Dual expression and anatomy lines allow simultaneous visualization of gene expression and anatomy
| Autor: | Darren M. Wells, George Janes, Alexander Ware, Nicola Leftley, Paul T. Tarr, Anthony Bishopp, Kevin A. Pyke, Britta M. C. Kümpers, Jingyi Han, Nicholas Redman, John Vaughan-Hirsch, Jonathan A. Atkinson, Ute Voß, Giuseppe Castiglione |
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| Rok vydání: | 2021 |
| Předmět: |
AcademicSubjects/SCI01280
Physiology Arabidopsis Cloning vector Organogenesis ORGANIZATION Plant Science Biology Genes Plant Plant Roots INITIATION Plant Growth Regulators Gene Expression Regulation Plant Genes Reporter Auxin Gene expression Genetics Arabidopsis thaliana Gene Ultrasonography chemistry.chemical_classification AcademicSubjects/SCI01270 Science & Technology AcademicSubjects/SCI02288 AcademicSubjects/SCI02287 Cell Membrane AcademicSubjects/SCI02286 Plant Sciences fungi Lateral root food and beverages Anatomy Genes Development and Evolution ARABIDOPSIS biology.organism_classification DIFFERENTIATION chemistry CELLS Breakthrough Technologies Tools and Resources Life Sciences & Biomedicine |
| Zdroj: | Plant Physiology |
| ISSN: | 1532-2548 0032-0889 |
| Popis: | 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. A vector system and flow cell set-up allow long-term imaging of both gene expression and anatomy in Arabidopsis primary and lateral roots. |
| Databáze: | OpenAIRE |
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