Single-cell-based system to monitor carrier driven cellular auxin homeostasis

BMC Plant Biology

Methodology Article

Elke Barbez¹  Jürgen Kleine-Vehn¹  Jiří Friml²  Alexis Maizel³  Martina Laňková⁴  Eva Zažímalová⁴  Markéta Pařezová⁴  Jan Petrášek⁴ 

[1] Department of Plant Systems Biology, VIB and Department of Plant Biotechnology and Genetics, Ghent University, 9052, Gent, Belgium;Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Vienna (BOKU), 1190, Vienna, Austria;Department of Plant Systems Biology, VIB and Department of Plant Biotechnology and Genetics, Ghent University, 9052, Gent, Belgium;Department of Functional Genomics and Proteomics, Faculty of Science, and CEITEC, Masaryk University, CZ-62500, Kamenice 5, Brno, Czech Republic;Department of Stem Cell Biology, Center for Organismal Studies, University of Heidelberg, 69120, Heidelberg, Germany;Institute of Experimental Botany, The Academy of Sciences of the Czech Republic, 16502, Praha 6, Czech Republic;

关键词: Auxin homeostasis;    DR5;    Auxin carrier;    Auxin transport;   

DOI  :  10.1186/1471-2229-13-20

 received in 2012-10-02, accepted in 2013-01-31,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundAbundance and distribution of the plant hormone auxin play important roles in plant development. Besides other metabolic processes, various auxin carriers control the cellular level of active auxin and, hence, are major regulators of cellular auxin homeostasis. Despite the developmental importance of auxin transporters, a simple medium-to-high throughput approach to assess carrier activities is still missing. Here we show that carrier driven depletion of cellular auxin correlates with reduced nuclear auxin signaling in tobacco Bright Yellow-2 (BY-2) cell cultures.ResultsWe developed an easy to use transient single-cell-based system to detect carrier activity. We use the relative changes in signaling output of the auxin responsive promoter element DR5 to indirectly visualize auxin carrier activity. The feasibility of the transient approach was demonstrated by pharmacological and genetic interference with auxin signaling and transport. As a proof of concept, we provide visual evidence that the prominent auxin transport proteins PIN-FORMED (PIN)2 and PIN5 regulate cellular auxin homeostasis at the plasma membrane and endoplasmic reticulum (ER), respectively. Our data suggest that PIN2 and PIN5 have different sensitivities to the auxin transport inhibitor 1-naphthylphthalamic acid (NPA). Also the putative PIN-LIKES (PILS) auxin carrier activity at the ER is insensitive to NPA in our system, indicating that NPA blocks intercellular, but not intracellular auxin transport.ConclusionsThis single-cell-based system is a useful tool by which the activity of putative auxin carriers, such as PINs, PILS and WALLS ARE THIN1 (WAT1), can be indirectly visualized in a medium-to-high throughput manner. Moreover, our single cell system might be useful to investigate also other hormonal signaling pathways, such as cytokinin.

【 授权许可】

Unknown   
© Barbez et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.