, Volume 122, Issue 1, pp 191–201 | Cite as

AFLP analysis of genetic diversity within and between populations of perennial ryegrass (Lolium perenne L.)

  • K.M. Guthridge
  • M.P. Dupal
  • R. Kölliker
  • E.S. Jones
  • K.F. Smith
  • J.W. Forster


Amplified fragment length polymorphism (AFLP) analysis has been used to measure genetic diversity in perennial ryegrass (Lolium perenne L.) and to relate intra- and interpopulation variation to breeding history. Cluster analysis of AFLP data from contrasting populations showed features consistent with the origins of these varieties. Significant differences in intrapopulation diversity were detected and partial separation of different cultivars was observed. Restricted base cultivars, derived from small numbers of foundation clones, were suitable for this type of study, allowing near complete discrimination of closely related cultivars. Analysis of bulked samples was based on the pooling of genomic DNA from 20 individuals from 6 selected populations. Cluster analysis of AFLP data from bulked samples produced a phenogram showing relationships consistent with the results of individual analysis. AFLP profiling provides an important tool for the detection and quantification of genetic variation in perennial ryegrass.

AFLP phenogram genetic diversity molecular marker Lolium perenne L. 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Anonymous, 1988. Perennial ryegrass (Lolium perenne) variety ‘Yatsyn 1’. Plant Var J 1: 5-7.Google Scholar
  2. Armstrong, C.S., 1977. 'Grasslands Nui' perennial ryegrass. NZ J Exp Agr 5: 381-384.Google Scholar
  3. Barker, J.H.A., M. Matthes, G.M. Arnold, K.J. Edwards, I. Ahman, S. Larsson & A. Karp, 1999. Characterisation of genetic diversity in potential biomass willows (Salix spp.) by RAPD and AFLP analyses. Genome 42: 173-183.PubMedCrossRefGoogle Scholar
  4. Barker, R.E. & S.E. Warnke, 1998. Characterization of grass cultivars by use of DNA markers. In: H. Nakagawa (Ed.), Proc. Int. Workshop on Utilization of Transgenic Plants and Genome Analysis in Forage Crops. National Grassland Research Institute, pp. 123-134, Nishinasuno, Japan.Google Scholar
  5. Barrett, B.A. & K.K. Kidwell, 1998. AFLP-based genetic diversity assessment among wheat cultivars from the Pacific Northwest. Crop Sci 38: 1261-1271.CrossRefGoogle Scholar
  6. Blumenthal, M.J., K. Prakash, A. Leonforte, P.J. Cunningham & H.I. Nicol, 1996. Characterisation of the Kangaroo Valley ecotype of perennial ryegrass. Aust J Agric Res 47: 1131-1142.CrossRefGoogle Scholar
  7. Cameron, N., 1994a. Perennial ryegrass (Lolium perenne) variety ‘Embassy’. Plant Var J 7: 10-11.Google Scholar
  8. Cameron, N., 1994b. Perennial ryegrass (Lolium perenne) variety ‘Banks’. Plant Var J 7: 14-15.Google Scholar
  9. Camlin, M.S., 1997. Plant breeding -- achievements and prospects, Grasses. In: J.R. Weddell (Ed.), Seeds of Progress. BGS Occasional Symposia 31: 2-14.Google Scholar
  10. Cornish, M.A., M.D. Hayward & M.J. Lawrence, 1979. Self-incompatibility in ryegrass. I. Genetic control in diploid Lolium perenne L. Heredity 43: 95-106.Google Scholar
  11. Divaret, I., E. Margale & G. Thomas, 1999. RAPD markers on seed bulks efficiently assess the genetic diversity of a Brassica oleracea L. collection. Theor Appl Genet 98: 1029-1035.CrossRefGoogle Scholar
  12. Easton, H.S., 1983. Ryegrasses. In: G.S. Wratt & C. Smith II (Eds.), Plant breeding in New Zealand, pp. 229-236. Butterworths, Wellington NZ.Google Scholar
  13. Excoffier, L., P.E. Smouse & J.M. Quattro, 1992. Analysis of molecular variance inferred from metric distances among DNA haplotypes: application to human mitochondrial restriction data. Genetics 131: 479-491.PubMedGoogle Scholar
  14. Fulton, T.M., J. Chunwongse & S.D. Tanksley, 1995. Microprep protocol for extraction of DNA from tomato and other herbaceous plants. Plant Molec Biol Rep 13: 207-209.Google Scholar
