Metabolomic approaches toward understanding nitrogen metabolism in plants.

Plants can assimilate inorganic nitrogen (N) sources to organic N such as amino acids. N is the most important of the mineral nutrients required by plants and its metabolism is tightly coordinated with carbon (C) metabolism in the fundamental processes that permit plant growth. Increased understanding of N regulation may provide important insights for plant growth and improvement of quality of crops and vegetables because N as well as C metabolism are fundamental components of plant life. Metabolomics is a global biochemical approach useful to study N metabolism because metabolites not only reflect the ultimate phenotypes (traits), but can mediate transcript levels as well as protein levels directly and/or indirectly under different N conditions. This review outlines analytical and bioinformatic techniques particularly used to perform metabolomics for studying N metabolism in higher plants. Examples are used to illustrate the application of metabolomic techniques to the model plants Arabidopsis and rice, as well as other crops and vegetables.

[1]  Johan Trygg,et al.  Integrated analysis of transcript, protein and metabolite data to study lignin biosynthesis in hybrid aspen. , 2009, Journal of proteome research.

[2]  T. Moritz,et al.  Metabolomic evaluation of pulsed electric field-induced stress on potato tissue , 2009, Planta.

[3]  P. Mendes,et al.  The origin of correlations in metabolomics data , 2005, Metabolomics.

[4]  Katsutoshi Takahashi,et al.  A strategy for the determination of the elemental composition by fourier transform ion cyclotron resonance mass spectrometry based on isotopic peak ratios. , 2010, Analytical chemistry.

[5]  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.

[6]  B. Hirel,et al.  Glutamine Synthetase in Rice: A COMPARATIVE STUDY OF THE ENZYMES FROM ROOTS AND LEAVES. , 1980, Plant physiology.

[7]  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.

[8]  Gloria M. Coruzzi,et al.  Light- and Carbon-Signaling Pathways. Modeling Circuits of Interactions1 , 2003, Plant Physiology.

[9]  R. Steuer,et al.  Metabolomic networks in plants: Transitions from pattern recognition to biological interpretation. , 2006, Bio Systems.

[10]  Masaru Tomita,et al.  Time-resolved metabolomics reveals metabolic modulation in rice foliage , 2008, BMC Systems Biology.

[11]  L. Willmitzer,et al.  13C isotope-labeled metabolomes allowing for improved compound annotation and relative quantification in liquid chromatography-mass spectrometry-based metabolomic research. , 2009, Analytical chemistry.

[12]  Yves Gibon,et al.  GC-EI-TOF-MS analysis of in vivo carbon-partitioning into soluble metabolite pools of higher plants by monitoring isotope dilution after 13CO2 labelling. , 2007, Phytochemistry.

[13]  Makoto Kobayashi,et al.  Unbiased characterization of genotype-dependent metabolic regulations by metabolomic approach in Arabidopsis thaliana , 2007, BMC Systems Biology.

[14]  Masanori Arita,et al.  Consolidating metabolite identifiers to enable contextual and multi-platform metabolomics data analysis , 2010, BMC Bioinformatics.

[15]  Kay S Tatsuoka,et al.  Multi-platform investigation of the metabolome in a leptin receptor defective murine model of type 2 diabetes. , 2008, Molecular bioSystems.

[16]  D. Schachtman,et al.  Nutrient sensing and signaling: NPKS. , 2007, Annual review of plant biology.

[17]  Björn H. Junker,et al.  Simulating Plant Metabolic Pathways with Enzyme-Kinetic Models1[C] , 2010, Plant Physiology.

[18]  D. Wells,et al.  Nitrate transport and signalling. , 2007, Journal of experimental botany.

[19]  Rodrigo A. Gutiérrez,et al.  VirtualPlant: A Software Platform to Support Systems Biology Research1[W][OA] , 2009, Plant Physiology.

[20]  Gabriel Krouk,et al.  A Systems View of Responses to Nutritional Cues in Arabidopsis: Toward a Paradigm Shift for Predictive Network Modeling1 , 2009, Plant Physiology.

