Homing in the Red-Eared Slider (Trachemys Scripta Elegans) in Illinois Authors: John K
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Homing in the Red-Eared Slider (Trachemys scripta elegans) in Illinois Authors: John K. Tucker, and James T. Lamer Source: Chelonian Conservation and Biology, 7(1) : 145-149 Published By: Chelonian Research Foundation and Turtle Conservancy URL: https://doi.org/10.2744/CCB-0669.1 BioOne Complete (complete.BioOne.org) is a full-text database of 200 subscribed and open-access titles in the biological, ecological, and environmental sciences published by nonprofit societies, associations, museums, institutions, and presses. Your use of this PDF, the BioOne Complete website, and all posted and associated content indicates your acceptance of BioOne’s Terms of Use, available at www.bioone.org/terms-of-use. Usage of BioOne Complete content is strictly limited to personal, educational, and non-commercial use. Commercial inquiries or rights and permissions requests should be directed to the individual publisher as copyright holder. BioOne sees sustainable scholarly publishing as an inherently collaborative enterprise connecting authors, nonprofit publishers, academic institutions, research libraries, and research funders in the common goal of maximizing access to critical research. Downloaded From: https://bioone.org/journals/Chelonian-Conservation-and-Biology on 08 Sep 2019 Terms of Use: https://bioone.org/terms-of-use Access provided by United States Fish & Wildlife Service National Conservation Training Center NOTES AND FIELD REPORTS Chelonian Conservation and Biology, 2008, 7(1): 88–95 Winokur (1968) could not determine the rate of hybrid- Ó 2008 Chelonian Research Foundation ization, concluded the identification of some specimens as hybrids to be uncertain, and verified pure A. atra. By 1979, The Status of Apalone atra Populations in Smith and Smith considered A. atra to be an extinct Cuatro Cie´negas, Coahuila, Me´xico: lineage due to hybridization with A. spinifera emoryi. In Preliminary Data 1983, a softshell resembling pure A. atra was noted in a field trip to the Cuatro Cie´negas basin (Ernst and Barbour 1 1992). SUZANNE E. MCGAUGH AND 1 In this study, we sought to illuminate the taxonomic FREDRIC J. JANZEN status of the endemic Cuatro Cie´negas black softshell, A. atra, by assessing its molecular distinction from the 1Department of Ecology, Evolution, and Organismal Biology, Iowa invasive congener, A. s. emoryi. We sampled all 5 State University, Ames, Iowa 50011 USA [[email protected]; drainages in the basin (Evans 2005) for turtles with A. [email protected]] atra morphological characteristics and used mitochondrial DNA sequence data from Cuatro Cie´negas softshell turtles ABSTRACT. – The species-level designation of the Mexican softshell turtle, Apalone atra,hasbeen and A. spinifera subspecies to reconstruct a haplotype repeatedly challenged, yet no DNA evidence has been network and a phylogenetic tree. In addition, we built collected. We conducted field studies of all the distance matrices with the mitochondrial DNA data and 3 drainages of the Cuatro Cie´negas basin, and the only nuclear loci. pure morphological population of A. atra found was in Methods. — Morphological surveys were extensive; Tı´o Candido, the type locality for the species. One we inspected all currently defined and flowing drainages in ´ nuclear intron known to show species-level divergence El Area de Proteccio´n de Flora y Fauna Cuatro Cie´negas, in the family Trionychidae, 2 nuclear genes, and a Coahuila, Me´xico, for A. atra (Fig. 1; Minckley 1969). mitochondrial gene revealed little molecular diver- This sampling included the type locality, Tı´o Candido, and gence for A. atra when compared with A. spinifera spanned 62 days of trapping from 16 May to 16 June 2003 emoryi from the Rio Grande. Further, no reciprocal and from 4 June to 5 July 2004 (Webb and Legler 1960). monophyly of the mitochondrial gene tree was seen Turtles were captured in lobster or hoop traps that were between A. atra and A. s. emoryi morphotypes. baited with sardines and checked every 12–14 hours. Each turtle was tattooed in a unique pattern on the plastron. Morphologically, A. atra is characterized by 5 main For many species, hybridization and high migration traits: 1) ‘‘blackish’’ pigmentation of dorsal surface, 2) rates are important components of an organism’s long- speckled pigmentation on ventral surfaces, 3) faint term evolutionary trajectory (Maddison 1997), but in a marginal bands and posterior white tubercles on the short time scale, these behaviors create difficulty in the carapace of males only, 4) longitudinal corrugations on the delimitation of these organisms as evolutionarily signifi- posterior of the carapace, and 5) ovoid adult carapaces cant units (ESU) for conservation protection (Moritz (Webb and Legler 1960; Winokur 1968). Apalone 1994). ESUs are defined in 2 ways: 1) populations that spinifera emoryi is defined by 1) tan to olive-brown have reciprocal monophyly at mitochondrial loci with carapace; 2) no ventral