Dans cette thèse, j'ai étudié le rôle des sRNAs dérivés de l'hôte et du virus lors de l'infection du colza (Brassica napus, Canola) par la souche UK1 du virus de la mosaïque du navet (TuMV-UK1). En utilisant un dérivé de TuMV fusionné avec un gène codant pour la protéine fluorescente verte (TuMV-GFP), deux cultivars de colza (‘Drakkar’ et ‘Tanto’) qui diffèrent par leur susceptibilité à ce virus ont été identifiés. Le profil transcriptionnel des foyers d'infection locale, dans les feuilles de Drakkar et de Tanto, par séquençage nouvelle génération (NGS) a révélé de nombreux gènes exprimés de manière différentielle. Les mêmes échantillons d'ARN provenant de feuilles de Drakkar et de Tanto, traitées par des virus ou utilisées en contrôle, ont également servi à établir le profil NGS des sRNAs (sRNAseq) et de leurs cibles potentielles d'ARN (PAREseq). Les analyses bioinformatiques et leur validation in vivo, ont permis d’identifier les événements de clivage de transcrits impliquant des micro ARN (miRNA) connus et encore inconnus. Fait important, les résultats indiquent que TuMV détourne la voie du RNA silencing de l’hôte avec des siRNAs issus de son propre génome (vsiRNA) pour cibler les gènes de l’hôtes. Le virus déclenche également le ciblage à grande échelle des ARN messagers (ARNm) de l’hôte par l’activation de la production de siRNAs secondaires en phase, à partir de locus PHAS. À leur tour, les vsiRNAs et les siRNAs dérivés de l'hôte (hsRNAs) ciblent et clivent l'ARN viral par le complexe RISC. Ces observations éclairent le rôle des siRNAs dérivés de l'hôte et du virus dans la coordination de l'infection virale. Un autre chapitre de cette thèse est consacré à l'analyse des maladies induites par des virus en utilisant comme modèle de plante Arabidopsis, infectée par un tobamovirus, le virus de la mosaïque du colza (ORMV). De plus, ces observations ont permis de proposer un modèle dans lequel cette guérison dépend d’un adressage important de vsiRNAs secondaires antiviraux depuis leur source de production jusqu’à leurs tissus de destination, et l'établissement d'un apport en vsiRNAs capable de bloquer l'activité VSR impliquée dans la formation des feuilles symptomatiques. / In this thesis, I investigated the role of host- and virus-derived sRNAs during infection of Rapeseed (Brassica napus, Canola) by the UK1 strain of Turnip mosaic virus (TuMV-UK1). By using a TuMV derivative tagged with a gene encoding green fluorescent protein (TuMV-GFP), two rapeseed cultivars (‘Drakkar’ and ‘Tanto’) that differ in susceptibility to this virus were identified. Transcriptional profiling of local infection foci in Drakkar and Tanto leaves by next generation sequencing (NGS) revealed numerous differentially expressed genes. The same RNA samples from mock- and virus- treated Drakkar and Tanto leaves were also used for the global NGS profiling of sRNAs (sRNAseq) and their potential RNA targets (PAREseq). The bioinformatic analysis and their in vivo validation led to the identification of transcript cleavage events involving known and yet unknown miRNAs. Importantly, the results indicate that TuMV hijacks the host RNA silencing pathway with siRNAs derived from its own genome (vsiRNAs) to target host genes. The virus also triggers the widespread targeting of host messenger RNAs (mRNAs) through activation of phased, secondary siRNA production from PHAS loci. In turn, both vsiRNAs and host-derived siRNAs (hsRNAs) target and cleave the viral RNA by the RISC-mediated pathway. These observations illuminate the role of host and virus-derived sRNAs in the coordination of virus infection. Another chapter of this thesis is dedicated to the analysis of virus-induced diseases by using Arabidopsis plants infected with the Oilseed rape mosaic tobamovirus (ORMV) as a model. Initially, the infected plants develop leaves with strong disease symptoms. However, at a later stage, disease-free, “recovered” leaves start to appear. Analysis of symptoms recovery led to the identification of a mechanism in which the VSR and virus derived-siRNAs play a central role. I used Arabidopsis mutants impaired in transcriptional and post-transcriptional silencing pathways (TGS and PTGS respectively) and a plant line carrying a promoter-driven GFP transgene silenced by PTGS (Arabidopsis line 8z2). Using various techniques able to monitor virus infection, small and long viral RNA molecules, VSR activity, as well as phloem-mediated transport with in these lines, this study led to the identification of genes required for disease symptoms and disease symptom recovery. Moreover, the observations allowed to propose a model in which symptoms recovery occurs upon robust delivery of antiviral secondary vsiRNAs from source to sink tissues, and establishment of a vsiRNA dosage able to block the VSR activity involved in the formation of disease symptoms.