[21]  O. Fiehn,et al.  Differential metabolic networks unravel the effects of silent plant phenotypes. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[22]  F. Mesnard,et al.  NMR analysis of plant nitrogen metabolism , 2004, Photosynthesis Research.

[23]  Nigel W. Hardy,et al.  The metabolomics standards initiative (MSI) , 2007, Metabolomics.

[24]  Royston Goodacre,et al.  An Introduction to Liquid Chromatography– Mass Spectrometry Instrumentation Applied in Plant Metabolomic Analyses † Untargeted Plant Metabolomics and the Potential of Liquid Chromatography Mass Spectrometry , 2022 .

[25]  M. Lehmann,et al.  The metabolic response of Arabidopsis roots to oxidative stress is distinct from that of heterotrophic cells in culture and highlights a complex relationship between the levels of transcripts, metabolites, and flux. , 2009, Molecular plant.

[26]  J. Araus,et al.  Gene expression, cellular localisation and function of glutamine synthetase isozymes in wheat (Triticum aestivum L.) , 2008, Plant Molecular Biology.

[27]  Guo Wangzhen,et al.  Molecular cloning and characterization of a cytosolic glutamine synthetase gene, a fiber strength-associated gene in cotton , 2008, Planta.

[28]  Henning Redestig,et al.  Integrative functional genomics of salt acclimatization in the model legume Lotus japonicus. , 2007, The Plant journal : for cell and molecular biology.

[29]  Mark Stitt,et al.  Use of reverse-phase liquid chromatography, linked to tandem mass spectrometry, to profile the Calvin cycle and other metabolic intermediates in Arabidopsis rosettes at different carbon dioxide concentrations. , 2009, The Plant journal : for cell and molecular biology.

[30]  U. Roessner,et al.  Metabolite profiling reveals distinct changes in carbon and nitrogen metabolism in phosphate-deficient barley plants (Hordeum vulgare L.). , 2008, Plant & cell physiology.

[31]  Thomas C R Williams,et al.  A Genome-Scale Metabolic Model Accurately Predicts Fluxes in Central Carbon Metabolism under Stress Conditions1[C][W] , 2010, Plant Physiology.

[32]  O. Fiehn,et al.  FiehnLib: mass spectral and retention index libraries for metabolomics based on quadrupole and time-of-flight gas chromatography/mass spectrometry. , 2009, Analytical chemistry.

[33]  K. Oda,et al.  Systematic approaches to using the FOX hunting system to identify useful rice genes. , 2009, The Plant journal : for cell and molecular biology.

[34]  B. M. Lange,et al.  A systems biology approach identifies the biochemical mechanisms regulating monoterpenoid essential oil composition in peppermint , 2008, Proceedings of the National Academy of Sciences.

[35]  Mark Stitt,et al.  Genome-Wide Reprogramming of Primary and Secondary Metabolism, Protein Synthesis, Cellular Growth Processes, and the Regulatory Infrastructure of Arabidopsis in Response to Nitrogen1[w] , 2004, Plant Physiology.

[36]  V. Tolstikov Metabolic analysis. , 2009, Methods in molecular biology.

[37]  Wenxu Zhou,et al.  Arabidopsis has a cytosolic fumarase required for the massive allocation of photosynthate into fumaric acid and for rapid plant growth on high nitrogen. , 2010, The Plant journal : for cell and molecular biology.

[38]  Matej Oresic,et al.  MZmine: toolbox for processing and visualization of mass spectrometry based molecular profile data , 2006, Bioinform..

[39]  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.

[40]  T. Ebbels,et al.  Recursive segment-wise peak alignment of biological (1)h NMR spectra for improved metabolic biomarker recovery. , 2009, Analytical chemistry.

[41]  J. Morgan,et al.  Mathematical modeling of plant metabolic pathways. , 2002, Metabolic engineering.

[42]  Michaela Scigelova,et al.  Coupling liquid chromatography to Orbitrap mass spectrometry. , 2010, Journal of chromatography. A.

