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Title: Dirigent Protein-Mediated Lignan and Cyanogenic Glucoside Formation in Flax Seed: Integrated Omics and MALDI Mass Spectrometry Imaging

Abstract

An integrated omics approach using genomics, transcriptomics, metabolomics (MALDI mass spectrometry imaging, MSI), and bioinformatics was employed to study spatiotemporal formation and deposition of health-protecting polymeric lignans and plant defense cyanogenic glucosides. Intact flax (Linum usitatissimum) capsules and seed tissues at different development stages were analyzed. Transcriptome analyses indicated distinct expression patterns of dirigent protein (DP) gene family members encoding (-)- and (+)-pinoresinol-forming DPs and their associated downstream metabolic processes, respectively, with the former expressed at early seed coat development stages. Genes encoding (+)-pinoresinol-forming DPs were, in contrast, expressed at later development stages. Recombinant DP expression and DP assays also unequivocally established their distinct stereoselective biochemical functions. Using MALDI MSI and ion mobility separation analyses, the pinoresinol downstream derivatives, secoisolariciresinol diglucoside (SDG) and SDG hydroxymethylglutaryl ester, were localized and detectable only in early seed coat development stages. SDG derivatives were then converted into higher molecular weight phenolics during seed coat maturation. By contrast, the plant defense cyanogenic glucosides, the monoglucosides linamarin/lotaustralin, were detected throughout the flax capsule, whereas diglucosides linustatin/neolinustatin only accumulated in endosperm and embryo tissues. A putative biosynthetic pathway to the cyanogens is proposed on the basis of transcriptome coexpression data. Localization of all metabolites was at ca.more » 20 μm resolution, with the web based tool OpenMSI enabling not only resolution enhancement but also an interactive system for real-time searching for any ion in the tissue under analysis.« less

