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.
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Acknowledgements
We gratefully acknowledge the support from the Région Centre (France, ABISAL grant, PhD fellowship to F.L., and postdoctoral fellowship to I. C.) and from Colciencias (Colombia, PhD fellowship to L.S). We thank Sarah O’Connor for providing us the R. serpentina seeds. We also thank Cédric Labarre, Evelyne Danos, and the Botanical Garden from Tours for the R. tetraphylla seeds and for the help in maintaining plants. We acknowledge Johan-Owen De Craene for critical reading of the manuscript.
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Handling Editor: Manfred Heinlein
Cyrielle Corbin, Florent Lafontaine, and Liuda Johana Sepúlved contributed equally to this work.
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Corbin, C., Lafontaine, F., Sepúlveda, L.J. et al. Virus-induced gene silencing in Rauwolfia species. Protoplasma 254, 1813–1818 (2017). https://doi.org/10.1007/s00709-017-1079-y
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DOI: https://doi.org/10.1007/s00709-017-1079-y