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

[44]  N. Sreenivasulu,et al.  Increasing Sucrose Uptake Capacity of Wheat Grains Stimulates Storage Protein Synthesis1[W] , 2009, Plant Physiology.

[45]  F. Carrari,et al.  Metabolic profiles of sunflower genotypes with contrasting response to Sclerotinia sclerotiorum infection. , 2010, Phytochemistry.

[46]  Kazuo Shinozaki,et al.  Characterization of the ABA-regulated global responses to dehydration in Arabidopsis by metabolomics. , 2009, The Plant journal : for cell and molecular biology.

[47]  D. Roby,et al.  Stress induces the expression of AtNADK-1, a gene encoding a NAD(H) kinase in Arabidopsis thaliana , 2005, Molecular Genetics and Genomics.

[48]  D. Scheel,et al.  The Multifunctional Enzyme CYP71B15 (PHYTOALEXIN DEFICIENT3) Converts Cysteine-Indole-3-Acetonitrile to Camalexin in the Indole-3-Acetonitrile Metabolic Network of Arabidopsis thaliana[W][OA] , 2009, The Plant Cell Online.

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

[50]  Masanori Arita,et al.  Metabolomic correlation-network modules in Arabidopsis based on a graph-clustering approach , 2011, BMC Systems Biology.

[51]  G. Krouk,et al.  Nitrate signaling: adaptation to fluctuating environments. , 2010, Current opinion in plant biology.

[52]  T. Yamaya,et al.  Assimilation of ammonium ions and reutilization of nitrogen in rice (Oryza sativa L.). , 2007, Journal of experimental botany.

[53]  P. Lundberg,et al.  Primary metabolism in N2-fixing Alnus incana–Frankia symbiotic root nodules studied with 15N and 31P nuclear magnetic resonance spectroscopy , 2004, Planta.

[54]  Marc-Thorsten Hütt,et al.  Consistency analysis of metabolic correlation networks , 2007, BMC Systems Biology.

[55]  Choon Nam Ong,et al.  A multi-analytical approach for metabolomic profiling of zebrafish (Danio rerio) livers. , 2009, Molecular bioSystems.

[56]  Yves Gibon,et al.  Steps towards an integrated view of nitrogen metabolism. , 2002, Journal of experimental botany.

[57]  J. L. Liu,et al.  Determination of metabolic fluxes in a non-steady-state system. , 2007, Phytochemistry.

[58]  J. Trygg,et al.  A cross-species transcriptomics approach to identify genes involved in leaf development , 2008, BMC Genomics.

[59]  Lijun Luo,et al.  Photosynthetic metabolism of C3 plants shows highly cooperative regulation under changing environments: A systems biological analysis , 2009, Proceedings of the National Academy of Sciences.

[60]  G M Coruzzi,et al.  Carbon and nitrogen sensing and signaling in plants: emerging 'matrix effects'. , 2001, Current opinion in plant biology.

[61]  G. Coruzzi,et al.  Nitrogen and carbon nutrient and metabolite signaling in plants. , 2001, Plant physiology.

[62]  W. Snedden,et al.  Identification, molecular cloning and functional characterization of a novel NADH kinase from Arabidopsis thaliana (thale cress). , 2005, The Biochemical journal.

[63]  Xiaorong Fan,et al.  Amino acids and nitrate as signals for the regulation of nitrogen acquisition. , 2007, Journal of experimental botany.

[64]  Atul J. Butte,et al.  Systematic survey reveals general applicability of "guilt-by-association" within gene coexpression networks , 2005, BMC Bioinformatics.

[65]  Richard A Dixon,et al.  Phytochemistry meets genome analysis, and beyond. , 2003, Phytochemistry.

[66]  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.

[67]  U. Sauer,et al.  Cross-platform comparison of methods for quantitative metabolomics of primary metabolism. , 2009, Analytical chemistry.

[68]  Ralf Steuer,et al.  Review: On the analysis and interpretation of correlations in metabolomic data , 2006, Briefings Bioinform..

