title: Molecular evolution of parsnip (Pastinaca sativa) membrane-bound prenyltransferases for linear and/or angular furanocoumarin biosynthesis.
abstract: In Apiaceae, furanocoumarins (FCs) are plant defence compounds that are present as linear or angular isomers. Angular isomers appeared during plant evolution as a protective response to herbivores that are resistant to linear molecules. Isomeric biosynthesis occurs through prenylation at the C6 or C8 position of umbelliferone. Here, we report cloning and functional characterization of two different prenyltransferases, Pastinaca sativa prenyltransferase 1 and 2 (PsPT1 and PsPT2), that are involved in these crucial reactions. Both enzymes are targeted to plastids and synthesize osthenol and demethylsuberosin (DMS) using exclusively umbelliferone and dimethylallylpyrophosphate (DMAPP) as substrates. Enzymatic characterization using heterologously expressed proteins demonstrated that PsPT1 is specialized for the synthesis of the linear form, demethylsuberosin, whereas PsPT2 more efficiently catalyses the synthesis of its angular counterpart, osthenol. These results are the first example of a complementary prenyltransferase pair from a single plant species that is involved in synthesizing defensive compounds. This study also provides a better understanding of the molecular mechanisms governing the angular FC biosynthetic pathway in apiaceous plants, which involves two paralogous enzymes that share the same phylogenetic origin.
authors.affiliation: Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Munakata
authors.affiliation: Laboratoire Agronomie et Environnement, INRA UMR 1121, 2 avenue de la forêt de Haye TSA 40602 54518, Vandœuvre-lès-Nancy, France.|Laboratoire Agronomie et Environnement, Université de Lorraine UMR 1121, 2 avenue de la forêt de Haye TSA 40602 54518, Vandœuvre-lès-Nancy, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Olry
authors.affiliation: Laboratoire Agronomie et Environnement, INRA UMR 1121, 2 avenue de la forêt de Haye TSA 40602 54518, Vandœuvre-lès-Nancy, France.|Laboratoire Agronomie et Environnement, Université de Lorraine UMR 1121, 2 avenue de la forêt de Haye TSA 40602 54518, Vandœuvre-lès-Nancy, France.
authors.forename: Fazeelat
authors.initials: F
authors.lastname: Karamat
authors.affiliation: EA2106 'Biomolécules et Biotechnologies Végétales', Université François-Rabelais de Tours, Tours, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Courdavault
authors.affiliation: Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.
authors.forename: Akifumi
authors.initials: A
authors.lastname: Sugiyama
authors.affiliation: Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Date
authors.affiliation: Laboratoire Agronomie et Environnement, INRA UMR 1121, 2 avenue de la forêt de Haye TSA 40602 54518, Vandœuvre-lès-Nancy, France.|Laboratoire Agronomie et Environnement, Université de Lorraine UMR 1121, 2 avenue de la forêt de Haye TSA 40602 54518, Vandœuvre-lès-Nancy, France.
authors.forename: Célia
authors.initials: C
authors.lastname: Krieger
authors.affiliation: Laboratoire Agronomie et Environnement, INRA UMR 1121, 2 avenue de la forêt de Haye TSA 40602 54518, Vandœuvre-lès-Nancy, France.|Laboratoire Agronomie et Environnement, Université de Lorraine UMR 1121, 2 avenue de la forêt de Haye TSA 40602 54518, Vandœuvre-lès-Nancy, France.
authors.forename: Prisca
authors.initials: P
authors.lastname: Silie
authors.affiliation: EA2106 'Biomolécules et Biotechnologies Végétales', Université François-Rabelais de Tours, Tours, France.
authors.forename: Emilien
authors.initials: E
authors.lastname: Foureau
authors.affiliation: EA2106 'Biomolécules et Biotechnologies Végétales', Université François-Rabelais de Tours, Tours, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Papon
authors.affiliation: Laboratoire Agronomie et Environnement, INRA UMR 1121, 2 avenue de la forêt de Haye TSA 40602 54518, Vandœuvre-lès-Nancy, France.|Laboratoire Agronomie et Environnement, Université de Lorraine UMR 1121, 2 avenue de la forêt de Haye TSA 40602 54518, Vandœuvre-lès-Nancy, France.
authors.forename: Jérémy
authors.initials: J
authors.lastname: Grosjean
authors.affiliation: Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611-0011, Japan.
authors.forename: Kazufumi
authors.initials: K
authors.lastname: Yazaki
authors.affiliation: Laboratoire Agronomie et Environnement, INRA UMR 1121, 2 avenue de la forêt de Haye TSA 40602 54518, Vandœuvre-lès-Nancy, France.|Laboratoire Agronomie et Environnement, Université de Lorraine UMR 1121, 2 avenue de la forêt de Haye TSA 40602 54518, Vandœuvre-lès-Nancy, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Bourgaud
authors.affiliation: Laboratoire Agronomie et Environnement, INRA UMR 1121, 2 avenue de la forêt de Haye TSA 40602 54518, Vandœuvre-lès-Nancy, France.|Laboratoire Agronomie et Environnement, Université de Lorraine UMR 1121, 2 avenue de la forêt de Haye TSA 40602 54518, Vandœuvre-lès-Nancy, France.
authors.forename: Alain
authors.identifier: 0000-0003-4507-8031
authors.initials: A
authors.lastname: Hehn
mesh_terms: D030008:Catharanthus / Q000235:genetics; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D003374:Coumarins / Q000378:metabolism; D004122:Dimethylallyltranstransferase / Q000235:genetics / Q000378:metabolism*; D019143:Evolution, Molecular; D011564:Furocoumarins / Q000096:biosynthesis*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D028529:Pastinaca / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D018087:Plastids / Q000378:metabolism; D013379:Substrate Specificity; D014026:Nicotiana / Q000235:genetics; D014468:Umbelliferones / Q000096:biosynthesis / Q000378:metabolism
chemical_list: D003374:Coumarins; D011564:Furocoumarins; D007527:Isoenzymes; D010940:Plant Proteins; D014468:Umbelliferones; C114745:osthenol; C031477:7-hydroxycoumarin; D004122:Dimethylallyltranstransferase
keywords: Pastinaca sativa; RNA sequencing; angular furanocoumarins; arms race; linear furanocoumarins; prenyltransferase