A set of species-specific repetitive DNA sequences was isolated from Lolium multiflorum and Festuca arundinacea. The degree of their species specificity as well as possible homologies among them were determined by dot-blot hybridization analysis. In order to understand the genomic organization of representative Lolium and Festuca-specific repetitive DNA sequences, we performed Southern blot hybridization and in situ hybridization to metaphase chromosomes.
Southern blot hybridization analysis of eight different repetitive DNA sequences of L. multiflorum and one of F. arundinacea indicated either tandem and clustered arrangements of partially dispersed localization in their respective genomes. Some of these sequences, e.g. LMB3, showed a similar genomic organization in F. arundinacea and F. pratensis, but a slightly different organization and degree of redundancy in L. multiflorum. Clones sequences varied in size between 100 bp and 1.2 kb. Estimated copy number in the corresponding haploid genomes varied between 300 and 2×104. Sequence analysis of the highly species-specific sequences from plasmids pLMH2 and pLMB4 (L. multiflorum specific) and from pFAH1 (F. arundinacea specific) revealed some internal repeats without higher order. No homologies between the sequences or to other repetitive sequences were observed. In situ hybridization with these latter sequences to metaphase chromosomes from L. multiflorum, F. arundinacea and from symmetric sexual Festulolium hybrid revealed their relatively even distribution in the corresponding genomes. The in situ hybridization thus also allowed a clearcut simple identification of parental chromosomes in the Festulolium hybrid.
The potential use of these species-specific clones as hybridization probes in quantitative dot-blot analysis of the genomic make-up of Festulolium (sexual and somatic) hybrids is also demonstrated.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Base pair (s)
nitroblue tetrazolium chloride
Ananiev EV, Bochkanov SS, Ryzhic MV, Sonina NV, Chernysehv AI, Schipkova NI (1986) Crosshybridization of clones: repeated sequences of barley genome with DNAs from wheat, rye and maize. Genetika 22:2021–2025
Appels R (1982) The molecular cytology of wheat-rye hybrids. Int Rev Cytol 80:93–132
Appels R, Dennis ES, Smyth DR, Peacock WJ (1981) Two repeated DNA sequences from the heterochromatic regions of rye (Secale cereale) chromosomes. Chromosoma 84:265–277
Bedbrook IR, Jones I, O'Dell M, Thompson RD, Flavell RB (1980) A molecular description of telomeric heterochromatin in Secale species. Cell 19:545–560
Bennett MD, Smith JB, Heslop-Harrison JS (1982) Nuclear DNA amounts in angiosperms. Proc R Soc London Ser B 216:179–199
Borrill M (1976) Temperate grasses. In: Simmonds NW (eds) Evolution of crop plants. Longman, London New York, pp 137–142
Bulinska-Radomska Z, Lester RN (1988) Intergeneric relationships of Lolium, Festuca and Vulpia (Poaceae) and their phylogeny. Plant Syst Evol 159:217–227
de Lautour G, Cooper BM (1971) Cold and chemical pretreatments to aid chromosome counts in a grass leaf squash technique incorporating hot pectinase maceration. Stain Technol 46:305–310
Dennis ES, Gerlach WL, Peacock WJ (1980) Identical polypyrimidine-polypurine satellite DNAs in wheat and barley. Heredity 44:349–366
Dretzen G (1981) A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal Biochem 112:295–298
Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–8
Flavell R (1980) The molecular characterization and organization of plant chromosomal DNA sequences. Annu Rev Plant Physiol 31:569–596
Franklin AL, Filion WG (1985) A new technique for retarding fading out fluorescence: DPX-BME. Stain Technol 60:125–135
Ganal MW, Lapitan NLV, Tanksley SD (1988) A molecular and cytogenetic survey of major repeated DNA sequences in tomato (Lycopersicon esculentum). Mol Gen Gent 213:262–268
Hutchinson J, Lonsdale DM (1982) The chromosomal distribution of cloned highly repetitive sequences from hexaploid wheat. Heredity 48:371–376
Hutchinson J, Rees H, Seal AG (1979) An assay of the activity of supplementary DNA in Lolium. Heredity 43:411–421
Imamura J, Saul MW, Potrykus I (1987) X-ray irradiation promoted asymmetric somatic hybridisation and molecular analysis of the products. Theor Appl Genet 74:445–450
