Ratiometric gibberellin biosensors for the analysis of signaling dynamics and metabolism in plant protoplasts.
Plant J
; 118(4): 927-939, 2024 May.
Article
em En
| MEDLINE
| ID: mdl-38525669
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.
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MEDLINE
Assunto principal:
Protoplastos
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Técnicas Biossensoriais
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Transdução de Sinais
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Arabidopsis
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Proteínas de Arabidopsis
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Giberelinas
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article