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Universidade Estadual De Campinas Instituto De Biologia Universidade Estadual de Campinas Instituto de Biologia Caracterização da diversidade genética molecular em germoplasma d e Brachiaria s p p . Tese apresentada à banca examinadora do Instituto de Biologia da Universidade Estadual de Campinas para obtenção do título de Doutor em Genética e Biologia Molecular, área de Genética Vegetal e Melhoramento Letícia Jungmann Cançado (Aluna) Dra. Anete Pereira de Souza (Orientadora) Dra. Cacilda Borges do Valle (co-Orientadora) C a m p i n a s – S ã o P a u l o – B r a s i l 2 0 0 9 i FICHA CATALOGRÁFICA ELABORADA PELA BIBLIOTECA DO INSTITUTO DE BIOLOGIA – UNICAMP Cançado, Letícia Jungmann C16c Caracterização da diversidade genética molecular em germoplasma de Brachiaria spp. / Letícia Jungmann Cançado. – Campinas, SP: [s.n.], 2009. Orientadores: Anete Pereira de Souza, Cacilda Borges do Valle. Tese (doutorado) – Universidade Estadual de Campinas, Instituto de Biologia. 1. Capim-brachiaria. 2. Poliplóide. 3. Variabilidade genética. 4. Marcadores moleculares. 5. Banco germoplasma. I. Souza, Anete Pereira de. II. Valle, Cacilda Borges do. III. Universidade Estadual de Campinas. Instituto de Biologia. IV. Título. Título em inglês: Molecular genetic diversity characterization in a germplasm collection of Brachiaria spp. Palavras-chave em inglês: Brachiaria; Polyploidy; Genetic variability; Molecular markers; Germplasm collection. Área de concentração: Genética Vegetal e Melhoramento. Titulação: Doutora em Genética e Biologia Molecular. Banca examinadora: Anete Pereira de Souza, José Francisco Montenegro Valls, Marcelo Mattos Cavalari, Mateus Mondin, Rosângela Maria Simeão Resende. Data da defesa: 15/05/2009. Programa de Pós-Graduação: Genética e Biologia Molecular. ii Campinas, 15 de maio de 2009 BANCA EXAMINADORA Dra. Anete Pereira de Souza Dr. José Francisco Montenegro Valls Dr. Mateus Mondin Dr. Marcelo Mattos Cavallari Dra. Rosângela Maria Simeão Resende Dr. Antônio Augusto Franco Garcia Dr. José Baldin Pinheiro Dr. Marcos Antônio Machado iii iv Agradecimentos À Universidade Estadual de Campinas, por tudo que aqui me foi oferecido, desde o conhecimento técnico a todos os demais aspectos da minha vida acadêmica. São incontáveis os aprendizados que vivenciei nestes quase 10 anos de estudos nesta instituição À Embrapa Gado de Corte, nas pessoas do Dr. Rafael Geraldo de Oliveira Alves, do Dr. Cleber Oliveira Soares, do Sr. Tenisson Waldow de Souza e da Sra. Lucia Gatto, pela oportunidade de realizar este trabalho de tese na Universidade Estadual de Campinas e pelo conhecimento que me foi passado nos últimos 6 anos de vida profissional À minha querida orientadora, Dra. Anete Pereira de Souza, por todos os ensinamentos, todas as oportunidades, todos os apoios e todos os carinhos. Não haverá um só dia em minha vida profissional que não me lembrarei de você como um exemplo a ser seguido À minha caríssima co-orientadora, Dra. Cacilda Borges do Valle, por ter me mostrado este feliz caminho por entre as Brachiarias. Pelos sábios ensinamentos e pelos conselhos sempre dados e sempre válidos. Pelas valiosíssimas informações, sempre disponibilizadas, fundamentais para o desenvolvimento deste trabalho À Fundação de Amparo à Pesquisa do Estado de São Paulo (Fapesp) pelos recursos que propiciaram a realização deste trabalho Ao Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) igualmente, pela disponibilização de recursos para viabilizar este trabalho À Fundação de Apoio ao Desenvolvimento do Ensino, Ciência e Tecnologia do Estado de Mato Grosso do Sul (Fundect) pela bolsa concedida durante a realização deste trabalho À Associação para o Fomento à Pesquisa de Melhoramento de Forrageiras Tropicais (Unipasto) pelo continuado apoio aos projetos com Forrageiras da Embrapa À minha mãe, Maria Inês, pela minha existência e pela sua. Você construiu em mim a base de tudo o que sou À minha avó, Lita, por ser a minha segunda mãe. A minha segunda base v Ao meu parceiro de vida, Vitor, presente que eu ganhei da vida. Amigo, irmão, cúmplice, amor... Pelo apoio incondicional para a realização deste trabalho A toda a minha família, pelo carinho com que sempre me apoiaram Às amigas, colegas e parceiras, jovens geneticistas, Bianca e Patrícia sem vocês este trabalho não teria sido tão prazeroso. Pelas horas e horas de dedicação e pela preciosa amizade que surgiu nesta jornada Às grandes companheiras Adna e Tatiana, pelos dias e dias passados lado a lado na bancada e pelas ótimas experiências compartilhadas ao longo destes anos. Esta amizade foi um presente Ao Juverlande Lugli, o Jú... o que seria de nossas teses e de nós mesmos sem você, hein, Jú? Aos companheiros de laboratório, todos... por terem feito parte de tantos bons momentos no laboratório e pelas trocas, infinitas À amiga e colega Maria Imaculada