New Forests

, Volume 31, Issue 3, pp 361–372 | Cite as

Use of Genetic Markers in the Management of Micropropagated Eucalyptus Germplasm

Article

Abstract

Clonal propagation through rooted cuttings and micropropagation is widely used for large-scale Eucalyptus plantation program because of its ability to fix the desirable traits of mature plus trees. However, when a large number of clones are handled, variations and mixings are commonly confronted which may go undetected in the absence of prominent morphological descriptors. Material from germplasm resources are also used in breeding program. The economic implications of such inadvertent variations could be serious as considerable time and money is spent before the mistakes are detected. This paper reports the identification of mislabeling in Eucalyptus clones maintained through tissue culture, and the reestablishment of the identity of the mislabeled clones using genetic markers viz., RAPDs/AFLPs. The in vitro propagated Eucalyptus plants from two groups, group 1 derived directly from SMD7 (a candidate plus tree), and group 2 derived from coppice shoots of trees of group 1, were assessed for their genetic uniformity using RAPD markers. The small intra-group genetic variations of 0.02 in the second group were attributed to somaclonal variations induced during long culture periods. However, the genetic distances of 0.20 and 0.31 between SMD7 and the two micropropagated groups were too high to be attributed to somaclonal variations as axillary bud culture was used for micropropagation. To test the possibility of inadvertent mixing, RAPD profiles of the micropropagated groups were compared with that of other clones in the tissue-cultured Eucalyptus germplasm. The RAPD profiles of group 2 plantlets matched with that of another unrelated clone in the germplasm. The authenticity of this donor was further re-established using AFLP markers.

Keywords

AFLPs clones DNA fingerprinting genetic fidelity RAPDs 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Belaj, A., Satovic, Z., Cipriani, G., Baldoni, L., Testolin, R., Rallo, L., Trujillo, I. 2003Comparative study of the discriminating capacity of RAPD, AFLP and SSR markers and of their effectiveness in establishing genetic relationships in oliveTheor. Appl. Genet.107736744CrossRefPubMedGoogle Scholar
  2. Brondani, R.P.V., Brondani, C., Tarchini, R., Grattapaglia, D. 1998Developmentcharacterization and mapping of microsatellite markers in Eucalyptus grandisE. urophyllaTheor. Appl. Genet.97816827CrossRefGoogle Scholar
  3. Caetano-Anollés and Gresshoff P.M.1994. Staining Nucleic Acids with Silver: An Alternative to Radioisotopic and Fluorescent Labeling. Promega Notes. No. 45. 13 p.Google Scholar
  4. Dice, L.R. 1945Measures of the amount of ecological association between speciesEcology26297302CrossRefGoogle Scholar
  5. Keil, M., Griffin, A.R. 1994Use of random amplified polymorphic DNA (RAPD) markers in the discrimination and verification of genotypes in EucalyptusTheor. Appl. Genet.89442450CrossRefGoogle Scholar
  6. Kretzschmar, U., Ewald, D. 1994Vegetative propagation of 140-year-old Larix decidua trees by different in vitro techniquesPlant Physiol.144627630Google Scholar
  7. Larkin, P.J., Scowcroft, W.R. 1981Somaclonal variation – a novel source of variability from cell cultures for plant improvementTheor. Appl. Genet.60197214CrossRefGoogle Scholar
  8. Mathish, N.V., Tripathi, S.B., Gurumurthi, K. 2001DNA-fingerprint database management using microsoft access – a simple strategy to corroborate fingerprints of clonesPlant Cell Biotechnol. Mol. Biol.2119124Google Scholar
  9. Meins, F.J. 1983Heritable variation in plant cell cultureAnnu. Rev. Plant Physiol.34327346CrossRefGoogle Scholar
  10. Muller, E., Brown, P.T.H., Hartke, S., Lorz, H. 1990DNA variation in tissue-culture-derived rice plantsTheor. Appl. Genet.80673679Google Scholar
  11. Murashige, T., Skoog, F. 1962A revised medium for rapid growth and bioassays with tobacco tissue culturesPhysiol. Plant.15473497CrossRefGoogle Scholar
  12. Murray, M.G., Thompson, W.F. 1980Rapid isolation of high molecular weight plant DNANucleic Acids Res.843214325PubMedGoogle Scholar
  13. Preetha N., Yasodha R., Madhavi S., Sumathi R., Stanley J. and Gurumurthi K. 1993. In: Vivekanandan K., Subramanian K.N., Zabala N.Q. and Gurumurthi K. (eds), Proceedings of Workshop on Production of Genetically Improved Planting Material for Afforestation Programmes. 18–25 June, Coimbatore. Food and Agriculture Organization of United Nations, Los Banos, Philippines pp. 128–149.Google Scholar
  14. Rani, V., Parida, A., Raina, S.N. 1995Random amplified polymorphic DNA (RAPD) markers for genetic analysis in micropropagated plants of Populus deltoides MarshPlant Cell Rep.14459462CrossRefGoogle Scholar
  15. Rohlf, F.J. 1997Numerical Taxonomy and Multivariate Analysis SystemNTSYSpc Version 2.0 Exter SoftwareSetauket, NYGoogle Scholar
  16. Sumathi, R., Malliga, P., Venketaraman, K.S., Jagadees, S.S., Yasodha, R. 1999In vitro regeneration of plants from mature clonal stock of Eucalyptus tereticornis SmSilva Plus.81617Google Scholar
  17. van der Nest, M.A., Steenkamp, E.T., Wingfield, B.D., Wingfield, M.J. 2000Development of simple sequence repeat (SSR) markers in Eucalyptus from amplified inter-simple sequence repeats (ISSR)Plant Breed.119433436CrossRefGoogle Scholar
  18. Vos, P., Hogers, R., Bleeker, M., Reijans, M., van de Lee, T., Hornes, M., Frijters, A., Pot, J., Peleman, J., Kuiper, , Zebeau, M. 1995AFLP: a new technique for DNA fingerprintingNucleic Acids Res.2344074414PubMedGoogle Scholar
  19. White, P.R. 1934Potentially unlimited growth of excised tomato root tips in liquid mediumPlant Physiol.9585600PubMedCrossRefGoogle Scholar
  20. Williams, J.G.K., Kubelik, A.R., Livak, K. J., Rafalski, J.A., Tingey, S.V. 1990DNA polymorphisms amplified by arbitrary primers are useful genetic markersNucleic Acids Res.1865316535PubMedGoogle Scholar

Copyright information

© Springer 2006

Authors and Affiliations

  • S. B. Tripathi
    • 1
    • 2
  • N. V. Mathish
    • 1
  • K. Gurumurthi
    • 1
  1. 1.Plant Biotechnology DivisionInstitute of Forest Genetics and Tree BreedingCoimbatoreIndia
  2. 2.The Energy and Resources Institute (TERI)New DelhiIndia

Personalised recommendations