Plant metabolomics: analytical platforms and integration with functional genomics

As the final downstream product of the genome, the plant metabolome is a highly complex, dynamic assortment of primary and secondary compounds. Although technological platforms to study genomes, transcriptomes and even proteomes are presently available, methods to pursue genuine metabolomics have not yet been developed due to the extensive chemical diversity of plant primary and secondary metabolites. No single analytical method can accurately survey the entire metabolome. However, recent technical, chemometric and bioinformatic advances promise to enhance our global understanding of plant metabolism. Separation-based mass spectrometry (MS) approaches, such as gas (GC) or liquid chromatography (LC)-MS, are relatively inexpensive, highly sensitive and provide excellent identifying capacity. However, Fourier transform-ion cyclotron resonance (FT-ICR)-MS is better suited for rapid, high-throughput applications and is currently the most sensitive method available. Unlike MS-based analyses, nuclear magnetic resonance (NMR) spectroscopy provides a large amount of information regarding molecular structure, and novel software innovations have facilitated the unequivocal identification and absolute quantification of compounds within composite samples. Due to the size and complexity of metabolomics datasets, numerous chemometric methods are used to extract and display systematic variation. Coupled with pattern recognition techniques and plant-specific metabolite databases, broad-scope metabolite analyses have emerged as functional genomics tools for novel gene discovery and functional characterization. In this review, key metabolomics technologies are compared and the applications of FT-ICR-MS and NMR to the study of benzylisoquinoline alkaloid metabolism in opium poppy are discussed.

[1]  Robert Verpoorte,et al.  Metabolomic Differentiation of Brassica rapa Following Herbivory by Different Insect Instars using Two-Dimensional Nuclear Magnetic Resonance Spectroscopy , 2006, Journal of Chemical Ecology.

[2]  M. Hirai,et al.  Integration of transcriptomics and metabolomics for understanding of global responses to nutritional stresses in Arabidopsis thaliana. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[3]  N. Kruger,et al.  Quantitative approaches for analysing fluxes through plant metabolic networks using NMR and stable isotope labelling , 2004, Phytochemistry Reviews.

[4]  John Skilling,et al.  Data analysis : a Bayesian tutorial , 1996 .

[5]  Jamin C. Hoggard,et al.  Recent advancements in comprehensive two-dimensional separations with chemometrics. , 2008, Journal of chromatography. A.

[6]  E Holmes,et al.  Chemometric contributions to the evolution of metabonomics: mathematical solutions to characterising and interpreting complex biological NMR spectra. , 2002, The Analyst.

[7]  A. J. Fist,et al.  Comparative qualitative and quantitative determination of alkaloids in narcotic and condiment Papaver somniferum cultivars. , 2005, Journal of natural products.

[8]  M. Greenacre Theory of Correspondence Analysis , 2007 .

[9]  T. Górecki,et al.  Recent Advances in Comprehensive Two‐Dimensional Gas Chromatography (GC×GC) , 2006 .

[10]  C. Enke,et al.  Practical implications of some recent studies in electrospray ionization fundamentals. , 2001, Mass spectrometry reviews.

[11]  L. Rabiner,et al.  An introduction to hidden Markov models , 1986, IEEE ASSP Magazine.

[12]  Benjamin P. MacLeod,et al.  Evidence for the monophyletic evolution of benzylisoquinoline alkaloid biosynthesis in angiosperms. , 2005, Phytochemistry.

[13]  R. Shellie Comprehensive Two-Dimensional Gas Chromatography–Mass Spectrometry and its Use in High-Resolution Metabolomics , 2005 .

[14]  Johan Trygg,et al.  Chemometrics in metabonomics. , 2007, Journal of proteome research.

[15]  W. Dunn,et al.  Measuring the metabolome: current analytical technologies. , 2005, The Analyst.

[16]  Joachim Selbig,et al.  The Metabolic Response of Heterotrophic Arabidopsis Cells to Oxidative Stress1[W] , 2006, Plant Physiology.

[17]  R. Ratcliffe,et al.  In vivo NMR Studies of Higher Plants and Algae , 1994 .

[18]  T. Kuromori,et al.  Top-down Phenomics of Arabidopsis thaliana , 2007, Journal of Biological Chemistry.

[19]  A. Aharoni,et al.  Terpenoid Metabolism in Wild-Type and Transgenic Arabidopsis Plants Online version contains Web-only data. Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.016253. , 2003, The Plant Cell Online.