[69]  Joachim Kopka,et al.  Current challenges and developments in GC-MS based metabolite profiling technology. , 2006, Journal of biotechnology.

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

[71]  Kazuki Saito,et al.  Phosphoenolpyruvate carboxylase intrinsically located in the chloroplast of rice plays a crucial role in ammonium assimilation , 2010, Proceedings of the National Academy of Sciences.

[72]  Rodrigo A Gutiérrez,et al.  A systems view of nitrogen nutrient and metabolite responses in Arabidopsis. , 2008, Current opinion in plant biology.

[73]  N. Kruger,et al.  Network flux analysis: impact of 13C-substrates on metabolism in Arabidopsis thaliana cell suspension cultures. , 2007, Phytochemistry.

[74]  O. Fiehn,et al.  Metabolite profiling for plant functional genomics , 2000, Nature Biotechnology.

[75]  Johann M Rohwer,et al.  Kinetic model of sucrose accumulation in maturing sugarcane culm tissue. , 2007, Phytochemistry.

[76]  S. Yanagisawa,et al.  Pleiotropic Modulation of Carbon and Nitrogen Metabolism in Arabidopsis Plants Overexpressing the NAD kinase2 Gene1[W] , 2009, Plant Physiology.

[77]  Kazuki Saito,et al.  Members of the LBD Family of Transcription Factors Repress Anthocyanin Synthesis and Affect Additional Nitrogen Responses in Arabidopsis[W][OA] , 2009, The Plant Cell Online.

[78]  Francesca Chiaromonte,et al.  Qualitative network models and genome-wide expression data define carbon/nitrogen-responsive molecular machines in Arabidopsis , 2007, Genome Biology.

[79]  S. Amâncio,et al.  Nitrogen Acquisition and Assimilation in Higher Plants , 2004, Plant Ecophysiology.

[80]  Kazuo Shinozaki,et al.  MS/MS spectral tag-based annotation of non-targeted profile of plant secondary metabolites , 2008, The Plant journal : for cell and molecular biology.

[81]  A. Hemerly,et al.  Characterization of glutamine synthetase genes in sugarcane genotypes with different rates of biological nitrogen fixation , 2005 .

[82]  Jun Kikuchi,et al.  Towards dynamic metabolic network measurements by multi-dimensional NMR-based fluxomics. , 2007, Phytochemistry.

[83]  Rodrigo A Gutiérrez,et al.  Systems Biology for the Virtual Plant1 , 2005, Plant Physiology.

[84]  Tomoyoshi Soga,et al.  Capillary electrophoresis-mass spectrometry for metabolomics. , 2007, Methods in molecular biology.

[85]  A. Scaloni,et al.  Leaf proteome analysis of transgenic plants expressing antiviral antibodies. , 2009, Journal of proteome research.

[86]  J. Rabinowitz,et al.  Absolute Metabolite Concentrations and Implied Enzyme Active Site Occupancy in Escherichia coli , 2009, Nature chemical biology.

[87]  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.

[88]  A. Fernie,et al.  Gas chromatography mass spectrometry–based metabolite profiling in plants , 2006, Nature Protocols.

[89]  S. Yanagisawa,et al.  Metabolome and Photochemical Analysis of Rice Plants Overexpressing Arabidopsis NAD Kinase Gene1[W][OA] , 2010, Plant Physiology.

[90]  B. Usadel,et al.  Arabidopsis and primary photosynthetic metabolism - more than the icing on the cake. , 2010, The Plant journal : for cell and molecular biology.

[91]  T. Yamaya,et al.  Metabolic Regulation of Ammonium uptake and Assimilation , 2004 .

[92]  T. Annesley Ion suppression in mass spectrometry. , 2003, Clinical chemistry.

[93]  Mayumi Tabuchi,et al.  Severe reduction in growth rate and grain filling of rice mutants lacking OsGS1;1, a cytosolic glutamine synthetase1;1. , 2005, The Plant journal : for cell and molecular biology.

