Plant Meiosis pp 281-300 | Cite as

Quantification of Recombination Rate and Segregation Distortion by Genotyping and Sequencing of Single Pollen Nuclei

  • Steven Dreissig
  • Jörg Fuchs
  • Axel Himmelbach
  • Martin MascherEmail author
  • Andreas HoubenEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2061)


Meiosis is a specialized cell division during which homologous chromosomes can exchange genetic material through recombination. This mechanism generates novel allelic combinations, which can be exploited by plant breeders to achieve crop improvement. Pollen grains are the haploid products of meiosis required in fertilization. Here, we describe two approaches to measure meiotic recombination in single haploid pollen nuclei. Pollen nuclei are first separated by fluorescence-activated cell-sorting. Afterwards, the DNA of single pollen nuclei can be amplified by multiple-displacement-amplification using Phi29 DNA polymerase and meiotic recombination events can be measured using KASP markers. Alternatively, the PicoPLEX DNA-seq kit can be used to amplify the DNA of single pollen nuclei followed by library preparation for whole-genome sequencing and subsequent bioinformatic analysis.

Key words

Pollen Single cell Recombination Segregation distortion Flow sorting Whole-genome amplification 



We thankfully acknowledge Petr Cápal for his advice on single-cell whole-genome-amplification. Furthermore, we like to acknowledge Nils Stein (IPK, Gatersleben) for providing “Morex” × “Barke” F1 seeds and Sandra Dreisslein as well as Ines Walde (IPK, Gatersleben) for their excellent technical assistance. This work was supported by IPK Gatersleben.


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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Leibniz Institute of Plant Genetics and Crop Plant Research (IPK)Stadt SeelandGermany

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