Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Wang, Peng, Guo, Longyun, Jaini, Rohit, Klempien, Antje, McCoy, Rachel M., Morgan, John A., Dudareva, Natalia, Chapple, Clint
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
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
_version_ 1783334419618070528
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
work_keys_str_mv AT wangpeng a13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT guolongyun a13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT jainirohit a13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT klempienantje a13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT mccoyrachelm a13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT morganjohna a13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT dudarevanatalia a13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT chappleclint a13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT wangpeng 13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT guolongyun 13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT jainirohit 13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT klempienantje 13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT mccoyrachelm 13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT morganjohna 13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT dudarevanatalia 13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems
AT chappleclint 13cisotopelabelingmethodforthemeasurementofligninmetabolicfluxinarabidopsisstems