Isolation and Characterization of 14 Polymorphic Microsatellite Loci in the Big-Headed Turtle (Platysternon megacephalum)


The big-headed turtle (Platysternon megacephalum) is critically endangered because of overharvesting, illegal trade, and habitat destruction. Assessment of genetic variability in existing populations becomes very important to the taxonomy and conservation of this species. Here we describe 14 microsatellite loci isolated from an enriched genomic library of the big-headed turtle, and the polymorphisms of these loci were assessed in 28 individuals from Huizhou, Heyuan, Zhaoqing, and Shaoguan of Guangdong, China. The range of polymorphism information content is 0.305–0.738, and no evidence of significant linkage disequilibrium was found among any pairs of loci. These 14 new polymorphic microsatellite loci can be used in population genetics, taxonomy, phylogeography, behavior ecology, and conservation efforts of Platysternon megacephalum.

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  1. Armour JAL, Neumann R, Gobert S, Jeffreys AJ (1994) Isolation of human simple repeat loci by hybridization selection. Hum Mol Genet 3:599–605

    CAS  PubMed  Article  Google Scholar 

  2. Ernst CH, Laemmerzahl AF (2002) Geographic variation in the Asian big-headed turtle, Platysternon megacephalum (Reptilia: Testudines: Platysternidae). Proc Biol Soc Wash 115:18–24

    Google Scholar 

  3. Fritz U, Havaš P (2007) Checklist of chelonians of the world. Vertebr Zool 57:149–368

    Google Scholar 

  4. Gong S, Shi H, Wang J, Fu Y, Xu R (2007) Report on the freshwater turtle survey on Hainan island. Living For 13:40–43

    Google Scholar 

  5. Gong S, Chow AT, Fong JJ, Shi H (2009) The chelonian trade in the largest pet market in China: scale, scope and impact on turtle conservation. Oryx 43:213–216

    Article  Google Scholar 

  6. Kandpal RP, Kandpal G, Weissman SM (1994) Construction of libraries enriched for sequence repeats and jumping clones, and hybridization selection for region-specific markers. Proc Natl Acad Sci USA 91:88–92

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  7. Kirkpatrick DT (1997) The family Platysternidae and its sole member: Platysternon megacephalum, the big-headed turtle. In: Ackerman L (ed) Biology, husbandry and health care of reptiles, vol 2. TFH Publication, Neptune, pp 438–446

    Google Scholar 

  8. Liu K, Muse SV (2005) PowerMarker: an integrated analysis environment for genetic marker analysis. Bioinformatics 21:2128–2129

    CAS  PubMed  Article  Google Scholar 

  9. Parham J, Feldman C, Boore J (2006) The complete mitochondrial genome of the enigmatic bigheaded turtle (Platysternon): description of unusual genomic features and the reconciliation of phylogenetic hypotheses based on mitochondrial and nuclear DNA. BMC Evol Biol 6:1–11

    Article  Google Scholar 

  10. Rice WR (1989) Analyzing tables of statistical tests. Evolution 43:223–225

    Article  Google Scholar 

  11. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  12. Shi YY, He L (2005) SHEsis, a powerful software platform for analyses of linkage disequilibrium, haplotype construction, and genetic association at polymorphism loci. Cell Res 15:97–98

    CAS  PubMed  Article  Google Scholar 

  13. Temnykh S, DeClerck G, Lukashova A, Lipovich L, Cartinhour S, McCouch S (2001) Computational and experimental analysis of microsatellites in rice (Oryza sativa L.): frequency, length variation, transposon associations, and genetic marker potential. Genome Res 11:1441–1452

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  14. Wang HJ, Luo TJ, Zhong XJ et al (2010) Survey of population status of Platysternon megacephalum in Guangdong. Chin J Wildl 31:110–112

    Google Scholar 

  15. Yeh F, Yang R, Boyle T, Ye Z, Xiyan J (2000) PopGene32, Microsoft Windows-based freeware for population genetic analysis. Version 1.32. Molecular Biology and Biotechnology Centre, University of Alberta, Edmonton, Alberta, Canada

  16. Zhang CL, Wang Y, Chen H, Lan XY, Lei CZ (2007) Enhance the efficiency of single-strand conformation polymorphism analysis by short polyacrylamide gel and modified silver staining. Anal Biochem 365:286–287

    CAS  PubMed  Article  Google Scholar 

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We thank Mr. Haijing Wang, Mr. Dongsheng Pan, Mr. Xiangjing Zhong, Mr. Taojun, and Mr. Chenyu for their help during the collection of tissue samples of turtles. This study was supported financially by the Zhujiang Scientific and Technological New Star Program (2011J2200025), the National Natural Science Foundation of China (No. 31071935), the Scientific and Technological Innovation and Guiding Program of the Guangdong Provincial Academy of Science (No. cx2011), the Guangdong Natural Science Foundation of China (No. S2011010000514), and the Scientific and Technological Program of Huizhou, Guangdong, China (No. 2008P09).

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Correspondence to Shiping Gong.

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L. Hua and F. Wang contributed equally to this work.

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Hua, L., Wang, F., Gong, S. et al. Isolation and Characterization of 14 Polymorphic Microsatellite Loci in the Big-Headed Turtle (Platysternon megacephalum). Biochem Genet 52, 203–208 (2014).

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  • Big-headed turtle
  • Platysternon megacephalum
  • Microsatellite loci
  • Isolation
  • Conservation