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

Dalisay, Doralyn S.; Kim, Kye Won; Lee, Choonseok; Yang, Hong; Rübel, Oliver; Bowen, Benjamin P.; Davin, Laurence B.; Lewis, Norman G.

doi:10.1021/acs.jnatprod.5b00023

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

Journal Article · Sun May 17 00:00:00 EDT 2015 · Journal of Natural Products

DOI:https://doi.org/10.1021/acs.jnatprod.5b00023· OSTI ID:1179830

Dalisay, Doralyn S. ^[1]; Kim, Kye Won ^[1]; Lee, Choonseok ^[1]; Yang, Hong ^[1]; Rübel, Oliver ^[2]; Bowen, Benjamin P. ^[3]; Davin, Laurence B. ^[1]; Lewis, Norman G. ^[1]

Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340, United States
Computational Research Division, Lawrence Berkeley National Laboratory, One Cyclotron Road, Berkeley, California 94720, United States
Life Sciences Division, Lawrence Berkeley National Laboratory, One Cyclotron Road, Berkeley, California 94720, United States

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.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: FG-0397ER20259; AC02-05CH11231

OSTI ID:: 1179830

Alternate ID(s):: OSTI ID: 1512206

Journal Information:: Journal of Natural Products, Journal Name: Journal of Natural Products Vol. 78 Journal Issue: 6; ISSN 0163-3864

Publisher:: American Chemical SocietyCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 77 works

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References (46)

