zweifels pattern classes of C. tesselatus in sympatry in Ninemile Valley of the Purgatoire River at Higbee, Otero Co., Colorado rather than two as indicated by Zweifel (1965), Densmore et al. (1989), and Wright (1993). These included diploid C. tesselatus C and D and triploid B, not triploid pattern class C as indicated by Parker and Selander (1976), Densmore et al. (1989), Wright (1993), and Leuck (1993). We found that the Ninemile Valley triploid population was morphologically indistinguishable from the pattern class B population described by Zweifel (1965) from Pueblo Co., Colorado. In samples collected in the valley since 1960 by various workers, diploid C. tesselatus outnumbered triploid C. tesselatus by a combined ratio of over 7:1. Moderate numbers of the gonochoristic species C. sexlineatus, the paternal parent of triploid C. tesselatus B (Wright and Lowe, 1967a; Parker and Selander, 1976), were also found in many parts of the area sampled. Four hybrid male C. tesselatus x C. sexlineatus were included in these collections. Three of these were described, and their probable parental forms identified as triploid C. tesselatus B x C. sexlineatus, by Walker et al. (1990). In the present study we describe the most recently collected hybrid male Cnemidophorus, infer its putative parentage on the basis of critical morphological characters, and evaluate the possibility of a hybrid origin for a female specimen displaying an ambiguous combination of classification characters. MATERIALS AND METHODS-Characters of scutellation that were quantitatively analyzed included: number of granules around midbody (GAB), number of granules from occiput to rump (OR), sum of left and right femoral pores (FP), number of subdigital lamellae on the longest toe of the left pes (SDL), sum of left and right circumorbital scales (COS), sum of This content downloaded from 207.46.13.57 on Mon, 08 Aug 2016 05:24:23 UTC All use subject to http://about.jstor.org/terms 236 The Southwestern Naturalist vol. 39, no. 3

PARTHENOGENETIC CNEMIDOPHORUS TESSELATUS COMPLEX AT HIGBEE, COLORADO : RESOLUTION OF 30 YEARS OF CONTROVERSY

At times over 30 years, various authors have expressed conflicting opinions on the composition of the parthenogenetic Cnemidophorus tesselatus complex at Higbee, Otero County, Colorado, the only known site of sympatry between diploid and triploid members of this complex. Recently, some workers have stated that two rather than three distinctive parthenogens occur at Higbee, one being an undescribed triploid species (tentatively designated pattern class C), which resulted from hybridization between C. tesselatus D(2n) and gonochoristic C. sexlineatus. Alternatively, our study identified two diploid pattern classes [C(2n) and D(2n)] and one triploid pattern class [B(3n)] among specimens of C. tesselatus from Higbee (including those with electrophoretically determined ploidy levels). We used two canonical variate analyses (CVA) to test hypotheses concerning the identity and origin of the triploid form. CVA1 suggested that specimens of the Higbee triploid form represented C. tesselatus B(3n), not an undescribed species. CVA2 suggested that C. tesselatus B(3n) was derived from a C. tesselatus C(2n) x C. sexlineatus hybrid, not a hybrid involving C. tesselatus D(2n).

Reproductive Characteristics and Body Size in the Parthenogenetic Teiid Lizard Cnemidophorus tesselatus: Comparison of Sympatric Color Pattern Classes C and E in De Baca County, New Mexico

Cnemidophorus tesselatus is an allodiploid, parthenogenetic lizard comprised of a number of distinctive tokogenetic arrays (Frost and Hillis, 1990) presently identified by four color pattern class descriptors: C, Colorado D, New Mexico D, and E (Taylor et al., 1996; Walker et al., 1997). The macrogeographic relationships of these pattern classes were outlined in Zweifels (1965) pioneering study of geographic variation in Cnemidophorus tesselatus. Pattern class C was known to exist isolated from the other pattern classes in parts of southeastern Colorado, northwestern Oklahoma, and northwestern Texas. Sympatry between pattern classes C and D was known to occur only in the vicinities of the historic townsite of Higbee, Otero County, Colorado, and Conchas Lake, San Miguel County, New Mexico. Pattern class E occurred to the south, with specimens from Fort Sumner, De Baca County, New Mexico, approximately 105 km south of the Conchas Lake locality, documenting the closest proximity of pattern class E to pattern classes C and D.