Metabolomic characterisation of the functional division of nitrogen metabolism in variegated leaves
Guillaume Tcherkez A B E , Florence Guérard C , Françoise Gilard C , Marlène Lamothe C , Caroline Mauve C , Elisabeth Gout D and Richard Bligny DA Institut de Biologie des Plantes, CNRS UMR8618, Université Paris-Sud, 91405 Orsay cedex, France.
B Institut Universitaire de France, 103 Boulevard Saint-Michel, 75005 Paris, France.
C Plateforme Métabolisme-Métabolome, IFR87, Batiment 630, Université Paris-Sud, 91405 Orsay cedex, France.
D Laboratoire de Physiologie Cellulaire Végétale, CEA-Grenoble, 17 rue des Martyrs, 38054 Grenoble cedex 9, France.
E Corresponding author. Email: guillaume.tcherkez@u-psud.fr
Functional Plant Biology 39(12) 959-967 https://doi.org/10.1071/FP12189
Submitted: 28 June 2012 Accepted: 30 August 2012 Published: 19 October 2012
Abstract
Many horticultural and natural plant species have variegated leaves, that is, patchy leaves with green and non-green or white areas. Specific studies on the metabolism of variegated leaves are scarce and although white (non-green) areas have been assumed to play the role of a ‘nitrogen store’, there is no specific studies showing the analysis of nitrogenous metabolites and the dynamics of nitrogen assimilation. Here, we examined the metabolism of variegated leaves of Pelargonium × hortorum. We show that white areas have a larger N : C ratio, more amino acids, with a clear accumulation of arginine. Metabolomic analyses revealed clear differences in the chemical composition, suggesting contrasted metabolic commitments such as an enhancement of alkaloid biosynthesis in white areas. Using isotopic labelling followed by nuclear magnetic resonance or liquid chromatography/mass spectrometry, we further showed that in addition to glutamine, tyrosine and tryptophan, N metabolism forms ornithine in green area and huge amounts of arginine in white areas. Fine isotopic measurements with isotope ratio mass spectrometry indicated that white and green areas exchange nitrogenous molecules but nitrogen export from green areas is quantitatively much more important. The biological significance of the metabolic exchange between leaf areas is briefly discussed.
Additional keywords: isotopes, nitrogen metabolism, Pelargonium, variegation.
References
Bacon JSD, Palmer MJ, DeKock PC (1961) The measurement of aconitase activity in the leaves of various normal and variegated plants. Biochemical Journal 78, 198–204.Borner T (1986) Chloroplast control of nuclear gene function. Endocytobiosis and Cell Research 3, 265–274.
Borner T, Mandel RR, Schiemann J (1986) Nitrate reductase is not accumulated in chloroplast-ribosome-deficient mutants of higher plants. Planta 169, 202–207.
| Nitrate reductase is not accumulated in chloroplast-ribosome-deficient mutants of higher plants.Crossref | GoogleScholarGoogle Scholar |
DeKock PC, Morrison RI (1958a) The metabolism of chlorotic leaves. 1. Amino acids. Biochemical Journal 70, 266–272.
DeKock PC, Morrison RI (1958b) The metabolism of chlorotic leaves. 2. Organic acids. Biochemical Journal 70, 272–277.
Downton WJS, Grant WJR (1994) Photosynthetic and growth responses of variegated ornamental species to elevated CO2. Australian Journal of Plant Physiology 21, 273–279.
| Photosynthetic and growth responses of variegated ornamental species to elevated CO2.Crossref | GoogleScholarGoogle Scholar |
Evenari M (1989) The history of research on white-green variegated plants. Botanical Review 55, 106–139.
| The history of research on white-green variegated plants.Crossref | GoogleScholarGoogle Scholar |
Godel H, Graser T, Földi P, Pfaender P, Fürst P (1984) Measurement of free amino acids in human biological fluids by high-performance liquid chromatography. Journal of Chromatography. A 297, 49–61.
| Measurement of free amino acids in human biological fluids by high-performance liquid chromatography.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL2cXltVKktro%3D&md5=c9d7538aac3a7dca8dd361ef848fac42CAS |
Guérard F, Petriacq P, Gakière B, Tcherkez G (2011) Liquid chromatography/time-of-flight mass spectrometry for the analysis of plant samples: a method for simultaneous screening of common cofactors or nucleotides and application to an engineered plant line. Plant Physiology and Biochemistry 49, 1117–1125.
| Liquid chromatography/time-of-flight mass spectrometry for the analysis of plant samples: a method for simultaneous screening of common cofactors or nucleotides and application to an engineered plant line.Crossref | GoogleScholarGoogle Scholar |
Houghton P, Lis-Balchin M (2002) Chemotaxonomy of Pelargonium based on alkaloids and essential oils. In ‘Geranium and Pelargonium, history of nomenclature, usage and cultivation’. (Ed. M Lis-Balchin) pp. 166–173. (CRC Press: London)
Ivanova TI, Sherstneva OA (1999) Dark respiration of variegated leaves in plants of different life forms. Russian Journal of Plant Physiology: a Comprehensive Russian Journal on Modern Phytophysiology 46, 666–673.
