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Title: A 13C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems

Abstract

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 13C 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, 13C 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. Moreover, the supplymore » of exogenous Phe leading to a higher lignin deposition rate indicates the availability of Phe is a determining factor for lignification rates.« less

Authors:
 [1];  [1];  [1];  [1];  [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]
  1. Purdue Univ., West Lafayette, IN (United States)
Publication Date:
Research Org.:
Purdue Univ., West Lafayette, IN (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1499868
Grant/Contract Number:  
SC0008628
Resource Type:
Accepted Manuscript
Journal Name:
Plant Methods
Additional Journal Information:
Journal Volume: 14; Journal Issue: 1; Journal ID: ISSN 1746-4811
Publisher:
BioMed Central
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Stable isotope labeling; Stem feeding; Lignin; Phenylpropanoids

Citation Formats

Wang, Peng, Guo, Longyun, Jaini, Rohit, Klempien, Antje, McCoy, Rachel M., Morgan, John A., Dudareva, Natalia, and Chapple, Clint. A 13C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems. United States: N. p., 2018. Web. doi:10.1186/s13007-018-0318-3.
Wang, Peng, Guo, Longyun, Jaini, Rohit, Klempien, Antje, McCoy, Rachel M., Morgan, John A., Dudareva, Natalia, & Chapple, Clint. A 13C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems. United States. doi:10.1186/s13007-018-0318-3.
Wang, Peng, Guo, Longyun, Jaini, Rohit, Klempien, Antje, McCoy, Rachel M., Morgan, John A., Dudareva, Natalia, and Chapple, Clint. Sat . "A 13C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems". United States. doi:10.1186/s13007-018-0318-3. https://www.osti.gov/servlets/purl/1499868.
@article{osti_1499868,
title = {A 13C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems},
author = {Wang, Peng and Guo, Longyun and Jaini, Rohit and Klempien, Antje and McCoy, Rachel M. and Morgan, John A. and Dudareva, Natalia and Chapple, Clint},
abstractNote = {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 13C 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, 13C 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. Moreover, the supply of exogenous Phe leading to a higher lignin deposition rate indicates the availability of Phe is a determining factor for lignification rates.},
doi = {10.1186/s13007-018-0318-3},
journal = {Plant Methods},
number = 1,
volume = 14,
place = {United States},
year = {2018},
month = {6}
}

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    Works referencing / citing this record:

    4-Coumarate 3-hydroxylase in the lignin biosynthesis pathway is a cytosolic ascorbate peroxidase
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