Caractérisation fonctionnelle de gènes impliqués dans la tolérance au stress métallique chez les champignons ectomycorhiziens par agrotransformation d'Hebeloma cylindrosporum

Abstract : The purpose of my work was to develop a method of genetic transformation through Agrobacterium tumefaciens (agrotransformation) of the ectomycorrhizal fungus Hebeloma cylindrosporum to characterize the function of target genes. These genes were the zinc transport protein HcZnT1 of H. cylindrosporum, the thioredoxin PiTrx and the metallothionein PiMt1, from the fungus Paxillus involutus. The method of agrotransformation, which has a relatively easy setup compared to other methods of transformation, has proved to be effective and it was possible to obtain stable transformants with reproducible rates of transformation. The cloning of genes in the plasmide pFAT3 revealed to be more favourable to carry out the transformation than the other binary vectors tested (pBGgHg, pBINplus) which could not be used to clone the target genes. For each gene it was possible to harvest stable transformants harboring the transgene. The analysis by RT-PCR of transcripts revealed that transgenes were expressed. The phenotypical analysis revealed that the PiMt1 gene is able to increase the tolerance capacity of H. cylindrosporum to copper, but not to the other tested metals (Zn, Cd, Ni).
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Marc Bellion. Caractérisation fonctionnelle de gènes impliqués dans la tolérance au stress métallique chez les champignons ectomycorhiziens par agrotransformation d'Hebeloma cylindrosporum. Sciences de la Terre. Université Henri Poincaré - Nancy 1, 2006. Français. ⟨NNT : 2006NAN10103⟩. ⟨tel-01747397⟩

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