Chromosome Research

, Volume 4, Issue 7, pp 491–499 | Cite as

Physical mapping of repetitive DNA sequences and 5S and 18S–26S rDNA in five wild species of the genusHordeum

  • A. de Bustos
  • A. Cuadrado
  • C. Soler
  • N. JouveEmail author


The genetic relationships between several wild species and subspecies of the genusHordeum were assessed using fluorescencein situ hybridization (FISH). Plant material included natural populations of wild barley growing in Spain of the annual species,H. marinum ssp.marinum (2n=14) andgussoneanum (2n=14), andH. murinum ssp.murinum (2n=28), andleporinum (2n=28) and the perennial speciesH. bulbosum (2n=14) andH. secalinum (2n=28), plus the South American perennial speciesH. chilense (2n=14). FISH was used to locate the chromosomal sites of two rDNA multigene families 5S and 18S–26S (pTa71 and pTa794) and three repetitive DNA sequences (pSc119.2, pAs1 and pHch950) isolated from different species and genera. The seven chromosomes of the diploid species were readily distinguished by their external morphology and hybridization patterns to pTa71, pTa794, pSc119.2 and pAs1. These DNA probes were also useful for the identification of homologous chromosomes and in differentiating these from unidentified chromosomes in the tetraploid taxa. The use of the probe pHch950 permitted intergenomic differentiation in tetraploids and supports the diphyletic origin ofH. murinum andH. secalinum. Thein situ experiments yielded the following conclusions: (1) differences between the subspeciesmarinum andgussoneanum; (2) close relationships between the subspeciesmurinum andLeporinum; and (3) major differences in physical mapping betweenH. bulbosum and the remaining taxa. The genomic and phylogenetic relationships between taxa, as inferred from the results, are discussed.

