TY - JOUR TI - Imaging plant germline differentiation within Arabidopsis flowers by light sheet microscopy AU - Valuchova, Sona AU - Mikulkova, Pavlina AU - Pecinkova, Jana AU - Klimova, Jana AU - Krumnikl, Michal AU - Bainar, Petr AU - Heckmann, Stefan AU - Tomancak, Pavel AU - Riha, Karel A2 - Hardtke, Christian S A2 - McCormick, Sheila A2 - Prunet, Nathanaƫl A2 - Dobritsa, Anna A VL - 9 PY - 2020 DA - 2020/02/11 SP - e52546 C1 - eLife 2020;9:e52546 DO - 10.7554/eLife.52546 UR - https://doi.org/10.7554/eLife.52546 AB - 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. KW - germline KW - meiosis KW - flower KW - live cell imaging KW - light sheet microscopy KW - SPIM JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -