Polyploidy breaks speciation barriers in Australian burrowing frogs Neobatrachus

Polyploidy has played an important role in evolution across the tree of life but it is still unclear how polyploid lineages may persist after their initial formation. While both common and well-studied in plants, polyploidy is rare in animals and generally less understood. The Australian burrowing frog genus Neobatrachus is comprised of six diploid and three polyploid species and offers a powerful animal polyploid model system. We generated exome-capture sequence data from 87 individuals representing all nine species of Neobatrachus to investigate species-level relationships, the origin and inheritance mode of polyploid species, and the population genomic effects of polyploidy on genus-wide demography. We describe rapid speciation of diploid Neobatrachus species and show that the three independently originated polyploid species have tetrasomic or mixed inheritance. We document higher genetic diversity in tetraploids, resulting from widespread gene flow between the tetraploids, asymmetric inter-ploidy gene flow directed from sympatric diploids to tetraploids, and isolation of diploid species from each other. We also constructed models of ecologically suitable areas for each species to investigate the impact of climate on differing ploidy levels. These models suggest substantial change in suitable areas compared to past climate, which correspond to population genomic estimates of demographic histories. We propose that Neobatrachus diploids may be suffering the early genomic impacts of climate-induced habitat loss, while tetraploids appear to be avoiding this fate, possibly due to widespread gene flow. Finally, we demonstrate that Neobatrachus is an attractive model to study the effects of ploidy on the evolution of adaptation in animals

Are 100 enough? Inferring acanthomorph teleost phylogeny using Anchored Hybrid Enrichment

BACKGROUND: The past decade has witnessed remarkable progress towards resolution of the Tree of Life. However, despite the increased use of genomic scale datasets, some phylogenetic relationships remain difficult to resolve. Here we employ anchored phylogenomics to capture 107 nuclear loci in 29 species of acanthomorph teleost fishes, with 25 of these species sampled from the recently delimited clade Ovalentaria. Previous studies employing multilocus nuclear exon datasets have not been able to resolve the nodes at the base of the Ovalentaria tree with confidence. Here we test whether a phylogenomic approach will provide better support for these nodes, and if not, why this may be. RESULTS: After using a novel method to account for paralogous loci, we estimated phylogenies with maximum likelihood and species tree methods using DNA sequence alignments of over 80,000 base pairs. Several key relationships within Ovalentaria are well resolved, including 1) the sister taxon relationship between Cichlidae and Pholidichthys, 2) a clade containing blennies, grammas, clingfishes, and jawfishes, and 3) monophyly of Atherinomorpha (topminnows, flyingfishes, and silversides). However, many nodes in the phylogeny associated with the early diversification of Ovalentaria are poorly resolved in several analyses. Through the use of rarefaction curves we show that limited phylogenetic resolution among the earliest nodes in the Ovalentaria phylogeny does not appear to be due to a deficiency of data, as average global node support ceases to increase when only 1/3rd of the sampled loci are used in analyses. Instead this lack of resolution may be driven by model misspecification as a Bayesian mixed model analysis of the amino acid dataset provided good support for parts of the base of the Ovalentaria tree. CONCLUSIONS: Although it does not appear that the limited phylogenetic resolution among the earliest nodes in the Ovalentaria phylogeny is due to a deficiency of data, it may be that both stochastic and systematic error resulting from model misspecification play a role in the poor resolution at the base of the Ovalentaria tree as a Bayesian approach was able to resolve some of the deeper nodes, where the other methods failed. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12862-015-0415-0) contains supplementary material, which is available to authorized users

Off-target capture data, endosymbiont genes and morphology reveal a relict lineage that is sister to all other singing cicadas

