title: Mapping the genetic and tissular diversity of 64 phenolic compounds in Citrus species using a UPLC-MS approach.
abstract: OBJECTIVE
  Phenolic compounds contribute to food quality and have potential health benefits. Consequently, they are an important target of selection for Citrus species. Numerous studies on this subject have revealed new molecules, potential biosynthetic pathways and linkage between species. Although polyphenol profiles are correlated with gene expression, which is responsive to developmental and environmental cues, these factors are not monitored in most studies. A better understanding of the biosynthetic pathway and its regulation requires more information about environmental conditions, tissue specificity and connections between competing sub-pathways. This study proposes a rapid method, from sampling to analysis, that allows the quantitation of multiclass phenolic compounds across contrasting tissues and cultivars.
  
  
  METHODS
  Leaves and fruits of 11 cultivated citrus of commercial interest were collected from adult trees grown in an experimental orchard. Sixty-four phenolic compounds were simultaneously quantified by ultra-high-performance liquid chromatography coupled with mass spectrometry.
  
  
  RESULTS
  Combining data from vegetative tissues with data from fruit tissues improved cultivar classification based on polyphenols. The analysis of metabolite distribution highlighted the massive accumulation of specific phenolic compounds in leaves and the external part of the fruit pericarp, which reflects their involvement in plant defence. The overview of the biosynthetic pathway obtained confirmed some regulatory steps, for example those catalysed by rhamnosyltransferases. The results suggest that three other steps are responsible for the different metabolite profiles in 'Clementine' and 'Star Ruby' grapefruit.
  
