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Methods Mol Biol

Title:		A Biolistic-Mediated Virus-Induced Gene Silencing in Apocynaceae to Map Biosynthetic Pathways of Alkaloids
Author(s):		Cruz PL; Restrepo MI; de Bernonville TD; Oudin A; Munsch T; Lanoue A; Besseau S; Atehortua L; Giglioli-Guivarc'h N; Papon N; Clastre M; Carqueijeiro I; Courdavault V;
Address:		"EA2106 Biomolecules et Biotechnologies Vegetales, Universite de Tours, Tours, France. Laboratorio de Biotecnologia, Universidad de Antioquia, Medellin, Colombia. EA3142 Groupe d'Etude des Interactions Hote-Pathogene, Universite d'Angers, Angers, France. EA2106 Biomolecules et Biotechnologies Vegetales, Universite de Tours, Tours, France. ines.carqueijeiro@univ-tours.fr. EA2106 Biomolecules et Biotechnologies Vegetales, Universite de Tours, Tours, France. vincent.courdavault@univ-tours.fr"
Journal Title:		Methods Mol Biol
Year:		2020
Volume:		2172
Issue:
Page Number:		93 - 110
DOI:		10.1007/978-1-0716-0751-0_8
ISSN/ISBN:		1940-6029 (Electronic) 1064-3745 (Linking)
Abstract:		"Monoterpene indole alkaloids (MIAs) are specialized metabolites synthesized in many plants of the Apocynaceae family including Catharanthus roseus and Rauvolfia sp. MIAs are part of the chemical arsenal that plants evolved to face pet and herbivore attacks, and their high biological activities also confer pharmaceutical properties exploited in human pharmacopeia. Developing robust and straightforward tools to elucidate each step of MIA biosynthetic pathways thus constitutes a prerequisite to the understanding of Apocynaceae defense mechanisms and to the exploitation of MIA cytotoxicity through their production by metabolic engineering. While protocols of virus-induced gene silencing (VIGS) based on Agrobacterium-based transformation have emerged, the recalcitrance of Apocynaceae to this type of transformation prompted us to develop an universal procedure of VIGS vector inoculation. Such procedure relies on the delivery of the transforming plasmids through a particle bombardment performed using a biolistic device and offers the possibility to overcome host specificity to silence genes in any plant species. Using silencing of geissoschizine oxidase as an example, we described the main steps of this biolistic mediated VIGS in C. roseus and R. tetraphylla"
Keywords:		"Alkaloids/*metabolism Apocynaceae/*genetics/*metabolism Biolistics Gene Expression Regulation, Plant/genetics/physiology Gene Silencing/physiology Plant Proteins/genetics/*metabolism Plasmids/genetics Alkaloids Apocynaceae Gene validation Silencing Vigs;"
Notes:		"MedlineCruz, Pamela Lemos Restrepo, Maria Isabel de Bernonville, Thomas Duge Oudin, Audrey Munsch, Thibaut Lanoue, Arnaud Besseau, Sebastien Atehortua, Lucia Giglioli-Guivarc'h, Nathalie Papon, Nicolas Clastre, Marc Carqueijeiro, Ines Courdavault, Vincent eng 2020/06/20 Methods Mol Biol. 2020; 2172:93-110. doi: 10.1007/978-1-0716-0751-0_8"