  15. Forster, J.W., E.S. Jones, R. Kölliker, M.C. Drayton, M.P. Dupal, K.M. Guthridge, K.F. Smith, 2001. DNA profiling in outbreeding forage species. In: R. Henry (Ed.), Plant Genotyping -- The DNA Fingerprinting of Plants. CABI Press, in press.Google Scholar
  16. Gilbert, J.E., R.V. Lewis, M.J. Wilkinson & P.D.S. Caligari, 1999. Developing an appropriate strategy to assess genetic variability in plant germplasm collections. Theor Appl Genet 98: 1125-1131.CrossRefGoogle Scholar
  17. Hartl, L. & S. Seefelder, 1998. Diversity of selected hop cultivars detected by fluorescent AFLPs. Theor Appl Genet 96: 112-116.CrossRefGoogle Scholar
  18. Hayward, M.D. & I.B. Abdullah, 1985. Selection and stability of synthetic varieties of Lolium perenne, I. The selected character and its expression over generations of multiplication. Theor Appl Genet 70: 48-51.CrossRefGoogle Scholar
  19. Hayward, M.D. & N.J. McAdam, 1977. Isozyme polymorphism as a measure of distinctiveness and stability in cultivars of Lolium perenne. Z Planzenzüchtg 79: 59-68.Google Scholar
  20. Huff, D.R., 1997. RAPD characterization of heterogeneous perennial ryegrass cultivars. Crop Sci 37: 557-564.CrossRefGoogle Scholar
  21. Jones, C.J., K.J. Edwards, S. Castiglione, M.O. Winfield, F. Sala, C. Vandewiel, G. Bredemeijer, B. Vosman, M. Matthes, A. Daly, R. Brettschneider, P. Bettini, M. Buiatti, E. Maestri, A. Malcevschi, N. Marmiroli, R. Aert, G. Volckaert, J. Rueda, R. Lincaero, A. Vasquez & A. Karp, 1997. Reproducibility testing of RAPD, AFLP and SSR markers in plants by a network of European laboratories. Mol Breed 3: 381-390.CrossRefGoogle Scholar
  22. Kemp, D.R., 1988. The effects of flowering and leaf area on sward growth in winter of temperate pasture grasses. Aust J Agric Res 39: 597-604.CrossRefGoogle Scholar
  23. Kölliker, R., F.J. Stadelmann, B. Reidy & J. Nösberger, 1999. Genetic variability of forage grass cultivars: a comparison of Festuca pratensis Huds., Lolium perenne L. and Dactylis glomerata L. Euphytica 106: 261-270.CrossRefGoogle Scholar
  24. Kölliker, R., E.S. Jones, M.Z.Z. Jahufer & J.W. Forster, 2001. Bulked AFLP analysis for the assessment of genetic diversity in white clover (Trifolium repens L.). Euphytica, in press.Google Scholar
  25. Kongkiatngam, P., M.J. Waterway, M.G. Fortin & B.E. Coulman, 1996. Genetic variation among cultivars of red clover (Trifolium pratense L.) detected by RAPD markers amplified from bulk genomic DNA. Euphytica 89: 355-361.Google Scholar
  26. Kraft, T. & T. Säll, 1999. An evaluation of the use of pooled samples in studies of genetic variation. Heredity 82: 488-494.PubMedCrossRefGoogle Scholar
  27. Liu, Z.-W., R.L. Jarret, R.R. Duncan & S. Kresovich, 1994. Genetic relationships and variation among ecotypes of seashore paspalum (Paspalum vaginatum Swartz.) determined by random amplified polymorphic DNA markers. Genome 37: 1011-1017.PubMedGoogle Scholar
  28. Marshall, D.R. & A.H. D. Brown, 1975. Optimum sampling strategies in genetic conservation. In: O.H. Frankel & J.G. Hawkes (Eds.), Crop Genetic Resources for Today and Tomorrow, pp. 53-80. Cambridge University Press, Cambridge.Google Scholar
  29. Maughan, P.J., M.A. Saghai-Maroof, G.R. Buss & G.M. Huestis, 1996. Amplified fragment length polymorphisms (AFLP) in soybean: species diversity, inheritance and near isogenic lines analysis. Theor Appl Genet 93: 392-401.CrossRefGoogle Scholar
  30. Michelmore, R.W., I. Paran & R.V. Kesseli, 1991. Identification of markers linked to disease-resistance genes by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations. Proc Natl Acad Sci USA 88: 9828-9832.PubMedCrossRefGoogle Scholar
  31. Morell, M.K., R. Peakall, R. Appels, L.R. Preston & H.L. Lloyd, 1995. DNA profiling techniques for plant variety identification. Aus J Exp Agr 35: 807-819.CrossRefGoogle Scholar
  32. Nei, M. & W.H. Li, 1979. Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc Natl Acad Sci USA 76: 5269-5273.PubMedCrossRefGoogle Scholar