Authors:
 [1];  [1];  [1];  [1];  [2];  [3];  [1];  [1]
+ Show Author Affiliations
  1. Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340, United States
  2. Computational Research Division, Lawrence Berkeley National Laboratory, One Cyclotron Road, Berkeley, California 94720, United States
  3. Life Sciences Division, Lawrence Berkeley National Laboratory, One Cyclotron Road, Berkeley, California 94720, United States
Publication Date:
Research Org.:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1179830
Alternate Identifier(s):
OSTI ID: 1512206
Grant/Contract Number:  
FG-0397ER20259; AC02-05CH11231
Resource Type:
Published Article
Journal Name:
Journal of Natural Products
Additional Journal Information:
Journal Name: Journal of Natural Products Journal Volume: 78 Journal Issue: 6; Journal ID: ISSN 0163-3864
Publisher:
American Chemical Society
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Dalisay, Doralyn S., Kim, Kye Won, Lee, Choonseok, Yang, Hong, Rübel, Oliver, Bowen, Benjamin P., Davin, Laurence B., and Lewis, Norman G. Dirigent Protein-Mediated Lignan and Cyanogenic Glucoside Formation in Flax Seed: Integrated Omics and MALDI Mass Spectrometry Imaging. United States: N. p., 2015. Web. doi:10.1021/acs.jnatprod.5b00023.
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Dalisay, Doralyn S., Kim, Kye Won, Lee, Choonseok, Yang, Hong, Rübel, Oliver, Bowen, Benjamin P., Davin, Laurence B., & Lewis, Norman G. Dirigent Protein-Mediated Lignan and Cyanogenic Glucoside Formation in Flax Seed: Integrated Omics and MALDI Mass Spectrometry Imaging. United States. https://doi.org/10.1021/acs.jnatprod.5b00023
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Dalisay, Doralyn S., Kim, Kye Won, Lee, Choonseok, Yang, Hong, Rübel, Oliver, Bowen, Benjamin P., Davin, Laurence B., and Lewis, Norman G. Sun . "Dirigent Protein-Mediated Lignan and Cyanogenic Glucoside Formation in Flax Seed: Integrated Omics and MALDI Mass Spectrometry Imaging". United States. https://doi.org/10.1021/acs.jnatprod.5b00023.
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@article{osti_1179830,
title = {Dirigent Protein-Mediated Lignan and Cyanogenic Glucoside Formation in Flax Seed: Integrated Omics and MALDI Mass Spectrometry Imaging},
author = {Dalisay, Doralyn S. and Kim, Kye Won and Lee, Choonseok and Yang, Hong and Rübel, Oliver and Bowen, Benjamin P. and Davin, Laurence B. and Lewis, Norman G.},
abstractNote = {An integrated omics approach using genomics, transcriptomics, metabolomics (MALDI mass spectrometry imaging, MSI), and bioinformatics was employed to study spatiotemporal formation and deposition of health-protecting polymeric lignans and plant defense cyanogenic glucosides. Intact flax (Linum usitatissimum) capsules and seed tissues at different development stages were analyzed. Transcriptome analyses indicated distinct expression patterns of dirigent protein (DP) gene family members encoding (-)- and (+)-pinoresinol-forming DPs and their associated downstream metabolic processes, respectively, with the former expressed at early seed coat development stages. Genes encoding (+)-pinoresinol-forming DPs were, in contrast, expressed at later development stages. Recombinant DP expression and DP assays also unequivocally established their distinct stereoselective biochemical functions. Using MALDI MSI and ion mobility separation analyses, the pinoresinol downstream derivatives, secoisolariciresinol diglucoside (SDG) and SDG hydroxymethylglutaryl ester, were localized and detectable only in early seed coat development stages. SDG derivatives were then converted into higher molecular weight phenolics during seed coat maturation. By contrast, the plant defense cyanogenic glucosides, the monoglucosides linamarin/lotaustralin, were detected throughout the flax capsule, whereas diglucosides linustatin/neolinustatin only accumulated in endosperm and embryo tissues. A putative biosynthetic pathway to the cyanogens is proposed on the basis of transcriptome coexpression data. Localization of all metabolites was at ca. 20 μm resolution, with the web based tool OpenMSI enabling not only resolution enhancement but also an interactive system for real-time searching for any ion in the tissue under analysis.},
doi = {10.1021/acs.jnatprod.5b00023},
journal = {Journal of Natural Products},
number = 6,
volume = 78,
place = {United States},
year = {Sun May 17 00:00:00 EDT 2015},
month = {Sun May 17 00:00:00 EDT 2015}
}
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Publisher's Version of Record
https://doi.org/10.1021/acs.jnatprod.5b00023
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Figures / Tables:

Figure 1 Figure 1: (A) Flax lignans (1−3) and cyanogenic glucosides (4−7). (B) Biochemical pathway to secoisolariciresinol diglucoside (SDG) (1).
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Works referenced in this record:

Structural characterization of flavonoid glycosides by multi-stage mass spectrometry: MS CHARACTERIZATION OF FLAVONOID GLYCOSIDES
journal, December 2008

  • Vukics, Viktoria; Guttman, András
  • Mass Spectrometry Reviews, Vol. 29, Issue 1
  • DOI: 10.1002/mas.20212

An improved RNA isolation method for succulent plant species rich in polyphenols and polysaccharides
journal, December 2000

  • Gehrig, H. H.; Winter, K.; Cushman, J.
  • Plant Molecular Biology Reporter, Vol. 18, Issue 4
  • DOI: 10.1007/BF02825065

Properties and functions of the cyanogenic system in higher plants
journal, June 1990

Opposite Stereoselectivities of Dirigent Proteins in Arabidopsis and Schizandra Species
journal, August 2012

  • Kim, Kye-Won; Moinuddin, Syed G. A.; Atwell, Kathleen M.
  • Journal of Biological Chemistry, Vol. 287, Issue 41
  • DOI: 10.1074/jbc.M112.387423

Seed coats: Structure, development, composition, and biotechnology
journal, September 2005

