TY - JOUR

T1 - Ratiometric gibberellin biosensors for the analysis of signaling dynamics and metabolism in plant protoplasts

AU - Andres, Jennifer

AU - Schmunk, Lisa J.

AU - Grau-Enguix, Federico

AU - Braguy, Justine

AU - Samodelov, Sophia L.

AU - Blomeier, Tim

AU - Ochoa-Fernandez, Rocio

AU - Weber, Wilfried

AU - Al-Babili, Salim

AU - Alabadí, David

AU - Blázquez, Miguel A.

AU - Zurbriggen, Matias D.

N1 - Publisher Copyright: © 2024 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.

PY - 2024

Y1 - 2024

N2 - 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.

AB - 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.

KW - gibberellin

KW - gibberellin metabolism

KW - phytohormone signaling

KW - plant protoplasts

KW - quantitative ratiometric biosensors

KW - technical advance

KW - gibberellin

KW - gibberellin metabolism

KW - phytohormone signaling

KW - plant protoplasts

KW - quantitative ratiometric biosensors

KW - technical advance

U2 - 10.1111/tpj.16725

DO - 10.1111/tpj.16725

M3 - Journal article

C2 - 38525669

AN - SCOPUS:85189206570

VL - 118

SP - 927

EP - 939

JO - Plant Journal

JF - Plant Journal

SN - 0960-7412

IS - 4

ER -