pigmentation; 3) defined pale significant divergence in nuclear loci (Moritz 1994) and 2) marginal band; 4) white, raised tubercles on the back third populations that are substantially reproductively isolated of the carapace; and 4) a clear triangular facial pattern and represent distinct evolutionary legacies (i.e., genetic (Ernst et al. 1994). In this study, A. atra were variability; Waples 1991, 1995). Here we investigate DNA distinguished from A. s. emoryi by dark pigmentation on evidence to determine if the turtle species Apalone atra, the carapace, speckled pigmentation on the plastron, and the endemic Cuatro Cie´negas black softshell, meets either corrugations on the posterior carapace. These traits of these criteria. showed no sexual dimorphism and little to no change The taxonomic and conservation standing of A. atra is through adult life and were exclusive to A. atra.See controversial. This turtle is currently listed on the CITES Appendix I for morphological characteristics and habitat (Convention on International Trade in Endangered Species types and Appendix II for photo voucher accession of Wild Flora and Fauna) Appendix I endangered species numbers for each specimen. list, but it has also been reported to be extinct (Smith and Less than 0.5 ml of blood were drawn from the caudal Smith 1979). Hybridization between A. atra and a south vein of the 26 field-collected animals, stored in buffer Texas species, Apalone spinifera emoryi, is thought to (0.01 M Tris, 10 mM EDTA, 0.01 M NaCl, and 1% SDS), have begun in the 1880s when irrigation canals were and frozen. DNA was extracted using Roche High Pure constructed, opening the Cuatro Cie´negas basin hydrolog- Template Preparation Kit (Cat. 1796828). DNA sequence ically (D. Hendrickson, pers. comm.; Webb 1973). data were obtained using an ABI 3730 DNA Analyzer. Downloaded From: https://bioone.org/journals/Chelonian-Conservation-and-Biology on 08 Sep 2019 Terms of Use: https://bioone.org/terms-of-use Access provided by United States Fish & Wildlife Service National Conservation Training Center NOTES AND FIELD REPORTS 89 Figure 1. Approximate localities sampled in the Cuatro Cie´negas basin. Map adapted from Moline et al. (2004). Populations include AM: Anteojo; AMG: Antiguos Mineros Grande; LG: Laguna Grande; LGa: Los Gatos; LI: Laguna Intermedia; LT: Posa de las Tortugas; TC: Tio Candido; RC: Rio Can˜o´n; RM: Rio Mesquites at Las Salinas; SM: San Marcos. Laguna Intermedia and Laguna Grande are connected by a short canal and considered 1 site. Rio Can˜o´n is located north of the town of Cuatro Cie´negas. Morphological but not molecular data were taken from drainage 4. This included 725 base pairs (bp) of cytochrome b Eight additional samples from a previous study were mitochondrial gene (CytbF: 50 ACAGGCGTAATCC- used for the total phylogenetic analysis (total n ¼ 34; TACTAC 30; DW1594; see Weisrock and Janzen 2000) Weisrock and Janzen 2000; LAcr1m, NMrg, ONtr1, from putative A. atra in the type locality (n ¼ 6), 8 other FLer1, GAsr, TXcc, TXki, and TXsc). Alignments were localities in the basin (n ¼ 19), and the pallid softshell (A. performed in CLUSTAL W (Thompson et al. 1994), and s. pallida; MidCon63). Additionally, 1638 bp of recom- sequences were visually reviewed and corrected in BioEdit bination activating gene-1 (RAG-1; see Krenz et al. 2005 7.0.0 (Hall 1999). Modeltest 3.7 (Posada and Crandall for primers), 886 bp of RNA fingerprint protein 35 nuclear 1998) was used to estimate parameters of sequence intron (R35; see Fujita et al. 2004 for primers), and 538 bp evolution for all genes using Akaike’s information of the oncogene C-mos (see Saint et al. 1998 for primers) criterion (Posada and Buckley 2004). No model of were used to construct distance matrices to compared the sequence evolution was proposed or tested in other studies type locality specimens (n ¼ 2) to A. s. pallida (CME63 of Apalone phylogeography (e.g., Weisrock and Janzen from Irion County, Texas), A. s. emoryi (TXsc from 2000), but the models for cytochrome b in this study were Valverde County, Texas, and NMrg from Socorro County, tested with representatives from major clades of Weisrock New Mexico; Weisrock and Janzen 2000), and A. mutica and Janzen (2000). These parameters were taken into (LAcrm1 from East Baton Rouge Parish, Baker, Louisi- account when constructing distance matrices in PHYLIP ana; Weisrock and Janzen 2000). The nuclear intron R35 3.62 (Felsenstein 2005). A maximum likelihood tree using has been successful in resolving species-level phylogenies cytochrome b was constructed using parameters estimated in the family Trionychidae (Engstrom et al. 2004), and in Modeltest 3.7 and bootstrapped for 100 replicates using RAG-1 has been successful in resolving species-level Paup 4.0b. Parsimony and neighbor-joining trees carried phylogenies in other Testudines (Krenz et al. 2005). The the same signature of weakly supported nodes and can be third nuclear marker, C-mos, has not been extensively used obtained from the first author. Nuclear DNA sequences for phylogenetics in turtles.