  • Moïse, Jaimie A.; Han, Shuyou; Gudynaitę-Savitch, Loreta
  • In Vitro Cellular & Developmental Biology - Plant, Vol. 41, Issue 5
  • DOI: 10.1079/IVP2005686

Next Generation Sequencing in Predicting Gene Function in Podophyllotoxin Biosynthesis
journal, January 2013

  • Marques, Joaquim V.; Kim, Kye-Won; Lee, Choonseok
  • Journal of Biological Chemistry, Vol. 288, Issue 1
  • DOI: 10.1074/jbc.M112.400689

Characterization of flavone and flavonol aglycones by collision-induced dissociation tandem mass spectrometry
journal, August 1997

Compartmentation of cyanogenic glucosides and their degrading enzymes
journal, November 1994

  • Gruhnert, Christiane; Biehl, B�le; Selmar, Dirk
  • Planta, Vol. 195, Issue 1
  • DOI: 10.1007/BF00206289

Non-host disease resistance response in pea (Pisum sativum) pods: Biochemical function of DRR206 and phytoalexin pathway localization
journal, May 2015

A multi-omics strategy resolves the elusive nature of alkaloids in Podophyllum species
journal, January 2014

  • Marques, Joaquim V.; Dalisay, Doralyn S.; Yang, Hong
  • Mol. BioSyst., Vol. 10, Issue 11
  • DOI: 10.1039/C4MB00403E

Recombinant Pinoresinol-Lariciresinol Reductases from Western Red Cedar (Thuja plicata) Catalyze Opposite Enantiospecific Conversions
journal, January 1999

  • Fujita, Masayuki; Gang, David R.; Davin, Laurence B.
  • Journal of Biological Chemistry, Vol. 274, Issue 2
  • DOI: 10.1074/jbc.274.2.618

Characterization of Arabidopsis thaliana Pinoresinol Reductase, a New Type of Enzyme Involved in Lignan Biosynthesis
journal, June 2008

  • Nakatsubo, Tomoyuki; Mizutani, Masaharu; Suzuki, Shiro
  • Journal of Biological Chemistry, Vol. 283, Issue 23
  • DOI: 10.1074/jbc.M801131200

Selection of reference genes for quantitative gene expression normalization in flax (Linum usitatissimum L.)
journal, January 2010

The genome of flax ( Linum usitatissimum ) assembled de novo from short shotgun sequence reads
journal, August 2012

Diversity of preferred nucleotide sequences around the translation initiation codon in eukaryote genomes
journal, November 2007

  • Nakagawa, S.; Niimura, Y.; Gojobori, T.
  • Nucleic Acids Research, Vol. 36, Issue 3
  • DOI: 10.1093/nar/gkm1102

OpenMSI: A High-Performance Web-Based Platform for Mass Spectrometry Imaging
journal, October 2013

  • Rübel, Oliver; Greiner, Annette; Cholia, Shreyas
  • Analytical Chemistry, Vol. 85, Issue 21
  • DOI: 10.1021/ac402540a

RNAi-mediated pinoresinol lariciresinol reductase gene silencing in flax (Linum usitatissimum L.) seed coat: Consequences on lignans and neolignans accumulation
journal, September 2014

  • Renouard, Sullivan; Tribalatc, Marie-Aude; Lamblin, Frederic
  • Journal of Plant Physiology, Vol. 171, Issue 15
  • DOI: 10.1016/j.jplph.2014.06.005

Structure and Development of Medicago truncatula Pod Wall and Seed Coat
journal, February 2005

Trimeric Structure of (+)-Pinoresinol-forming Dirigent Protein at 1.95 Å Resolution with Three Isolated Active Sites
journal, November 2014

  • Kim, Kye-Won; Smith, Clyde A.; Daily, Michael D.
  • Journal of Biological Chemistry, Vol. 290, Issue 3
  • DOI: 10.1074/jbc.M114.611780

Flax seed lignan in disease prevention and health promotion
journal, October 2003