[94]  M. Hirai,et al.  A Chloroplastic UDP-Glucose Pyrophosphorylase from Arabidopsis Is the Committed Enzyme for the First Step of Sulfolipid Biosynthesis[W][OA] , 2009, The Plant Cell Online.

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

[96]  Johan Lindberg,et al.  Predictive metabolite profiling applying hierarchical multivariate curve resolution to GC-MS data--a potential tool for multi-parametric diagnosis. , 2006, Journal of proteome research.

[97]  K. Edwards,et al.  Two Cytosolic Glutamine Synthetase Isoforms of Maize Are Specifically Involved in the Control of Grain Production[W][OA] , 2006, The Plant Cell Online.

[98]  M. Hirai,et al.  Comparative metabolomics charts the impact of genotype-dependent methionine accumulation in Arabidopsis thaliana , 2010, Amino Acids.

[99]  Oliver Fiehn,et al.  Divergent metabolome and proteome suggest functional independence of dual phloem transport systems in cucurbits , 2010, Proceedings of the National Academy of Sciences.

[100]  Kazuki Saito,et al.  Impact of clock-associated Arabidopsis pseudo-response regulators in metabolic coordination , 2009, Proceedings of the National Academy of Sciences.

[101]  Marcel Dicke,et al.  Combined Transcript and Metabolite Analysis Reveals Genes Involved in Spider Mite Induced Volatile Formation in Cucumber Plants1 , 2004, Plant Physiology.

[102]  M. Stitt,et al.  Adjustment of growth and central metabolism to a mild but sustained nitrogen-limitation in Arabidopsis. , 2009, Plant, cell & environment.

[103]  Eiichiro Fukusaki,et al.  In Vivo 15N‐Enrichment of Metabolites in Suspension Cultured Cells and Its Application to Metabolomics , 2006, Biotechnology progress.

[104]  M. Hirai,et al.  Decoding genes with coexpression networks and metabolomics - 'majority report by precogs'. , 2008, Trends in plant science.

[105]  B. M. Lange,et al.  Experimental and mathematical approaches to modeling plant metabolic networks. , 2007, Phytochemistry.

[106]  William Stafford Noble,et al.  The effect of replication on gene expression microarray experiments , 2003, Bioinform..

[107]  Nigel W. Hardy,et al.  The Metabolomics Standards Initiative , 2007, Nature Biotechnology.

[108]  Mark Stitt,et al.  Accumulation of nitrate in the shoot acts as a signal to regulate shoot‐root allocation in tobacco† , 1997 .

[109]  G. Coruzzi,et al.  THE MOLECULAR-GENETICS OF NITROGEN ASSIMILATION INTO AMINO ACIDS IN HIGHER PLANTS. , 1996, Annual review of plant physiology and plant molecular biology.

[110]  Kazuki Saito,et al.  Metabolomics for functional genomics, systems biology, and biotechnology. , 2010, Annual review of plant biology.

[111]  M. Hirai,et al.  Widely Targeted Metabolomics Based on Large-Scale MS/MS Data for Elucidating Metabolite Accumulation Patterns in Plants , 2008, Plant & cell physiology.

[112]  Xin-Guang Zhu,et al.  Optimizing the Distribution of Resources between Enzymes of Carbon Metabolism Can Dramatically Increase Photosynthetic Rate: A Numerical Simulation Using an Evolutionary Algorithm1[W][OA] , 2007, Plant Physiology.

[113]  A. Fukushima,et al.  Metabolomic screening applied to rice FOX Arabidopsis lines leads to the identification of a gene-changing nitrogen metabolism. , 2010, Molecular plant.

[114]  R. Ramautar,et al.  CE‐MS in metabolomics , 2009, Electrophoresis.

[115]  O. Fiehn,et al.  Interpreting correlations in metabolomic networks. , 2003, Biochemical Society transactions.

[116]  Jing Li,et al.  New Bioinformatics Resources For Metabolomics , 2006, Pacific Symposium on Biocomputing.

[117]  Shigehiko Kanaya,et al.  Application of Fourier-transform ion cyclotron resonance mass spectrometry to metabolic profiling and metabolite identification. , 2010, Current opinion in biotechnology.