  
  CONCLUSIONS
  The method described provides a high-throughput method to study the distribution of phenolic compounds across contrasting tissues and cultivars in Citrus, and offers the opportunity to investigate their regulation and physiological roles. The method was validated in four different tissues and allowed the identification and quantitation of 64 phenolic compounds in 20 min, which represents an improvement over existing methods of analysing multiclass polyphenols.
authors.affiliation: CIRAD, UMR AGAP, F-20230 San Giuliano, France, INRA, UMR AGAP, F-20230 San Giuliano, France, Université de Lorraine, UMR 1121 Laboratoire Agronomie et Environnement Nancy-Colmar, 2 avenue de la forêt de Haye, TSA 40602, F-54518 Vandœuvre-lès-Nancy, France, INRA, UMR AGAP, Place P. Viala, F-34060 Montpellier, France, Université Montpellier II, UMR DIADE, F-34394 Montpellier, France and INRA, UR 1115, Plantes et Systèmes de Culture Horticoles, Domaine St-Paul - Site Agroparc, F-84914 Avignon, France CIRAD, UMR AGAP, F-20230 San Giuliano, France, INRA, UMR AGAP, F-20230 San Giuliano, France, Université de Lorraine, UMR 1121 Laboratoire Agronomie et Environnement Nancy-Colmar, 2 avenue de la forêt de Haye, TSA 40602, F-54518 Vandœuvre-lès-Nancy, France, INRA, UMR AGAP, Place P. Viala, F-34060 Montpellier, France, Université Montpellier II, UMR DIADE, F-34394 Montpellier, France and INRA, UR 1115, Plantes et Systèmes de Culture Horticoles, Domaine St-Paul - Site Agroparc, F-84914 Avignon, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Durand-Hulak
authors.affiliation: CIRAD, UMR AGAP, F-20230 San Giuliano, France, INRA, UMR AGAP, F-20230 San Giuliano, France, Université de Lorraine, UMR 1121 Laboratoire Agronomie et Environnement Nancy-Colmar, 2 avenue de la forêt de Haye, TSA 40602, F-54518 Vandœuvre-lès-Nancy, France, INRA, UMR AGAP, Place P. Viala, F-34060 Montpellier, France, Université Montpellier II, UMR DIADE, F-34394 Montpellier, France and INRA, UR 1115, Plantes et Systèmes de Culture Horticoles, Domaine St-Paul - Site Agroparc, F-84914 Avignon, France.
authors.forename: Audray
authors.initials: A
authors.lastname: Dugrand
authors.affiliation: CIRAD, UMR AGAP, F-20230 San Giuliano, France, INRA, UMR AGAP, F-20230 San Giuliano, France, Université de Lorraine, UMR 1121 Laboratoire Agronomie et Environnement Nancy-Colmar, 2 avenue de la forêt de Haye, TSA 40602, F-54518 Vandœuvre-lès-Nancy, France, INRA, UMR AGAP, Place P. Viala, F-34060 Montpellier, France, Université Montpellier II, UMR DIADE, F-34394 Montpellier, France and INRA, UR 1115, Plantes et Systèmes de Culture Horticoles, Domaine St-Paul - Site Agroparc, F-84914 Avignon, France.
authors.forename: Thibault
authors.initials: T
authors.lastname: Duval
authors.affiliation: CIRAD, UMR AGAP, F-20230 San Giuliano, France, INRA, UMR AGAP, F-20230 San Giuliano, France, Université de Lorraine, UMR 1121 Laboratoire Agronomie et Environnement Nancy-Colmar, 2 avenue de la forêt de Haye, TSA 40602, F-54518 Vandœuvre-lès-Nancy, France, INRA, UMR AGAP, Place P. Viala, F-34060 Montpellier, France, Université Montpellier II, UMR DIADE, F-34394 Montpellier, France and INRA, UR 1115, Plantes et Systèmes de Culture Horticoles, Domaine St-Paul - Site Agroparc, F-84914 Avignon, France.
authors.forename: Luc P R
authors.initials: LP
authors.lastname: Bidel
authors.affiliation: CIRAD, UMR AGAP, F-20230 San Giuliano, France, INRA, UMR AGAP, F-20230 San Giuliano, France, Université de Lorraine, UMR 1121 Laboratoire Agronomie et Environnement Nancy-Colmar, 2 avenue de la forêt de Haye, TSA 40602, F-54518 Vandœuvre-lès-Nancy, France, INRA, UMR AGAP, Place P. Viala, F-34060 Montpellier, France, Université Montpellier II, UMR DIADE, F-34394 Montpellier, France and INRA, UR 1115, Plantes et Systèmes de Culture Horticoles, Domaine St-Paul - Site Agroparc, F-84914 Avignon, France.
authors.forename: Christian
authors.initials: C
authors.lastname: Jay-Allemand
authors.affiliation: CIRAD, UMR AGAP, F-20230 San Giuliano, France, INRA, UMR AGAP, F-20230 San Giuliano, France, Université de Lorraine, UMR 1121 Laboratoire Agronomie et Environnement Nancy-Colmar, 2 avenue de la forêt de Haye, TSA 40602, F-54518 Vandœuvre-lès-Nancy, France, INRA, UMR AGAP, Place P. Viala, F-34060 Montpellier, France, Université Montpellier II, UMR DIADE, F-34394 Montpellier, France and INRA, UR 1115, Plantes et Systèmes de Culture Horticoles, Domaine St-Paul - Site Agroparc, F-84914 Avignon, France froelicher@cirad.fr.
authors.forename: Yann
authors.initials: Y
authors.lastname: Froelicher
authors.affiliation: CIRAD, UMR AGAP, F-20230 San Giuliano, France, INRA, UMR AGAP, F-20230 San Giuliano, France, Université de Lorraine, UMR 1121 Laboratoire Agronomie et Environnement Nancy-Colmar, 2 avenue de la forêt de Haye, TSA 40602, F-54518 Vandœuvre-lès-Nancy, France, INRA, UMR AGAP, Place P. Viala, F-34060 Montpellier, France, Université Montpellier II, UMR DIADE, F-34394 Montpellier, France and INRA, UR 1115, Plantes et Systèmes de Culture Horticoles, Domaine St-Paul - Site Agroparc, F-84914 Avignon, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Bourgaud
authors.affiliation: CIRAD, UMR AGAP, F-20230 San Giuliano, France, INRA, UMR AGAP, F-20230 San Giuliano, France, Université de Lorraine, UMR 1121 Laboratoire Agronomie et Environnement Nancy-Colmar, 2 avenue de la forêt de Haye, TSA 40602, F-54518 Vandœuvre-lès-Nancy, France, INRA, UMR AGAP, Place P. Viala, F-34060 Montpellier, France, Université Montpellier II, UMR DIADE, F-34394 Montpellier, France and INRA, UR 1115, Plantes et Systèmes de Culture Horticoles, Domaine St-Paul - Site Agroparc, F-84914 Avignon, France CIRAD, UMR AGAP, F-20230 San Giuliano, France, INRA, UMR AGAP, F-20230 San Giuliano, France, Université de Lorraine, UMR 1121 Laboratoire Agronomie et Environnement Nancy-Colmar, 2 avenue de la forêt de Haye, TSA 40602, F-54518 Vandœuvre-lès-Nancy, France, INRA, UMR AGAP, Place P. Viala, F-34060 Montpellier, France, Université Montpellier II, UMR DIADE, F-34394 Montpellier, France and INRA, UR 1115, Plantes et Systèmes de Culture Horticoles, Domaine St-Paul - Site Agroparc, F-84914 Avignon, France.
authors.forename: Anne-Laure
authors.initials: AL
authors.lastname: Fanciullino
mesh_terms: D053898:Biosynthetic Pathways; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002957:Citrus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016000:Cluster Analysis; D005419:Flavonoids / Q000737:chemistry / Q000378:metabolism*; D005638:Fruit / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D009928:Organ Specificity; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D018515:Plant Leaves / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D059808:Polyphenols / Q000737:chemistry / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D005419:Flavonoids; D010936:Plant Extracts; D059808:Polyphenols
keywords: Citrus; Phenolic compounds; UPLC–MS; biosynthetic pathway.; coumarins; flavonoids; high-throughput method; phylogenetic relationships; tissue diversity