CaltechAUTHORS
  A Caltech Library Service

Dual expression and anatomy lines allow simultaneous visualization of gene expression and anatomy

Kümpers, Britta M. C. and Han, Jingyi and Vaughan-Hirsch, John and Redman, Nicholas and Ware, Alexander and Atkinson, Jonathan A. and Leftley, Nicola and Janes, George and Castiglione, Giuseppe and Tarr, Paul T. and Pyke, Kevin and Voß, Ute and Wells, Darren M. and Bishopp, Anthony (2022) Dual expression and anatomy lines allow simultaneous visualization of gene expression and anatomy. Plant Physiology, 188 (1). pp. 56-69. ISSN 1532-2548. PMCID PMC8774739. doi:10.1093/plphys/kiab503. https://resolver.caltech.edu/CaltechAUTHORS:20211118-223714489

[img] PDF - Published Version
Creative Commons Attribution.

1MB
[img] Archive (ZIP) (Supplementary data) - Supplemental Material
Creative Commons Attribution.

32MB

Use this Persistent URL to link to this item: https://resolver.caltech.edu/CaltechAUTHORS:20211118-223714489

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.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1093/plphys/kiab503DOIArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/pmc8774739/PubMed CentralArticle
ORCID:
AuthorORCID
Kümpers, Britta M. C.0000-0001-9931-6985
Han, Jingyi0000-0003-1106-2461
Tarr, Paul T.0000-0003-2977-3683
Wells, Darren M.0000-0002-4246-4909
Bishopp, Anthony0000-0003-2962-9542
Alternate Title:Dual expression anatomy lines (DEAL) allow simultaneous visualization of gene expression and anatomy, Dual visualization of gene expression and anatomy
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).
Funders:
Funding AgencyGrant Number
Biotechnology and Biological Sciences Research Council (BBSRC)BB/L023555/1
Biotechnology and Biological Sciences Research Council (BBSRC)BB/M019837/1
Biotechnology and Biological Sciences Research Council (BBSRC)BB/M008770/1
Royal SocietyUF160729
University of NottinghamUNSPECIFIED
Paper Enhancement FundUNSPECIFIED
Future Food Beacon of ExcellenceUNSPECIFIED
Issue or Number:1
PubMed Central ID:PMC8774739
DOI:10.1093/plphys/kiab503
Record Number:CaltechAUTHORS:20211118-223714489
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20211118-223714489
Official Citation:Britta M C Kümpers, Jingyi Han, John Vaughan-Hirsch, Nicholas Redman, Alexander Ware, Jonathan A Atkinson, Nicola Leftley, George Janes, Giuseppe Castiglione, Paul T Tarr, Kevin Pyke, Ute Voß, Darren M Wells, Anthony Bishopp, Dual expression and anatomy lines allow simultaneous visualization of gene expression and anatomy, Plant Physiology, Volume 188, Issue 1, January 2022, Pages 56–69, https://doi.org/10.1093/plphys/kiab503
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:111951
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:18 Nov 2021 22:48
Last Modified:25 Jan 2022 17:48

Repository Staff Only: item control page