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Conservation Genetics

, Volume 11, Issue 3, pp 1177–1179 | Cite as

A molecular screening method for unisexual Ambystoma using mitochondrial DNA

  • Elizabeth A. RhoadsEmail author
  • Carissa M. Krane
  • P. Kelly Williams
Technical Note

Abstract

The geographical range of unisexual Ambystoma overlaps with four bisexual species that also breed in spring ponds. Several of these species are of conservation concern, and both adults and larvae can be difficult to distinguish morphologically from unisexuals. Here we present a rapid molecular method for screening unisexuals, whose mtDNA is most similar to Ambystoma barbouri. A 258 bp segment of the cytochrome b gene was amplified in six Ambystoma species and exemplar unisexuals by PCR using taxon-specific primers. An internal 113 bp segment was amplified only in unisexuals and A. barbouri using Universal forward and Hybrid reverse primers. Multisequence alignment comparing the nucleotide sequence where Hybrid reverse primer anneals revealed nucleotide diversity in this region among Ambystoma species. This simple method for discriminating between unisexuals and bisexuals, excluding A. barbouri, can be applied prior to further research on these declining species.

Keywords

Mole salamanders Larval identification Cytochrome b 

Abbreviations

LJJ

A. laterale, (2) jeffersonianum

LTJTi

A. laterale, texanum, jeffersonianum, tigrinum

Notes

Acknowledgments

The authors wish to thank Dr. J.P. Bogart for providing voucher DNA, and to thank both Dr. Bogart and Dr. Emily Latch for careful review of the manuscript.

References

  1. Bogart JP (2003) Genetics and systematics of hybrid species. In: Sever DM (ed) Reproductive biology and phylogeny of Urodela. Science Publishers, Enfield, NH, pp 109–134Google Scholar
  2. Bogart JP, Elinson RP, Licht LE (1989) Temperature and sperm incorporation in polyploidy salamanders. Science 246:1032–1034. doi: 10.1126/science.2587986 CrossRefPubMedGoogle Scholar
  3. Bogart JP, Bi K, Fu J, Noble DWA, Niedzwiecki J (2007) Unisexual salamanders (genus Ambystoma) present a new reproductive mode for eukaryotes. Genome 50:119–136. doi: 10.1139/G06-152 CrossRefPubMedGoogle Scholar
  4. Brandon RA (1961) A comparison of the larvae of five northeastern species of Ambystoma (Amphibia, Caudata). Copeia 4:377–383. doi: 10.2307/1439576 CrossRefGoogle Scholar
  5. DNA Analysis LLC 3921 Montgomery Rd. Cincinnati, OH 45212 www.agctsequencing.com
  6. Guggenbiller MG (1998) Distributional ecology of unisexual populations of the mole salamander Ambystoma jeffersonianum complex (Amphibia: Caudata) in western Ohio. University of Dayton, DaytonGoogle Scholar
  7. Hebert PDN, Penton EH, Burns JM, Janzen DH, Hallwachs W (2004) Ten species in one: DNA barcoding reveals cryptic species in the neotropical skipper butterfly Astraptes fulgerator. Proc Natl Acad Sci USA 41:14812–14817. doi: 10.1073/pnas.0406166101 CrossRefGoogle Scholar
  8. Hedges SB, Bogart JP, Maxson LR (1992) Ancestry of unisexual salamanders. Nature 356:708–710. doi: 10.1038/356708a0 CrossRefPubMedGoogle Scholar
  9. Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) ClustalW and ClustalX version 2. Bioinformatics 23(21):2947–2948. doi: 10.1093/bioinformatics/btm404 CrossRefPubMedGoogle Scholar
  10. NatureServe (2008) Natureserve explorer: an online encyclopedia of life [web application]. Version 7.0 NatureServe, Arlington, Virginia. Available from http://www.natureserve.org/explorer. Accessed 23 Oct 2008
  11. Noel S, Dumoulin J, Ouellet M, Galois P, Lapointe FJ (2008) Rapid identification of salamanders from the Jefferson complex with taxon-specific primers. Copeia 1:158–161. doi: 10.1643/CG-06-255 CrossRefGoogle Scholar
  12. Petranka JW (1998) Salamanders of the United States and Canada. Smithsonian Institution Press, Washington DCGoogle Scholar
  13. Robertson AV, Ramsden C, Niedzwiecki J, Fu J, Bogart JP (2006) An unexpected recent ancestor of unisexual Ambystoma. Mol Ecol 15:3339–3351. doi: 10.1111/j.1365-294X.2006.03005.x CrossRefPubMedGoogle Scholar
  14. Stephen FA, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402. doi: 10.1093/nar/25.17.3389 CrossRefGoogle Scholar
  15. Thomas M, Raharivololoniaina L, Glaw F, Vences M, Vieites DR (2005) Montane tadpoles in Madagascar: molecular identification and description of the larval stages of Mantidactylus elegans, Mantidactylus madecassus, and Boophis laurenti from the Andringitra Massif. Copeia 1:174–183CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Elizabeth A. Rhoads
    • 1
    Email author
  • Carissa M. Krane
    • 1
  • P. Kelly Williams
    • 1
  1. 1.University of DaytonDaytonUSA

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