Skip to main content
Log in

Efficiency of phenotypic and DNA markers for a genetic diversity study of alfalfa

  • Plant Genetics
  • Published:
Russian Journal of Genetics Aims and scope Submit manuscript

Abstract

Information on genetic diversity and germplasm characterization is essential for successful crop improvement. Diverse data sets (pedigree, morphological, biochemical, DNA based-markers) are employed in various aspects of plant analysis. The objective of this study was to determine the efficiency of phenotypic and RAPD markers in diversity assessment of ten alfalfa (Medicago spp.) accessions from Europe, North America and Australia. Field experiment was designed as a randomised complete block with three replications over two consecutive years (2004, 2005) at one location. Twelve morpho-agronomic traits were recorded on 50 plants per each accession. Genomic DNA’s from 16–20 randomly selected individual plants per accession were used for RAPD analysis. Six primers selected in this study generated a total of 93 polymorphic RAPD bands. The number of polymorphic bands detected per primer ranged from 11 to 20. Genetic distances (GD) among investigated accessions and two-dimensional principal coordinate analysis (2D PCoA) based on phenotypic and molecular data were obtained. The average GD between (0.283–0.416) and within (0.247–0.332) accessions based on RAPD data was higher than GD values obtained by morpho-agronomic traits (0.171–0.354 and 0.157–0.261, respectively). 2D PCoA based on GD from RAPD data grouped most of the studied individual plants to four clusters according to their geographical or taxonomy origin. 2D PCoA based only on morpho-agronomic data did not group plants congruently to their origin, probably due to a strong environmental influence on studied traits. Our results indicated that the RAPD markers were effective in assessing genetic diversity within and between studied alfalfa accessions. In addition, the obtained results suggested that the RAPD markers might be useful for grouping of germplasm with similar genetic background and for pre-screening of potential heterotic groups in our breeding programme.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Mohammadi, S.A. and Prasanna, B.M., Analysis of Genetic Diversity in Crop Plants-Salient Statistical Tools and Considerations, Crop Sci., 2003, vol. 43, pp. 1235–1248.

    Article  Google Scholar 

  2. Barcaccia, G., Albertini, E., Tavoletti, S., et al., AFLP Fingerprinting in Medicago spp.: Its Development and Application in Linkage Mapping, Plant Breed., 1999, vol. 118, pp. 335–340.

    Article  CAS  Google Scholar 

  3. Crea, F., Calderini, O., Nenz, E., et al., Chromosomal and Molecular Rearrangements in Somatic Hybrids between Tetraploid Medicago sativa and Diploid Medicago falcata, Theor. Appl. Genet., 1997, vol. 95, pp. 1112–1118.

    Article  CAS  Google Scholar 

  4. Riday, H. and Brummer, E.C., Forage Yield Heterosis in Alfalfa, Crop Sci., 2002, vol. 42, pp. 716–723.

    Article  Google Scholar 

  5. Christopher, M., Mace, E., Jordan, D., et al., Applications of Pedigree-Based Genome Mapping in Wheat and Barley Breeding Programs, Euphytica, 2007, vol. 154, pp. 307–316.

    Article  Google Scholar 

  6. Ceolin, A.C.G. and Goncalves-Vidigal, M.C., Vidigal Filho, P.S., et al., Genetic Divergence of the Common Bean (Phaseolus vulgaris L.) Group Carioca Using Morpho-Agronomic Traits by Multivariate Analysis, Hereditas, 2007, vol. 144, pp. 1–9.

    Article  Google Scholar 

  7. Prosperi, J.M., Jenczewski, E., Angevain, M., and Ronfort, J., Morphologic and Agronomic Diversity of Wild Genetic Resources of Medicago sativa L. Collected in Spain, Genet. Resour. Crop Evol., 2006, pp. 1–14.

  8. Mosjidis, J.A., Greene, S.L., Klingler, K.A., and Afonin, A., Isozyme Diversity in Wild Red Clover Populations from the Caucasus, Crop Sci., 2004, vol. 44, pp. 665–670.

    Article  CAS  Google Scholar 

  9. Maras, M., Sunik, S., Vozli, J., and Megli, V., Temporal Changes in Genetic Diversity of Common Bean (Phaseolus vulgaris L.) Accessions Cultivated between 1800 and 2000, Russ. J. Genet., 2006, vol. 42, pp. 775–782.

    Article  CAS  Google Scholar 

  10. Savo Sardaro, M.L., Atallah, M., Tavakol, E., et al., Diversity for AFLP and SSR in Natural Populations of Lotus corniculatus L. from Italy, Crop Sci., 2008, vol. 48, pp. 1080–1089.

    Article  Google Scholar 

  11. Fufa, H., Baenziger, P.S., Beecher, B.S., et al., Comparison of Phenotypic and Molecular Marker-Based Classifications of Hard Red Winter Wheat Cultivars, Euphytica, 2005, vol. 145, pp. 133–146.

    Article  CAS  Google Scholar 

  12. Taran, B., Zhang, C., Warkentin, T., et al., Genetic Diversity among Varieties and Wild Species Accessions of Pea (Pisum sativum L.) Based on Molecular Markers, and Morphological and Physiological Characters, Genome, 2005, vol. 48, pp. 257–272.

    CAS  Google Scholar 

  13. Geleta, N., Labuschagne, M.L., and Viljoen, C.D., Genetic Diversity Analysis in Sorghum Germplasm as Estimated by AFLP, SSR and Morpho-Agronomical Markers, Biodivers. Conserv., 2006, vol. 15, pp. 3251–3265.

