Comparative analysis of metabolites profile in spinach (Spinacia oleracea L.) affected by different concentrations of Gly and nitrate.

Abstract The ability of plants to uptake amino acids has been confirmed by extensive research; however, the subsequent post-uptake metabolism of absorbed amino acids is poorly characterized. With the aim of elucidating the different effects of inorganic and organic nitrogen sources on primary metabolic pathways in spinach, a gas chromatography–mass spectrometry (GC–MS) based metabolic approach was used to identify differentially abundant compounds in spinach ( Spinacia oleracea ) leaves after 2 days hydroponic-culture in one of three concentrations of glycine or nitrate. Principal component analysis (PCA) revealed distinct clusters for different treatments. The first and second components of PCA accounted for 63.6% and 13.6% of total variance, and seemed to be mainly influenced by the nitrogen concentration and amount of Gly input, respectively. Compared to control treatment, most identified metabolites increased in dose-dependent manners as nitrogen availability increased; however, distinct patterns were observed for NO 3 − - and Gly-treated leaves. When nitrogen was deficient, NO 3 − -treated leaves had lower relative contents of carbohydrates, organic acids and amino acids than Gly treated leaves, this trend reversed under N-sufficient conditions. Furthermore, the metabolites identified were profiled to highlight the overall metabolic shifts between NO 3 − and Gly as N sources at different concentrations.

[1]  Danfeng Huang,et al.  Metabolic Response of Pakchoi Leaves to Amino Acid Nitrogen , 2014 .

[2]  Bowei Xi,et al.  Principal component directed partial least squares analysis for combining nuclear magnetic resonance and mass spectrometry data in metabolomics: application to the detection of breast cancer. , 2011, Analytica chimica acta.

[3]  Danfeng Huang,et al.  Nitrate/Gly ratios in nutrition influenced the growth and amino acid composition in Spinach (Spinacia oleracea L.) , 2014 .

[4]  A. Keys,et al.  An investigation into the interaction between nitrogen nutrition, photosynthesis and photorespiration , 1984, Photosynthesis Research.

[5]  B. Forde,et al.  Glutamate in plants: metabolism, regulation, and signalling. , 2007, Journal of experimental botany.

[6]  Jonathan I. Watkinson,et al.  Physiological and molecular adaptations to drought in Andean potato genotypes , 2008, Journal of experimental botany.

[7]  O. Fiehn.  Metabolomics – the link between genotypes and phenotypes , 2004, Plant Molecular Biology.

[8]  F. Chapin,et al.  Preferential use of organic nitrogen for growth by a non-mycorrhizal arctic sedge , 1993, Nature.

[9]  R. Molinié,et al.  Osmotic stress alters the balance between organic and inorganic solutes in flax (Linum usitatissimum). , 2014, Journal of plant physiology.

[10]  Martin Parry,et al.  Markers and signals associated with nitrogen assimilation in higher plants. , 2003, Journal of experimental botany.

[11]  U. Roessner,et al.  Metabolomics of capsicum ripening reveals modification of the ethylene related-pathway and carbon metabolism , 2014 .

[12]  C. Foyer,et al.  Nitrate activation of cytosolic protein kinases diverts photosynthetic carbon from sucrose to amino Acid biosynthesis: basis for a new concept. , 1992, Plant physiology.

[13]  Zhaohui Li,et al.  Effects of Nitrogen and Calcium Nutrition on Oxalate Contents, Forms, and Distribution in Spinach , 2009 .

[14]  K. Okada,et al.  Differential responses of growth and nitrogen uptake to organic nitrogen in four gramineous crops. , 2004, Journal of experimental botany.

[15]  M. Stitt,et al.  Nitrate Acts as a Signal to Induce Organic Acid Metabolism and Repress Starch Metabolism in Tobacco. , 1997, The Plant cell.

[16]  D. Schomburg,et al.  GC–MS libraries for the rapid identification of metabolites in complex biological samples , 2005, FEBS letters.

[17]  Yves Gibon,et al.  GMD@CSB.DB: the Golm Metabolome Database , 2005, Bioinform..

[18]  O. Ueno,et al.  Ammonia emission from rice leaves in relation to photorespiration and genotypic differences in glutamine synthetase activity. , 2011, Annals of botany.

[19]  T. Näsholm,et al.  Regulation of amino acid uptake in conifers by exogenous and endogenous nitrogen , 2002, Planta.

[20]  Ute Roessner,et al.  What is metabolomics all about? , 2009, BioTechniques.

[21]  D. H. Jennings.  MINERAL NUTRITION IN HIGHER PLANTS (Book). , 1988 .

[22]  C. Bellini,et al.  Comprehensive Screening of Arabidopsis Mutants Suggests the Lysine Histidine Transporter 1 to Be Involved in Plant Uptake of Amino Acids1[W] , 2007, Plant Physiology.

[23]  Mark Stitt,et al.  The interaction between elevated carbon dioxide and nitrogen nutrition: the physiological and molecular background , 1999 .

[24]  D. Appling.  Compartmentation of folate‐mediated one‐carbon metabolism in eukaryotes , 1991, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[25]  F. Chapin,et al.  Tundra Plant Uptake of Amino Acid and NH4+ Nitrogen in Situ: Plants Complete Well for Amino Acid N , 1996 .

[26]  L. Willmitzer,et al.  Transgenic potato plants reveal the indispensable role of cystathionine beta-lyase in plant growth and development. , 2000, The Plant journal : for cell and molecular biology.

[27]  J. Reganold,et al.  Glycine, nitrate, and ammonium uptake by classic and modern wheat varieties in a short-term microcosm study , 2009, Biology and Fertility of Soils.

[28]  R. Azevedo,et al.  The aspartic acid metabolic pathway, an exciting and essential pathway in plants , 2006, Amino Acids.

[29]  H. Marschner.  Mineral Nutrition of Higher Plants , 1988 .

[30]  Bingru Huang,et al.  Differential metabolic responses of perennial grass Cynodon transvaalensis×Cynodon dactylon (C₄) and Poa Pratensis (C₃) to heat stress. , 2011, Physiologia plantarum.

[31]  M. Osaki,et al.  Differences in the metabolite profiles of spinach (Spinacia oleracea L.) leaf in different concentrations of nitrate in the culture solution. , 2008, Plant & cell physiology.

[32]  N. Crawford,et al.  Nitrate: nutrient and signal for plant growth. , 1995, The Plant cell.

[33]  Yves Gibon,et al.  Deficiency of mitochondrial fumarase activity in tomato plants impairs photosynthesis via an effect on stomatal function. , 2007, The Plant journal : for cell and molecular biology.

[34]  K. Kielland,et al.  Uptake of organic nitrogen by plants. , 2009, The New phytologist.

[35]  F. Stuart Chapin,et al.  Integrated Responses of Plants to Stress , 1991 .

[36]  P. Gardeström,et al.  Uptake, metabolism and distribution of organic and inorganic nitrogen sources by Pinus sylvestris. , 2006, Journal of experimental botany.

[37]  M. Osaki,et al.  Metabolite profiling of spinach (Spinacia oleracea L.) leaves by altering the ratio of NH+ 4/NO− 3 in the culture solution , 2009 .

[38]  A. El‐naggar,et al.  Simultaneous Uptake of Multiple Amino Acids by Wheat , 2009 .

[39]  F. Rébeillé,et al.  Glycine and serine catabolism in non-photosynthetic higher plant cells: their role in C1 metabolism. , 1999, The Plant journal : for cell and molecular biology.

[40]  S. Schmidt,et al.  Transporters for uptake and allocation of organic nitrogen compounds in plants , 2007, FEBS letters.