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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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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