Phylogenetic asymmetry is common throughout the tree of life and results from contrasting patterns of speciation and extinction in the paired descendant lineages of ancestral nodes. On the depauperate side of a node, we find extant ´relict´ taxa that sit atop long, unbranched lineages. Here, we show that a tiny, pale green, inconspicuous and poorly known cicada in the genus Derotettix, endemic to degraded salt-plain habitats in arid regions of central Argentina, is a relict lineage that is sister to all other modern cicadas. Nuclear and mitochondrial phylogenies of cicadas inferred from probe-based genomic hybrid capture data of both target and non-target loci and a morphological cladogram support this hypothesis. We strengthen this conclusion with genomic data from one of the cicada nutritional bacterial endosymbionts, Sulcia, an ancient and obligate endosymbiont of the larger plant-sucking bugs (Auchenorrhyncha) and an important source of maternally inherited phylogenetic data. We establish Derotettiginae subfam. nov. as a new, monogeneric, fifth cicada subfamily, and compile existing and new data on the distribution, ecology and diet of Derotettix. Our consideration of the palaeoenvironmental literature and host-plant phylogenetics allows us to predict what might have led to the relict status of Derotettix over 100 Myr of habitat change in South America.Fil: Simon, Chris. University of Connecticut; Estados UnidosFil: Gordon, Eric R. L.. University of Connecticut; Estados UnidosFil: Moulds, M.S.. Australian Museum Research Institute; AustraliaFil: Cole, Jeffrey A.. Pasadena City College; Estados UnidosFil: Haji, Diler. University of Connecticut; Estados UnidosFil: Lemmon, Alan R.. Florida State University; Estados UnidosFil: Lemmon, Emily Moriarty. Florida State University; Estados UnidosFil: Kortyna, Michelle. Florida State University; Estados UnidosFil: Nazario, Katherine. University of Connecticut; Estados UnidosFil: Wade, Elizabeth J.. Curry College. Department of Natural Sciences and Mathematics; Estados Unidos. University of Connecticut; Estados UnidosFil: Meister, Russell C.. University of Connecticut; Estados UnidosFil: Goemans, Geert. University of Connecticut; Estados UnidosFil: Chiswell, Stephen M.. National Institute of Water and Atmospheric Research; Nueva ZelandaFil: Pessacq, Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Centro de Investigación Esquel de Montaña y Estepa Patagónica. Universidad Nacional de la Patagonia "San Juan Bosco". Centro de Investigación Esquel de Montaña y Estepa Patagónica; ArgentinaFil: Veloso, Claudio. Universidad de Chile; ChileFil: McCutcheon, John P.. University of Montana; Estados UnidosFil: Lukasik, Piotr. University of Montana; Estados Unidos. Swedish Museum of Natural History. Department of Bioinformatics and Genetics; Sueci

Phylogenomics Reveals Ancient Gene Tree Discordance in the Amphibian Tree of Life

The Author(s) 2020. Published by Oxford University Press, on behalf of the Society of Systematic Biologists. Molecular phylogenies have yielded strong support for many parts of the amphibian Tree of Life, but poor support for the resolution of deeper nodes, including relationships among families and orders. To clarify these relationships, we provide a phylogenomic perspective on amphibian relationships by developing a taxon-specific Anchored Hybrid Enrichment protocol targeting hundreds of conserved exons which are effective across the class. After obtaining data from 220 loci for 286 species (representing 94% of the families and 44% of the genera), we estimate a phylogeny for extant amphibians and identify gene tree-species tree conflict across the deepest branches of the amphibian phylogeny. We perform locus-by-locus genealogical interrogation of alternative topological hypotheses for amphibian monophyly, focusing on interordinal relationships. We find that phylogenetic signal deep in the amphibian phylogeny varies greatly across loci in a manner that is consistent with incomplete lineage sorting in the ancestral lineage of extant amphibians. Our results overwhelmingly support amphibian monophyly and a sister relationship between frogs and salamanders, consistent with the Batrachia hypothesis. Species tree analyses converge on a small set of topological hypotheses for the relationships among extant amphibian families. These results clarify several contentious portions of the amphibian Tree of Life, which in conjunction with a set of vetted fossil calibrations, support a surprisingly younger timescale for crown and ordinal amphibian diversification than previously reported. More broadly, our study provides insight into the sources, magnitudes, and heterogeneity of support across loci in phylogenomic data sets.[AIC; Amphibia; Batrachia; Phylogeny; gene tree-species tree discordance; genomics; information theory.].This work was supported by grants from a graduate student research award from the Society of Systematic Biologists and the University of Kentucky G.F. Ribble Endowment (to P.M.H.), by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES/BEX 2806/09-6 to P.L.V.P.), and by the National Science Foundation (DEB-0949532 and DEB-1355000 to D.W.W., DEB-1120516 to E.M.L., IIP-1313554 to A.R.L. and E.M.L, DEB-1355071 to J.M.B., DEB-1441719 to R.A.P., DEB-1311442 to P.L.V.P., DEB-1354506 to R.C.T., DEB-1021247 to E.P. and C.J.R., DEB-1021299 to K.M. Kjer, and DEB-1257610, DEB-0641023, DEB-0423286, and DEB-9984496 to C.J.R.), and the Australian Research Council (DP120104146 to J.S.K. and S.C.D.). S.R.R. thanks SENESCYT (Arca de Noé Initiative; SRR and O. Torres-Carvajal principal investigators) for funding for tissue collection. J.L. was supported by the Systematics Association and the Linnean Society Systematics Research Fund. This material is based upon work supported by the National Science Foundation Graduate Research Fellowship Program (DGE-3048109801 to P.M.H.) and by the National Science Foundation-supported National Center for Supercomputing Applications Blue Waters Graduate Research Fellowship Program (under Grant No. 0725070, subaward 15836, to P.M.H.). Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation

Whole genome duplication potentiates inter-specific hybridisation and niche shifts in Australian burrowing frogs Neobatrachus

Polyploidy plays an important role in evolution because it can lead to increased genetic complexity and speciation. It also provides an extra copy buffer and increases genetic novelty. While both common and well-studied in plants, polyploidy is rare in animals, and most polyploid animals reproduce asexually. Amphibians represent a dramatic vertebrate exception, with multiple independent sexually reproducing polyploid lineages, but very few cases have been studied in any detail. The Australian burrowing frog genus Neobatrachus is comprised of six diploid and three polyploid species and offers a powerful model animal polyploid system. We generated exome-capture sequence data from 87 individuals representing all nine species of Neobatrachus to investigate species-level relationships, the origin of polyploid species, and the population genomic effects of polyploidy on genus-wide demography. We resolve the phylogenetic relationships among Neobatrachus species and provide further support that the three polyploid species have independent origins. We document higher genetic diversity in tetraploids, resulting from widespread gene flow specifically between the tetraploids, asymmetric inter-ploidy gene flow directed from sympatric diploids to tetraploids, and current isolation of diploid species from each other. We also constructed models of ecologically suitable areas for each species to investigate the impact of climate variation on frogs with differing ploidy levels. These models suggest substantial change in suitable areas compared to past climate, which in turn corresponds to population genomic estimates of demographic histories. We propose that Neobatrachus diploids may be suffering the early genomic impacts of climate-induced habitat loss, while tetraploids appear to be avoiding this fate, possibly due to widespread gene flow into tetraploid lineages specifically. Finally, we demonstrate that Neobatrachus is an attractive model to study the effects of ploidy on evolution of adaptation in animals

Phylogenomics Reveals Ancient Gene Tree Discordance in the Amphibian Tree of Life

Molecular phylogenies have yielded strong support for many parts of the amphibian Tree of Life, but poor support for the resolution of deeper nodes, including relationships among families and orders. To clarify these relationships, we provide a phylogenomic perspective on amphibian relationships by developing a taxon-specific Anchored Hybrid Enrichment protocol targeting hundreds of conserved exons which are effective across the class. After obtaining data from 220 loci for 286 species (representing 94% of the families and 44% of the genera), we estimate a phylogeny for extant amphibians and identify gene tree–species tree conflict across the deepest branches of the amphibian phylogeny. We perform locus-by-locus genealogical interrogation of alternative topological hypotheses for amphibian monophyly, focusing on interordinal relationships. We find that phylogenetic signal deep in the amphibian phylogeny varies greatly across loci in a manner that is consistent with incomplete lineage sorting in the ancestral lineage of extant amphibians. Our results overwhelmingly support amphibian monophyly and a sister relationship between frogs and salamanders, consistent with the Batrachia hypothesis. Species tree analyses converge on a small set of topological hypotheses for the relationships among extant amphibian families. These results clarify several contentious portions of the amphibian Tree of Life, which in conjunction with a set of vetted fossil calibrations, support a surprisingly younger timescale for crown and ordinal amphibian diversification than previously reported. More broadly, our study provides insight into the sources, magnitudes, and heterogeneity of support across loci in phylogenomic data sets

