In higher plants, germline differentiation occurs during a relatively short period within developing flowers. Understanding of the mechanisms that govern germline differentiation lags behind other plant developmental processes. This is largely because the germline is restricted to relatively few cells buried deep within floral tissues, which makes them difficult to study. To overcome this limitation, we have developed a methodology for live imaging of the germ cell lineage within floral organs of Arabidopsis using light sheet fluorescence microscopy. We have established reporter lines, cultivation conditions, and imaging protocols for high-resolution microscopy of developing flowers continuously for up to several days. We used multiview imagining to reconstruct a three-dimensional model of a flower at subcellular resolution. We demonstrate the power of this approach by capturing male and female meiosis, asymmetric pollen division, movement of meiotic chromosomes, and unusual restitution mitosis in tapetum cells. This method will enable new avenues of research into plant sexual reproduction.

Central European Institute of Technology Masaryk University Brno Czech Republic

Department of Computer Science FEECS VSB Technical University of Ostrava Ostrava Czech Republic

IT4Innovations VSB Technical University of Ostrava Ostrava Czech Republic

Leibniz Institute of Plant Genetics and Crop Plant Research Seeland Germany

Max Planck Institute of Molecular Cell Biology and Genetics Dresden Germany

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Elife. 2020 Feb 11;9:e54984. doi: 10.7554/eLife.54984. PubMed

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