  33. Paul, S., F.N. Wachira, W. Powell & R. Waugh, 1997. Diversity and genetic differentiation among populations of Indian and Kenyan tea (Camellia sinensis [L.] O. Kuntze) revealed by AFLP markers. Theor Appl Genet 94: 255-263.CrossRefGoogle Scholar
  34. Perera, L., J.R. Russell, J. Provan, J.W. McNicol & W. Powell. 1998. Evaluating genetic relationships between indigenous coconut (Cocos nucifera L.) accessions from Sri Lanka by means of AFLP profiling. Theor Appl Genet 96: 545-550.CrossRefGoogle Scholar
  35. Preston, L.R., N. Harker, T. Holton & M.K. Morell, 1999. Plant cultivar identification using DNA analysis. Plant Var Seeds 12: 191-205.Google Scholar
  36. Rohlf, F.J., 1993. NTSYS-pc: Numerical taxonomy and multivariate analysis system. Exeter Publishers, Setauket, NY.Google Scholar
  37. Roldán-Ruiz, I., J. Dendauw, E. Van Bockstaele, A. Depicker & M. De Loose, 2000. AFLP markers reveal high polymorphic rates in ryegrasses (Lolium spp.). Mol Breed 6: 125-134.CrossRefGoogle Scholar
  38. Sanders, P.M. & D.J. Barker, 1988. Pasture plant identification using enzyme electrophoresis. Proc NZ Grassl Assoc 49: 77-80.Google Scholar
  39. Sanders, P.M., D.J. Barker & G.S. Wewala, 1989. Phosphoglucoisomerase-2 allozymes for distinguishing perennial ryegrass cultivars in binary mixtures. J Ag Sci Camb 112: 179-184.CrossRefGoogle Scholar
  40. Sato, M., M. Yamashita, S. Kato & T. Mikami & Y. Shimamoto, 1995. Mitochondrial DNA analysis reveals cytoplasmic variation within a single cultivar of perennial ryegrass (Lolium perenne L.). Euphytica 83: 205-208.CrossRefGoogle Scholar
  41. Sneath, P.H.A. & R.R. Sokal, 1973. Numerical Taxonomy. Freeman, San Francisco, USA. pp. 573.Google Scholar
  42. Strang, J., 1961. Kangaroo Valley perennial ryegrass. Ag Gazette NSW 72: 75-133.Google Scholar
  43. Sweeney, P.M. & T.K. Danneberger, 1994. Random amplified polymorphic DNA in perennial ryegrass: a comparison of bulk samples vs. individuals. HortScience 29: 624-626.Google Scholar
  44. Sweeney, P.M. & T.K. Danneberger, 1997. RAPD markers from perennial ryegrass DNA extracted from seeds. HortScience 32: 1212-1215.Google Scholar
  45. Vogel, K.P. & J.F. Pedersen, 1993. Breeding systems for cross-pollinated forage grasses. Plant Breed Revs 11: 251-274.Google Scholar
  46. Vos, P., R. Hogers, M. Bleeker, M. Reijans, T. Vandelee, M. Hornes, A. Frijters, J. Pot, J. Peleman, M. Kuiper & M. Zabeau, 1995. AFLP-a new technique for DNA fingerprinting. Nucl Acids Res 23: 4407-4414.PubMedGoogle Scholar
  47. Warpeha, K.M.F., I. Capesius & T.J. Gilliland, 1998. Genetic diversity in perennial ryegrass (Lolium perenne) evaluated by hybridisation with ribosomal DNA: implications for cultivar identification and breeding. J Ag Sci Camb 131: 23-30.CrossRefGoogle Scholar
  48. Wilkins, P.W., 1991. Breeding perennial ryegrass for agriculture. Euphytica 52: 201-214.CrossRefGoogle Scholar
  49. Williams, J.G.K., A.R. Kubelik, K.J. Livak, J.A. Rafalski & S.V. Tingey, 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucl Acids Res 18: 6531-6535.PubMedGoogle Scholar
  50. Yu, K. & K.P. Pauls, 1993. Rapid estimation of genetic relatedness among heterogeneous populations of alfalfa by random amplification of bulked genomic DNA samples. Theor Appl Genet 86: 788-794.Google Scholar
  51. Zhu, L., M.D. Gale, S. Quarrie, M.T. Jackson & G.J. Bryan, 1998. AFLP markers for the study of rice biodiversity. Theor Appl Genet 96: 602-611.CrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • K.M. Guthridge
    • 1
    • 2
  • M.P. Dupal
    • 1
    • 2
  • R. Kölliker
    • 1
  • E.S. Jones
    • 1
    • 2
  • K.F. Smith
    • 3
    • 2
  • J.W. Forster
    • 1
    • 2
  1. 1.Plant Biotechnology Centre, Agriculture VictoriaLa Trobe UniversityBundooraAustralia
  2. 2.CRC for Molecular Plant BreedingAustralia
  3. 3.Pastoral and Veterinary InstituteAgriculture Victoria HamiltonHamiltonAustralia

Personalised recommendations