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Plant Germline Development pp 193–210Cite as

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

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Part of the book series: Methods in Molecular Biology ((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.

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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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Authors and Affiliations

  1. Instituto Gulbenkian de Ciência, Rua da Quinta Grande Nº6, 2780-156, Oeiras, Portugal

    Mário R. Santos, Cláudia Bispo & Jörg D. Becker

  2. UCSF Flow Cytometry Core Facility, 513 Parnassus Ave., San Francisco, CA, 94143, USA

    Cláudia Bispo

Authors
  1. Mário R. Santos
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  2. Cláudia Bispo
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  3. Jörg D. Becker
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Corresponding author

Correspondence to Jörg D. Becker .

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Editors and Affiliations

  1. Centre for Organismal Studies, University of Heidelberg, Heidelberg, Germany

    Anja Schmidt

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Santos, M.R., Bispo, C., Becker, J.D. (2017). Isolation of Arabidopsis Pollen, Sperm Cells, and Vegetative Nuclei by Fluorescence-Activated Cell Sorting (FACS). In: Schmidt, A. (eds) Plant Germline Development. Methods in Molecular Biology, vol 1669. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7286-9_16

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  • DOI: https://doi.org/10.1007/978-1-4939-7286-9_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7285-2

  • Online ISBN: 978-1-4939-7286-9

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