    Article  Google Scholar 

  14. Liu, Z.P., Liu, G.S., and Yang, Q.C., A Novel Statistical Method for Assessing SSR Variation in Autotetraploid Alfalfa (Medicago sativa), Genet. Mol. Biol., 2007, vol. 30, pp. 385–391.

    Google Scholar 

  15. Flajoulot, S., Ronfort, J., Baudouin, P., et al., Genetic Diversity among Alfalfa (Medicago sativa) Cultivars Coming from a Breeding Program, Using SSR Markers, Theor. Appl. Genet., 2005, vol. 111, pp. 1420–1429.

    Article  CAS  PubMed  Google Scholar 

  16. Gherardi, M., Mangin, B., Goffinet, B., et al., A Method to Measure Genetic Distance between Allogamous Populations of Alfalfa (Medicago sativa) Using RAPD Molecular Markers, Theor. Appl. Genet., 1998, vol. 96, pp. 406–412.

    Article  CAS  Google Scholar 

  17. Segovia-Lerma, A., Cantrell, R.G., Conway, J.M., and Ray, I.M., AFLP-Based Assessment of Genetic Diversity among Nine Alfalfa Germplasms Using Bulk DNA Templates, Genome, 2003, vol. 46, pp. 51–58.

    Article  CAS  PubMed  Google Scholar 

  18. SAS Institute, SAS/STAT Users Guide Version 9.1, Cary: SAS Inst., 2003.

  19. Roldan-Ruiz, I., Eeuwijk, F.A., Gilliland, T.J., et al., A Comparative Study of Molecular and Morphological Methods of Describing Relationships between Perennial Ryegrass (Lolium perenne L.) Varieties, Theor. Appl. Genet., 2001, vol. 103, pp. 1138–1150.

    Article  CAS  Google Scholar 

  20. Rohlf, F.J., NTSYS-pc: Numerical Taxonomy and Multivariate Analysis System, Version 2.2.b, New York: Applied Biostatistics, 2005.

    Google Scholar 

  21. Yu, K. and Pauls, K.P., Rapid Estimation of Genetic Relatedness among Heterogeneous Populations of Alfalfa by Random Amplification of Bulked Genomic DNA Samples, Theor. Appl. Genet., 1993, vol. 86, pp. 788–794.

    CAS  Google Scholar 

  22. Mengoni, A., Gori, A., and Bazzicalupo, M., Use of RAPD and Microsatellite (SSR) Variation to Assess Genetic Relationships among Populations of Tetraploid Alfalfa, Medicago sativa, Plant Breed., 2000, vol. 119, pp. 311–317.

    Article  CAS  Google Scholar 

  23. Horn, P. and Rafalski, A., Non-Destructive RAPD Genetic Diagnostics of Microspore-Derived Brassica Embryos, Plant Mol. Biol. Reporter, 1992, vol. 10, pp. 285–293.

    Article  CAS  Google Scholar 

  24. Mantel, N., The Detection of Disease Clustering and a Generalized Regression Approach, Cancer Res., 1967, vol. 27, pp. 209–220.

    CAS  PubMed  Google Scholar 

  25. IBPGR/84/191, International Board for Plant Genetic Resources: Forage Legume Descriptors, Rome: Commission of European Communities, Committee on Disease Resistance Breeding and Use of Gene Banks, 1984.

  26. Johns, M.A., Skroch, P.W., Nienhuis, J., et al., Gene Pool Classification of Common Bean Landraces from Chile Based on RAPD and Morphological Data, Crop Sci., 1997, vol. 37, pp. 605–613.

    Article  Google Scholar 

  27. Steiner, J.J. and Ios Santos, G.G., Adaptive, Ecology of Lotus corniculatus Genotypes: I. Plant Morphology and RAPD Marker Characterizations, Crop Sci., 2001, vol. 41, pp. 552–563.

    Article  Google Scholar 

  28. Musial, J.M., Basford, K.E., and Irwin, J.A.G., Analysis of Genetic Diversity within Australian Lucerne Cultivars and Implications for Future Genetic Improvement, Aust. J. Agric. Res., 2002, vol. 53, pp. 629–636.

    Article  Google Scholar 

  29. Maureira, I.J., Ortega, F., Campos, H., and Osborn, T.C., Population Structure and Combining Ability of Diverse Medicago sativa Germplasms, Theor. Appl. Genet., 2004, vol. 109, pp. 775–782.

    Article  CAS  PubMed  Google Scholar 

  30. Mari, S., Bolari, S., Martini, J., et al., Genetic Diversity of Hexaploid Wheat Cultivars Estimated by RAPD Markers, Morphological Traits and Coefficients of Parentage, Plant Breed., 2004, vol. 123, pp. 366–369.

    Article  Google Scholar 

  31. Greene, S.L., Gritsenko, M., and Vandemark, G., Relating Morphologic and RAPD Marker Variation to Collection Site Environment in Wild Populations of Red Clover (Trifolium pratense L.), Genet. Resour. Crop Evol., 2004, vol. 51, pp. 643–653.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to M. Tucak.

Additional information

The article is published in the original.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tucak, M., Popović, S., Čupić, T. et al. Efficiency of phenotypic and DNA markers for a genetic diversity study of alfalfa. Russ J Genet 46, 1314–1319 (2010). https://doi.org/10.1134/S1022795410110062

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S1022795410110062

Keywords

Navigation