[20]  Kazuo Shinozaki,et al.  Stable isotope labeling of Arabidopsis thaliana for an NMR-based metabolomics approach. , 2004, Plant & cell physiology.

[21]  Dong Xu,et al.  Bioinformatics and its applications in plant biology. , 2006, Annual review of plant biology.

[22]  W. El-Deredy,et al.  Pattern recognition approaches in biomedical and clinical magnetic resonance spectroscopy: a review , 1997, NMR in biomedicine.

[23]  I. Wilson,et al.  Application of directly coupled HPLC-NMR-MS/MS to the identification of metabolites of 5-trifluoromethylpyridone (2-hydroxy-5-trifluoromethylpyridine) in hydroponically grown plants. , 2000, Journal of agricultural and food chemistry.

[24]  Akira Oikawa,et al.  Differential metabolomics unraveling light/dark regulation of metabolic activities in Arabidopsis cell culture , 2007, Planta.

[25]  Heekuck Oh,et al.  Neural Networks for Pattern Recognition , 1993, Adv. Comput..

[26]  Erin E. Carlson,et al.  Targeted profiling: quantitative analysis of 1H NMR metabolomics data. , 2006, Analytical chemistry.

[27]  John C. Lindon,et al.  Pattern recognition methods and applications in biomedical magnetic resonance , 2001 .

[28]  Christoph Seger,et al.  Analytical aspects of plant metabolite profiling platforms: current standings and future aims. , 2007, Journal of proteome research.

[29]  Y. Shachar-Hill,et al.  Revealing metabolic phenotypes in plants: inputs from NMR analysis , 2005, Biological reviews of the Cambridge Philosophical Society.

[30]  Joachim Kopka,et al.  Transcript and metabolite profiling during cold acclimation of Arabidopsis reveals an intricate relationship of cold-regulated gene expression with modifications in metabolite content. , 2007, The Plant journal : for cell and molecular biology.

[31]  N. Kruger,et al.  Metabolite fingerprinting and profiling in plants using NMR. , 2004, Journal of experimental botany.

[32]  Aalim M Weljie,et al.  Quantitative 1H NMR metabolomics reveals extensive metabolic reprogramming of primary and secondary metabolism in elicitor-treated opium poppy cell cultures , 2008, BMC Plant Biology.

[33]  A. Marshall,et al.  Characterization of vegetable oils: detailed compositional fingerprints derived from electrospray ionization fourier transform ion cyclotron resonance mass spectrometry. , 2004, Journal of agricultural and food chemistry.

[34]  M. Hirai,et al.  Elucidation of Gene-to-Gene and Metabolite-to-Gene Networks in Arabidopsis by Integration of Metabolomics and Transcriptomics* , 2005, Journal of Biological Chemistry.

[35]  Daniel Raftery,et al.  Comparing and combining NMR spectroscopy and mass spectrometry in metabolomics , 2007, Analytical and bioanalytical chemistry.

[36]  M. Viant Improved methods for the acquisition and interpretation of NMR metabolomic data. , 2003, Biochemical and biophysical research communications.

[37]  M. Zenk,et al.  Poppy alkaloid profiling by electrospray tandem mass spectrometry and electrospray FT-ICR mass spectrometry after [ring-13C6]-tyramine feeding. , 2007, Phytochemistry.

[38]  Y. Choi,et al.  Classification of Ilex species based on metabolomic fingerprinting using nuclear magnetic resonance and multivariate data analysis. , 2005, Journal of agricultural and food chemistry.

[39]  S. Stein,et al.  Deconvolution gas chromatography/mass spectrometry of urinary organic acids--potential for pattern recognition and automated identification of metabolic disorders. , 1999, Rapid communications in mass spectrometry : RCM.

[40]  S. Kanaya,et al.  KNApSAcK: A Comprehensive Species-Metabolite Relationship Database , 2006 .

[41]  L. Angenot,et al.  Metabolomic analysis of Strychnos nux-vomica, Strychnos icaja and Strychnos ignatii extracts by 1H nuclear magnetic resonance spectrometry and multivariate analysis techniques. , 2004, Phytochemistry.

[42]  A. Marshall,et al.  Mass-selective ion accumulation and fragmentation in a linear octopole ion trap external to a fourier transform ion cyclotron resonance mass spectrometer , 2000 .

