Improved virus-induced gene silencing allows discovery of a serpentine synthase gene in Catharanthus roseus.

Autor:	Yamamoto K; Department of Natural Product Biosynthesis, Max Planck Institute for Chemical Ecology, Jena 07745, Germany.; Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 263-8522, Japan., Grzech D; Department of Natural Product Biosynthesis, Max Planck Institute for Chemical Ecology, Jena 07745, Germany., Koudounas K; EA2106 'Biomolécules et Biotechnologies Végétales', Université de Tours, Tours 37200, France., Stander EA; EA2106 'Biomolécules et Biotechnologies Végétales', Université de Tours, Tours 37200, France., Caputi L; Department of Natural Product Biosynthesis, Max Planck Institute for Chemical Ecology, Jena 07745, Germany., Mimura T; Department of Biology, Graduate School of Science, Kobe University, Kobe, Hyogo 657-8501, Japan., Courdavault V; EA2106 'Biomolécules et Biotechnologies Végétales', Université de Tours, Tours 37200, France., O'Connor SE; Department of Natural Product Biosynthesis, Max Planck Institute for Chemical Ecology, Jena 07745, Germany.
Jazyk:	angličtina
Zdroj:	Plant physiology [Plant Physiol] 2021 Oct 05; Vol. 187 (2), pp. 846-857.
DOI:	10.1093/plphys/kiab285
Abstrakt:	Specialized metabolites are chemically complex small molecules with a myriad of biological functions. To investigate plant-specialized metabolite biosynthesis more effectively, we developed an improved method for virus-induced gene silencing (VIGS). We designed a plasmid that incorporates fragments of both the target gene and knockdown marker gene (phytoene desaturase, PDS), which identifies tissues that have been successfully silenced in planta. To demonstrate the utility of this method, we used the terpenoid indole alkaloid (TIA) pathway in Madagascar periwinkle (Catharanthus roseus) as a model system. Catharanthus roseus is a medicinal plant well known for producing many bioactive compounds, such as vinblastine and vincristine. Our VIGS method enabled the discovery of a previously unknown biosynthetic enzyme, serpentine synthase (SS). This enzyme is a cytochrome P450 (CYP) that produces the β-carboline alkaloids serpentine and alstonine, compounds with strong blue autofluorescence and potential pharmacological activity. The discovery of this enzyme highlights the complexity of TIA biosynthesis and demonstrates the utility of this improved VIGS method for discovering unidentified metabolic enzymes in plants. (© The Author(s) 2021. Published by Oxford University Press on behalf of American Society of Plant Biologists.)
Databáze:	MEDLINE
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