Characterization of Meiotic Crossovers in Pollen from Arabidopsis thaliana

  • Jan DrouaudEmail author
  • Christine Mézard
Part of the Methods in Molecular Biology book series (MIMB, volume 745)


Homologous recombination processes, which occur during the prophase of the first meiotic division, while generating new allelic combinations, are mechanistically important for the regular segregation of homologous chromosomes. They generate either crossovers, which are reciprocal exchanges between chromosome segments, or gene conversions. Both kinds of events occur in narrow regions (less than 10 kb) called hotspots, which are distributed along chromosomes. Classical genetic methods for CO characterization, which rely on the building of large populations and require appropriately located markers, are not well suited to the study of meiotic recombination hotspots. Here, we present a method based on allele-specific PCR amplification of single molecules from pollen genomic DNA. It allows detection, quantification and characterization of CO events arising at low frequencies in recombination hotspots.

Key words

Meiosis crossover pollen DNA allele-specific PCR 



We are grateful to Wayne Crismani, Mathilde Grelon, Anouchka Guyon, Arnaud Ronceret and Nathalie Vrielynck for critical reading of the manuscript and helpful comments.

This work was supported by grants from INRA and ANR (COMEREC1 and COPATH).


  1. 1.
    Hunter, N. (2007) Meiotic recombination. In Molecular Genetics of Recombination. A. Aguilera, R. Rothstein, eds. (Berlin: Springer), pp. 381–442.CrossRefGoogle Scholar
  2. 2.
    Whitby, M.C. (2005) Making crossovers during meiosis. Biochem Soc Trans 33, 1451–1455.PubMedCrossRefGoogle Scholar
  3. 3.
    Mezard, C., Vignard, J., Drouaud, J., and Mercier, R. (2007) The road to crossovers: plants have their say. Trends Genet 23, 91–99.PubMedCrossRefGoogle Scholar
  4. 4.
    Anderson, L.K., and Stack, S.M. (2005) Recombination nodules in plants. Cytogenet Genome Res 109, 198–204.PubMedCrossRefGoogle Scholar
  5. 5.
    Gabriel, S.B., Schaffner, S.F., Nguyen, H., Moore, J.M., Roy, J., Blumenstiel, B., Higgins, J., DeFelice, M., Lochner, A., Faggart, M., Liu-Cordero, S.N., Rotimi, C., Adeyemo, A., Cooper, R., Ward, R., Lander, E.S., Daly, M.J., and Altshuler, D. (2002) The structure of haplotype blocks in the human genome. Science 296, 2225–2229.PubMedCrossRefGoogle Scholar
  6. 6.
    McVean, G.A., Myers, S.R., Hunt, S., Deloukas, P., Bentley, D.R., and Donnelly, P. (2004) The fine-scale structure of recombination rate variation in the human genome. Science 304, 581–584.PubMedCrossRefGoogle Scholar
  7. 7.
    HapMap (2005) A haplotype map of the human genome. Nature 437, 1299–1320.CrossRefGoogle Scholar
  8. 8.
    Myers, S., Bottolo, L., Freeman, C., McVean, G., and Donnelly, P. (2005) A fine-scale map of recombination rates and hotspots across the human genome. Science 310, 321–324.PubMedCrossRefGoogle Scholar
  9. 9.
    Petes, T.D. (2001) Meiotic recombination hot spots and cold spots. Nat Rev Genet 2, 360–369.PubMedCrossRefGoogle Scholar
  10. 10.
    Kauppi, L., Jeffreys, A.J., and Keeney, S. (2004) Where the crossovers are: recombination distributions in mammals. Nat Rev Genet 5, 413–424.PubMedCrossRefGoogle Scholar
  11. 11.
    Jeffreys, A.J., Murray, J., and Neumann, R. (1998) High-resolution mapping of crossovers in human sperm defines a minisatellite-associated recombination hotspot. Mol Cell 2, 267–273.PubMedCrossRefGoogle Scholar
  12. 12.
    Baudat, F., and de Massy, B. (2009) Parallel detection of crossovers and noncrossovers in mouse germ cells. Methods Mol Biol 557, 305–322.PubMedCrossRefGoogle Scholar
  13. 13.
    Kauppi, L., May, C.A., and Jeffreys, A.J. (2009) Analysis of meiotic recombination products from human sperm. Methods Mol Biol 557, 323–355.PubMedCrossRefGoogle Scholar
  14. 14.
    Bennett, M.D., Leitch, I.J., Price, H.J., and Johnston, J.S. (2003) Comparisons with Caenorhabditis (approximately 100 Mb) and Drosophila (approximately 175 Mb) using flow cytometry show genome size in Arabidopsis to be approximately 157 Mb and thus approximately 25% larger than the Arabidopsis genome initiative estimate of approximately 125 Mb. Ann Bot 91, 547–557.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Institut Jean-Pierre Bourgin, UMR1318 INRA-AgroParisTechVersailles CedexFrance
  2. 2.Institut National de Recherche, Agronomique, Centre de Versailles-Grignon Route de St-Cyr (RD10)Versailles CedexFrance
  3. 3.Institut Jean-Pierre BourginVersailles CedexFrance

Personalised recommendations