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Isolation of Arabidopsis Pollen, Sperm Cells, and Vegetative Nuclei by Fluorescence-Activated Cell Sorting (FACS)

  • Mário R. Santos
  • Cláudia Bispo
  • Jörg D. BeckerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1669)

Abstract

Efficient methods to isolate highly purified Arabidopsis thaliana pollen and the subcellular components of the male gametophyte (the vegetative nucleus and two sperm cells) have enabled genome-scale studies revealing a highly dynamic reprogramming of the transcriptome and epigenome during pollen development. Here, we describe the isolation of uninucleate microspores, mature pollen, as well as sperm cells and vegetative nuclei by Fluorescence-Activated Cell Sorting.

Key words

Pollen Microspore Sperm cell Vegetative nucleus Arabidopsis FACS Transcriptomics Epigenetics 

Notes

Acknowledgement

We thank Marta Monteiro, Mariana Fernandes (Flow Cytometry Unit), and Vera Nunes (Plant Facility) for technical support. Leonor Boavida is acknowledged for providing invaluable insight and comments. This work was supported by project EVOREPRO within the framework of the 2nd call ERA-NET for Coordinating Plant Sciences, with funding from Fundacao para a Ciencia e a Tecnologia-FCT (Portugal) through grant ERA-CAPS/0001/2014. J.D.B. received salary support from FCT through an “Investigador FCT” position.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Mário R. Santos
    • 1
  • Cláudia Bispo
    • 1
    • 2
  • Jörg D. Becker
    • 1
    Email author
  1. 1.Instituto Gulbenkian de CiênciaOeirasPortugal
  2. 2.UCSF Flow Cytometry Core FacilitySan FranciscoUSA

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