Abstract
Unraveling the mechanisms that govern division plane orientation is a major challenge to understand plant development. In this respect, the Arabidopsis early embryo is a model system of choice since embryogenesis is relatively simple and cell division planes orientation is highly predictable. Here, we present an integrated set of protocols to study 3D cell division patterns in early-stage Arabidopsis embryos that combine both cellular and sub-cellular localization of selected protein markers with spatial organization of cells, cytoskeleton, and nuclei.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Li R, Bowerman B (2010) Symmetry breaking in biology. Cold Spring Harb Perspect Biol 2:a003475
De Smet I, Beeckman T (2011) Asymmetric cell division in land plants and algae: the driving force for differentiation. Nat Rev Mol Cell Biol 12:177–188
Bouchez D, Van Damme D, Boruc J, Schaefer E, Pastuglia M (2014) Cell division plane determination in plant development. In: Assmann S, Liu B (eds) The plant sciences - cell biology. Springer, Heidelberg
Weijers D, Jurgens G (2005) Auxin and embryo axis formation: the ends in sight? Curr Opin Plant Biol 8:32–37
Webb MC, Gunning BE (1991) The microtubular cytoskeleton during development of the zygote, proemhryo and free-nuclear endosperm in Arabidopsis thaliana (L.) Heynh. Planta 184:187–195
Lauber MH, Waizenegger I, Steinmann T, Schwarz H, Mayer U, Hwang I, Lukowitz W, Jürgens G (1997) The Arabidopsis KNOLLE protein is a cytokinesis-specific syntaxin. J Cell Biol 139:1485–1493
Sauer M, Paciorek T, Benková E, Friml J (2006) Immunocytochemical techniques for whole-mount in situ protein localization in plants. Nat Protoc 1:98–103
Smertenko AP, Hussey PJ (2008) Immunolocalization of proteins in somatic embryos: applications for studies on the cytoskeleton. Methods Mol Biol 427:157–171
Spinner L, Gadeyne A, Belcram K, Goussot M, Moison M, Duroc Y, Eeckhout D, De Winne N, Schaefer E, Van De Slijke E, Persiau G, Witters E, Gevaert K, De Jaeger G, Bouchez D, Van Damme D, Pastuglia M (2013) A protein phosphatase 2A complex spatially controls plant cell division. Nat Commun 4:1863
De Rybel B, Moller B, Yoshida S, Grabowicz I, de Reuille PB, Boeren S, Smith RS, Borst JW, Weijers D (2013) A bHLH complex controls embryonic vascular tissue establishment and indeterminate growth in Arabidopsis. Dev Cell 24:426–437
Enugutti B, Schneitz K (2013) Genetic analysis of ectopic growth suppression during planar growth of integuments mediated by the Arabidopsis AGC protein kinase UNICORN. BMC Plant Biol 13:2
Haecker A, Gross-Hardt R, Geiges B, Sarkar A, Breuninger H, Herrmann M, Laux T (2004) Expression dynamics of WOX genes mark cell fate decisions during early embryonic patterning in Arabidopsis thaliana. Development 131(3):657–668
Ditengou FA, Tealea WD, Kochersperger P, Flittner KA, Kneuper I, van der Graaff E, Nziengui H, Pinosa F, Li X, Nitschke R, Laux T, Palme K (2008) Mechanical induction of lateral root initiation in Arabidopsis thaliana. Proc Natl Acad Sci U S A 105:18818–18823
Miart F, Desprez T, Biot E, Morin H, Belcram K, Höfte H, Gonneau M, Vernhettes S (2014) Spatio-temporal analysis of cellulose synthesis during cell plate formation in Arabidopsis. Plant J 77:71–84
Truernit E, Bauby H, Dubreucq B, Grandjean O, Runions J et al (2008) High-resolution whole-mount imaging of three-dimensional tissue organization and gene expression enables the study of phloem development and structure in Arabidopsis. Plant Cell 20:1494–1503
Acknowledgements
We thank David Legland and Ignacio Arganda-Carreras for the development of the morphological segmentation plugin in Fiji software. We are grateful to Daniel Vezon for taking care of the plants, to Gerd Jürgens (Tüebingen, Germany) for the gift of the anti-KNOLLE antibody and to Marie-Cécile Caillaud (ENS Lyon, France) for helpful advices with the immunolocalization protocol. We thank the “Région Île-de-France” and “Conseil Général des Yvelines” for supporting our microscopy platform.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Belcram, K., Palauqui, JC., Pastuglia, M. (2016). Studying Cell Division Plane Positioning in Early-Stage Embryos. In: Caillaud, MC. (eds) Plant Cell Division. Methods in Molecular Biology, vol 1370. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3142-2_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3142-2_14
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3141-5
Online ISBN: 978-1-4939-3142-2
eBook Packages: Springer Protocols