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Plant Meiosis pp 331-346 | Cite as

Induction and Characterization of Diploid Pollen Grains in Arabidopsis thaliana

  • Nico De Storme
  • Danny GeelenEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2061)

Abstract

Polyploidization or whole genome duplication (WGD) is one of the main forces driving plant genome evolution and biodiversity with major implications for plant breeding and crop improvement. In nature, de novo formation of polyploid plant genomes most likely occurs through a modification of the sexual reproductive pathway. By interfering with reproductive genome stability, for example, via induction of meiotic restitution, diploid or polyploid gametes are ectopically formed that may participate in fertilization to yield polyploid offspring. This mechanism of WGD is generally referred to as sexual polyploidization. Considering the central role of sexual polyploidization in speciation, genome evolution and crop breeding, we provide here a set of methodologies to induce and characterize 2n pollen grain formation in plants. Using Arabidopsis thaliana as a model, we outline two different methods, that is, one chemical and one environmental, to induce male meiotic restitution and high frequency 2n pollen grain formation. In addition, we provide a set of simple and straightforward techniques to characterize alterations in male meiotic cell division and gametophytic ploidy stability underpinning 2n pollen formation. This comprehensive toolbox is applicable in a broad range of plant species to enable quick induction and assessment of 2n gamete formation during plant male reproduction.

Key words

2n pollen Meiotic restitution Temperature GA and Arabidopsis thaliana 

Notes

Acknowledgments

We would like to thank C. Petit and P. Vandenbroucke for practical assistance with the growing and maintenance of Arabidopsis plants. Also many thanks to Prof. M. Höfte (Phytopathology Lab, UGent) for the use of the light microscopy platform.

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Copyright information

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

Authors and Affiliations

  1. 1.Horticell Lab, Department of Plants and Crops, Faculty of Bioscience EngineeringGhent University (UGent)GhentBelgium

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