Phylogenomic Reconstruction Sheds Light on New Relationships and Timescale of Rails (Aves: Rallidae) Evolution

The integration of state-of-the-art molecular techniques and analyses, together with a broad taxonomic sampling, can provide new insights into bird interrelationships and divergence. Despite their evolutionary significance, the relationships among several rail lineages remain unresolved as does the general timescale of rail evolution. Here, we disentangle the deep phylogenetic structure of rails using anchored phylogenomics. We analysed a set of 393 loci from 63 species, representing approximately 40% of the extant familial diversity. Our phylogenomic analyses reconstruct the phylogeny of rails and robustly infer several previously contentious relationships. Concatenated maximum likelihood and coalescent species-tree approaches recover identical topologies with strong node support. The results are concordant with previous phylogenetic studies using small DNA datasets, but they also supply an additional resolution. Our dating analysis provides contrasting divergence times using fossils and Bayesian and non-Bayesian approaches. Our study refines the evolutionary history of rails, offering a foundation for future evolutionary studies of birds

Pseudacris fouquettei Lemmon, Lemmon, Collins & Cannatella, 2008, sp. nov.

Pseudacris fouquettei sp. nov. (Figs. 2 and 3) Cajun Chorus Frog Holotype: (Fig. 2) TNHC 62265 (Texas Natural History Collection; field no. ECM 0029), adult male from the United States: Louisiana: East Baton Rouge Parish: (NW of Baywood on Lee Price Road, 1.4 mi W of jct. with SR 37; N 30.7147 W 90.8919), collected by Emily Moriarty Lemmon and David C. Cannatella on 11 March 2001. Paratypes: TNHC 62266–62267, same data as holotype and TNHC 63471–63479, same data as holotype except collected 21 February 2003 between 0.3–0.6 mi W of jct. with SR 37 on Lee Price Road. Etymology: The specific epithet is a patronym for Martin J. “Jack” Fouquette, Jr., who studied Pseudacris in the 1960 s and 1970 s. His extensive unpublished field data were instrumental in efforts to elucidate the species diversity of chorus frogs. Synonymy: A detailed history of Pseudacris nomenclature is available on the Amphibian Species of the World website (Frost 2007). Diagnosis: Pseudacris fouquettei is distinguished from other chorus frogs by 1) genetic data (Lemmon et al. 2007 b; Gartside 1980), 2) geographic distribution (Fig. 1), 3) advertisement call (Figs. 4–6), and 4) to a lesser degree by morphological data (Figs. 6 –7). This small slender species, with a subacuminate snout, has a dorsal pattern of three medium to dark brown longitudinal stripes or rows of spots on a pale tan or gray ground color; a white labial stripe is present. Pseudacris fouquettei can be distinguished from three broadly sympatric chorus frogs in the south-central United States using color pattern, morphology, and the terminal discs on the digits. Pseudacris crucifer typically has an “X” pattern on the dorsal surface, larger terminal discs, and is more arboreal. Pseudacris streckeri is larger and heavier-bodied and lacks terminal discs. In addition, both of these species have unpulsed singlenote advertisement calls compared to the pulsed call of P. fouquettei. Pseudacris clarkii typically has green spots or stripes on the dorsal surface, an interorbital triangle, and produces a much faster pulse-rate call (Conant and Collins 1998; E. Moriarty Lemmon, unpub. data). Pseudacris fouquettei can also be distinguished from three taxa with parapatric distributions: P. feriarum, P. maculata, and P. n i g r i t a (Fig. 1; Lemmon et al. 2007 b). Genetic data show that Pseudacris fouquettei is not closely related to the species in which it was formerly included (P. feriarum) or to