, Volume 128, Issue 2, pp 145–151 | Cite as

Intraspecific phenotypic variation associated with nuclear DNA content in Lolium perenne L.

  • Shuichi Sugiyama
  • Kenichi Yamaguchi
  • Toshihiko Yamada


The variation in nuclear DNA content and its association with phenotypic traits were examined in 15 cultivated populations of perennial ryegrass (Lolium perenne L.), including herbage and turf populations. Nuclear DNA contents of 15 populations were measured by a flow cytometric method using DAPI (4′-6-diamidino-2-phenylindole) as a fluorochrome. DNA contents were also measured using PI (propidium iodide) for six populations that showed large differences in DAPI values to confirm the difference. There were significant differences in nuclear DNA contents among the populations for both dyes. Of the total variation in nuclear DNA content measured with DAPI, 29% was ascribed to the inter-population variation, 46% was ascribed to the intra-population variation, 24% was ascribed to the random error variation. Herbage populations tended to have larger DNA content than did turf populations. 2C DNA content was positively correlated with cell size, seed size and single leaf size, although significant correlations were mainly due to a small number of populations with large and small DNA contents. These results suggest that intraspecific variation in nuclear DNA content plays important roles in determining phenotypic differences between cultivated populations of L. perenne.

flow cytometry nuclear DNA content intraspecificvariation Lolium perenne L. nucleotype effect 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Arabidopsis Genome Initiative, 2000. Analysis of the genome sequence of the flowering plant Arabidopsis thaliana. Nature 48: 796–813.CrossRefGoogle Scholar
  2. Bennett, M.D., 1972. Nuclear DNA content and minimum generation time in herbaceous plants. Proc R Soc Lond, B 181: 109–135.CrossRefGoogle Scholar
  3. Bennett, M.D., 1976. DNA amount, latitude, and crop plant distribution. Env Exp Bot 16: 93–108.CrossRefGoogle Scholar
  4. Bennett, M.D. & J.B. Smith, 1976. Nuclear DNA amounts in angiosperm. Philos T Roy Soc B 274: 227–274.Google Scholar
  5. Bennett, M.D. & I.J. Leitch, 1995. Nuclear DNA amounts in angiosperms. Ann Bot 76: 113–176.CrossRefGoogle Scholar
  6. Bennett, M.D., P. Bhandol & I.J. Leitch, 2000. Nuclear DNA amounts in Angiosperms and their modern uses - 807 new estimates. Ann Bot 86: 859–909.CrossRefGoogle Scholar
  7. Campbell, B.D., J.R. Caradus & C.L. Hunt, 1999. Temperature responses and nuclear DNA amounts of seven white clover populations which differ in early spring growth rates. New Zeal J Agric Res 42: 9–17.Google Scholar
  8. Cavalier-Smit, T., 1985. Cell volume and the evolution of eukaryoto genome size. In: T. Cavalier-Smith (Ed.), The Evolution of Genome Size, pp. 105–184. John Wiley & Sons, London.Google Scholar
  9. Creber, H.M.C., M.S. Davies, D. Francis & H.D. Walker, 1994. Variation in DNA C value in natural populations of Dactylis glomerata L. New Phytol 128: 555–561.CrossRefGoogle Scholar
  10. Chung, J., J.H. Lee, K. Arumuganathan, G.L. Graef & J.E. Specht, 1998. Relationship between nuclear DNA content and seed and leaf size in soybean. Theor Appl Genet 96: 1064–1068.CrossRefGoogle Scholar
  11. Dolezel, J., J. Greilhuber, S. Lucretti, M.A. Lyska & L. Nardi, 1998. Plant genome size estimation by flow cytometry: inter-laboratory comparison. Ann Bot 82 (suppl.): 17–26.Google Scholar
  12. Doolittle, W.F. & C. Spienza, 1980. Selfish genes, the phenotype paradigm and genome evolution. Nature 284: 601–603.PubMedCrossRefGoogle Scholar
  13. Greilhuber, J., 1988. 'self-tanning' - A new and important source of stoichiometric error in cytophotometric determination of nuclear DNA content in plants. Plant Syst Evol 158: 87–96.CrossRefGoogle Scholar
  14. Greilhuber, J., 1998. Intraspecific variation in genome size: a critical reassessment. Ann Bot 82 (Suppl.): 27–35.Google Scholar
  15. Grime, J.P. & M.A. Mowforth, 1982. Variation in genome size-an ecological interpretation. Nature 299: 151–153.CrossRefGoogle Scholar
  16. Johnston, J.S., D. Bennett, A.L. Rayburn, D.W. Galbraith & H.J. Price, 1999. Reference standards for determination of DNA content of plant nuclei. Am J Bot 86: 609–613.PubMedCrossRefGoogle Scholar
  17. Marshall, H.G., 1966. A technique of freezing plant crowns to determine the cold resistance of winter oats. Crop Sci 6: 173–176.CrossRefGoogle Scholar
  18. McMurphy, L.M. & A.L. Rayburn, 1992. Chromosomal and cell size analysis of cold tolerant maize. Theor Appl Genet 84: 798–802.CrossRefGoogle Scholar
  19. Minelli, S., P. Moscariello, M. Ceccarelli & P.G. Cionini, 1996. Nucleotype and phenotype in Vicia faba. Heredity 76: 524–530.Google Scholar
  20. Mowforth, M.A. & J.P. Grime, 1989. Intra-population variation in nuclear DNA amount, cell size and growth rate in Poa annua L. Funct Ecol 3: 289–295.CrossRefGoogle Scholar
  21. Ohr, D., 1998. Genome size variation and plant systematics. Ann Bot 82 (Suppl.): 75–83.Google Scholar
  22. Orgel, L.E. & F.H.C. Crick, 1980. Selfish DNA: the ultimate parasite. Nature 284: 604–607.PubMedCrossRefGoogle Scholar
  23. Price, H.J., K. Bachman, K.L. Chambers & J. Riggs, 1980. Detection of intraspecific variation in nuclear DNA content in Microseris douglastii. Bot Gaz 12: 156–159.Google Scholar
  24. Price, H.J., 1988. Nuclear DNA content variation within angiosperm species. Evol Trend Plant 2: 53–60.Google Scholar
  25. Rayburn, A.L., J.A. Auger, A. Benzinger & A.G. Hepburn 1989. Detection of intraspecific DNA content variation in Zea mays L. by flow cytometry. J Exp Bot 40: 1179–1183.Google Scholar
  26. Sokal, R.R. & F.J. Rohlf, 1981. Biometry, 2nd edn. Freeman, New York.Google Scholar
  27. Stebbins, G.L., 1971 Chromosomal Evolution in Higher Plants. Edward Arnold, New York.Google Scholar
  28. Sugiyama, S., 1999. Differentiation in competitive ability and cold tolerance between diploid and tetraploid cultivars in Lolium perenne. Euphytica 103: 55–59.CrossRefGoogle Scholar
  29. Yamashita, M. & Y. Shimamoto, 1996. Differentiation in freezing hardiness among cultivar groups of perennial ryegrass (Lolium perenne). Grassland Sci 42: 57–62 (in Japanese with English summary).Google Scholar

Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Shuichi Sugiyama
    • 1
  • Kenichi Yamaguchi
    • 1
  • Toshihiko Yamada
    • 2
  1. 1.Faculty of Agriculture and Life ScienceHirosaki UniversityHirosakiJapan
  2. 2.Yamanashi Prefectual Dairy Experimental StationYamanashiJapan

Personalised recommendations