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Conservation Genetics Resources

, Volume 5, Issue 3, pp 887–890 | Cite as

Isolation, via 454 sequencing, characterization and transferability of microsatellites for Grevillea thelemanniana subsp. thelemanniana and cross-species amplification in the Grevillea thelemanniana complex (Proteaceae)

  • Tanya H. Hevroy
  • Michael L. Moody
  • Siegfried L. Krauss
  • Michael G. Gardner
Microsatellite Letters

Abstract

A set of 12 polymorphic microsatellite markers were developed for Grevillea thelemanniana subsp. thelemanniana, a rare and threatened shrub, endemic to southwest Western Australia. Genomic sequences were obtained from next generation (454) sequencing in the target species. Primer pairs for a total of 30 microsatellite loci were designed from these of which 13 were successfully amplified in 25 individuals from Kenwick Nature Reserve, Perth. Twelve loci were polymorphic with observed heterozygosity ranging from 0.12 to 0.6 and the number of alleles per locus ranged from 2 to 8. These markers were trialed in all species within the G. thelemanniana species complex. Amplification success and level of polymorphism varied among loci and taxa, but most (96 %) were successful. These loci will be useful in understanding the genetic variation, molecular ecology and phylogenetic relationships in the G. thelemanniana complex.

Keywords

Grevillea Microsatellites 454 sequencing Cross amplification 

References

  1. Doyle J, Doyle J (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15Google Scholar
  2. England PR, Ayre DJ, Whelan RJ (1999) Microsatellites in the Australian shrub Grevillea macleayana (Proteaceae). Mol Ecol 8:685–702CrossRefGoogle Scholar
  3. Gardner MG, Fitch AJ, Bertozzi T, Lowe AJ (2011) Rise of the machines-recommendations for ecologists when using next generation sequencing for microsatellite development. Mol Ecol Resour 11:1093–1101PubMedCrossRefGoogle Scholar
  4. Hoebee SE (2011) Development and cross-species amplification of microsatellites markers from the endangered Wee Jasper Grevillea (Grevillea iaspicula, Proteaceae). Muelleria 29(1):93–96Google Scholar
  5. James EA, Brown GK, Citroen R, Blacket MJ (2012) Microsatellite development for two species of holly-leafed Grevillea and cross-species amplification in the Asplendiifolia/Hookeriana subgroup (Proteaceae). Cons Gen Res 4:137–140CrossRefGoogle Scholar
  6. Makinson RO (2000) Grevillea. Flora of Australia 17A. Australian Biological Resources Study: Canberra/CSIRO, MelbourneGoogle Scholar
  7. Mantello C, Kestring DR, Sousa VA, Aguiar AV, Sousa AP (2011) Development and characterization of microsatellite loci in Grevillea robusta. BMC Proc 5:16CrossRefGoogle Scholar
  8. Meglécz E, Costedoat C, Dubut V, Gilles A, Malausa T, Pech N, Martin J-F (2010) QDD: a user-friendly program to select microsatellite markers and design primers from large sequencing projects. Bioinformatics 26:403–404PubMedCrossRefGoogle Scholar
  9. Meglécz E, Nève G, Biffin E, Gardner MG (2012) Breakdown of phylogenetic signal: a survey of microsatellite densities in 454 shotgun sequences from 154 non model eukaryote species. PLoS One 7(7):e40861PubMedCrossRefGoogle Scholar
  10. Oosterhout CV, Hutchinson WF, Wills DPM, Shipley P (2004) Micro-checker: software for identifying and correcting genotyping errors in microsatellite data. Mol Ecol Notes 4:535–538CrossRefGoogle Scholar
  11. Peakall R, Smouse PE (2006) GENALEX 6: genetic analysis in Excel. Population genetic software for teaching and research. Mol Ecol Notes 6:288–295CrossRefGoogle Scholar
  12. Raymond M, Rousset F (1995) GENEPOP (version 1.2): population genetics software for exact tests and ecumenicism. J Hered 86:248–249Google Scholar
  13. Rousset F (2008) Genepop ‘007: a complete re-implementation of the GENEPOP software for Windows and Linux. Mol Ecol Res 8:103–106CrossRefGoogle Scholar
  14. Rozen S, Skaletsky HJ (2000) Primer3 on the WWW for general users and for biologist programmers. In: Krawetz S, Misener S (eds) Bioinformatics methods and protocols: methods in molecularbiology. Humana Press, Totowa, pp 365–386Google Scholar
  15. Smith MG (2012) Threatened and Priority Flora List for Western Australia, 27 April 2012. Department of Environment and Conservation. Como, WAGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Tanya H. Hevroy
    • 1
  • Michael L. Moody
    • 2
  • Siegfried L. Krauss
    • 3
  • Michael G. Gardner
    • 4
    • 5
    • 6
  1. 1.School of Plant BiologyUniversity of Western AustraliaCrawleyAustralia
  2. 2.Department of Biological SciencesUniversity of Texas at El PasoEl PasoUSA
  3. 3.Kings Park and Botanic GardenBotanic Gardens and Parks AuthorityWest PerthAustralia
  4. 4.School of Biological SciencesFlinders UniversityAdelaideAustralia
  5. 5.Australian Centre for Evolutionary Biology and BiodiversityUniversity of AdelaideAdelaideAustralia
  6. 6.Evolutionary Biology Unit South Australian MuseumAdelaideAustralia

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