P. maculata or P. triseriata. The new species instead forms the sister clade to P. nigrita (Fig. 8; Lemmon et al. 2007 b). Pseudacris fouquettei (referred to as P. triseriata feriarum by Gartside [1980]) is known to hybridize with P. nigrita in a narrow <20 km zone in the Pearl River floodplain along the border between Louisiana and Mississippi (Gartside 1980). The two species are fixed for alternative alleles at two or more allozyme loci outside the hybrid zone, however, indicating species-specific differences between these taxa (Gartside 1980). In addition, these taxa differ at 38 diagnostic SNPs in the 12 S/ 16 S mitochondrial gene region (27 P. fouquettei and 17 P. n i g r i t a were examined; Lemmon et al. 2007 b). Average pairwise sequence divergence between the two species is comparable to genetic distances of other Pseudacris species pairs (Fig. 8; GTR+G+I corrected p-distances for the 12 S/ 16 S mitochondrial region, with parameter settings derived from the mean of the posterior distribution from the Lemmon et al. [2007 b] Bayesian analysis). Pseudacris fouquettei and P. nigrita also show a sharp cline in color pattern across this contact zone and are easily distinguished using this character outside of the zone (Gartside 1980; EML unpub. data). Advertisement call data indicate that P. fouquettei differs from the three parapatric species with respect to several variables. The new species has a slower call rate than P. feriarum and P. m a c u l a t a (0.34 ± 0.06 s.d. vs. 0.49 ± 0.05 and 0.42 ± 0.07 calls/sec, respectively), a higher call duty cycle than P. nigrita (0.36 ± 0.05 vs. 0.31 ± 0.04), a longer call length than all three species (1115.42 ± 150.34 vs. 745.95 ± 79.25, 906.56 ± 127.48, and 892.73 ± 106.43 ms, respectively), and an intermediate pulse number between P. feriarum, P. maculata, and P. nigrita (13.07 ± 1.63 vs. 17.04 ± 2.25, 17.09 ± 1.64 and 9.56 ± 1.67, respectively). There is broad overlap among species with regard to dominant frequency (Figs. 4 –5; Table 1). Holotype P. fouquettei P. feriarum P. nigrita P. maculata (n = 26) (n = 19) (n = 19) (n= 15) DF 3273.05 3138.80 ± 205.79 2952.28 ± 312.93 3044.63 ± 156.84 3078.81 ± 135.96 2845.97–3712.94 2583.98–3583.74 2767.02–3294.58 2855.63–3283.81 CDC 0.37 0.36 ± 0.05 0.38 ± 0.05 0.31 ± 0.04 0.37 ± 0.04 0.26–0.44 0.31–0.49 0.25–0.40 0.31–0.44 CL 910.31 1115.42 ± 150.34 745.95 ± 79.25 906.56 ± 127.48 892.73 ± 106.43 867.15–1554.53 599.08–908.53 701.39–1161.68 728.39–1183.05 CR 0.41 0.34 ± 0.06 0.49 ± 0.05 0.34 ± 0.05 0.42 ± 0.07 0.14–0.43 0.42–0.59 0.26–0.46 0.24–0.55 PN 13.50 13.07 ± 1.63 17.04 ± 2.25 9.56 ± 1.67 17.09 ± 1.64 9.93–15.69 12.45–22.43 6.40–12.92 13.45 –20.00 Principal component analyses of call variables indicate that P. fouquettei does not overlap with P. feriarum along PCI (explains 47 % of variance), which has high loadings of call rate and pulse number. The new species overlaps to a small degree with P. maculata and to a greater degree with P. n i g r i t a along this axis. Pseudacris fouquettei overlaps with all three species along PCII (explains 27 % of variance), which has high loadings of call duty cycle and call length (Fig. 6; Table 2). In congruence with previous studies, Pseudacris fouquettei overlaps morphologically with the parapatric taxa P. feriarum and P. nigrita. These three species are morphologically distinct from their more distant relative, P. maculata, with respect to head width, head length, eye width, tibia length, and femur length (Fig. 7). Pseudacris fouquettei is more similar to its sister species, P. nigrita, in terms of head width and femur length, more similar to P. feriarum with regard to head length, intermediate between the two species with respect to snout angle and foot length, and nearly identical to both species in terms of