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Characterization of Meiotic Crossovers in Pollen from Arabidopsis thaliana

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

Abstract

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 

Notes

Acknowledgements

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).

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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

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