[43]  R. Hall,et al.  Plant metabolomics: from holistic hope, to hype, to hot topic. , 2006, The New phytologist.

[44]  A. Makarov,et al.  The Orbitrap: a new mass spectrometer. , 2005, Journal of mass spectrometry : JMS.

[45]  David I. Ellis,et al.  Metabolomics: Current analytical platforms and methodologies , 2005 .

[46]  Y. Choi,et al.  Metabolic characterization of Brassica rapa leaves by NMR spectroscopy. , 2007, Journal of agricultural and food chemistry.

[47]  J. Drozd Chemical Derivatization in Gas Chromatography , 1981 .

[48]  Anthony C. Davison,et al.  Rapid Classification of Phenotypic Mutants of Arabidopsis via Metabolite Fingerprinting1[W][OA] , 2007, Plant Physiology.

[49]  W. A. Anderson,et al.  A Proton Magnetic Resonance Study of the Hydration of Deoxyribonucleic Acid , 1954, Nature.

[50]  M. Bizzarri,et al.  NMR-based metabonomic study of transgenic maize. , 2004, Phytochemistry.

[51]  Nigel W. Hardy,et al.  Hierarchical metabolomics demonstrates substantial compositional similarity between genetically modified and conventional potato crops. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[52]  P. Facchini,et al.  Molecular Cloning and Characterization of Tetrahydroprotoberberine cis-N-Methyltransferase, an Enzyme Involved in Alkaloid Biosynthesis in Opium Poppy* , 2007, Journal of Biological Chemistry.

[53]  Y. Choi,et al.  Identification of phenylpropanoids in methyl jasmonate treated Brassica rapa leaves using two-dimensional nuclear magnetic resonance spectroscopy. , 2006, Journal of chromatography. A.

[54]  H. K. Kim,et al.  NMR-based metabolomics at work in phytochemistry , 2007, Phytochemistry Reviews.

[55]  B. Hammock,et al.  Mass spectrometry-based metabolomics. , 2007, Mass spectrometry reviews.

[56]  W. Weckwerth Metabolomics in systems biology. , 2003, Annual review of plant biology.

[57]  Robert Verpoorte,et al.  Metabolic fingerprinting of Ephedra species using 1H-NMR spectroscopy and principal component analysis. , 2005, Chemical & pharmaceutical bulletin.

[58]  Marianne Defernez,et al.  Factors affecting the robustness of metabolite fingerprinting using 1H NMR spectra. , 2003, Phytochemistry.

[59]  Y. Choi,et al.  Metabolic differentiation of Arabidopsis treated with methyl jasmonate using nuclear magnetic resonance spectroscopy , 2006 .

[60]  Christoph W. Sensen,et al.  Gene transcript and metabolite profiling of elicitor-induced opium poppy cell cultures reveals the coordinate regulation of primary and secondary metabolism , 2007, Planta.

[61]  D. Goodenowe,et al.  Nontargeted metabolome analysis by use of Fourier Transform Ion Cyclotron Mass Spectrometry. , 2002, Omics : a journal of integrative biology.

[62]  Robert D. Hall,et al.  Breakthrough Technologies A Novel Approach for Nontargeted Data Analysis for Metabolomics . Large-Scale Profiling of Tomato Fruit Volatiles 1 [ w ] , 2005 .

[63]  M. Orešič,et al.  Data processing for mass spectrometry-based metabolomics. , 2007, Journal of chromatography. A.

[64]  L. Tseng,et al.  Separation and identification of phenolic compounds in olive oil by coupling high-performance liquid chromatography with postcolumn solid-phase extraction to nuclear magnetic resonance spectroscopy (LC-SPE-NMR). , 2005, Journal of agricultural and food chemistry.

[65]  Å. Thureson-Klein Observations on the Development and Fine Structure of the Articulated Laticifers of Papaver somniferum , 1970 .

[66]  C. Ammer,et al.  Comparative transcript and alkaloid profiling in Papaver species identifies a short chain dehydrogenase/reductase involved in morphine biosynthesis. , 2006, The Plant journal : for cell and molecular biology.

[67]  G. Glish,et al.  The basics of mass spectrometry in the twenty-first century , 2003, Nature Reviews Drug Discovery.

[68]  K. Robards,et al.  Metabolomics: The greatest omics of them all? , 2006, Analytical chemistry.