snout length, eye width, tympanum diameter, and tibia-fibula length (Fig. 7; Table 3). Multivariate analyses of morphometric data indicate that P. fouquettei is essentially identical to P. f e r i - arum and P. nigrita along PC 1 (explains 53 % of variance), which is dominated by head size and leg length variables. The three species are distinct from P. m a c u l a t a along this axis. Pseudacris fouquettei is intermediate between P. feriarum and P. nigrita, however, along PC 2 (explains 18 % of variance), which is dominated by snout angle and foot length (Fig. 6; Table 4). FIGURE 5. Box and whisker plots (median = central black bar, boxes = 25 th– 75 th quartiles, whiskers = maximum and minimum values after excluding outliers) showing advertisement call variation among Pseudacris fouquettei (fou), P. feriarum (fer), P. nigrita (nig), and P. maculata (mac). Five variables are presented: call rate, call length, dominant frequency, pulse number, and call duty cycle. Individuals analyzed are listed in Appendix 2. Holotype P. fouquettei P. feriarum P. nigrita P. maculata (n = 117) (n = 202) (n = 78) (n = 74) SVL 27.38 26.39 ± 1.71 25.54 ± 1.90 25.73 ± 1.75 24.35 ± 2.80 22.20–29.79 19.98–30.28 21.26–29.50 19.95–30.56 SA 1.04 0.99 ± 0.05 1.02 ± 0.06 0.96 ± 0.05 0.96 ± 0.06 0.84–1.13 0.81–1.20 0.86–1.08 0.79–1.11 HW 9.30 8.72 ± 0.63 8.81 ± 0.63 8.56 ± 0.61 7.36 ± 1.07 7.01–10.14 6.67–10.18 6.80–10.15 5.34–10.03 HL 9.36 9.17 ± 0.54 9.07 ± 0.62 9.27 ± 0.57 7.99 ± 0.99 7.74–10.44 7.36–10.73 7.97–10.70 6.28–10.21 TD 1.72 1.34 ± 0.20 1.36 ± 0.20 1.30 ± 0.13 1.22 ± 0.23 0.77–1.80 0.81–1.83 1.01–1.62 0.87–1.81 EW 3.07 2.88 ± 0.25 2.91 ± 0.27 2.93 ± 0.23 2.48 ± 0.29 2.18–3.55 2.23–3.65 2.35–3.55 1.92–3.18 Snout 2.29 2.48 ± 0.25 2.41 ± 0.26 2.49 ± 0.24 2.17 ± 0.33 1.86–2.97 1.64–3.05 1.94–3.16 1.32–2.95 FeL 13.14 11.69 ± 0.94 11.96 ± 1.02 11.68 ± 0.86 9.75 ± 1.46 9.38–14.18 9.33–14.74 9.38–13.68 7.73–13.44 TL 14.07 13.08 ± 0.91 13.05 ± 1.04 12.87 ± 0.98 10.14 ± 1.50 10.97–15.11 10.42–15.37 10.82–15.06 8.08–13.50 FoL 13.60 12.87 ± 0.92 12.21 ± 1.04 12.94 ± 1.08 11.60 ± 1.59 10.14–14.85 9.84–15.17 10.54–15.54 8.97–15.36 The color pattern of P. fouquettei closely resembles that of P. feriarum in terms of the three longitudinal stripes along the dorsal surface, although there is high inter-population variation in this character (Fig. 3). Pseudacris fouquettei can be easily distinguished from P. n i g r i t a, however, based on color pattern. The latter species has generally darker markings including a broken stripe or spotted pattern on the dorsal surface and wider, darker (tending to black), transverse bars on the legs (Figs. 2 and 3). Description: Male Pseudacris fouquettei attain a maximum snout-vent length of 30 mm, and females reach at least 27 mm. The head is slightly narrower than the body, and the top of the head is barely convex. In dorsal profile, the snout is acuminate and in ventral profile, it projects well beyond the tip of the lower jaw. The snout is long with slightly protuberant nostrils situated at a point about two-thirds of the distance from the anterior corner of the eye to the tip of the snout. The eyes are of moderate size and not protuberant. The canthus rostralis is rounded, and the loreal region is barely concave; the lips are moderately thick and not flared. A thin supratympanic fold extends posteriorly from the eye, above the tympanum, and downward to a point above the insertion of the arm. The fold barely obscures the upper edge of the tympanum, which otherwise is distinct and separated from the eye by a distance equal to about two-thirds of the diameter of the tympanum. The arms are moderately long and robust; an axillary membrane is absent. A slight ulnar fold is present, with no rows of tubercles, and a distinct dermal fold is present