Abstract
The protocol allows to define and characterize mitosis distribution patterns in the plant root meristem. The method does not require genetic markers, which makes it applicable to plants of different non-transgenic genotypes, including ecotypes, mutants, and non-model plant species. Computer analysis of the mitosis distribution in three dimensions with iRoCS Toolbox identifies statistically significant changes in proliferation activity within specific root tissues and cell lineages.
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
Similar content being viewed by others
References
Bishopp A, Lehesranta S, Vaten A, Help H, El-Showk S et al (2011) Phloem-transported cytokinin regulates polar auxin transport and maintains vascular pattern in the root meristem. Curr Biol 21:927–932
Lavrekha VV, Pasternak T, Ivanov VB, Palme K, Mironova VV (2017) 3D analysis of mitosis distribution defines the longitudinal zonation and bilateral symmetry of the Arabidopsis thaliana root meristem. Plant J 92(5):834–845
Casamitjana-Martınez E, Hofhuis HF, Xu J, Liu CM, Heidstra R, Scheres B (2003) Root-specific CLE19 overexpression and the sol1/2 suppressors implicate a CLV-like pathway in the control of Arabidopsis root meristem maintenance. Curr Biol 13(16):1435–1441
Blein T, Duerr J, Pasternak T, Haser T, Falk T et al (2018) Light dynamically regulates growth rate and cellular organisation of the Arabidopsis root meristem. bioRxiv 1:353987
Colon-Carmona A, You R, Haimovitch-Gal T, Doerner P (1999) Technical advance: spatio-temporal analysis of mitotic activity with a labile cyclin-GUS fusion protein. Plant J 20(4):503–508
Hayashi K, Hasegawa J, Matsunaga S (2013) The boundary of the meristematic and elongation zones in roots: endoreduplication precedes rapid cell expansion. Sci Rep 3:2723
Pasternak T, Haser T, Falk T, Ronneberger O, Palme K, Otten L (2017) A 3D digital atlas of the Nicotiana tabacum root tip and its use to investigate changes in the root apical meristem induced by the Agrobacterium 6b oncogene. Plant J 92(1):31–42
Pasternak TP, Rudas VA, Lörz H, Kumlehn J (1999) Embryogenic callus formation and plant regeneration from leaf base segments of barley (Hordeum vulgare L.). J Plant Physiol 155:371–375
Pasternak T, Tietz O, Rapp K, Begheldo M, Nitschke R, Ruperti B et al (2015) Protocol: an improved and universal procedure for whole-mount immunolocalization in plants. Plant Methods 11:1
Emmenlauer M, Ronneberger O, Ponti A, Schwarb P, Griffa A, Filippi A et al (2009) XuvTools: free, fast and reliable stitching of large 3D datasets. J Microsc 233:42–60
Schmidt T, Pasternak T, Liu K, Blein T, Aubry-Hivet D et al (2014) The iRoCS Toolbox—3D analysis of the plant root apical meristem at cellular resolution. Plant J 77:806–814
Truernit E, Bauby H, Belcram K, Barthélémy J, Palauqui J-C (2012) OCTOPUS, a polarly localised membrane-associated protein, regulates phloem differentiation entry in Arabidopsis thaliana. Development 139:1306–1315
Acknowledgment
We thank Roland Nitschke and the staff of the Life Imaging Center (LIC) in the Center for Biological Systems Analysis (ZBSA) of the Albert-Ludwigs-University Freiburg for help with their confocal microscopy resources. This work was supported by the Russian State Budget Project (0324-2019-0040), by the Russian Federation President Grant for young scientists (MK-1297.2017.4), and by the Russian Foundation for Basic Research (RFBR-18-34-00485). This work was also supported by Bundesministerium für Bildung und Forschung (BMBF Microsystems, Haploswitch), the German Research Foundation (DFG SFB746, INST 39/839,840,841), and Deutsches Zentrum für Luft und Raumfahrt (DLR 50WB1022). The authors declare that no competing interests exist.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Lavrekha, V.V., Pasternak, T., Palme, K., Mironova, V.V. (2020). 3D Analysis of Mitosis Distribution Pattern in the Plant Root Tip with iRoCS Toolbox. In: Naseem, M., Dandekar, T. (eds) Plant Stem Cells. Methods in Molecular Biology, vol 2094. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0183-9_13
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0183-9_13
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0182-2
Online ISBN: 978-1-0716-0183-9
eBook Packages: Springer Protocols