[69]  A. Marshall,et al.  Resolution of 11,000 compositionally distinct components in a single electrospray ionization Fourier transform ion cyclotron resonance mass spectrum of crude oil. , 2002, Analytical chemistry.

[70]  Corey D Broeckling,et al.  MET-IDEA: data extraction tool for mass spectrometry-based metabolomics. , 2006, Analytical chemistry.

[71]  B. Denoyes-Rothan,et al.  Quantitative metabolic profiling by 1-dimensional 1H-NMR analyses: application to plant genetics and functional genomics. , 2004, Functional plant biology : FPB.

[72]  Jean-Marc Nuzillard,et al.  NMR metabolomics to revisit the tobacco mosaic virus infection in Nicotiana tabacum leaves. , 2006, Journal of natural products.

[73]  H. Hotelling The most predictable criterion. , 1935 .

[74]  Johan Lindberg,et al.  A comparison of methods for alignment of NMR peaks in the context of cluster analysis. , 2005, Journal of pharmaceutical and biomedical analysis.

[75]  Peter McCourt,et al.  Systems approaches to understanding cell signaling and gene regulation. , 2004, Current opinion in plant biology.

[76]  A. Segre,et al.  Nuclear Magnetic Resonance Spectroscopy-Based Metabolite Profiling of Transgenic Tomato Fruit Engineered to Accumulate Spermidine and Spermine Reveals Enhanced Anabolic and Nitrogen-Carbon Interactions1[W][OA] , 2006, Plant Physiology.

[77]  M. Zanor,et al.  Integrated Analysis of Metabolite and Transcript Levels Reveals the Metabolic Shifts That Underlie Tomato Fruit Development and Highlight Regulatory Aspects of Metabolic Network Behavior1[W] , 2006, Plant Physiology.

[78]  P. Fraser,et al.  Chemical derivatization and mass spectral libraries in metabolic profiling by GC/MS and LC/MS/MS. , 2005, Journal of experimental botany.

[79]  T. Veenstra,et al.  Design and performance of an ESI interface for selective external ion accumulation coupled to a Fourier transform ion cyclotron mass spectrometer. , 2001, Analytical chemistry.

[80]  D. Lightfoot,et al.  Metabolite Fingerprinting in Transgenic Nicotiana tabacum Altered by the Escherichia coli Glutamate Dehydrogenase Gene , 2005, Journal of biomedicine & biotechnology.

[81]  Hyung-Kyoon Choi,et al.  Metabolic fingerprinting of wild type and transgenic tobacco plants by 1H NMR and multivariate analysis technique. , 2004, Phytochemistry.

[82]  Dirk Inzé,et al.  Plant cell factories in the post-genomic era: new ways to produce designer secondary metabolites. , 2004, Trends in plant science.

[83]  A. Marshall,et al.  Fourier Transform Ion Cyclotron Resonance Spectroscopy , 1974 .

[84]  Erik Johansson,et al.  Using chemometrics for navigating in the large data sets of genomics, proteomics, and metabonomics (gpm) , 2004, Analytical and bioanalytical chemistry.

[85]  Dudley H. Williams,et al.  Influence of Deoxyribonucleic Acid on the Intermolecular Structure of Water , 1962, Nature.

[86]  P. Facchini,et al.  Phloem-Specific Expression of Tyrosine/Dopa Decarboxylase Genes and the Biosynthesis of Isoquinoline Alkaloids in Opium Poppy. , 1995, The Plant cell.

[87]  Yair Shachar-Hill,et al.  Towards the plant metabolome and beyond: Innovations , 2007 .

[88]  J. Gershenzon,et al.  Biosynthesis and Emission of Terpenoid Volatiles from Arabidopsis Flowers Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.007989. , 2003, The Plant Cell Online.

[89]  Y. Choi,et al.  Metabolomic analysis of methyl jasmonate treated Brassica rapa leaves by 2-dimensional NMR spectroscopy. , 2006, Phytochemistry.

[90]  P. Schnier,et al.  Multiple ion isolation applications in FT-ICR MS: exact-mass MSn internal calibration and purification/interrogation of protein-drug complexes. , 2002, Analytical chemistry.

[91]  S. Kanaya,et al.  Clarification of Pathway-Specific Inhibition by Fourier Transform Ion Cyclotron Resonance/Mass Spectrometry-Based Metabolic Phenotyping Studies1[W] , 2006, Plant Physiology.