Structural characterization of flavonoid glycosides by multi-stage mass spectrometry: MS CHARACTERIZATION OF FLAVONOID GLYCOSIDES Vukics, Viktoria; Guttman, András Mass Spectrometry Reviews, Vol. 29, Issue 1 https://doi.org/10.1002/mas.20212	journal	December 2008
An improved RNA isolation method for succulent plant species rich in polyphenols and polysaccharides Gehrig, H. H.; Winter, K.; Cushman, J. Plant Molecular Biology Reporter, Vol. 18, Issue 4 https://doi.org/10.1007/BF02825065	journal	December 2000
Properties and functions of the cyanogenic system in higher plants Kakes, P. Euphytica, Vol. 48, Issue 1 https://doi.org/10.1007/BF00028958	journal	June 1990
Identification and functional characterization of a flax UDP-glycosyltransferase glucosylating secoisolariciresinol (SECO) into secoisolariciresinol monoglucoside (SMG) and diglucoside (SDG) Ghose, Kaushik; Selvaraj, Kumarakurubaran; McCallum, Jason BMC Plant Biology, Vol. 14, Issue 1 https://doi.org/10.1186/1471-2229-14-82	journal	January 2014
Opposite Stereoselectivities of Dirigent Proteins in Arabidopsis and Schizandra Species Kim, Kye-Won; Moinuddin, Syed G. A.; Atwell, Kathleen M. Journal of Biological Chemistry, Vol. 287, Issue 41 https://doi.org/10.1074/jbc.M112.387423	journal	August 2012
Seed coats: Structure, development, composition, and biotechnology Moïse, Jaimie A.; Han, Shuyou; Gudynaitę-Savitch, Loreta In Vitro Cellular & Developmental Biology - Plant, Vol. 41, Issue 5 https://doi.org/10.1079/IVP2005686	journal	September 2005
Next Generation Sequencing in Predicting Gene Function in Podophyllotoxin Biosynthesis Marques, Joaquim V.; Kim, Kye-Won; Lee, Choonseok Journal of Biological Chemistry, Vol. 288, Issue 1 https://doi.org/10.1074/jbc.M112.400689	journal	January 2013
Characterization of flavone and flavonol aglycones by collision-induced dissociation tandem mass spectrometry Ma, Y. L.; Li, Q. M.; Van den Heuvel, H. Rapid Communications in Mass Spectrometry, Vol. 11, Issue 12 https://doi.org/10.1002/(SICI)1097-0231(199708)11:12<1357::AID-RCM983>3.0.CO;2-9	journal	August 1997
Compartmentation of cyanogenic glucosides and their degrading enzymes Gruhnert, Christiane; Biehl, B�le; Selmar, Dirk Planta, Vol. 195, Issue 1 https://doi.org/10.1007/BF00206289	journal	November 1994
Non-host disease resistance response in pea (Pisum sativum) pods: Biochemical function of DRR206 and phytoalexin pathway localization Seneviratne, Herana Kamal; Dalisay, Doralyn S.; Kim, Kye-Won Phytochemistry, Vol. 113 https://doi.org/10.1016/j.phytochem.2014.10.013	journal	May 2015
Pinoresinol–lariciresinol reductase gene expression and secoisolariciresinol diglucoside accumulation in developing flax (Linum usitatissimum) seeds Hano, C.; Martin, I.; Fliniaux, O. Planta, Vol. 224, Issue 6 https://doi.org/10.1007/s00425-006-0308-y	journal	May 2006
A multi-omics strategy resolves the elusive nature of alkaloids in Podophyllum species Marques, Joaquim V.; Dalisay, Doralyn S.; Yang, Hong Mol. BioSyst., Vol. 10, Issue 11 https://doi.org/10.1039/C4MB00403E	journal	January 2014
Recombinant Pinoresinol-Lariciresinol Reductases from Western Red Cedar (Thuja plicata) Catalyze Opposite Enantiospecific Conversions Fujita, Masayuki; Gang, David R.; Davin, Laurence B. Journal of Biological Chemistry, Vol. 274, Issue 2 https://doi.org/10.1074/jbc.274.2.618	journal	January 1999
Characterization of Arabidopsis thaliana Pinoresinol Reductase, a New Type of Enzyme Involved in Lignan Biosynthesis Nakatsubo, Tomoyuki; Mizutani, Masaharu; Suzuki, Shiro Journal of Biological Chemistry, Vol. 283, Issue 23 https://doi.org/10.1074/jbc.M801131200	journal	June 2008
Characterization and expression profile of two UDP-glucosyltransferases, UGT85K4 and UGT85K5, catalyzing the last step in cyanogenic glucoside biosynthesis in cassava: Manihot esculenta cyanohydrin glucosyltransferases Kannangara, Rubini; Motawia, Mohammed S.; Hansen, Natascha K. K. The Plant Journal, Vol. 68, Issue 2 https://doi.org/10.1111/j.1365-313X.2011.04695.x	journal	August 2011
