Abstract
Considerable progress has been made in understanding the influence of physical and genetic factors on the patterns of cell division in various model systems. However, how each of these factors directs changes in subcellular structures has remained unclear. Generic machineries for the execution of cell expansion and division have been characterized, but how these are influenced by genetic regulators and physical cell properties remains an open question. To a large degree, the complexity of growing post-embryonic tissues and a lack of precise predictability have prevented the extraction of rigid correlations between subcellular structures and future orientation of cell division. The Arabidopsis embryo offers an exquisitely predictable and simple model for studying such correlations, but so far the tools and methodology for studying subcellular structures in the early embryo have been lacking. Here, we describe a set of markers to visualize a range of subcellular structures in the early Arabidopsis embryo. We have designed a series of fluorescent cellular reporters optimized for embryos, and demonstrate the effectiveness of using these ‘ACE’ reporters with simple three-dimensional imaging procedures that preserve delicate cellular structures. We describe the ontogeny of subcellular structures in the early embryo and find that central/peripheral cell polarity is established much earlier than suspected. In addition, we show that the actin and microtubule cytoskeleton has distinct topologies in the embryo. These tools and methods will allow detailed analysis of the events of cellular reorganization that underlie morphogenesis in the Arabidopsis embryo.
| Original language | English |
|---|---|
| Pages (from-to) | 963-976 |
| Number of pages | 14 |
| Journal | Plant Journal |
| Volume | 93 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - Mar 2018 |
| Externally published | Yes |
Bibliographical note
Funding Information:We thank Thomas Laux (Universita€t Freiburg) for the pGII/N plasmid backbone, Niko Geldner (University of Lausanne) for the mCherry-NIP5;1 and BOR1-Cit reporting cassettes, and Tijs Kete-laar (Wageningen University) for the GFP-TUA6 reporting cassette and seeds of the p35S:: GFP: TUA6 reporter line. This research was supported by the European Research Council (CELLPATTERN; contract no. 281573).
Funding Information:
We thank Thomas Laux (Universität Freiburg) for the pGII/N plasmid backbone, Niko Geldner (University of Lausanne) for the mCherry-NIP5;1 and BOR1-Cit reporting cassettes, and Tijs Ketelaar (Wageningen University) for the GFP-TUA6 reporting cassette and seeds of the p35S:: GFP: TUA6 reporter line. This research was supported by the European Research Council (CELLPATTERN; contract no. 281573).
Publisher Copyright:
© 2018 The Authors The Plant Journal published by John Wiley & Sons Ltd and Society for Experimental Biology.
Funding
We thank Thomas Laux (Universita€t Freiburg) for the pGII/N plasmid backbone, Niko Geldner (University of Lausanne) for the mCherry-NIP5;1 and BOR1-Cit reporting cassettes, and Tijs Kete-laar (Wageningen University) for the GFP-TUA6 reporting cassette and seeds of the p35S:: GFP: TUA6 reporter line. This research was supported by the European Research Council (CELLPATTERN; contract no. 281573). We thank Thomas Laux (Universität Freiburg) for the pGII/N plasmid backbone, Niko Geldner (University of Lausanne) for the mCherry-NIP5;1 and BOR1-Cit reporting cassettes, and Tijs Ketelaar (Wageningen University) for the GFP-TUA6 reporting cassette and seeds of the p35S:: GFP: TUA6 reporter line. This research was supported by the European Research Council (CELLPATTERN; contract no. 281573).
Keywords
- Arabidopsis thaliana
- cell biology
- embryogenesis
- imaging
- oriented cell division