Le développement des plantes repose sur le fonctionnement des méristèmes qui sont à la base de la production des organes durant toute la vie post-embryonnaire de la plante. Ce développement repose également sur la définition de frontières d'une part entre les méristèmes et les organes et d'autre part entre les organes. Les gènes NAM/CUC3 de la famille de facteur de transcription à domaine NAC et leur microARN régulateur miR164, jouent un rôle important dans de tels mécanismes chez les eudicotylédones. En plus de leur rôle dans la définition de frontières, ces gènes sont nécessaires à la formation du méristème apical durant l'embryogenèse, et au contrôle de l'architecture de la plante et de ses organes. Sur la base des connaissances acquises chez les eudicotylédones, se posait la question de la conservation de ces gènes et de leur régulation ainsi que leur implication dans le contrôle du développement chez les palmiers (Arecales, Arecaceae) et, de façon plus large, chez les monocotylédones. Dans ce contexte, trois gènes similaires aux gènes NAM/CUC3 ont été isolés chez le palmier à huile (Elaeis guineensis L.), EgNAM1, EgNAM2 et EgCUC3. Ces gènes sont exprimés dans les tissus méristématiques végétatifs et reproducteurs. Notre analyse a révélée une conservation du module de régulation NAM-miR164 chez cette espèce et une divergence en terme de domaine d'expression entre monocotylédones et eudicotylédones, qui pourrait être associée à des différences majeures des régions cis-régulatrices des gènes NAM. L'analyse des profils d'expression des gènes NAM/CUC3 au cours de l'embryogenèse somatique précoce indique des similarités entre le palmier à huile, le maïs et le riz, ainsi qu'une conservation de la régulation via l'auxine comme observé chez Arabidopsis thaliana. Cependant, même si une régulation post-transcriptionnelle via miR164 a été détectée au cours de l'embryogenèse somatique, la répression par l'auxine semble essentiellement transcriptionelle. / Plant development depends on functioning of meristems, which are at the base of organ production during the post-embryonic phase. The development depends also on the definition of boundaries between meristem and primordia but also between organs. NAM/CUC3 genes belonging to the NAC domain transcription factor family and their microRNA regulator miR164 play an important role in these mechanisms in eudicot species. In addition to their role in boundary definition, they are involved in the establishment of the shoot apical meristem during embryogenesis and in the control of the plant and organ architectures. On the basis of data from eudicot species, the conservation of these genes and their regulation, and their involvement in meristem functioning in palm species (Arecales, Arecaceae) and more generally in monocot species was still an open question. In this context, three NAM/CUC3-related genes have been isolated in oil palm (Elaeis guineensis L.), EgNAM1, EgNAM2 and EgCUC3. Theses genes are expressed in both vegetative and reproductive meristematic tissues. Our analysis revealed the conservation of the NAM-miR164 regulatory module in this species and a divergence in term of expression pattern between monocots and eudicots, which may be related to differences in cis-regulatory regions of NAM genes. In contrast the expression pattern of NAM/CUC3 during somatic embryogenesis indicates similarity in the timing of expression between oil palm, maize and rice and also a conservation of the auxin-dependent regulation of NAM genes during this developmental phase as observed in Arabidopsis thaliana. However, even if miR164-dependent post-transcriptional regulation of NAM genes was detected during somatic embryogenesis, auxin-dependent repression seems to be essentially through transcriptional regulation.