on the dorsal surface of the wrist. The fingers are long and slender and bear discs that are only slightly wider than the fingers. The subarticular tubercles are moderately large and round, and none are bifid. The supernumerary tubercles are absent. A large almost bifid palmar tubercle is present. The prepollex is not enlarged and in breeding males does not bear a nuptial excrescence. No webbing is present on the hands. The legs are of moderate length and slender. A well-developed, flaplike inner tarsal fold extends the full length of the tarsus and connects to the inner metatarsal tubercle. An outer tarsal fold is lacking. The inner metatarsal tubercle is small, elliptical, and elevated. A smaller, conical outer metatarsal tubercle is present. The toes are long and slender; the small toe discs are slightly wider than the digits. The subarticular tubercles are large, round, and flattened in profile. A few supernumerary tubercles are barely evident on the proximal segments of the outer digits. The toes are webbed only basally between digits III and IV and between IV and V. FIGURE 7. Box and whisker plots (median = central black bar, boxes = 25 th– 75 th quartiles, whiskers = maximum and minimum values after excluding outliers) showing morphological variation among Pseudacris fouquettei (fou), P. f e r i - arum (fer), P. nigrita (nig), and P. m a c u l a t a (mac). Nine variables are presented: residual head width, residual head length, residual snout length, residual snout angle, residual eye width, residual tympanum diameter, residual tibia-fibula length, residual femur length, and residual foot length. Individuals analyzed are listed in Appendix 1. The cloacal opening is directed posteriorly near the mid level of the thighs; a short transverse flap lies dorsal to the opening, and partially covers it. The skin on the dorsum is weakly granular, whereas that on the venter is strongly granular. The tongue is cordiform, shallowly notched posteriorly, and barely free behind. The dentigerous processes of the vomers are small rounded elevations that are widely separated medially and lie between the ovoid choanae. Two or three teeth are present on each process. The short elliptical vocal slits extend along the posterior two-thirds of the tongue to the angle of the jaws. The vocal sac is single, median, subgular, and greatly distensible. Measurements of holotype: Adult male, morphometric data: SVL 27.38; SA 1.04; HW 9.30; HL 9.36; TD 1.72; EW 3.07; Snout 2.29; FeL 13.14; TL 14.07; FoL 13.60 mm; advertisement call data: DF 3273.05 Hz; CDC 0.37; CL 910.31 ms; CR 0.41 calls per sec; PN 13.50; genetic data: mitochondrial haplotype of the Pseudacris fouquettei clade (Lemmon et al. 2007 b). Color in preservative: The general coloration of Pseudacris fouquettei is light brown above with three darker brown stripes or three sets of elongate spots forming rows on the back. The dorsal surface ranges from light gray to tan. The markings on the back and transverse bars on the limbs vary from medium to dark brown. There is a dark brown to reddish-brown stripe from the nostril to the eye, which extends to the mid or posterior flank region. A white to cream labial stripe is present, and extends beneath the eye to just posterior to the tympanum. The venter is creamy white and may have some brown flecking in the pectoral and mid abdominal region. The eye has a dark pupil with a bronze-gold iris. Color in life: In life, the coloration is similar to that in preservative except the labial stripe is a bright iridescent white, the ground dorsal color may have a very slight pinkish hue, and the dorsal surface may have occasional brassy or gold flecking. Based on color photographs before preservation, paratypes TNHC 63471 and TNHC 63473 are tan to medium brown on the dorsal