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The adjustment plant arrangement is one of the fundamental management techniques for canola show positive reaction in the ability to absorb light energy, nutrients and water, which reflects on maximizing the production of photosynthate directed to the plant and grains. The aim of this study was to assess the interference of arrangement plant on direct and indirect components of yield in canola, and includes determining the effects produced in oil content. The study was conducted at the Instituto Regional de Desenvolvimento Rural do Departamento de Estudos Agrários da Universidade Regional do Noroeste do Estado do Rio Grande do Sul (IRDeR/DEAg/UNIJUÍ) in Augusto Pestana, RS, involving the execution of three experiments for different spacing of 0.20, 0.40 and 0.60m. The experimental design consisted of a factorial arrangement in randomized blocks with four replicates of 2 x 2 x 4, for the crop year (2008 and 2009), genotype (Hyola 432 and 61) and plant density (20, 40, 60 and 80 plants m-2) in each set line spacing. Components were determined direct and indirect grain yield, oil content, morphological and adaptive, as well as the thermal and solar radiation during the grain filling. The year of cultivation was more effective in changing the grain yield and other traits in canola direct production and oil content, followed by the genetic potential to grow and smaller share of the planting density. The cv. Hyola 432 shows greater effectiveness in maximizing grain yield over Hyola 61 regardless of year, row spacing and planting density. Moreover, in the shortest space of culture, the character of direct and indirect yield high values of heritability and character number of tertiary branches shows stability when subjected to strong variations in the plants arrangement. The thermal shows no correlation with grain yield and oil content. On the other hand, variables related to the crop cycle are associated with the thermal and solar radiation accumulated in the grain filling stage of canola. / O ajuste do arranjo de plantas representa um dos termos de manejo fundamental para a canola mostrar reação positiva na capacidade de absorção de energia luminosa, nutrientes e água, o que reflete em maximizar a produção de fotoassimilados direcionados a planta e aos grãos. O objetivo deste trabalho foi verificar a interferência do arranjo de plantas sobre os componentes diretos e indiretos do rendimento de grãos em canola, incluindo também, a determinação dos efeitos proporcionados no conteúdo de óleo. Nesse sentido, foi desenvolvido um experimento nos anos agrícolas de 2008 e 2009 no Instituto Regional de Desenvolvimento Rural, pertencente ao Departamento de Estudos Agrários da Universidade Regional do Noroeste do Rio Grande do Sul (IRDeR/DEAg/UNIJUÍ), localizada no município de Augusto Pestana, RS. Para o estudo dos diferentes arranjos de população, foram utilizados três espaçamentos entrelinhas (0,20, 0,40 e 0,60 m), quatro densidades de plantas por metro quadrado (20, 40, 60 e 80 plantas m-2) e utilizando dois híbridos de canola (Hyola 432 e Hyola 61). O delineamento experimental foi o de blocos completamente casualizados com quatro repetições, num esquema fatorial 2 x 3 x 4, para ano, genótipo e densidade de cultivo em três distintos espaçamentos entre linhas, respectivamente. Foram determinados os componentes direto e indiretos do rendimento de grãos, o conteúdo de óleo, caracteres morfológicos e adaptativos, bem como a soma térmica e a insolação durante o período de enchimento de grãos. O ano de cultivo foi mais efetivo em alterar o rendimento de grãos e demais caracteres diretos de produção em canola, bem como o teor de óleo, seguido do potencial genético de cultivar e de menor participação da densidade de cultivo. A cultivar Hyola 432 mostra maior efetividade em maximizar a produção de grãos em relação a Hyola 61, independente do ano, espaçamento entre linhas e densidade de cultivo. Além disso, no menor espaçamento de cultivo, os caracteres diretos e indiretos do rendimento de grãos apresentam elevados valores de herdabilidade e o caráter número de ramos terciários evidencia forte estabilidade quando submetido a variações no arranjo de plantas. A soma térmica não apresenta correlação com o rendimento de grãos e teor de óleo. Por outro lado, variáveis relacionadas ao ciclo da cultura apresentam associação com a soma térmica e com a insolação acumulada na fase de enchimento de grãos de canola.