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Modification de la synthèse des furocoumarines chez Ruta graveolens L. par une approche de génie métabolique

Abstract : Garden Rue (Ruta graveolens L.) is a plant known as being particularly rich in secondary metabolites and in particular producing molecules of pharmaceutical interest like furocoumarines. By the use of a metabolic engineering approach, we tried to increase the content of furocoumarines produced in these plants but also to better understand the regulation mechanisms of the phenylpropanoïd biosynthesis pathway. To achieve these goals we transformed Ruta plants with various genes placed under the control of a strong constitutive promoter, CaMV 35S promoter. The plants we obtained were analyzed for their ability to overproduce furocoumarines but also other phenylpropanoïds like ferulate, umbelliferone, scopoletine or rutin. Using Real Time PCR experiments, a correlation was carried out with the level of expression of each transgene and several endogenous genes. Plants overexpressing either the Ruta or the Arabidopsis Coumaroyl ester 3 '-Hydroxylase (CYP98A22 and CYP98A3 respectively) display both a significant increase (3 time level) of the furocoumarin. However if the S-98A22 plants are characterized by a reduction in the production of rutin and umbelliferone, S-98A3 transgenic plants display a significant increase scopoletine and umbelliferone content. These results suggest the coexistence of two C3'H having different functions in Ruta. One of them might be involved more specifically in the synthesis of scopoletine. If the transformation of Ruta with genes belonging to the CYP98A family generates an enlarged of the secondary metabolism, we also showed that the overexpression of a gene belonging to the furocoumarins biosynthesis pathway (CYP71AJ1, the psoralen synthase) allowed a specific stimulation. Indeed a 4 time increase of the content of furocouramins was noticed in these transgenic plant lines. This work made it possible to make evidence of the interest of a metabolic engineering approach to generate plants of interest for the production of pharmaceutical molecules
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Sébastien Doerper. Modification de la synthèse des furocoumarines chez Ruta graveolens L. par une approche de génie métabolique. Sciences agricoles. Institut National Polytechnique de Lorraine, 2008. Français. ⟨NNT : 2008INPL070N⟩. ⟨tel-01753051⟩



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