surface with three dark brown stripes that run longitudinally down the back of the frog. A broad dark reddish brown stripe runs laterally from the tip of the snout through the eye and tympanum to just anterior to the rear legs. A narrow bright white line runs laterally from the tip of the snout to the posterior end of the jaw just below the brown lateral stripe. Front and rear legs have dark brown transverse bars on a tan to medium brown background. The ventral surface is cream with several dark flecks. The throat is yellowish-brown. Tadpoles: The tadpoles of this species have been described by Siekmann (1949; referred to as Pseudacris triseriata feriarum). Trauth et al. (2004) show multiple photographs of P. fouquettei tadpoles (referred to as P. triseriata) at different developmental stages. Variation: There is marked variation in color pattern types in our sample of twelve Pseudacris fouquettei from the type locality (East Baton Rouge Parish, Louisiana). Four exhibit a strong three-stripe pattern on the dorsal surface (TNHC 62267, 63471, 63477, and 63478), two show a three-stripe pattern with dark dots bounding the stripes (TNHC 63473 and 63475), four show a broken three-stripe pattern (TNHC 62265, 62266, 63472, and 63479), and two are patternless, except for markings on the legs (TNHC 63474 and 63476). An interorbital triangle is not present in any specimens. Dark transverse bars are present on the legs and vary in number from 2 to 15 among specimens. A dark brown stripe runs laterally from anterior to the nares to mid-flank, and a white labial stripe is present on all specimens. The ventral surfaces are generally cream, but some specimens have venters with scattered flecks of gray pigment. The vocal sac area is yellowish-orange with dark gray pigment (in males). Other Pseudacris fouquettei populations are similar in color pattern, except that the stripe pattern is more consistent. In ten specimens from Craighead Co., Arkansas, all except one exhibit the solid three-stripe pattern (TNHC 62255–62264, not TNHC 62259, which shows a faint broken-stripe pattern). In twelve specimens from Newton Co., Texas, all specimens show a strong three-stripe pattern (TNHC 20691–20696 and TNHC 20698–20704; Appendix 1). Ecology and natural history: Pseudacris fouquettei is a winter or early spring breeder that can be heard chorusing in temporary bodies of water from January to May. Breeding activity depends on amenable temperatures (4 °C to 21 °C; nocturnal temperature of 10–18 °C is optimal) and recent rainfall. Pseudacris fouquettei congregate to breed in ephemeral pools and ponds in a variety of habitats, ranging from forested areas to open fields. The species has successfully colonized wet roadside ditches throughout its range. Little is known about the activity of the species outside of the breeding season. Frogs disperse from breeding sites and presumably forage on small invertebrates like other trilling chorus frogs (Whitaker 1971) and range within an area of about 200 m from the breeding pool (Kramer 1973, 1974). Pseudacris fouquettei is not ecologically limited to pine forest, as is its sister species, P. nigrita. Rather, the new species appears to tolerate a much broader range of environmental conditions. Distribution: Pseudacris fouquettei is distributed along the coast of the Gulf of Mexico from western Mississippi, Louisiana, and eastern Texas north to eastern Oklahoma (nearly to Kansas), Arkansas, and barely into southern Missouri (Fig. 1; Lemmon et al. 2007 b).Published as part of Lemmon, Emily Moriarty, Lemmon, Alan R., Collins, Joseph T. & Cannatella, David C., 2008, A new North American chorus frog species (Amphibia: Hylidae: Pseudacris) from the south-central United States, pp. 1-30 in Zootaxa 1675 on pages 4-14, DOI: 10.5281/zenodo.18028