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.
Highlights
The primary and lateral roots of Arabidopsis (Arabidopsis thaliana) provide well-studied systems for cell fate acquisition
We developed a variant of this destination vector which outlines cells within the lateral root primordia (LRP) and the stele suitable for longterm imaging of lateral root organogenesis
An efficient method for dual visualization of gene expression and root anatomy We sought to develop a vector system where we could introduce a single plasmid to plants to simultaneously report gene expression alongside root anatomy
Summary
The primary and lateral roots of Arabidopsis (Arabidopsis thaliana) provide well-studied systems for cell fate acquisition. Proliferative cell divisions in the root meristem lead to a stereotypic anatomical pattern in which a diarch vascular cylinder is surrounded by radially symmetric layers of outer cells (Dolan et al, 1993). Decades of research into the mechanisms underlying cell fate specification have provided us with a broad set of cell-type specific promoters that can be used to investigate identity of individual files. While we have a good understanding of the mechanistic processes through which a selection of cell identities is established, there are Received April 20, 2021.
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