High resolution FISH to delineate contiguous and small DNA sequences

  • U. C. Lavania

Abstract

Somatic and meiotic metaphase, and pachytene chromosomes were subjected to DNA: DNA in situ hybridization to elucidate relative resolution of FISH signals for weak/contiguous hybridization sites. Hybridization with a ‘350 family’ rye repetitive DNA probe pSc 200 characteristically differentiated the rye chromosome 5 from the rest of the complement on account of two small terminal homologous sites in the long arm, resolution of which is substantially improved using pachytene. Higher resolution of the two weak hybridization sites; a very small distal and a small proximal, is unequivocally demonstrated in the FISH painted 5RL examined at pachytene in the 5AS/5RL wheat background. Additionally this probe exhibits a large block of distal telomeric hybridization site in 5RS, followed by a more prominent proximal site homologous to ‘610 family’ rye repetitive probe pSc 250. Precise denaturation — hybridization incubation and post hybridization stringency washing facilitates spatial resolution of contiguous repetitive rye probes pSc 200 and pSc 250, and physical localisation of small RFLP probe xpr 115 of wheat on barley chromosomes.

Key words

Chromosome painting High resolution FISH Pachytene FISH Rye chromosome 5 Rye repetitive DNA probe 

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References

  1. 01.
    Appels R, Dennis ES, Smith DR, Peacock WJ (1981). Two repeated DNA sequences from heteromorphic regions of rye (Secale cereale) chromosomes. Chromosoma 84: 265–277.CrossRefGoogle Scholar
  2. 02.
    Bedbrook JR, Jones J, O’Neil M, Thompson RD, Flavell RB (1980). Molecular characterisation of telomeric heterochromatin in Secale species. Cell 19: 545–560.PubMedCrossRefGoogle Scholar
  3. 03.
    Brandis A, Thompson H, Dean C, Heslop-Harrison JS (1997). Multiple repetitive DNA sequences in the paracentromeric regions of Arabidopsis thaliana L. Chromosome Res 5: 238–246.CrossRefGoogle Scholar
  4. 04.
    Buckle VJ, Kearney L (1994). New methods in cytogenetics. Curr Opinion Genet Development 4: 374–382.CrossRefGoogle Scholar
  5. 05.
    de Jong JH, Fransz P, Zabel P (1999). High resolution FISH in plants — techniques and applications. Trends Plant Sci 4: 258–263.CrossRefGoogle Scholar
  6. 06.
    Fransz PF, Stam M, Montijn B, ten Hoopen R, Wiegant J, Koorter JM, Oud O, Nanninga N (1996). Detection of single copy genes and chromosome rearrangement in Petunia hybrida by fluorescence in situ hybridization. Plant J 9: 767–774.CrossRefGoogle Scholar
  7. 07.
    Heslop-Harrison JS (2000). Comparative genome organization in plants: from sequence and markers to chromatin and chromosomes. Plant Cell 12: 617–635.PubMedGoogle Scholar
  8. 08.
    Jiang J, Gill BS (1994). Nonisotopic in situ hybridization and plant genome mapping: the first 10 years. Genome 37: 717–725.PubMedCrossRefGoogle Scholar
  9. 09.
    Jiang J, Gill BS, Wang G, Ronald PC, Ward DC (1995). Metaphase and interphase fluorescence in situ hybridization mapping of the rice genome with bacterial artificial chromosomes. Proc Natl Acad Sci USA 92: 4487–4491.PubMedCrossRefGoogle Scholar
  10. 10.
    Lapitan NLV, Brown SE, Kennard W, Stephens JL, Knudson DL (1997). FISH physical mapping with barley BAC clones. Plant J 11: 149–156.CrossRefGoogle Scholar
  11. 11.
    Lavania UC (1998). Fluorescence in situ hybridization in genome, chromosome and gene identification in plants. Curr Sci 74: 126–133.Google Scholar
  12. 12.
    Lavania UC (1999). Large-scale chromosome organization in plants underpin genome homogenization, characteristic distribution of repetitive DNAs and occurrence of genes in discrete clusters. Curr Sci 77: 216–218.Google Scholar
  13. 13.
    Moscone EA, Matzke MA, Matzke AJM (1996). The use of combined FISH/GISH in conjunction with DAPI counterstaining to identify chromosomes containing transgene inserts in amphidiploid tobaco. Chromosoma 105: 231–236.CrossRefGoogle Scholar
  14. 14.
    Ohimido N, Aklyama Y, Fukui K (1998). Physical mapping of unique nucleotide sequences on identified rice chromosomes. Plant Mol Biol 38: 1043–1052.CrossRefGoogle Scholar
  15. 15.
    Schmidt T, Heslop-Harrison JS (1998). Genomes, genes and junk — the large-scale organization of plant chromosomes. Trends Plant Sci 3: 195–199.CrossRefGoogle Scholar
  16. 16.
    Schwarzacher T, Heslop-Harrison JS (2000). Practical in situ hybridization. Oxford: BIOS Scientific Publishers.Google Scholar
  17. 17.
    Sybenga J (1983). Rye chromosome nomenclature and homoeology relationships. Z Pflanzenzuchtg 90: 297–304.Google Scholar
  18. 18.
    ten Hoopen R, Robbins TP, Fransz PF, Montijn BM, Oud O, Gerats AGM, Nanninga N (1996). Localization of T-DNA insertions in Petunia by fluorescence in situ hybridization: physical evidence for suppression of recombination. Plant Cell 8: 823–830.PubMedGoogle Scholar
  19. 19.
    Vershinin AV, Schwarzacher T, Heslop-Harrison JS (1995). The large scale genome organization of repetitive DNA families at the telomeres of rye chromosomes. Plant Cell 7: 1823–1833.PubMedGoogle Scholar
  20. 20.
    Zhong X-B, Bodean J, Fransz PF, Williamson VM, van Kammen A, de Jong JH, Zabel P (1999). FISH to meiotic pachytene chromosomes of tomato indicates the root-knot nematode resistance gene Mi-1 and the acid phosphatase gene Aps-1 near the junction of euchromatin and pericentromeric heterochromatin of chromosome arms 6S and 6L, respectively. Theor Appl Genet 98: 365–370.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2001

Authors and Affiliations

  • U. C. Lavania
    • 1
  1. 1.Cytogenetics DivisionCentral Institute of Medicinal and Aromatic PlantsLucknowIndia

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