Selection of reference genes for quantitative gene expression normalization in flax (Linum usitatissimum L.) Huis, Rudy; Hawkins, Simon; Neutelings, Godfrey BMC Plant Biology, Vol. 10, Issue 1 https://doi.org/10.1186/1471-2229-10-71	journal	January 2010
The genome of flax ( Linum usitatissimum ) assembled de novo from short shotgun sequence reads Wang, Zhiwen; Hobson, Neil; Galindo, Leonardo The Plant Journal, Vol. 72, Issue 3 https://doi.org/10.1111/j.1365-313X.2012.05093.x	journal	August 2012
Diversity of preferred nucleotide sequences around the translation initiation codon in eukaryote genomes Nakagawa, S.; Niimura, Y.; Gojobori, T. Nucleic Acids Research, Vol. 36, Issue 3 https://doi.org/10.1093/nar/gkm1102	journal	November 2007
OpenMSI: A High-Performance Web-Based Platform for Mass Spectrometry Imaging Rübel, Oliver; Greiner, Annette; Cholia, Shreyas Analytical Chemistry, Vol. 85, Issue 21 https://doi.org/10.1021/ac402540a	journal	October 2013
RNAi-mediated pinoresinol lariciresinol reductase gene silencing in flax (Linum usitatissimum L.) seed coat: Consequences on lignans and neolignans accumulation Renouard, Sullivan; Tribalatc, Marie-Aude; Lamblin, Frederic Journal of Plant Physiology, Vol. 171, Issue 15 https://doi.org/10.1016/j.jplph.2014.06.005	journal	September 2014
Biosynthesis of the Cyanogenic Glucosides Linamarin and Lotaustralin in Cassava: Isolation, Biochemical Characterization, and Expression Pattern of CYP71E7, the Oxime-Metabolizing Cytochrome P450 Enzyme Jørgensen, Kirsten; Morant, Anne Vinther; Morant, Marc Plant Physiology, Vol. 155, Issue 1 https://doi.org/10.1104/pp.110.164053	journal	November 2010
Structure and Development of Medicago truncatula Pod Wall and Seed Coat Wang, H. L. Annals of Botany, Vol. 95, Issue 5 https://doi.org/10.1093/aob/mci080	journal	February 2005
Trimeric Structure of (+)-Pinoresinol-forming Dirigent Protein at 1.95 Å Resolution with Three Isolated Active Sites Kim, Kye-Won; Smith, Clyde A.; Daily, Michael D. Journal of Biological Chemistry, Vol. 290, Issue 3 https://doi.org/10.1074/jbc.M114.611780	journal	November 2014
Pattern of the Cyanide-Potential in Developing Fruits: Implications for Plants Accumulating Cyanogenic Monoglucosides ( Phaseolus lunatus ) or Cyanogenic Diglucosides in Their Seeds ( Linum usitatissimum, Prunus amygdalus ) Frehner, Marco; Scalet, Mario; Conn, Eric E. Plant Physiology, Vol. 94, Issue 1 https://doi.org/10.1104/pp.94.1.28	journal	September 1990
Flax seed lignan in disease prevention and health promotion Westcott, Neil D.; Muir, Alister D. Phytochemistry Reviews, Vol. 2, Issue 3 https://doi.org/10.1023/B:PHYT.0000046174.97809.b6	journal	October 2003
Transgenic Hybrid Poplar for Sustainable and Scalable Production of the Commodity/Specialty Chemical, 2-Phenylethanol Costa, Michael A.; Marques, Joaquim V.; Dalisay, Doralyn S. PLoS ONE, Vol. 8, Issue 12 https://doi.org/10.1371/journal.pone.0083169	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. Schmidt, Thomas J.; Hemmati, Shiva; Fuss, Elisabeth Phytochemical Analysis, Vol. 17, Issue 5 https://doi.org/10.1002/pca.918	journal	January 2006
Isolation and characterization of two cyanogenic β-glucosidases from flax seeds Fan, Teresa W. -M.; Conn, Eric E. Archives of Biochemistry and Biophysics, Vol. 243, Issue 2 https://doi.org/10.1016/0003-9861(85)90513-2	journal	December 1985
An Enantiocomplementary Dirigent Protein for the Enantioselective Laccase-Catalyzed Oxidative Coupling of Phenols Pickel, Benjamin; Constantin, Mihaela-Anca; Pfannstiel, Jens Angewandte Chemie International Edition, Vol. 49, Issue 1 https://doi.org/10.1002/anie.200904622	journal	November 2009
Cytochromes P-450 from Cassava ( Manihot esculenta Crantz) Catalyzing the First Steps in the Biosynthesis of the Cyanogenic Glucosides Linamarin and Lotaustralin: CLONING, FUNCTIONAL EXPRESSION IN PICHIA PASTORIS , AND SUBSTRATE SPECIFICITY OF THE ISOLATED RECOMBINANT ENZYMES Andersen, Mette Dahl; Busk, Peter Kamp; Svendsen, Ib Journal of Biological Chemistry, Vol. 275, Issue 3 https://doi.org/10.1074/jbc.275.3.1966	journal	January 2000
