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A (13)C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems
BACKGROUND: Metabolic fluxes represent the functional phenotypes of biochemical pathways and are essential to reveal the distribution of precursors among metabolic networks. Although analysis of metabolic fluxes, facilitated by stable isotope labeling and mass spectrometry detection, has been applie...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6015466/ https://www.ncbi.nlm.nih.gov/pubmed/29977324 http://dx.doi.org/10.1186/s13007-018-0318-3 |
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author | Wang, Peng Guo, Longyun Jaini, Rohit Klempien, Antje McCoy, Rachel M. Morgan, John A. Dudareva, Natalia Chapple, Clint |
author_facet | Wang, Peng Guo, Longyun Jaini, Rohit Klempien, Antje McCoy, Rachel M. Morgan, John A. Dudareva, Natalia Chapple, Clint |
author_sort | Wang, Peng |
collection | PubMed |
description | BACKGROUND: Metabolic fluxes represent the functional phenotypes of biochemical pathways and are essential to reveal the distribution of precursors among metabolic networks. Although analysis of metabolic fluxes, facilitated by stable isotope labeling and mass spectrometry detection, has been applied in the studies of plant metabolism, we lack experimental measurements for carbon flux towards lignin, one of the most abundant polymers in nature. RESULTS: We developed a feeding strategy of excised Arabidopsis stems with (13)C labeled phenylalanine (Phe) for the analysis of lignin biosynthetic flux. We optimized the feeding methods and found the stems continued to grow and lignify. Consistent with lignification profiles along the stems, higher levels of phenylpropanoids and activities of lignin biosynthetic enzymes were detected in the base of the stem. In the feeding experiments, (13)C labeled Phe was quickly accumulated and used for the synthesis of phenylpropanoid intermediates and lignin. The intermediates displayed two different patterns of labeling kinetics during the feeding period. Analysis of lignin showed rapid incorporation of label into all three subunits in the polymers. CONCLUSIONS: Our feeding results demonstrate the effectiveness of the stem feeding system and suggest a potential application for the investigations of other aspects in plant metabolism. The supply of exogenous Phe leading to a higher lignin deposition rate indicates the availability of Phe is a determining factor for lignification rates. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13007-018-0318-3) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6015466 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-60154662018-07-05 A (13)C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems Wang, Peng Guo, Longyun Jaini, Rohit Klempien, Antje McCoy, Rachel M. Morgan, John A. Dudareva, Natalia Chapple, Clint Plant Methods Research BACKGROUND: Metabolic fluxes represent the functional phenotypes of biochemical pathways and are essential to reveal the distribution of precursors among metabolic networks. Although analysis of metabolic fluxes, facilitated by stable isotope labeling and mass spectrometry detection, has been applied in the studies of plant metabolism, we lack experimental measurements for carbon flux towards lignin, one of the most abundant polymers in nature. RESULTS: We developed a feeding strategy of excised Arabidopsis stems with (13)C labeled phenylalanine (Phe) for the analysis of lignin biosynthetic flux. We optimized the feeding methods and found the stems continued to grow and lignify. Consistent with lignification profiles along the stems, higher levels of phenylpropanoids and activities of lignin biosynthetic enzymes were detected in the base of the stem. In the feeding experiments, (13)C labeled Phe was quickly accumulated and used for the synthesis of phenylpropanoid intermediates and lignin. The intermediates displayed two different patterns of labeling kinetics during the feeding period. Analysis of lignin showed rapid incorporation of label into all three subunits in the polymers. CONCLUSIONS: Our feeding results demonstrate the effectiveness of the stem feeding system and suggest a potential application for the investigations of other aspects in plant metabolism. The supply of exogenous Phe leading to a higher lignin deposition rate indicates the availability of Phe is a determining factor for lignification rates. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13007-018-0318-3) contains supplementary material, which is available to authorized users. BioMed Central 2018-06-23 /pmc/articles/PMC6015466/ /pubmed/29977324 http://dx.doi.org/10.1186/s13007-018-0318-3 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Wang, Peng Guo, Longyun Jaini, Rohit Klempien, Antje McCoy, Rachel M. Morgan, John A. Dudareva, Natalia Chapple, Clint A (13)C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems |
title | A (13)C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems |
title_full | A (13)C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems |
title_fullStr | A (13)C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems |
title_full_unstemmed | A (13)C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems |
title_short | A (13)C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems |
title_sort | (13)c isotope labeling method for the measurement of lignin metabolic flux in arabidopsis stems |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6015466/ https://www.ncbi.nlm.nih.gov/pubmed/29977324 http://dx.doi.org/10.1186/s13007-018-0318-3 |
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