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Novel markers for high-throughput protoplast-based analyses of phytohormone signaling

Phytohormones mediate most diverse processes in plants, ranging from organ development to immune responses. Receptor protein complexes perceive changes in intracellular phytohormone levels and trigger a signaling cascade to effectuate downstream responses. The in planta analysis of elements involved...

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Autores principales: Lehmann, Silke, Dominguez-Ferreras, Ana, Huang, Wei-Jie, Denby, Katherine, Ntoukakis, Vardis, Schäfer, Patrick
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7272087/
https://www.ncbi.nlm.nih.gov/pubmed/32497144
http://dx.doi.org/10.1371/journal.pone.0234154
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author Lehmann, Silke
Dominguez-Ferreras, Ana
Huang, Wei-Jie
Denby, Katherine
Ntoukakis, Vardis
Schäfer, Patrick
author_facet Lehmann, Silke
Dominguez-Ferreras, Ana
Huang, Wei-Jie
Denby, Katherine
Ntoukakis, Vardis
Schäfer, Patrick
author_sort Lehmann, Silke
collection PubMed
description Phytohormones mediate most diverse processes in plants, ranging from organ development to immune responses. Receptor protein complexes perceive changes in intracellular phytohormone levels and trigger a signaling cascade to effectuate downstream responses. The in planta analysis of elements involved in phytohormone signaling can be achieved through transient expression in mesophyll protoplasts, which are a fast and versatile alternative to generating plant lines that stably express a transgene. While promoter-reporter constructs have been used successfully to identify internal or external factors that change phytohormone signaling, the range of available marker constructs does not meet the potential of the protoplast technique for large scale approaches. The aim of our study was to provide novel markers for phytohormone signaling in the Arabidopsis mesophyll protoplast system. We validated 18 promoter::luciferase constructs towards their phytohormone responsiveness and specificity and suggest an experimental setup for high-throughput analyses. We recommend novel markers for the analysis of auxin, abscisic acid, cytokinin, salicylic acid and jasmonic acid responses that will facilitate future screens for biological elements and environmental stimuli affecting phytohormone signaling.
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spelling pubmed-72720872020-06-09 Novel markers for high-throughput protoplast-based analyses of phytohormone signaling Lehmann, Silke Dominguez-Ferreras, Ana Huang, Wei-Jie Denby, Katherine Ntoukakis, Vardis Schäfer, Patrick PLoS One Research Article Phytohormones mediate most diverse processes in plants, ranging from organ development to immune responses. Receptor protein complexes perceive changes in intracellular phytohormone levels and trigger a signaling cascade to effectuate downstream responses. The in planta analysis of elements involved in phytohormone signaling can be achieved through transient expression in mesophyll protoplasts, which are a fast and versatile alternative to generating plant lines that stably express a transgene. While promoter-reporter constructs have been used successfully to identify internal or external factors that change phytohormone signaling, the range of available marker constructs does not meet the potential of the protoplast technique for large scale approaches. The aim of our study was to provide novel markers for phytohormone signaling in the Arabidopsis mesophyll protoplast system. We validated 18 promoter::luciferase constructs towards their phytohormone responsiveness and specificity and suggest an experimental setup for high-throughput analyses. We recommend novel markers for the analysis of auxin, abscisic acid, cytokinin, salicylic acid and jasmonic acid responses that will facilitate future screens for biological elements and environmental stimuli affecting phytohormone signaling. Public Library of Science 2020-06-04 /pmc/articles/PMC7272087/ /pubmed/32497144 http://dx.doi.org/10.1371/journal.pone.0234154 Text en © 2020 Lehmann et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Lehmann, Silke
Dominguez-Ferreras, Ana
Huang, Wei-Jie
Denby, Katherine
Ntoukakis, Vardis
Schäfer, Patrick
Novel markers for high-throughput protoplast-based analyses of phytohormone signaling
title Novel markers for high-throughput protoplast-based analyses of phytohormone signaling
title_full Novel markers for high-throughput protoplast-based analyses of phytohormone signaling
title_fullStr Novel markers for high-throughput protoplast-based analyses of phytohormone signaling
title_full_unstemmed Novel markers for high-throughput protoplast-based analyses of phytohormone signaling
title_short Novel markers for high-throughput protoplast-based analyses of phytohormone signaling
title_sort novel markers for high-throughput protoplast-based analyses of phytohormone signaling
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7272087/
https://www.ncbi.nlm.nih.gov/pubmed/32497144
http://dx.doi.org/10.1371/journal.pone.0234154
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