Transgenic Hybrid Poplar for Sustainable and Scalable Production of the Commodity/Specialty Chemical, 2-Phenylethanol
journal, December 2013

A combined HPLC-UV and HPLC-MS method for the identification of lignans and its application to the lignans ofLinum usitatissimum L. andL. bienne Mill.
journal, January 2006

  • Schmidt, Thomas J.; Hemmati, Shiva; Fuss, Elisabeth
  • Phytochemical Analysis, Vol. 17, Issue 5
  • DOI: 10.1002/pca.918

Isolation and characterization of two cyanogenic β-glucosidases from flax seeds
journal, December 1985

An Enantiocomplementary Dirigent Protein for the Enantioselective Laccase-Catalyzed Oxidative Coupling of Phenols
journal, November 2009

  • Pickel, Benjamin; Constantin, Mihaela-Anca; Pfannstiel, Jens
  • Angewandte Chemie International Edition, Vol. 49, Issue 1
  • DOI: 10.1002/anie.200904622

A systematic nomenclature for carbohydrate fragmentations in FAB-MS/MS spectra of glycoconjugates
journal, January 1988

  • Domon, Bruno; Costello, Catherine E.
  • Glycoconjugate Journal, Vol. 5, Issue 4
  • DOI: 10.1007/BF01049915

(+)-Pinoresinol/(+)-Lariciresinol Reductase from Forsythia intermedia : PROTEIN PURIFICATION, cDNA CLONING, HETEROLOGOUS EXPRESSION AND COMPARISON TO ISOFLAVONE REDUCTASE
journal, November 1996

  • Dinkova-Kostova, Albena T.; Gang, David R.; Davin, Laurence B.
  • Journal of Biological Chemistry, Vol. 271, Issue 46
  • DOI: 10.1074/jbc.271.46.29473

A small-scale procedure for the rapid isolation of plant RNAs
journal, January 1989

  • Verwoerd, Theo C.; Dekker, Ben M. M.; Hoekema, Andre
  • Nucleic Acids Research, Vol. 17, Issue 6
  • DOI: 10.1093/nar/17.6.2362

Stereoselective Bimolecular Phenoxy Radical Coupling by an Auxiliary (Dirigent) Protein Without an Active Center
journal, January 1997

Pinoresinol–lariciresinol reductases with different stereospecificity from Linum album and Linum usitatissimum
journal, June 2005

Metabolism of Hydrogen Cyanide by Higher Plants
journal, June 1980

Development of antibodies against secoisolariciresinol – Application to the immunolocalization of lignans in Linum usitatissimum seeds
journal, December 2010

On the stereoselective synthesis of (+)-pinoresinol in Forsythia suspensa from its achiral precursor, coniferyl alcohol
journal, January 1992

Analysis of the developmental regulation of the cyanogenic compounds in seedlings of two lines of Linum usitatissimum L.
journal, October 2003

  • Krech, M. J.; Fieldes, M. A.
  • Canadian Journal of Botany, Vol. 81, Issue 10
  • DOI: 10.1139/b03-097

Cyanogenic Glucosides In Linum Usitatissimum
journal, September 1998

Mobilization and Utilization of Cyanogenic Glycosides: The Linustatin Pathway
journal, March 1988

  • Selmar, Dirk; Lieberei, Reinhard; Biehl, Böle
  • Plant Physiology, Vol. 86, Issue 3
  • DOI: 10.1104/pp.86.3.711

An oligomer from flaxseed composed of secoisolariciresinoldiglucoside and 3-hydroxy-3-methyl glutaric acid residues
journal, October 2001

Investigation of lignan accumulation in developing Linum usitatissimum seeds by immunolocalization and HPLC
journal, June 2011

  • Attoumbré, Jacques; Mahamane Laoualy, Abarchi Boubé; Bienaimé, Christophe
  • Phytochemistry Letters, Vol. 4, Issue 2
  • DOI: 10.1016/j.phytol.2011.03.004
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