A systematic nomenclature for carbohydrate fragmentations in FAB-MS/MS spectra of glycoconjugates Domon, Bruno; Costello, Catherine E. Glycoconjugate Journal, Vol. 5, Issue 4 https://doi.org/10.1007/BF01049915	journal	January 1988
(+)-Pinoresinol/(+)-Lariciresinol Reductase from Forsythia intermedia : PROTEIN PURIFICATION, cDNA CLONING, HETEROLOGOUS EXPRESSION AND COMPARISON TO ISOFLAVONE REDUCTASE Dinkova-Kostova, Albena T.; Gang, David R.; Davin, Laurence B. Journal of Biological Chemistry, Vol. 271, Issue 46 https://doi.org/10.1074/jbc.271.46.29473	journal	November 1996
A small-scale procedure for the rapid isolation of plant RNAs Verwoerd, Theo C.; Dekker, Ben M. M.; Hoekema, Andre Nucleic Acids Research, Vol. 17, Issue 6 https://doi.org/10.1093/nar/17.6.2362	journal	January 1989
Stereoselective Bimolecular Phenoxy Radical Coupling by an Auxiliary (Dirigent) Protein Without an Active Center Davin, L. B.; Wang, H. -B.; Crowell, A. L. Science, Vol. 275, Issue 5298 https://doi.org/10.1126/science.275.5298.362	journal	January 1997
Pinoresinol–lariciresinol reductases with different stereospecificity from Linum album and Linum usitatissimum von Heimendahl, Cosima B. I.; Schäfer, Katrin M.; Eklund, Patrik Phytochemistry, Vol. 66, Issue 11 https://doi.org/10.1016/j.phytochem.2005.04.026	journal	June 2005
Metabolism of Hydrogen Cyanide by Higher Plants Miller, Jacqueline M.; Conn, Eric E. Plant Physiology, Vol. 65, Issue 6 https://doi.org/10.1104/pp.65.6.1199	journal	June 1980
The chain length of lignan macromolecule from flaxseed hulls is determined by the incorporation of coumaric acid glucosides and ferulic acid glucosides Struijs, Karin; Vincken, Jean-Paul; Doeswijk, Timo G. Phytochemistry, Vol. 70, Issue 2 https://doi.org/10.1016/j.phytochem.2008.12.015	journal	January 2009
Biosynthetic Pathway to the Cancer Chemopreventive Secoisolariciresinol Diglucoside−Hydroxymethyl Glutaryl Ester-Linked Lignan Oligomers in Flax ( Linum u sitatissimum) Seed ^† Ford, Joshua D.; Huang, Kai-Sheng; Wang, Huai-Bin Journal of Natural Products, Vol. 64, Issue 11 https://doi.org/10.1021/np010367x	journal	November 2001
Development of antibodies against secoisolariciresinol – Application to the immunolocalization of lignans in Linum usitatissimum seeds Attoumbré, Jacques; Bienaimé, Christophe; Dubois, Frédéric Phytochemistry, Vol. 71, Issue 17-18 https://doi.org/10.1016/j.phytochem.2010.09.002	journal	December 2010
Pinoresinol-Lariciresinol Reductases with Opposite Enantiospecificity Determine the Enantiomeric Composition of Lignans in the Different Organs of Linum usitatissimum L. Hemmati, Shiva; Heimendahl, Cosima; Klaes, Michael Planta Medica, Vol. 76, Issue 09 https://doi.org/10.1055/s-0030-1250036	journal	May 2010
On the stereoselective synthesis of (+)-pinoresinol in Forsythia suspensa from its achiral precursor, coniferyl alcohol Davin, Laurence B.; Bedgar, Diana L.; Katayama, Takeshi Phytochemistry, Vol. 31, Issue 11 https://doi.org/10.1016/S0031-9422(00)97544-7	journal	January 1992
Analysis of the developmental regulation of the cyanogenic compounds in seedlings of two lines of Linum usitatissimum L. Krech, M. J.; Fieldes, M. A. Canadian Journal of Botany, Vol. 81, Issue 10 https://doi.org/10.1139/b03-097	journal	October 2003
Cyanogenic Glucosides In Linum Usitatissimum Niedźwiedź-Siegień, Irena Phytochemistry, Vol. 49, Issue 1 https://doi.org/10.1016/S0031-9422(97)00953-9	journal	September 1998
Mobilization and Utilization of Cyanogenic Glycosides: The Linustatin Pathway Selmar, Dirk; Lieberei, Reinhard; Biehl, Böle Plant Physiology, Vol. 86, Issue 3 https://doi.org/10.1104/pp.86.3.711	journal	March 1988
An oligomer from flaxseed composed of secoisolariciresinoldiglucoside and 3-hydroxy-3-methyl glutaric acid residues Kamal-Eldin, Afaf; Peerlkamp, Nienke; Johnsson, Pernilla Phytochemistry, Vol. 58, Issue 4 https://doi.org/10.1016/S0031-9422(01)00279-5	journal	October 2001
Investigation of lignan accumulation in developing Linum usitatissimum seeds by immunolocalization and HPLC Attoumbré, Jacques; Mahamane Laoualy, Abarchi Boubé; Bienaimé, Christophe Phytochemistry Letters, Vol. 4, Issue 2 https://doi.org/10.1016/j.phytol.2011.03.004	journal	June 2011