title: Virus-induced gene silencing in Rauwolfia species.
abstract: Elucidation of the monoterpene indole alkaloid biosynthesis has recently progressed in Apocynaceae through the concomitant development of transcriptomic analyses and reverse genetic approaches performed by virus-induced gene silencing (VIGS). While most of these tools have been primarily adapted for the Madagascar periwinkle (Catharanthus roseus), the VIGS procedure has scarcely been used on other Apocynaceae species. For instance, Rauwolfia sp. constitutes a unique source of specific and valuable monoterpene indole alkaloids such as the hypertensive reserpine but are also well recognized models for studying alkaloid metabolism, and as such would benefit from an efficient VIGS procedure. By taking advantage of a recent modification in the inoculation method of the Tobacco rattle virus vectors via particle bombardment, we demonstrated that the biolistic-mediated VIGS approach can be readily used to silence genes in both Rauwolfia tetraphylla and Rauwolfia serpentina. After establishing the bombardment conditions minimizing injuries to the transformed plantlets, gene downregulation efficiency was evaluated at approximately a 70% expression decrease in both species by silencing the phytoene desaturase encoding gene. Such a gene silencing approach will thus constitute a critical tool to identify and characterize genes involved in alkaloid biosynthesis in both of these prominent Rauwolfia species.
authors.affiliation: EA 2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, 37200, Tours, France.
authors.forename: Cyrielle
authors.initials: C
authors.lastname: Corbin
authors.affiliation: EA 2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, 37200, Tours, France.
authors.forename: Florent
authors.initials: F
authors.lastname: Lafontaine
authors.affiliation: EA 2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, 37200, Tours, France.|Laboratorio de Biotecnología, Sede de Investigación Universitaria, Universidad de Antioquia, Antioquia, Medellin, Colombia.
authors.forename: Liuda Johana
authors.initials: LJ
authors.lastname: Sepúlveda
authors.affiliation: EA 2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, 37200, Tours, France.
authors.forename: Ines
authors.initials: I
authors.lastname: Carqueijeiro
authors.affiliation: EA 2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, 37200, Tours, France.
authors.forename: Martine
authors.initials: M
authors.lastname: Courtois
authors.affiliation: EA 2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, 37200, Tours, France.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Lanoue
authors.affiliation: EA 2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, 37200, Tours, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Dugé de Bernonville
authors.affiliation: EA 2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, 37200, Tours, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Besseau
authors.affiliation: EA 2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, 37200, Tours, France.
authors.forename: Gaëlle
authors.initials: G
authors.lastname: Glévarec
authors.affiliation: EA 3142 "Groupe d'Etude des Interactions Hôte-Pathogène", Université d'Angers, Angers, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Papon
authors.affiliation: Laboratorio de Biotecnología, Sede de Investigación Universitaria, Universidad de Antioquia, Antioquia, Medellin, Colombia.
authors.forename: Lucia
authors.initials: L
authors.lastname: Atehortúa
authors.affiliation: EA 2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, 37200, Tours, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Giglioli-Guivarc'h
authors.affiliation: EA 2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, 37200, Tours, France.
authors.forename: Marc
authors.initials: M
authors.lastname: Clastre
authors.affiliation: EA 2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, 37200, Tours, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: St-Pierre
authors.affiliation: EA 2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, 37200, Tours, France.
authors.forename: Audrey
authors.initials: A
authors.lastname: Oudin
authors.affiliation: EA 2106 "Biomolécules et Biotechnologies Végétales", UFR Sciences et Techniques, Université François-Rabelais de Tours, 37200, Tours, France. vincent.courdavault@univ-tours.fr.
authors.forename: Vincent
authors.identifier: http://orcid.org/0000-0001-8902-4532
authors.initials: V
authors.lastname: Courdavault
mesh_terms: D019470:Biolistics; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D005822:Genetic Vectors; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D010942:Plant Viruses / Q000235:genetics; D011926:Rauwolfia / Q000201:enzymology / Q000235:genetics*
chemical_list: D010940:Plant Proteins; D010088:Oxidoreductases; C022398:phytoene dehydrogenase
keywords: Medicinal plant; Phytoene desaturase; Rauwolfia; Virus-induced gene silencing