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A Biolistic-Mediated Virus-Induced Gene Silencing in Apocynaceae to Map Biosynthetic Pathways of Alkaloids

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Virus-Induced Gene Silencing in Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2172))

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.

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Authors and Affiliations

  1. EA2106 Biomolécules et Biotechnologies Végétales, Université de Tours, Tours, France

    Pamela Lemos Cruz, Thomas Dugé de Bernonville, Audrey Oudin, Thibaut Munsch, Arnaud Lanoue, Sébastien Besseau, Nathalie Giglioli-Guivarc’h, Marc Clastre, Inês Carqueijeiro & Vincent Courdavault

  2. Laboratorio de Biotecnología, Universidad de Antioquia, Medellin, Colombia

    María Isabel Restrepo & Lucia Atehortùa

  3. EA3142 Groupe d’Etude des Interactions Hôte-Pathogène, Université d’Angers, Angers, France

    Nicolas Papon

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  1. Pamela Lemos Cruz
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Corresponding authors

Correspondence to Inês Carqueijeiro or Vincent Courdavault .

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Editors and Affiliations

  1. EA2106 Biomolécules et Biotechnologies Végétales, Université de Tours, Tours, France

    Vincent Courdavault

  2. EA2106 Biomolécules et Biotechnologies Végétales, Université de Tours, Tours, France

    Sébastien Besseau

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Cruz, P.L. et al. (2020). A Biolistic-Mediated Virus-Induced Gene Silencing in Apocynaceae to Map Biosynthetic Pathways of Alkaloids. In: Courdavault, V., Besseau, S. (eds) Virus-Induced Gene Silencing in Plants. Methods in Molecular Biology, vol 2172. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0751-0_8

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  • DOI: https://doi.org/10.1007/978-1-0716-0751-0_8

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0750-3

  • Online ISBN: 978-1-0716-0751-0

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