Cellular and Subcellular Compartmentation of the 2 C -Methyl-D-Erythritol 4-Phosphate Pathway in the Madagascar Periwinkle
Guirimand, Grégory (Kobe University. Graduate School of Science, Technology & Innovation)
Guihur, Anthony (University of Lausanne. Department of Plant Molecular Biology)
Perello, Catalina (Centre de Recerca en Agrigenòmica)
Phillips, Michael (University of Toronto-Mississauga. Department of Biology)
Mahroug, Samira (University of Sidi-Bel-Abbes. Department of Environment Sciences)
Oudin, Audrey (Université de Tours. Département of Agronomie, productions animale et végétale et agro-alimentaire)
Dugé de Bernonville, Thomas (Université de Tours. Département of Agronomie, productions animale et végétale et agro-alimentaire)
Besseau, Sébastien (Université de Tours. Département of Agronomie, productions animale et végétale et agro-alimentaire)
Lanoue, Arnaud (Université de Tours. Département of Agronomie, productions animale et végétale et agro-alimentaire)
Giglioli-Guivarc'h, Nathalie (Université de Tours. Département of Agronomie, productions animale et végétale et agro-alimentaire)
Papon, Nicolas (Université d'Angers)
St-Pierre, Benoit (Université de Tours. Département of Agronomie, productions animale et végétale et agro-alimentaire)
Rodríguez Concepción, Manuel (Centre de Recerca en Agrigenòmica)
Burlat, Vincent (Université de Toulouse. Laboratoire de Recherche en Sciences Végétales)
Courdavault, Vincent (Université de Tours. Département of Agronomie, productions animale et végétale et agro-alimentaire)

Fecha:	2020
Resumen:	The Madagascar periwinkle (Catharanthus roseus) synthesizes the highly valuable monoterpene indole alkaloids (MIAs) through a long metabolic route initiated by the 2 C -methyl-D-erythritol 4-phosphate (MEP) pathway. In leaves, a complex compartmentation of the MIA biosynthetic pathway occurs at both the cellular and subcellular levels, notably for some gene products of the MEP pathway. To get a complete overview of the pathway organization, we cloned four genes encoding missing enzymes involved in the MEP pathway before conducting a systematic analysis of transcript distribution and protein subcellular localization. RNA in situ hybridization revealed that all MEP pathway genes were coordinately and mainly expressed in internal phloem-associated parenchyma of young leaves, reinforcing the role of this tissue in MIA biosynthesis. At the subcellular level, transient cell transformation and expression of fluorescent protein fusions showed that all MEP pathway enzymes were targeted to plastids. Surprisingly, two isoforms of 1-deoxy-D-xylulose 5-phosphate synthase and 1-deoxy-D-xylulose 5-phosphate reductoisomerase initially exhibited an artifactual aggregated pattern of localization due to high protein accumulation. Immunogold combined with transmission electron microscopy, transient transformations performed with a low amount of transforming DNA and fusion/deletion experiments established that both enzymes were rather diffuse in stroma and stromules of plastids as also observed for the last six enzymes of the pathway. Taken together, these results provide new insights into a potential role of stromules in enhancing MIA precursor exchange with other cell compartments to favor metabolic fluxes towards the MIA biosynthesis.
Derechos:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Lengua:	Anglès
Documento:	Article ; recerca ; Versió publicada
Materia:	2 C -methyl-D-erythritol 4-phosphate pathway ; Compartmentation ; Catharanthus roseus ; Stromules ; GFP imaging ; Localization artifact ; Alkaloids
Publicado en:	Plants, Vol. 9, Issue 4 (April 2020) , art. 462, ISSN 2223-7747

DOI: 10.3390/plants9040462
PMID: 32272573

25 p, 10.1 MB

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