[118]  Manuel Desco,et al.  A novel R-package graphic user interface for the analysis of metabonomic profiles , 2009, BMC Bioinformatics.

[119]  Kazuki Saito,et al.  Integrated omics approaches in plant systems biology. , 2009, Current opinion in chemical biology.

[120]  A. Kendall,et al.  Barley mutants lacking chloroplast glutamine synthetase-biochemical and genetic analysis. , 1987, Plant physiology.

[121]  Chung-Jui Tsai,et al.  Metabolic Profiling of the Sink-to-Source Transition in Developing Leaves of Quaking Aspen1 , 2004, Plant Physiology.

[122]  Rui An,et al.  NADK2, an Arabidopsis Chloroplastic NAD Kinase, Plays a Vital Role in Both Chlorophyll Synthesis and Chloroplast Protection , 2005, Plant Molecular Biology.

[123]  D E Shasha,et al.  Using combinatorial design to study regulation by multiple input signals. A tool for parsimony in the post-genomics era. , 2001, Plant physiology.

[124]  R. Motohashi,et al.  The FOX hunting system: an alternative gain-of-function gene hunting technique. , 2006, The Plant journal : for cell and molecular biology.

[125]  Arjen Lommen,et al.  MetAlign: interface-driven, versatile metabolomics tool for hyphenated full-scan mass spectrometry data preprocessing. , 2009, Analytical chemistry.

[126]  Y. Choi,et al.  NMR-based metabolomic analysis of plants , 2010, Nature Protocols.

[127]  A. Fernie,et al.  Metabolic profiling reveals altered nitrogen nutrient regimes have diverse effects on the metabolism of hydroponically-grown tomato (Solanum lycopersicum) plants , 2005 .

[128]  J. W. Allwood,et al.  1H NMR, GC-EI-TOFMS, and data set correlation for fruit metabolomics: application to spatial metabolite analysis in melon. , 2009, Analytical chemistry.

[129]  Hideyuki Takahashi,et al.  Chloroplast NAD kinase is essential for energy transduction through the xanthophyll cycle in photosynthesis. , 2006, Plant & cell physiology.

[130]  Xiao-Jiang Feng,et al.  Metabolomics-driven quantitative analysis of ammonia assimilation in E. coli , 2009, Molecular systems biology.

[131]  S. Naito,et al.  Mutation in the threonine synthase gene results in an over-accumulation of soluble methionine in Arabidopsis. , 2000, Plant physiology.

[132]  F. Sato,et al.  Nitrogen Recycling and Remobilization Are Differentially Controlled by Leaf Senescence and Development Stage in Arabidopsis under Low Nitrogen Nutrition1 , 2008, Plant Physiology.

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

[134]  Takayuki Tohge,et al.  Web-based resources for mass-spectrometry-based metabolomics: a user's guide. , 2009, Phytochemistry.

[135]  J. Baker,et al.  Establishing substantial equivalence: metabolomics. , 2009, Methods in molecular biology.

[136]  R. Molinié,et al.  Metabolic profiling of maize mutants deficient for two glutamine synthetase isoenzymes using 1H-NMR-based metabolomics. , 2010, Phytochemical analysis : PCA.

[137]  Yasushi Noguchi,et al.  Network analysis of plasma and tissue amino acids and the generation of an amino index for potential diagnostic use. , 2006, The American journal of clinical nutrition.

[138]  Athel Cornish-Bowden,et al.  Understanding the regulation of aspartate metabolism using a model based on measured kinetic parameters , 2009, Molecular systems biology.

[139]  Oliver Sawodny,et al.  Mathematical Modeling of the Central Carbohydrate Metabolism in Arabidopsis Reveals a Substantial Regulatory Influence of Vacuolar Invertase on Whole Plant Carbon Metabolism1[W] , 2010, Plant Physiology.

[140]  G. Coruzzi Primary N-assimilation into Amino Acids in Arabidopsis , 2003, The arabidopsis book.