A 13C 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

doi:10.1186/s13007-018-0318-3

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

Journal Article · Sat Jun 23 00:00:00 EDT 2018 · Plant Methods

DOI:https://doi.org/10.1186/s13007-018-0318-3· OSTI ID:1618906

Wang, Peng; Guo, Longyun; Jaini, Rohit; Klempien, Antje; McCoy, Rachel M.;

;

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.

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Research Organization:: Purdue Univ., West Lafayette, IN (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0008628

OSTI ID:: 1618906

Alternate ID(s):: OSTI ID: 1499868

Journal Information:: Plant Methods, Journal Name: Plant Methods Vol. 14 Journal Issue: 1; ISSN 1746-4811

Publisher:: Springer Science + Business MediaCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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Cited by: 19 works

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59 BASIC BIOLOGICAL SCIENCES
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MOESM1 of A 13C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems Wang, Peng; Guo, Longyun; Jaini, Rohit https://doi.org/10.6084/m9.figshare.6663260.v1 The current record is supplemented by this text	text	January 2018
A 13C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems Wang, Peng; Guo, Longyun; Jaini, Rohit https://doi.org/10.6084/M9.FIGSHARE.C.4143608 The current record is supplemented by this collection	collection	January 2018
A 13C isotope labeling method for the measurement of lignin metabolic flux in Arabidopsis stems Wang, Peng; Guo, Longyun; Jaini, Rohit https://doi.org/10.6084/m9.figshare.c.4143608.v1 The current record is supplemented by this collection	collection	January 2018