Zucchi, pela grande ajuda em todos os momentos necessários, pelas aulas concedidas, as metodologias discutidas. Tudo, sempre, com muito carinho À amiga e colega Rosângela M. S. Resende, por compartilhar comigo todos os sabores deste trabalho, sempre disponível a contribuir com o que fosse necessário Ao amigo e colega Marcelo Mattos Cavallari, pelo enorme apoio na realização das análises, pela disponibilidade e boa vontade sempre demonstradas A toda a equipe de Melhoramento de Forrageiras Tropicais da Embrapa Gado de Corte, cujo trabalho tem gerado frutos de valor inestimável para o Brasil e resultou na produção de informações usadas nesta Tese de Doutorado Às colegas Lucimara Chiari e Gisele O. C. Leguizamon, pelas horas passadas sob Sol para coletar o material de Brachiaria no campo À Universidade Estadual de Maringá, nas pessoas da Dra. Maria Suely Pagliarini e da Dra. Kellen R. Boldrini, pela gentileza em ceder informações citogenéticas Às colegas da Secretaria do Centro de Biologia Molecular e Engenharia Genética da Unicamp: Tânia, Sandra e Andreza, pelo profissionalismo e pelo carinho ao longo destes anos vi Índice Geral PÁGINA RESUMO ........................................................................................................................ ix ABSTRACT ..................................................................................................................... x APRESENTAÇÃO .......................................................................................................... 1 INTRODUÇÃO GERAL .................................................................................................. 4 Posicionamento taxonômico do gênero Brachiaria ................................................... 4 Importância econômica de Brachiaria ......................................................................... 6 Coleções de germoplasma ex situ .............................................................................. 7 Biologia básica em Brachiaria ..................................................................................... 9 Descrição das espécies envolvidas neste estudo.................................................... 12 Brachiaria brizantha ................................................................................................. 12 Brachiaria decumbens .............................................................................................. 14 Brachiaria dictyoneura .............................................................................................. 16 Brachiaria humidicola ............................................................................................... 17 Brachiaria ruziziensis ............................................................................................... 18 Marcadores moleculares ............................................................................................ 19 Caracterização morfológica e molecular do germoplasma de Brachiaria da Embrapa Gado de Corte ......................................... 21 JUSTIFICATIVA ........................................................................................................... 23 OBJETIVOS ................................................................................................................. 24 CAPÍTULO 1 Isolation and characterization of microsatellite markers for Brachiaria brizantha (Hochst. ex A. Rich.) Stapf ...................................................... 25 CAPÍTULO 2 Use of microsatellites developed for Brachiaria brizantha (Hochst. ex A. Rich.) Stapf for revealing inter- and intraspecific genetic diversity in Brachiaria spp. ..................................................... 30 Introduction ................................................................................................................... 31 Materials and Methods .................................................................................................. 32 Results ......................................................................................................................... 37 Discussion .................................................................................................................... 42 References ................................................................................................................... 46 vii CAPÍTULO 3 Development of microsatellite markers for Brachiaria humidicola (Rendle) Schweick. ............................................................... 49 CAPÍTULO 4 A survey of the Brazilian germplasm collection of the tropical pasture Brachiaria humidicola (Rendle) Schweick. [Koroniviagrass] using microsatellite markers, cytogenetics, morphological traits and geographical origin....................................................................................................
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