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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]
  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:
Journal Article: 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.
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
Dalisay, Doralyn S., Kim, Kye Won, Lee, Choonseok, Yang, Hong, Rübel, Oliver, Bowen, Benjamin P., Davin, Laurence B., and Lewis, Norman G. 2015. "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.
@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},
url = {https://www.osti.gov/biblio/1179830}, journal = {Journal of Natural Products},
issn = {0163-3864},
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}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record at https://doi.org/10.1021/acs.jnatprod.5b00023

Citation Metrics:
Cited by: 77 works
Citation information provided by
Web of Science

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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Figures/Tables have been extracted from DOE-funded journal article accepted manuscripts.