[92]  Jacques Vervoort,et al.  LC-UV-solid-phase extraction-NMR-MS combined with a cryogenic flow probe and its application to the identification of compounds present in Greek oregano. , 2003, Analytical chemistry.

[93]  Yair Shachar-Hill,et al.  PROBING PLANT METABOLISM WITH NMR. , 2003, Annual review of plant physiology and plant molecular biology.

[94]  Vladimir Shulaev,et al.  Metabolomics technology and bioinformatics , 2006, Briefings Bioinform..

[95]  Tadashi Eguchi,et al.  Metabolite profiling of plant carotenoids using the matrix-assisted laser desorption ionization time-of-flight mass spectrometry. , 2007, The Plant journal : for cell and molecular biology.

[96]  J. Vervoort,et al.  Analytical procedure for the in-vial derivatization--extraction of phenolic acids and flavonoids in methanolic and aqueous plant extracts followed by gas chromatography with mass-selective detection. , 2004, Journal of chromatography. A.

[97]  R. Goodacre,et al.  Metabolic Profiling: Its Role in Biomarker Discovery and Gene Function Analysis , 2003, Springer US.

[98]  P. Facchini,et al.  Uncoupled Defense Gene Expression and Antimicrobial Alkaloid Accumulation in Elicited Opium Poppy Cell Cultures , 1996, Plant physiology.

[99]  T. V. van Beek,et al.  Development of a triple hyphenated HPLC-radical scavenging detection-DAD-SPE-NMR system for the rapid identification of antioxidants in complex plant extracts. , 2005, Journal of chromatography. A.

[100]  Alisdair R Fernie,et al.  Plant metabolomics: towards biological function and mechanism. , 2006, Trends in plant science.

[101]  Tomoyoshi Soga,et al.  Metabolome analysis by capillary electrophoresis-mass spectrometry. , 2007, Journal of chromatography. A.

[102]  R. Abagyan,et al.  XCMS: processing mass spectrometry data for metabolite profiling using nonlinear peak alignment, matching, and identification. , 2006, Analytical chemistry.

[103]  S. Zeeman,et al.  Arabidopsis mutants Atisa1 and Atisa2 have identical phenotypes and lack the same multimeric isoamylase, which influences the branch point distribution of amylopectin during starch synthesis. , 2005, The Plant journal : for cell and molecular biology.

[104]  Robert Verpoorte,et al.  Metabolic Discrimination of Catharanthus roseus Leaves Infected by Phytoplasma Using 1H-NMR Spectroscopy and Multivariate Data Analysis1 , 2004, Plant Physiology.

[105]  K. Pallett,et al.  Combined use of 13C- and 19F-NMR to analyse the mode of action and the metabolism of the herbicide isoxaflutole , 2000 .

[106]  John C Lindon,et al.  Automatic alignment of individual peaks in large high-resolution spectral data sets. , 2004, Journal of magnetic resonance.

[107]  Robert Verpoorte,et al.  Metabolomic differentiation of Cannabis sativa cultivars using 1H NMR spectroscopy and principal component analysis. , 2004, Journal of natural products.

[108]  R. Dixon,et al.  Plant metabolomics: large-scale phytochemistry in the functional genomics era. , 2003, Phytochemistry.

[109]  F. Lottspeich,et al.  (R,S)-Reticuline 7-O-methyltransferase and (R,S)-norcoclaurine 6-O-methyltransferase of Papaver somniferum - cDNA cloning and characterization of methyl transfer enzymes of alkaloid biosynthesis in opium poppy. , 2003, The Plant journal : for cell and molecular biology.

[110]  G. Kruppa,et al.  Metabolomics applications of FT-ICR mass spectrometry. , 2005, Mass spectrometry reviews.

[111]  David C Muddiman,et al.  Analysis of the low molecular weight fraction of serum by LC-dual ESI-FT-ICR mass spectrometry: precision of retention time, mass, and ion abundance. , 2004, Analytical chemistry.

[112]  M. Hirai,et al.  Functional genomics by integrated analysis of metabolome and transcriptome of Arabidopsis plants over-expressing an MYB transcription factor. , 2005, The Plant journal : for cell and molecular biology.

[113]  Jane L. Ward,et al.  NMR Spectroscopy in Plant Metabolomics , 2006 .