Jones H, Eagles JE (1962) Translocation of 14C-carbon within and between leaves. Annals of Botany 26, 505–510.
Lis-Balchin M (1997) A chemotaxonomic study of the Pelargonium (Geraniaceae) species and their modern cultivars. Journal of Horticultural Science 72, 791–795.
Madore MA (1990) Carbohydrate metabolism in photosynthetic and non-photosynthetic tissues of variegated leaves of Coleus blumei Benth. Plant Physiology 93, 617–622.
| Carbohydrate metabolism in photosynthetic and non-photosynthetic tissues of variegated leaves of Coleus blumei Benth.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3cXkslCqurY%3D&md5=58df0f33467eb0a1d7a0bfe3759d9addCAS |
Pringsheim EG, Schwarz W (1933) Das Auftretenweissbunter(panaschierter) Pflanzen in der Natur. Flora 28, 111–122.
Sawhney SK, Prakash V, Naik MS (1972) Nitrate reductase and nitrite reductase activities in induced chlorophyll mutants of barley. FEBS Letters 22, 200–202.
| Nitrate reductase and nitrite reductase activities in induced chlorophyll mutants of barley.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaE38XksFyrurs%3D&md5=68d29fa7042a99f056124a5cf3635530CAS |
Schlee D, Reinbothe H (1965) A relation of adenine to serine biosynthesis in chlorophyll-deficient leaves of Pelargonium zonale. Phytochemistry 4, 311–315.
| A relation of adenine to serine biosynthesis in chlorophyll-deficient leaves of Pelargonium zonale.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaF2MXktVCqu7Y%3D&md5=71f130bbe2e2925929685891b0cf07acCAS |
Seltmann H (1955) Comparative physiology of green and albino corn seedlings. Plant Physiology 30, 258–263.
| Comparative physiology of green and albino corn seedlings.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaG2MXosFWguw%3D%3D&md5=a0fcc7085f2212e73088284828a55679CAS |
Slocum RD (2005) Genes, enzymes and regulation of arginine biosynthesis in plants. Plant Physiology and Biochemistry 43, 729–745.
| Genes, enzymes and regulation of arginine biosynthesis in plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXhtFWhtr3K&md5=ab912471b148e6e09abc83f83f767117CAS |
Smith HB (1999) Photosynthetic pigmentation – variegation on a theme. The Plant Cell 11, 1–3.
| Photosynthetic pigmentation – variegation on a theme.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXhtVGrsL0%3D&md5=7db3074c5981e2008abdda812dea497dCAS |
Tanabe Y, Kawashima N (1982) Comparison of free and bound amino acids in white and green tissues of variegated tobacco. Plant & Cell Physiology 23, 433–439.
Tanabe Y, Sano M, Kawashima N (1982) Changes in free amino acids in white and green tissues of variegated tobacco leaves during water stress. Plant & Cell Physiology 23, 1229–1235.
Taylor WC (1989) Regulatory interactions between nuclear and plastid genomes. Annual Review of Plant Physiology and Plant Molecular Biology 40, 211–233.
| Regulatory interactions between nuclear and plastid genomes.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL1MXktlKmurk%3D&md5=dcffef67ee7319edd19eb6dbdea70bddCAS |
Tcherkez G, Mahé A, Gauthier P, Mauve C, Gout E, Bligny R, Cornic G, Hodges M (2009) In folio respiratory fluxomics revealed by 13C isotopic labeling and H/D isotope effects highlight the non-cyclic nature of the tricarboxylic acid ‘cycle’ in illuminated leaves. Plant Physiology 151, 620–630.
| In folio respiratory fluxomics revealed by 13C isotopic labeling and H/D isotope effects highlight the non-cyclic nature of the tricarboxylic acid ‘cycle’ in illuminated leaves.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXht12qu7fK&md5=e8fd2894fe28ffbff8b425980ca6944eCAS |
Tcherkez G, Mauve C, Lamothe M, Le Bras C, Grapin A (2011) The 13C/12C isotopic signal of day and night respired CO2 in variegated leaves of Pelargonium × hortorum. Plant, Cell & Environment 34, 270–283.
| The 13C/12C isotopic signal of day and night respired CO2 in variegated leaves of Pelargonium × hortorum.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXisVGjsb8%3D&md5=ac780a22318141e1a25c1db0366bccebCAS |
Toshoji H, Katsumata T, Takusagawa M, Yusa Y, Sakai A (2011) Effects of chloroplast dysfunction on mitochondria: white sectors in variegated leaves have higher mitochondrial DNA levels and lower dark respiration rates than green sectors. Protoplasma 249, 805–817.
| Effects of chloroplast dysfunction on mitochondria: white sectors in variegated leaves have higher mitochondrial DNA levels and lower dark respiration rates than green sectors.Crossref | GoogleScholarGoogle Scholar |
Vaughn KC, Stewart KD (1978) Light-harvesting pigment-protein complex deficiency in Hosta (Liliaceae). Planta 143, 275–278.
| Light-harvesting pigment-protein complex deficiency in Hosta (Liliaceae).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaE1MXjs1Cgtw%3D%3D&md5=ac80b144de858eec26a026b7d254b6bcCAS |
