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Identification et validation de gènes pour l’amélioration du sorgho : de l’analyse transcriptomique à la validation fonctionnelle

Abstract : Plants Secondary Cell Walls (SCW) represent most of the renewable biomass and participate in some plant physiological functions such as nutrient transport, stress resistance and stiffness. Sorghum is a C4 grass well adapted to nutrient and water limiting conditions. Improving knowledge SCW establishment in sorghum aims at contributing to the development of varieties adapted to biomass sector including animal feed, energy production and the development of bio-materials. The objectives of the thesis presented here are to contribute to the understanding of the molecular mechanisms of SCW regulation and biosynthesis by identifying genes networks involved in this process and validating their role using reverse genetics. As a first step, a phylogenetic analysis including 9 species, allowed us to identify homologies within families of NAC and MYB TF, which are the main regulators of SCW establishment in Arabidopsis and other species. We then built gene co-expression networks of sorghum developing internode and focused on NAC and MYB networks as well as on networks with enrichment in SCW related genes Then, genes identified by these approaches were compared to those located in the chromosomal regions impacting the variability of SCW composition. In a second step we have developed genetic transformation approaches to characterize the role of identified genes by reverse genetics.We have identified 6 modules of co-expressed genes enriched with genes and gene ontologies related to cell walls and in particular 2 modules whose genes are potentially involved in the establishment of SCW. In addition, 122 NAC and 135 MYB were identified in sorghum. Among them, 14 NAC and 25 MYB are involved in the 6 co-expression networks highly enriched in cell wall genes. Of these, 19 are homologues of genes already validated in other species for their role SCW establishment and 20 have no known role. The comparison of the genes belonging to the 6 modules involved in cell wall establishment and chromosomal regions associated with the composition of the biomass revealed an intersect of 70 genes and some structural genes involved in the biosynthesis of the walls but little convergence concerning NAC and MYB TF.Although the optimization of stable genetic transformation protocols was unsuccessful, an efficient protoplast-based transient transformation protocol was optimized. This tool will be mobilized to test the roles of identified TF in networks enriched with cell wall genes.
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Lauriane Hennet. Identification et validation de gènes pour l’amélioration du sorgho : de l’analyse transcriptomique à la validation fonctionnelle. Sciences agricoles. Université Montpellier, 2019. Français. ⟨NNT : 2019MONTG090⟩. ⟨tel-02868696⟩

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