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Bioluminescent imaging of Arabidopsis thaliana using an enhanced Nano-lantern luminescence reporter system

Bioluminescent detection has become a powerful method that is used extensively in numerous areas in life science research. Given that fluorescence detection in plant cells is difficult owing to the autofluorescence of chlorophyll, the use of a luciferin–luciferase system should be effective in plant...

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Autores principales: Furuhata, Yuichi, Sakai, Ayako, Murakami, Tomi, Nagasaki, Akira, Kato, Yoshio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6941820/
https://www.ncbi.nlm.nih.gov/pubmed/31899919
http://dx.doi.org/10.1371/journal.pone.0227477
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author Furuhata, Yuichi
Sakai, Ayako
Murakami, Tomi
Nagasaki, Akira
Kato, Yoshio
author_facet Furuhata, Yuichi
Sakai, Ayako
Murakami, Tomi
Nagasaki, Akira
Kato, Yoshio
author_sort Furuhata, Yuichi
collection PubMed
description Bioluminescent detection has become a powerful method that is used extensively in numerous areas in life science research. Given that fluorescence detection in plant cells is difficult owing to the autofluorescence of chlorophyll, the use of a luciferin–luciferase system should be effective in plant biology. However, the suitable optical window for a luminescence system in plants remains unexplored. In this study, we sought to determine the optical window and optimal luciferase reporter system for terrestrial plant analyses using Arabidopsis thaliana as a model organism. We compared six different luciferase systems and found the green enhanced Nano-lantern (GeNL)–furimazine combination to be the optimal luciferase reporter. Spectral measurements of GeNL–furimazine showed that its luminescence peak falls within the range of optical transparency for chlorophyll and, therefore, enables greater penetration through a layer of cultured A. thaliana cells. Moreover, A. thaliana plants expressing GeNL with furimazine emitted strong luminescence, which could be detected even with the naked eye. Thus, the GeNL–furimazine combination should facilitate biological analyses of genes and cellular functions in A. thaliana and all other terrestrial plants.
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spelling pubmed-69418202020-01-10 Bioluminescent imaging of Arabidopsis thaliana using an enhanced Nano-lantern luminescence reporter system Furuhata, Yuichi Sakai, Ayako Murakami, Tomi Nagasaki, Akira Kato, Yoshio PLoS One Research Article Bioluminescent detection has become a powerful method that is used extensively in numerous areas in life science research. Given that fluorescence detection in plant cells is difficult owing to the autofluorescence of chlorophyll, the use of a luciferin–luciferase system should be effective in plant biology. However, the suitable optical window for a luminescence system in plants remains unexplored. In this study, we sought to determine the optical window and optimal luciferase reporter system for terrestrial plant analyses using Arabidopsis thaliana as a model organism. We compared six different luciferase systems and found the green enhanced Nano-lantern (GeNL)–furimazine combination to be the optimal luciferase reporter. Spectral measurements of GeNL–furimazine showed that its luminescence peak falls within the range of optical transparency for chlorophyll and, therefore, enables greater penetration through a layer of cultured A. thaliana cells. Moreover, A. thaliana plants expressing GeNL with furimazine emitted strong luminescence, which could be detected even with the naked eye. Thus, the GeNL–furimazine combination should facilitate biological analyses of genes and cellular functions in A. thaliana and all other terrestrial plants. Public Library of Science 2020-01-03 /pmc/articles/PMC6941820/ /pubmed/31899919 http://dx.doi.org/10.1371/journal.pone.0227477 Text en © 2020 Furuhata 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
Furuhata, Yuichi
Sakai, Ayako
Murakami, Tomi
Nagasaki, Akira
Kato, Yoshio
Bioluminescent imaging of Arabidopsis thaliana using an enhanced Nano-lantern luminescence reporter system
title Bioluminescent imaging of Arabidopsis thaliana using an enhanced Nano-lantern luminescence reporter system
title_full Bioluminescent imaging of Arabidopsis thaliana using an enhanced Nano-lantern luminescence reporter system
title_fullStr Bioluminescent imaging of Arabidopsis thaliana using an enhanced Nano-lantern luminescence reporter system
title_full_unstemmed Bioluminescent imaging of Arabidopsis thaliana using an enhanced Nano-lantern luminescence reporter system
title_short Bioluminescent imaging of Arabidopsis thaliana using an enhanced Nano-lantern luminescence reporter system
title_sort bioluminescent imaging of arabidopsis thaliana using an enhanced nano-lantern luminescence reporter system
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6941820/
https://www.ncbi.nlm.nih.gov/pubmed/31899919
http://dx.doi.org/10.1371/journal.pone.0227477
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