Key words

barley, fluorescencein situ hybridization Hordeum bulbosum Hordeum chilense Hordeum marinum Hordeum murinum Hordeum secalinum repetitive DNA 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Anamthwat-Jónsson K, Heslop-Harrison JS (1993) Isolation and characterization of genome-specific DNA sequences in Triticeae species.Mol Gen Genet 240: 151–158.Google Scholar
  2. Bedbrook JR, Jones J, O'Neill M, Thompson RD, Flavell RB (1980) A molecular description of telomeric heterochromatin inSecale species.Cell,19: 545–560.Google Scholar
  3. Bernardo A, Montero M, Cuadrado A, Jouve N (1992). Image analysis of C-banded chromosomes and pairing regionalization in wheat.Genome 35: 1062–1067.Google Scholar
  4. Bothmer R, Jacobsen N (1985) Origin, taxonomy and related species. In: Rasmusson, D, ed.Barley. Agronomy Monograph No. 26, ASA, pp 19–26.Google Scholar
  5. Bothmer R, Flink J, Landström T (1986) Meiosis inHordeum interspecific hybrids. I. Diploid combinations.Can J Genet Cytol 28: 525–535.Google Scholar
  6. Bothmer R, Flink J, Landström T (1987) Meiosis inHordeum interspecific hybrids. II. Triploid combinations.Evol Trend Plants 1: 41–50.Google Scholar
  7. Bothmer R, Flink J, Landström T (1988) Meiosis in interspecificHordeum hybrids. IV. Tetraploid (4x 4x) hybrids.Genome 30: 47–485.Google Scholar
  8. Bothmer R, Flink J, Jacobsen N, Jorgensen RB (1989) Variation and differentiation inHordeum marinum (Poaceae).Nord J Bot 9: 1–10.Google Scholar
  9. Bothmer R, Jacobsen N, Baden C, Jorgensen RB, Linde-Laursen I (1991). AnEcogeographical Study of the Genus Hordeum. Systematic and Ecogeographic Studies on Crop Genepools. IBPGR: Rome.Google Scholar
  10. Cabrera A, Friebe B, Jiang J, Gill BS (1995) Characterization ofHordeum chilense chromosomes by C-banding andin situ hybridization using highly repeated DNA probes.Genome 38: 435–442.Google Scholar
  11. Castilho A, Heslop-Harrison JS (1995) Physical mapping of 5S and 18S26S rDNA and repetitive DNA sequences inAegilops umbellulata.Genome 38: 91–96.Google Scholar
  12. Covas G (1949) Taxonomic observations on the North American species ofHordeum.Madroño 10: 1–21.Google Scholar
  13. Cuadrado A, Jouve N (1994) Mapping and organization of highly-repeated DNA sequences by means of simultaneous and sequential FISH and C-banding in 6x-Triticale.Chrom Res 2: 231–338.Google Scholar
  14. Cuadrado A, Jouve N (1995) Fluorescencein situ hybridization and C-banding analyses of highly repetitive DNA sequences in the heterochromatin of rye (Secale montanum Guss) and wheat incorporatingS. montanum chromosome segments.Genome 38: 795–802Google Scholar
  15. Cuadrado A, Ceoloni C, Jouve N (1995) Variation in highly repetitive DNA composition of heterochromatin in rye studied by FISH.Genome 38: 1061–1069.Google Scholar
  16. Doebley, J, Bothmer R von, Larson S (1992) Chloroplast DNA variation and the phylogeny ofHordeum (Poaceae).Am J Bot 79: 576–584.Google Scholar
  17. Fernández JA, Jouve N (1984) Giemsa C-banding of the chromosomes ofHordeum chilense and its amphiploid ×Triticum turgidum conv.durum.Z. Pflanzenzüchtg 93: 212–221.Google Scholar
  18. Ferrer E, Loarce Y, Hueros G (1995) Molecular characterization and chromosome location of repeated DNA sequences inHordeum species and in the amphiploid tritordeum (×Tritodeum Ascherson et Graebner).Genome 38: 850–857.Google Scholar
  19. Gerlach WL, Bedbrook JR (1979) Cloning and characterization of ribosomal RNA genes from wheat and barley.Nucleic Acid Res 7: 1869–1885.Google Scholar
  20. Gerlach WL, Dyer TA (1980) Sequence organization of the repeating units in the nucleus of wheat which contain 5S rRNA genes.Nucleic Acid Res 8: 4851–4865.Google Scholar
  21. Gupta PK, Fedak G, Molnar SJ, Wheatcroft R (1989) Distribution of aSecale cereale sequence among 25Hordeum species.Genome 32: 383–388.Google Scholar
  22. Heslop-Harrison JS, Schwarzacher T, Anamthwat-Jónsson K, Leitch AR, Shi M, Leitch IJ. (1991)In situ hybridization with automated chromosome denaturation.J Methods Cell Mol Biol 3: 109–116.Google Scholar
  23. Hueros G, Loarce Y, Ferrer E (1993) A structural and evolutionary analysis of a dispersed repetitive sequence.Plant Mol Genet 22: 635–643.Google Scholar
  24. Jaaska V, Jaaska V (1986) Isozyme variation in the barley genus Hordeum L. 1. Alcohol dehydrogenase and superoxide dismutase.Biochem Physiol Pflanzen 181: 301–320.Google Scholar
  25. Jacobsen N, Bothmer R (1992) Supraspecific groups in the genusHordeum.Hereditas 116: 21–24.Google Scholar
  26. Jones JDG, Flavell, RB (1982) The structure, amount and chromosomal location of defined repeated sequences in species of the genusSecale.Chromosoma 86: 613–641.Google Scholar
  27. Jorgensen RB (1986) Relationships in the barley genus (Hordeum): and electrophoretic examination of proteins.Hereditas 104: 273–291.Google Scholar
  28. Kasha KJ, Sadasivaiah RS (1971) Genome relationships betweenHordeum vulgare L. andH. bulbosum L.Chromosoma 35: 264–287.Google Scholar
  29. Lapitan NLV, Gill BS, Sears RG (1987) Genomic and phylogenetic relationships among rye and perennial speices in the Triticeae.Crop Sci 27: 682–687.Google Scholar
  30. Leitch IJ, Heslop-Harrison JS (1992) Physical mapping of the 18S-5.8S-26S rRNA genes in barley byin situ hybridization.Genome 35: 1013–1018.Google Scholar
