Ratiometric gibberellin biosensors for the analysis of signaling dynamics and metabolism in plant protoplasts
Abstract: Gibberellins (GAs) are major regulators of developmental and growth processes in plants. Using the degradation-based signaling mechanism of GAs, we have built transcriptional regulator (DELLA)-based, genetically encoded ratiometric biosensors as proxies for hormone quantification at high temporal resolution and sensitivity that allow dynamic, rapid and simple analysis in a plant cell system, i.e. Arabidopsis protoplasts. These ratiometric biosensors incorporate a DELLA protein as a degradation target fused to a firefly luciferase connected via a 2A peptide to a renilla luciferase as a co-expressed normalization element. We have implemented these biosensors for all five Arabidopsis DELLA proteins, GA-INSENSITIVE, GAI; REPRESSOR-of-ga1-3, RGA; RGA-like1, RGL1; RGL2 and RGL3, by applying a modular design. The sensors are highly sensitive (in the low pm range), specific and dynamic. As a proof of concept, we have tested the applicability in three domains: the study of substrate specificity and activity of putative GA-oxidases, the characterization of GA transporters, and the use as a discrimination platform coupled to a GA agonists' chemical screening. This work demonstrates the development of a genetically encoded quantitative biosensor complementary to existing tools that allow the visualization of GA in planta
- Standort
-
Deutsche Nationalbibliothek Frankfurt am Main
- Umfang
-
Online-Ressource
- Sprache
-
Englisch
- Anmerkungen
-
The plant journal. - 118, 4 (2024) , 927-939, ISSN: 1365-313X
- Ereignis
-
Veröffentlichung
- (wo)
-
Freiburg
- (wer)
-
Universität
- (wann)
-
2024
- Urheber
-
Andres, Jennifer
Schmunk, Lisa J.
Grau‐Enguix, Federico
Braguy, Justine
Samodelov, Sophia L.
Blomeier, Tim
Ochoa-Fernandez, Rocio
Weber, Wilfried
Al-Babili, Salim
Alabadí, David
Vega, Miguel
Zurbriggen, Matias D.
- DOI
-
10.1111/tpj.16725
- URN
-
urn:nbn:de:bsz:25-freidok-2533015
- Rechteinformation
-
Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
- Letzte Aktualisierung
-
14.08.2025, 10:56 MESZ
Datenpartner
Deutsche Nationalbibliothek. Bei Fragen zum Objekt wenden Sie sich bitte an den Datenpartner.
Beteiligte
- Andres, Jennifer
- Schmunk, Lisa J.
- Grau‐Enguix, Federico
- Braguy, Justine
- Samodelov, Sophia L.
- Blomeier, Tim
- Ochoa-Fernandez, Rocio
- Weber, Wilfried
- Al-Babili, Salim
- Alabadí, David
- Vega, Miguel
- Zurbriggen, Matias D.
- Universität
Entstanden
- 2024