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Organização Genômica De Genótipos Diploides E Alotetraploides Espontâneos E Induzidos De Arachis L

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Organização Genômica De Genótipos Diploides E Alotetraploides Espontâneos E Induzidos De Arachis L Universidade de Brasília – UnB Instituto de Ciências Biológicas Departamento de Botânica Organização genômica de genótipos diploides e alotetraploides espontâneos e induzidos de Arachis L. revelada por citogenética Eliza Fabricio de Melo Bellard do Nascimento Orientadora: Dra. Ana Cláudia Guerra de Araújo Brasília – DF Fevereiro/ 2021 Universidade de Brasília – UnB Instituto de Ciências Biológicas Departamento de Botânica Organização genômica de genótipos diploides e alotetraploides espontâneos e induzidos de Arachis L. revelada por citogenética Eliza Fabricio de Melo Bellard do Nascimento Orientadora: Dra. Ana Cláudia Guerra de Araújo Tese apresentada ao Departamento de Botânica do Instituto de Ciências Biológicas, da Universidade de Brasília, como requisito parcial para obtenção do grau de doutora em Botânica. Brasília – DF Fevereiro/ 2021 2 Tese de autoria de Eliza Fabricio de Melo Bellard do Nascimento, intitulada “Organização genômica de genótipos diploides e alotetraploides espontâneos e induzidos de Arachis L. revelada por citogenética”, realizada junto ao Departamento de Botânica do Instituto de Ciências Biológica da Universidade de Brasília e à Embrapa Recursos Genéticos e Biotecnologia, sob orientação da Dra. Ana Cláudia Guerra de Araújo, com apoio financeiro da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). Banca examinadora: Orientadora: Dra. Ana Cláudia Guerra de Araújo Embrapa Recursos Genéticos e Biotecnologia. Membro interno: Dra. Regina Célia de Oliveira Departamento de Botânica, Universidade de Brasília - UnB. Membro externo: Dr. José Francisco Montenegro Valls Embrapa Recursos Genéticos e Biotecnologia. Membro externo: Dra. Eliana Regina Forni Martins Universidade Estadual de Campinas - UNICAMP. Suplente: Dra. Marisa Toniolo Pozzobon Embrapa Recursos Genéticos e Biotecnologia. 3 À Deus, aos meus pais, à minha família e à minha sobrinha, Ana Sofia, dedico. 4 AGRADECIMENTOS A Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) pela concessão da bolsa de estudo durante os quatro anos de doutorado. Ao Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) pelo apoio financeiro para execução do projeto. A Fundação de Apoio à Pesquisa do Distrito Federal (FAP/DF) pelo apoio financeiro para execução do projeto. A Embrapa Recursos Genéticos e Biotecnologia pelo espaço, infraestrutura e recursos operacionais disponibilizados para execução de todo meu projeto. A minha querida orientadora, Dra. Ana Cláudia Guerra Araújo, que me acompanhou durante esses longos anos de formação acadêmica, da graduação ao doutorado, sempre com muito carinho, incentivo e aprendizado. Obrigada por acreditar e confiar em mim! Sentirei saudades. A Dra. Ana Cristina Miranda Brasileiro e a Dra. Patrícia Messenberg Guimarães, pelo acolhimento e carinho de sempre, pelos constantes ensinamentos, correções e sugestões e pelos momentos de descontração. A Ana Cristina Meneses Mendes Gomes (Aninha), pelo carinho de mãe com o qual me ensinou e me ajudou durante esse caminho. Aos membros da minha da banca, pelo interesse e disposição em me ajudar na melhora do meu trabalho final. Ao Leandro Mesquita, por sempre cuidar com tanto carinho dos nossos amendoins nas casas de vegetação. Aos meus queridos amigos que integram ou de alguma forma já integraram o Laboratório de Interação Planta-Praga III da Embrapa Recursos Genéticos e Biotecnologia (Bruna Medeiros, Andressa Martins, Andreza Vidal, Artur Fonseca, Bruna Vidigal, Lara Marques, Ana Zotta e Larissa Arrais), muitíssimo obrigada por todo apoio e ajuda emocional, operacional e experimental. Cada um de vocês me ajudou a chegar aqui. Ao meu noivo, Danillo Rabelo, pela paciência e carinho. Te amo! A minha família, que torceu, que se preocupou, que incentivou e que vibrou com essa conquista. Muito obrigada pela ajuda e pela compreensão nos momentos difíceis. Amo vocês. 5 A Deus e minha mãezinha, Nossa Senhora, por serem tão bons comigo! Obrigada por sempre estarem comigo, me fortalecendo, me guiando e me abençoando. “Tudo tem o seu tempo determinado, e há tempo para todo o propósito debaixo do céu ”. Eclesiastes 3:1. A todos que me ajudaram, me incentivaram de alguma forma e torceram por mim, de perto ou de longe, muito obrigada! 6 Sumário AGRADECIMENTOS .................................................................................................................................. 5 RESUMO ..................................................................................................................................................... 10 ABSTRACT ................................................................................................................................................. 11 LISTA DE FIGURAS .................................................................................................................................. 12 LISTA DE TABELAS ................................................................................................................................. 16 LISTA DE ABREVIATURAS .................................................................................................................... 17 INTRODUÇÃO GERAL ............................................................................................................................. 19 REVISÃO DA LITERATURA ................................................................................................................... 23 1. O amendoim (Arachis hypogaea L.) ................................................................................................... 23 2. Classificação taxonômica do gênero Arachis ..................................................................................... 25 3. A origem do amendoim ...................................................................................................................... 28 4. O cultivo do amendoim por índios no Parque Indígena do Xingu ..................................................... 30 5. A importância de espécies silvestres e alotetraploides induzidos de Arachis .................................... 31 6. Conteúdo genômico de Arachis - elementos de transposição e sequências gênicas ........................... 33 JUSTIFICATIVA ........................................................................................................................................ 39 OBJETIVOS ................................................................................................................................................ 41 1. Geral ............................................................................................................................................. 41 2. Específicos .................................................................................................................................... 41 CAPÍTULO I ............................................................................................................................................... 42 Amendoim cultivado por indígenas brasileiros: origem, diversidade morfológica e seleção ..................... 42 1. INTRODUÇÃO ................................................................................................................................... 43 2. MATERIAL E MÉTODOS ................................................................................................................. 46 2.1. Local de realização dos experimentos ............................................................................................. 46 2.2. Material vegetal ............................................................................................................................... 46 2.3. Obtenção de cromossomos metafásicos ........................................................................................... 48 2.4. Bandamento CMA3/DAPI ............................................................................................................... 48 2.5. Obtenção de sondas genômicas para GISH ..................................................................................... 49 2.5.1. Extração de DNA genômico ..................................................................................................... 49 2.5.2. Síntese de sondas genômicas .................................................................................................... 50 2.6. Extração e síntese de sondas de DNAr para FISH ........................................................................... 50 2.7. Incorporação de nucleotídeos marcados - Dot blot .......................................................................... 50 2.8. Hibridização in situ - FISH e GISH ................................................................................................. 51 2.8.1. Pré-hibridização ........................................................................................................................ 51 2.8.2. Hibridização in situ ................................................................................................................... 52 2.8.3. Pós-hibridização ........................................................................................................................ 52 2.8.4. Detecção dos sítios de hibridização .......................................................................................... 52 2.8.5. Montagem e análise das lâminas ............................................................................................... 53 3. RESULTADOS ......................................................................................................................................
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