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

Kümpers, Britta M C; Han, Jingyi; Vaughan-Hirsch, John; Redman, Nicholas; Ware, Alexander; Atkinson, Jonathan A; Leftley, Nicola; Janes, George; Castiglione, Giuseppe; Tarr, Paul T; Pyke, Kevin; Voß, Ute; Wells, Darren M; Bishopp, Anthony

Abstract

Studying the developmental genetics of plant organs, requires following gene expression in specific tissues. To facilitate this, we have developed the Dual Expression Anatomy Lines (DEAL), 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 24h,. and cCoupled with the development of a flow cell and perfusion system, we follow changes in activity of the DII auxin sensor following application of auxin.

Citation

Kümpers, B. M. C., Han, J., Vaughan-Hirsch, J., Redman, N., Ware, A., Atkinson, J. A., Leftley, N., Janes, G., Castiglione, G., Tarr, P. T., Pyke, K., Voß, U., Wells, D. M., & Bishopp, A. (2022). Dual expression and anatomy lines allow simultaneous visualization of gene expression and anatomy. Plant Physiology, 188(1), 56-69. https://doi.org/10.1093/plphys/kiab503

• Journal Article Type: Article
• Acceptance Date: Oct 4, 2021
• Online Publication Date: Oct 29, 2021
• Publication Date: 2022-01
• Deposit Date: Oct 28, 2021
• Publicly Available Date: Nov 1, 2021
• Journal: Plant Physiology
• Print ISSN: 0032-0889
• Electronic ISSN: 1532-2548
• Publisher: American Society of Plant Biologists
• Peer Reviewed: Peer Reviewed
• Volume: 188
• Issue: 1
• Pages: 56-69
• DOI: https://doi.org/10.1093/plphys/kiab503
• Keywords: Plant Science; Genetics; Physiology
• Public URL: https://nottingham-repository.worktribe.com/output/6541866
• Publisher URL: https://academic.oup.com/plphys/article/188/1/56/6414209