Itoh K, Iwabuchi M, Shimamoto K (1991) In situ hybridization with species specific DNA probes gives evidence for asymmetric nature of Brassica hybrids obtained by X-ray fusion. Theor Appl Genet 81:356–362
Iwabuchi M, Itoh K, Shimamoto K (1991) Molecular and cytological characterization of repetitive DNA sequences in Brassica. Theor Appl Genet 81:349–355
Junghans H, Metzlaff M (1988) Genome specific, highly repeated sequences of Hordeum vulgare: cloning, sequencing and squash dot test. Theor Appl Genet 76:728–732
Kato A, Iida Y, Yakura K, Tanifuji S (1985) Sequence analysis of Vicia faba highly repeated DNA: the BamHI repeated sequence families. Plant Mol Biol 5:41–53
Leclerc RF, Siegel A (1987) Characterization of repetitive elements in several Cucurbita species. Plant Mol Biol 8:497–507
Lichtenstein C, Draper J (1985) Genetic engineering of plants. In: Glover DM (eds) DNA cloning, vol II, IRL Press, Oxford Washington, pp 67–119
Macfarlane TD (1986) Poaceae subfamily Pooideae. In: Soderstrom TR et al. (eds) An international symposium held at the Smithsonian Institution. Smithsonian Institution Press, Washington D.C. London, pp 265–276
Malik CP, Thomas PT (1966) Karyotypic studies in some Lolium and Festuca species. Caryologia 19:167–196
Metzlaff M, Tröbner W, Baldauf F, Schlegel R, Cullum J (1986) Wheat specific repetitive DNA-sequences-construction and characterization of four different genomic clones. Theor Appl Genet 72:207–210
Peacock WJ, Dennis ES, Rhoades MM, Pryor (1981) Highly repeated DNA sequences limit knob heterochromatin in maize. Proc Natl Acad Sci USA 78:4490–4494
Piastuch WC, Bates GW (1990) Chromosomal analysis of Nicotiana asymmetric somatic hybrids by dot blotting and in situ hybridization. Mol Gen Genet 222:97–103
Rayburn AL, Gill BS (1986) Molecular identification of the D-genome chromosomes of wheat. Heredity 77:253–255
Sakowicz T, Galazka G, Konarzewska A, Kwinkowski M, Klysik J (1986) An usually high number of direct repeats detected by sequence analysis of the dispersed EcoRI-family fragments in Lupinus luteus L. Planta 168:207–213
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning — a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
Sanger F, Coulson AR, Barrel BG, Smith AJH, Roe BA (1980) Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol 143:161–178
Saul MW, Potrykus I (1984) Species-specific repetitive DNA used to identify interspecific somatic hybrids. Plant Cell Rep 3:65–67
Sleper DA, Nelson CJ (1990) Breeding and genetics: potential use of haploids and doubled haploids. In: Kasperbauer MJ (eds) Biotechnology in fall fescue improvement, CRC Press, Boca Raton Ann Arbor Boston, pp 167–191
Smyth DR (1991) Dispersed repeats in plant genomes. Chromosoma 100:355–359
Smyth DR, Kalitsis P, Joseph JL, Sentry JW (1989) Plant retrotransposon from Lilium henryi is related to Ty3 of yeast and the gypsy group of Drosophila. Proc Natl Acad Sci USA 86:5015–5019
Takamizo T, Spangenberg G, Suginobu K, Potrykus I (1991) Intergeneric somatic hybridization in Gramineae: somatic hybrid plants between tall fescue (Festuca arundinacea Schreb.) and Italian ryegrass (Lolium multiflorum Lam.). Mol Gen Gent 231:1–6
Viotti A, Privitera AE, Pogna N, Sala E (1985) Distribution and clustering of two highly repetitive sequences in the A and B chromosomes of maize. Theor Appl Genet 70:234–240
Wu TY, Wu R (1987) A new rice repetitive DNA shows sequence homology to both 5S RNA and tRNA. Nucleic Acids Res 15:5913–5923
Zamir D, Tanskley SD (1988) Tomato genome is comprised largely of fast evolving low copy number sequences. Mol Gen Genet 213:254–261
Zhao X, Wu T, Xie Y, Wu R (1989) Genome-specific repetitive sequences in the genus Oryza. Theor Appl Genet 78:201–209
Communicated by G. Wenzel
About this article
Cite this article
Perez-Vicente, R., Petris, L., Osusky, M. et al. Molecular and cytogenetic characterization of repetitive DNA sequences from Lolium and Festuca: applications in the analysis of Festulolium hybrids. Theoret. Appl. Genetics 84, 145–154 (1992). https://doi.org/10.1007/BF00223994
- Lolium multiflorum
- Festuca arundinacea
- Repetitive DNA sequences
- In situ hybridization
- Festulolium hybrids