Submitted by LUANA JANDHY MANTOVANINI (lumantovanini@gmail.com) on 2017-08-29T19:11:30Z No. of bitstreams: 1 Dissertação_Luana_Jandhy_Mantovanini.pdf: 1646514 bytes, checksum: 4c536d5f2d4ac50ce5a852c9b10ce27c (MD5) / Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-08-29T19:18:01Z (GMT) No. of bitstreams: 1 mantovanini_lj_me_jabo.pdf: 1646514 bytes, checksum: 4c536d5f2d4ac50ce5a852c9b10ce27c (MD5) / Made available in DSpace on 2017-08-29T19:18:01Z (GMT). No. of bitstreams: 1 mantovanini_lj_me_jabo.pdf: 1646514 bytes, checksum: 4c536d5f2d4ac50ce5a852c9b10ce27c (MD5) Previous issue date: 2017-05-24 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A cana-de-açúcar é atualmente uma das principais culturas da agroindústria mundial. Devido à ampla expansão de seu plantio é submetida constantemente a solos não produtivos. A presença de moléculas tóxicas no solo, como o alumínio (Al3+), interfere diretamente no desenvolvimento radicular ocasionando baixa absorção de água e nutrientes levando a pouca produtividade e desenvolvimento das plantas. Os microRNAs tem sido descritos como um dos fatores responsáveis pela regulação gênica e a descoberta dessas moléculas abre um novo caminho para a elucidação da tolerância e adaptação das plantas aos estresses abióticos. Este estudo visou avaliar em duas cultivares de cana-de-açúcar (CTC-2 e RB855453) a expressão dos microRNAs miR159, miR164 e miR168, associados à resposta ao alumínio em espécies como Arabidopsis thaliana, arroz (Oriza sativa) e tabaco (Nicotiana tabacum). Algumas características foram avaliadas, como densidade de raízes (DS), área foliar (AR), produção de massa seca (MS) e teor de prolina nas folhas, em quatro cultivares de cana-de-açúcar submetidas a diferentes concentrações de alumínio. A cultivar CTC 2 foi classificada como tolerante e a RB855453 como sensível ao estresse. Ambas foram selecionadas e em sistema de hidroponia submetidas novamente ao estresse pela toxidez de alumínio na concentração de 221 μmol L -1 . Parâmetros fisiológicos foram mensurados (área foliar, potencial osmótico, taxa de fotossíntese, transpiração, condutância estomática) e a expressão dos miRNAs 159, 164 e 168 avaliada por PCR em tempo real. Plantas das duas cultivares apresentaram alterações fisiológicas e morfológicas ao longo do estresse, com redução significativa para a área foliar da cultivar RB855453. O miR164 e 159 foram induzidos nas duas cultivares, principalmente após 72 horas de estresse, e o miR168 diferencialmente expresso. Esses miRNAs regulam genes e fatores de transcrição que estão envolvidos na resposta e desenvolvimento da planta diante ao estresse por alumínio. / The sugarcane is currently one of the main crops of global agribusiness. Due to the wide expansion of its plantation is constantly subjected unproductive soils. The presence of toxic molecules in the soil, such as aluminum (Al3+), directly affects root development, leading to poor absorption of nutrients and water leading to low productivity and development of plants. Studies of the interactions of plants with the environment are being conducted to clarify the resistance or susceptibility of various cultures, favoring the discovery of important mechanisms that participate in physiological and molecular responses to environmental stresses. MicroRNAs have been described as one of the factors responsible for gene regulation and the discovery of these molecules opens a new path for the elucidation of tolerance and adaptation of plants to abiotic stresses. This study evaluated in two sugarcane varieties the expression of microRNA miR159, miR164 and miR168, associated with the response to the aluminum species such as Arabidopsis thaliana, rice (Oryza sativa) and tobacco (Nicotiana tabacum). Some characteristics were evaluated, such as density (DS), area (AR), dry mass production (DM) and proline content, in four sugarcane cultivars submitted to different concentrations of aluminum. CTC 2 cultivar was classified as tolerant and RB855453 as stress sensitive. Both were selected and in a hydroponics system again submitted to stress by the aluminum toxicity in the concentration of 221 μmol L-1. Physiological parameters were measured (leaf area, osmotic potential, photosynthesis rate, transpiration, stomatal conductance) and the expression of miRNAs 159, 164 and 168 evaluated by real-time PCR. Plants of both cultivars presented physiological and morphological changes along the stress, with a significant reduction for the leaf area of cultivar RB855453. The miR164 and 159 were induced in the two cultivars, mainly after 72 hours of stress, and the miR168 differentially expressed. These miRNAs regulate genes and transcription factors that are involved in the response and development of the plant in the face of aluminum stress.