Ratiometric gibberellin biosensors for the analysis of signaling dynamics and metabolism in plant protoplasts
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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.
Original language | English |
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Journal | Plant Journal |
Volume | 118 |
Issue number | 4 |
Pages (from-to) | 927-939 |
Number of pages | 13 |
ISSN | 0960-7412 |
DOIs | |
Publication status | Published - 2024 |
Bibliographical note
Publisher Copyright:
© 2024 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.
- gibberellin, gibberellin metabolism, phytohormone signaling, plant protoplasts, quantitative ratiometric biosensors, technical advance
Research areas
ID: 392107929