Advertisement

Using Sequential Fluorescence and Genomic In Situ Hybridization (FISH and GISH) to Distinguish the A and C Genomes in Brassica napus

  • Elaine C. Howell
  • Susan Armstrong
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 990)

Abstract

We have developed a sequential procedure with fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH) that enables us to distinguish between the A and C genomes in Brassica napus and to identify certain individual chromosomes or chromosome groups within a genome. Our modified GISH technique uses a repetitive sequence in addition to the whole genome in the blocking DNA, and it is effective on meiotic and mitotic cells present in the anther material that we use.

Keywords

Brassica A genome C genome Genomic in situ hybridization GISH Fluorescence in situ hybridization FISH Meiosis BAC 

Notes

Acknowledgments

E.C. Howell was supported by BBSRC. We thank S. Price for technical assistance and I.A. Parkin (AAFC, Saskatoon, Canada) for B. napus seed.

References

  1. 1.
    Howell EC, Kearsey MJ, Jones GJ, King GJ, Armstrong SJ (2008) A and C genome distinction and chromosome identification in Brassica napus by sequential fluorescence in situ hybridization and genomic in situ hybridization. Genetics 180:1849–1857PubMedCrossRefGoogle Scholar
  2. 2.
    Leflon M, Eber F, Letanneur JC, Chelysheva L, Coriton O, Huteau V et al (2006) Pairing and recombination at meiosis of Brassica rapa (AA) x Brassica napus (AACC) hybrids. Theor Appl Genet 113:1467–1480PubMedCrossRefGoogle Scholar
  3. 3.
    Xiong ZY, Pires JC (2011) Karyotype and identification of all homoeologous chromosomes of allopolyploid Brassica napus and its diploid progenitors. Genetics 187:37–49PubMedCrossRefGoogle Scholar
  4. 4.
    Alix K, Joets J, Ryder CD, Moore J, Barker GC, Bailey JP et al (2008) The CACTA transposon Bot1 played a major role in Brassica genome divergence and gene proliferation. Plant J 56:1030–1044PubMedCrossRefGoogle Scholar
  5. 5.
    Gerlach WL, Bedrock JR (1979) Cloning and characterization of ribosomal RNA genes from wheat and barley. Nucleic Acids Res 7:1869–1885PubMedCrossRefGoogle Scholar
  6. 6.
    Zwick MS, Hanson RE, Islam-Faridi MN, Stelly DM, Wing RA, Price HJ et al (1997) A rapid procedure for the isolation of C0 t-1 DNA from plants. Genome 40:138–142PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Elaine C. Howell
    • 1
  • Susan Armstrong
    • 1
  1. 1.School of BiosciencesThe University of BirminghamBirminghamUK

Personalised recommendations