Inventory of metal complexes circulating in plant fluids: a reliable method based on HPLC coupled with dual elemental and high-resolution molecular mass spectrometric detection.
Description of metal species in plant fluids such as xylem, phloem or related saps remains a complex challenge usually addressed either by liquid chromatography-mass spectrometry, X-ray analysis or computational prediction. To date, none of these techniques has achieved a complete and true picture of metal-containing species in plant fluids, especially for the least concentrated complexes. Here, we present a generic analytical methodology for a large-scale (> 10 metals, > 50 metal complexes) detection, identification and semiquantitative determination of metal complexes in the xylem and embryo sac liquid of the green pea, Pisum sativum. The procedure is based on direct injection using hydrophilic interaction chromatography with dual detection by elemental (inductively coupled plasma mass spectrometry) and molecular (high-resolution electrospray mass spectrometry) mass spectrometric detection. Numerous and novel complexes of iron(II), iron(III), copper(II), zinc, manganese, cobalt(II), cobalt(III), magnesium, calcium, nickel and molybdenum(IV) with several ligands including nicotianamine, citrate, malate, histidine, glutamine, aspartic acid, asparagine, phenylalanine and others are observed in pea fluids and discussed. This methodology provides a large inventory of various types of metal complexes, which is a significant asset for future biochemical and genetic studies into metal transport/homeostasis.
Laboratoire de Chimie Analytique Bio-Inorganique et Environnement (LCABIE), Institut des Sciences Analytiques et de Physico-chimie pour l'Environnement et les Matériaux (UMR5254), Centre National de la Recherche Scientifique, Université de Pau et des Pays de l'Adour, Pau Cedex 9, F-64063, France.
Paulina
P
Flis
Laboratoire de Chimie Analytique Bio-Inorganique et Environnement (LCABIE), Institut des Sciences Analytiques et de Physico-chimie pour l'Environnement et les Matériaux (UMR5254), Centre National de la Recherche Scientifique, Université de Pau et des Pays de l'Adour, Pau Cedex 9, F-64063, France.
Laurent
L
Ouerdane
Laboratoire de Biochimie et Physiologie Moléculaire des Plantes (BPMP), Institut de Biologie Intégrative des Plantes, Centre National de la Recherche Scientifique (UMR5004), Institut National de la Recherche Agronomique, Université Montpellier II, Ecole Nationale Supérieure d'Agronomie, Montpellier Cedex 2, F-34060, France.
Louis
L
Grillet
Laboratoire de Biochimie et Physiologie Moléculaire des Plantes (BPMP), Institut de Biologie Intégrative des Plantes, Centre National de la Recherche Scientifique (UMR5004), Institut National de la Recherche Agronomique, Université Montpellier II, Ecole Nationale Supérieure d'Agronomie, Montpellier Cedex 2, F-34060, France.
Catherine
C
Curie
Laboratoire de Biochimie et Physiologie Moléculaire des Plantes (BPMP), Institut de Biologie Intégrative des Plantes, Centre National de la Recherche Scientifique (UMR5004), Institut National de la Recherche Agronomique, Université Montpellier II, Ecole Nationale Supérieure d'Agronomie, Montpellier Cedex 2, F-34060, France.
Stéphane
S
Mari
Laboratoire de Chimie Analytique Bio-Inorganique et Environnement (LCABIE), Institut des Sciences Analytiques et de Physico-chimie pour l'Environnement et les Matériaux (UMR5254), Centre National de la Recherche Scientifique, Université de Pau et des Pays de l'Adour, Pau Cedex 9, F-64063, France.
Ryszard
R
Lobinski
D000596:Amino Acids / Q000378:metabolism; D001692:Biological Transport; D002264:Carboxylic Acids / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056831:Coordination Complexes / Q000032:analysis*; D006706:Homeostasis; D057927:Hydrophobic and Hydrophilic Interactions; D013058:Mass Spectrometry / Q000379:methods*; D055442:Metabolome; D008670:Metals / Q000032:analysis; D018532:Pisum sativum / Q000378:metabolism; D010944:Plants / Q000737:chemistry*; D015203:Reproducibility of Results; D012639:Seeds / Q000378:metabolism; D052584:Xylem / Q000378:metabolism
D000596:Amino Acids; D002264:Carboxylic Acids; D056831:Coordination Complexes; D008670:Metals
Pisum sativum; embryo sac liquid; hydrophilic interaction chromatography (HILIC); mass spectrometry; metal complexes; metal speciation; nicotianamine (NA); xylem