Advertisement

Development and characterization of 14 microsatellite loci from an enriched genomic library of Eucalyptus camaldulensis Dehnh

  • Janete Motta da Silva
  • Adna Cristina Barbosa de  Sousa
  • Anete Pereira de Souza
  • Edson Seizo Mori
  • Miguel Luiz Menezes Freitas
  • Alexandre Magno Sebbenn
  • Mario Luiz Teixeira de MoraesEmail author
Technical Note

Abstract

Eucalyptus camaldulensis Dehnh is an Australian tree species which occurs in various climatic and environmental conditions and show large genetic diversity. Twenty five microsatellite markers were developed from a CT8–GT8 enriched genomic library of E. camaldulensis. The number of alleles ranged from 4 to 13 (average of 8). The polymorphism information content (PIC) and the discriminating power (D) of each primer ranged from 0.37 to 0.88 (average of 0.72) and 0.48 to 0.99 (average of 0.84), respectively. The observed and expected heterozygosities ranged from 0.28 to 0.84 and 0.25 to 0.90, respectively. Four loci showed statistically significant from Hardy-Weinberg equilibrium after Bonferroni correction (P, (5%) < 0.0038). All polymorphic markers were tested for cross-amplification in 25 different Eucalyptus species. Those microsatellite loci will be useful to investigate questions of genetic diversity and structure, gene flow, mating system and ex situ genetic conservation of E. camaldulensis.

Keywords

Conservation genetic Hardwoods Molecular marker Microsatellites Cross-species amplification 

Notes

Acknowledgments

This work was financed by a grant from the São Paulo State Government to M.L.T. Moraes and J.M. Silva: Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) 05/58721-9 and 05/59217-2. The study is part of the graduate program of the first author. We thank Alonso A. Silva, José Cambuim, Manoel F.R. Bonfim and Flávio C. Arantes for sample collection; Ana P.S.C. Gaino, Cecília L. Dourado, Juliana P. Moreira, Laila T. Cardin, Patrícia F. Alves and Ricardo O. Manoel for DNA extractions; Selma M.B. Moraes for technical advices.

References

  1. Billotte N, Lagoda PJR, Risterucci AM, Baurens FC (1999) Microsatellite enriched libraries: applied methodology for the development of SSR markers in tropical crops. Fruits 54:277–288Google Scholar
  2. Brondani RPV, Brondani C, Tarchini R, Grattapaglia D (1998) Development, characterization and mapping of microsatellite markers in Eucalyptus grandis and E. urophylla. Theor Appl Genet 97:816–827CrossRefGoogle Scholar
  3. Butcher PA, Otero A, McDonald MW, Moran GF (2002) Nuclear RFLP variation in Eucalyptus camaldulensis Dehnh from northern Australia. Heredity 88:402–412CrossRefPubMedGoogle Scholar
  4. Butcher PA, McDonald MW, Bell JC (2009) Congruence between environmental parameters, morphology and genetic structure in Australia’s most widely distributed eucalypt, Eucalyptus camaldulensis. Tree Genet Genomes 5:189–210CrossRefGoogle Scholar
  5. Cordeiro GM, Taylor GO, Henry RJ (2000) Characterization of microsatellite markers from sugarcane (Saccharum sp.), a highly polyploidy species. Plant Sci 155:161–168CrossRefPubMedGoogle Scholar
  6. Creste S, Tulmann Neto A, Figueira A (2001) Detection of single sequence repeat polymorphisms in denaturing polyacrylamide sequencing gels by silver staining. Plant Mol Biol Rep 19:299–306CrossRefGoogle Scholar
  7. Dexter BD, Rose HJ, Davies N (1986) River regulation and associate forest management problems in the River Murray red gum forests. Aust For 49:16–27Google Scholar
  8. Don RH, Cox PT, Wainwright BJ, Baker K, Mattick JS (1991) “Touchdown” PCR to circumvent spurious priming during gene amplification. Nucleic Acids Res 19:4008CrossRefPubMedGoogle Scholar
  9. Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15Google Scholar
  10. Eldridge K, Davidson J, Harwood C, Wyk G (1993) Eucalypt domestication and breeding. Clarendon, Oxford, pp 60–71Google Scholar
  11. Goudet J (1995) Fstat (Version 2.9.3.2.): a computer program to calculate F-statistics. J Hered 86:485–486Google Scholar
  12. Kalinowski ST, Taper ML, Marshall TC (2007) Revising how computer program Cervus accommodates genotyping error increase success in paternity assignment. Mol Ecol 16:1099–1106CrossRefPubMedGoogle Scholar
  13. Lewis P, Zaykin D (2002) Genetic data analysis (GDA): computer program for the analysis of allelic data (software), version 1.1 (d12). http://alleyn.eeb.uconn.edu/gda/
  14. Nieto VM, Rodriguez J (2003) Eucalyptus camaldulensis Dehnh. In: Vozzo JA (ed) Tropical tree seed manual. Part II–species descriptions. United States Department of Agriculture, Forest Service, Washington, pp 466–467Google Scholar
  15. Pryor LD (1976) Biology of Eucalyptus. Esward Arnold, LondonGoogle Scholar
  16. Temnykh S, DeClerck G, Lukashova A, Lipovich L, Catinhour 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 11:1441–1452CrossRefGoogle Scholar
  17. Tessier C, David J, This P, Boursiquot JM, Charrier A (1999) Optimization of the choice of molecular markers for varietal identification in Vitis vinifera L. Theor Appl Genet 98:171–177CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Janete Motta da Silva
    • 1
  • Adna Cristina Barbosa de  Sousa
    • 2
  • Anete Pereira de Souza
    • 2
    • 3
  • Edson Seizo Mori
    • 4
  • Miguel Luiz Menezes Freitas
    • 5
  • Alexandre Magno Sebbenn
    • 5
  • Mario Luiz Teixeira de Moraes
    • 1
    • 6
    Email author
  1. 1.Faculdade de EngenhariaUniversidade Estadual Paulista (UNESP)Ilha SolteiraBrazil
  2. 2.Centro de Biologia Molecular e Engenharia GenéticaUniversidade Estadual de Campinas (Unicamp)CampinasBrazil
  3. 3.Departamento de Biologia VegetalUniversidade Estadual de Campinas (Unicamp)CampinasBrazil
  4. 4.Faculdade de Ciências AgronômicasUniversidade Estadual Paulista (UNESP)BotucatuBrazil
  5. 5.Instituto Florestal de São PauloSão PauloBrazil
  6. 6.Departamento de Fitotecnia, Tecnologia de Alimentos e Sócio-Economia, Faculdade de Engenharia de Ilha SolteiraUniversidade Estadual Paulista (UNESP)Ilha SolteiraBrazil

Personalised recommendations