  31. Leitch IJ, Heslop-Harrison JS (1993) Physical mapping of four sites of 5S rDNA sequences and one site of theamylase-2 gene in barley (Hordeum vulgare).Genome 36: 517–523.Google Scholar
  32. Leitch, IJ, Leitch AR, Heslop-Harrison JS (1991) Physical mapping of plant DNA sequences by simultaneousin situ hybridization of two differently labelled fluorescent probes.Genome 34: 329–333.Google Scholar
  33. Linde-Laursen I, Bothmer R, Jacobsen N (1986) Giemsa C-banded karyotypes ofHordeum secalinum, H. capense and their interspecific hybrids withH. vulgare.Hereditas 105: 179–185.Google Scholar
  34. Linde-Laursen I, Bothmer R, Jacobsen N (1989) Giemsa C-banded karyotypes ofHordeum marinum, andH. murinum.Genome 32: 629–639.Google Scholar
  35. Linde-Laursen I, Bothmer R, Jacobsen N (1990) Giemsa C-banded karyotypes of diploid and tetraploidHordeum bulbosum (Poaceae).Plant Syst Evol 172: 141–150Google Scholar
  36. Linde-Laursen I, Ibsen E, Bothmer R von, Giese H (1992a) Physical localization of active and inactive rRNA gene loci inHordeum marinum ssp.gussoneanum (4x) by in situ hybridization.Genome 35: 1032–1036.Google Scholar
  37. Linde-Laursen I, Bothmer R von, Jacobsen N (1992b). Relationships in the genusHordeum: Giemsa C-banded karyotypes.Hereditas 116: 111–116.Google Scholar
  38. McIntyre CL, Clarke BC, Apples R (1988) Amplification and dispersion of repeated DNA sequences in the Triticeae.Plant Syst Evol 160: 39–59.Google Scholar
  39. McIntyre CL, Pereira S, Moran LB, Appels R (1990) NewSecale cereale (rye) DNA derivatives for the detection of rye chromosome segments in wheat.Genome 33: 635–640.Google Scholar
  40. Molnar SJ, Gupta PK, Fedak G, Wheatcroft R (1989) Ribosomal DNA repeat unit polymorphism in 25 Hordeum species.Theor Appl Genet 78: 387–392.Google Scholar
  41. Molnar SJ, Wheatcroft R, Fedak G (1992) RFLP analysis ofHordeum species relationships.Hereditas 116: 87–91.Google Scholar
  42. Monte JV, McIntyre CL, Gustafson JP (1993) Analysis of phylogenetic relationships in the Triticeae tribe using RFLPs.Theor Appl Genet 86: 649–655.Google Scholar
  43. Morrison JW (1958) Hordeum murinum in Holland.Acta Bot Neerl 3: 654–664.Google Scholar
  44. Mukai Y, Endo TR, Gill BS (1990) Physical mapping of the 5S rRNA multigene family in common wheat.J Hered 81: 290–295.Google Scholar
  45. Mukai Y, Endo TR, Gill BS. (1991) Physical mapping of the 18S.26S rRNA multigene family in common wheat: identification of a new locus.Chromosoma 100: 71–78.Google Scholar
  46. Mukai Y, Nakahara Y, Yamamoto M (1993) Simultaneous discrimination of the three genomes in hexaploid wheat by multicolor fluorescencein situ hybridization using total genomic and highly repeated DNA probes.Genome 36: 489–494.Google Scholar
  47. Nevski SA (1941) Beiträge zur Kenntnis der wildwachsenden Gersten in Zusammenhang mit der Frage über den Ursprung vonHordeum vulgare 1. (Versuch einer Monographie der GattungHordeum) (in Russian).Acta Inst Botnum V L. Komarovii Acad Sci USSR Sec 1, Fasc5: 64–255.Google Scholar
  48. Orgaard M, Heslop-Harrison JS (1994) Relationships between species of Leymus, Psathyrostachys andHordeum (Poaceae, Tritidceae) inferred from Southern hybridization of genomic DNA and cloned DNA probes.Plant Syst Evol 189: 217–231.Google Scholar
  49. Pedersen C, Linde-Laursen I (1994) Chromosomal locations of four minor rDNA loci and a marker microsatellite sequence in barley.Chrom Res 2: 65–71.Google Scholar
  50. Rajhathy T, Morrison JW (1962) Cytogenetic studies in the genus Hordeum. VI. Themurinum complex.Can J Genet Cytol 4: 240–247.Google Scholar
  51. Rayburn AL, Gill BS (1985) Use of biotin-labelled probes to map specific DNA sequences on wheat chromosomes.J Hered 76: 78–81.Google Scholar
  52. Shcherban AB, Vershinin AV (1992) The stretched BamHI-fragment of barley genome containing richly repetitive DNA sequences.Genetika 28: 15–21Google Scholar
  53. Svitashev S, Bryngelsson T, Vershinin A, Pedersen C, Säll T, von Bothmer R (1994) Phylogenetic analysis of the genusHordeum using repetitive DNA sequences.Theor Appl Genet 89: 801–810.Google Scholar
  54. Symeonidis L, Moustakas M (1986) Biosystematic study of the Hordeum murinum group in Greece.Flora 178: 177–182.Google Scholar
  55. Tercero JA, Bernardo A, Jouve N (1991) Encoding genes for endosperm proteins inHordeum chilense.Theor Appl Genet 81: 127–132.Google Scholar
  56. Vershinin AV, Salina EA, Solovyov VV, Timofeeva LL (1990) Genomic organization, evolution, and structural peculiarities of highly repetitive DNA ofHordeum vulgare.Genome 33: 441–449Google Scholar
  57. Vershinin AV, Svitashev SK, Gummesson PO, Salomon B, von Bothmer R, Bryngelsson T (1994) Characterization of a family of tandemly repeated DNA sequences in Triticeae.Theor Appl Genet 89: 217–225Google Scholar
  58. Vosa CG (1976) Chromosome banding patterns in cultivated and wild barleys (Hordeum spp).Heredity 37: 395–403.Google Scholar
  59. Xu J, Procunier JD, Kasha KJ (1990) Species-specificin situ hybridization ofHordeum bulbosum chromosomes.Genome 33: 628–634.Google Scholar

Copyright information

© Rapid Science Publishers 1996

Authors and Affiliations

  • A. de Bustos
    • 1
  • A. Cuadrado
    • 2
  • C. Soler
    • 1
  • N. Jouve
    • 2
    Email author
  1. 1.Department of Plant BreedingCIT, INIAAlcalá de Henares (Madrid)Spain
  2. 2.Department of Cell Biology and GeneticsUniversity of Alcalá de HenaresAlcalá de Henares (Madrid)Spain

Personalised recommendations