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

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Publikace nespadá pod Pedagogickou fakultu, ale pod Středoevropský technologický institut. Oficiální stránka publikace je na webu muni.cz.
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BARBEZ Elke LANKOVA Martina PAREZOVA Marketa MAIZEL Alexis ZAŽÍMALOVÁ Eva PETRASEK Jan FRIML Jiří KLEINE-VEHN Jurgen

Rok publikování 2013
Druh Článek v odborném periodiku
Časopis / Zdroj BMC Plant Biology
Fakulta / Pracoviště MU

Středoevropský technologický institut

Citace
www http://www.biomedcentral.com/1471-2229/13/20
Doi http://dx.doi.org/10.1186/1471-2229-13-20
Obor Genetika a molekulární biologie
Klíčová slova Auxin homeostasis; DR5; Auxin carrier; Auxin transport
Přiložené soubory
Popis Background: Abundance 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. Results: We 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. Conclusions: This 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.
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