Dual expression and anatomy lines allow simultaneous visualization of gene expression and anatomy
Abstract
Studying the developmental genetics of plant organs requires following gene expression in specific tissues. To facilitate this, we have developed dual expression anatomy lines, which incorporate a red plasma membrane marker alongside a fluorescent reporter for a gene of interest in the same vector. Here, we adapted the GreenGate cloning vectors to create two destination vectors showing strong marking of cell membranes in either the whole root or specifically in the lateral roots. This system can also be used in both embryos and whole seedlings. As proof of concept, we follow both gene expression and anatomy in Arabidopsis (Arabidopsis thaliana) during lateral root organogenesis for a period of over 24 h. Coupled with the development of a flow cell and perfusion system, we follow changes in activity of the DII auxin sensor following application of auxin.
Additional Information
© The Author(s) 2021. Published by Oxford University Press on behalf of American Society of Plant Biologists. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. Received: 20 April 2021; Accepted: 04 October 2021; Published: 29 October 2021; Corrected and typeset: 10 November 2021. This work was supported by the Biotechnology and Biological Sciences Research Council [grant numbers BB/L023555/1 (BMCK and AB), BB/M019837/1 (D.M.W. and U.V.) and (BB/M008770/1] (K.P., N.R., J.V.-H.)]; the Royal Society [grant number UF160729 (A.B.)] and the University of Nottingham via the Nottingham Research Fellowship Scheme (U.V.), the Paper Enhancement Fund (A.W., D.M.W., and U.V.) and the Future Food Beacon of Excellence (J.A.A. and D.M.W.). Conflict of interest statement. None declared. B.M.C.K., J.A.A., D.W., and A.B. designed the concept; B.M.K., J.V.-H. and G.C. built the destination vectors; B.M.K., J.H., A.W., J.V.-H., and A.B. imaged the lines; N.R., J.A.A., and D.W. designed and built the flow cell; G.J., P.T.T., and N.L. donated material or provided entry clones used within this project; K.P. and U.V. supervised parts of this project; B.M.C.K., D.M.W., and A.B. wrote the manuscript with input from all the authors. The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (https://academic.oup.com/plphys/pages/general-instructions) is: Anthony Bishopp (anthony.bishopp@nottingham.ac.uk).Attached Files
Published - kiab503.pdf
Supplemental Material - kiab503_supplementary_data.zip
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Additional details
- Alternative title
- Dual expression anatomy lines (DEAL) allow simultaneous visualization of gene expression and anatomy
- Alternative title
- Dual visualization of gene expression and anatomy
- PMCID
- PMC8774739
- Eprint ID
- 111951
- DOI
- 10.1093/plphys/kiab503
- Resolver ID
- CaltechAUTHORS:20211118-223714489
- BB/L023555/1
- Biotechnology and Biological Sciences Research Council (BBSRC)
- BB/M019837/1
- Biotechnology and Biological Sciences Research Council (BBSRC)
- BB/M008770/1
- Biotechnology and Biological Sciences Research Council (BBSRC)
- UF160729
- Royal Society
- University of Nottingham
- Paper Enhancement Fund
- Future Food Beacon of Excellence
- Created
-
2021-11-18Created from EPrint's datestamp field
- Updated
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2022-01-25Created from EPrint's last_modified field