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VARIABILIDADE GENÉTICA DE GUARIROBA (Syagrus Oleracea UNIVERSIDADE FEDERAL DE GOIÁS CAMPUS JATAÍ PROGRAMA DE PÓS-GRADUAÇÃO EM AGRONOMIA VARIABILIDADE GENÉTICA DE GUARIROBA ( Syagrus oleracea Becc.) DETERMINADAS POR DESCRITORES MORFOLÓGICOS E MARCADORES RAPD Jefferson Fernando Naves Pinto Biólogo JATAÍ – GOIÁS – BRASIL Julho de 2009 UNIVERSIDADE FEDERAL DE GOIÁS CAMPUS JATAÍ PROGRAMA DE PÓS-GRADUAÇÃO EM AGRONOMIA VARIABILIDADE GENÉTICA DE GUARIROBA ( Syagrus oleracea Becc.) DETERMINADAS POR DESCRITORES MORFOLÓGICOS E MARCADORES RAPD Jefferson Fernando Naves Pinto Biólogo Orientador: Prof. Dr. Edésio Fialho dos Reis Co-orientador: Dr. Fábio Gelape Faleiro Dissertação apresentada à Universidade Federal de Goiás (UFG), Campus Jataí, como parte das exigências para a obtenção do título de Mestre em Agronomia (Produção Vegetal). JATAÍ – GOIÁS – BRASIL Julho de 2009 Dados Internacionais de Catalogação-na-Publicação (CIP) (GPT/BC/UFG) Pinto, Jefferson Fernando Naves. P659v Variabilidade genética de guariroba ( Syagrus oleracea Becc .) determinadas por descritores morfológicos e marcadores RAPD [manuscrito] / Jefferson Fernando Naves Pinto. – 2009. xii, 81 f. : il., color., figs. Orientador: Prof. Dr. Edésio Fialho dos Reis; Co-Orientador: Prof. Dr. Fábio Gelape Faleiro. Dissertação (Mestrado) – Universidade Federal de Goiás, Campus Jataí, Programa de Pós-Graduação em Agronomia, 2009. Bibliografia: f. 70-81. Inclui lista de quadros, figuras e tabelas. 1. Guariroba – Variabilidade Genética – Estado de Goiás 2. Marcadores Moleculares RAPD 3. Syagrus oleracea Becc I. Reis, Edésio Fialho dos II. Faleiro, Fábio Gelape. III. Universidade Federal de Goiás, Campus Jataí, Programa de Pós- Graduação em Agronomia IV. Título. CDU: 582.545(817.3) DADOS CURRICULARES DO AUTOR JEFFERSON FERNANDO NAVES PINTO, filho de Luiz Fernando Alves Pinto e Stael Naves Ferreira Pinto, nascido em Jataí - Goiás a 22 de janeiro de 1982. Graduou-se em Ciências Biológicas na UFG em dezembro de 2004. Iniciou o curso Pós-graduação strictu-sensu em nível de Mestrado em Agronomia, área de concentração em Fitotecnia/Produção Vegetal, na Universidade Federal de Goiás (UFG), em março de 2007, concluindo-a em julho de 2009. Tudo na vida é possível, basta saber como, e o que fazemos nela ecoa na eternidade. Autor desconhecido Aos meus amados Pais e queridos irmãos. Os quais sem eles, nada seria. À minha namorada Eliane Alves dos Santos pelo amor, carinho, companheirismo, incentivo e principalmente pela enorme compreensão em todos momentos de nossa jornada. Ofereço e Dedico AGRADECIMENTOS À Universidade Federal de Goiás (UFG) pela oportunidade de realização do curso. Ao Dr. Edésio Fialho dos Reis a quem devo a orientação desta dissertação e, particularmente pela confiança, pelo estímulo e pela grande atenção amiga sempre oferecida. Ao Dr. Fábio Gelape Faleiro, da Embrapa Cerrados, por ter-me co-orientado e ajudado em todos os momentos com apoio científico, tecnológico e, sempre que necessário, com palavras de incentivo. Ao Dr. Marco Aurelio Carbone Carneiro, Coordenandor do Curso de Pós Graduação em Agronomia da UFG – CAJ, pelo constante incentivo durante todo o curso. A meus pais Luiz Fernando Alves Pinto e Stael Ferreira Naves Pinto pelo o amor, a confiança, o estimulo e apoio durante a minha vida. Aos meus irmãos Jenniffer Fernanda Naves Pinto e Jeeder Fernando Naves Pinto pelo companheirismo em todos os momentos de minha jornada. Às minhas avós pelo constante incentivo para continuar sempre a estudar. A todos os colegas de curso que juntos superamos várias dificuldades. E a minha namorada, Eliane Alves dos Santos, por ser o meu ponto de apoio. Muito obrigado a todos!!!!! i SUMÁRIO Página RESUMO ................................................................................................................... viii ABSTRACT ............................................................................................................... x 1 – INTRODUÇÃO .................................................................................................... 11 2 – REFERENCIAL TEÓRICO ................................................................................. 13 2.1 – Aspectos gerais das palmeiras ........................................................................ 13 2.2 – Gênero Syagrus ............................................................................................... 14 2.2.1 – Syagrus oleracea (Mart.) Becc. .................................................................... 15 2.2.1.1 – Taxonomia ................................................................................................. 15 2.2.1.2 – Distribuição geográfica .............................................................................. 16 2.2.1.3. – Características morfológicas .................................................................... 17 2.2.1.4 – Características do palmito .......................................................................... 19 2.2.1.5 – Importância econômica ............................................................................. 20 2.2.1.6 – Características genéticas .......................................................................... 21 2.3 – Marcadores moleculares ................................................................................. 23 2.3.1 – Marcadores moleculares em Arecaceas ...................................................... 24 2.3.2 – RAPD ( Random Amplified Polymorphic DNA ) ............................................... 26 2.4 – Divergência genética por análise multivariada ................................................. 28 2.4.1 – Análise de agrupamento ................................................................................ 29 3 – MATERIAL E MÉTODOS .................................................................................. 31 3.1 – Coletas dos acessos ........................................................................................ 33 3.2 – Análise de variância univariada ........................................................................ 34 3.3 – Análise multivariada ......................................................................................... 34 3.3.1 – Distância de Mahalanobis ............................................................................. 34 3.3.2 – Variáveis canônicas e descarte de variáveis ................................................. 35 3.3.3 – Análise de agrupamento ................................................................................ 35 ii 3.3.4 – Correlação fenotípica entre os caracteres estudados ................................... 36 3.4 – Caracterização molecular ................................................................................. 36 3.4.1 – Material utilizado ........................................................................................... 36 3.4.2 – Extração do DNA .......................................................................................... 37 3.4.3 – Quantificação do DNA .................................................................................. 37 3.4.4 – Reação de RAPD ......................................................................................... 38 3.4.5 – Análise estatística dos dados moleculares .................................................... 39 3.4.5.1 – Análises moleculares ................................................................................. 39 3.4.5.2 – Porcentagem de polimorfismo .................................................................... 40 3.4.5.3 – Coeficiente de similaridade (Sij) ................................................................. 40 3.4.5.4 – Coeficiente de dissimilaridade ................................................................... 41 3.4.5.5 – BOOTSTRAP ............................................................................................ 41 4 - RESULTADOS E DISCUSSÃO ........................................................................... 42 4.1 – Análise morfológica ......................................................................................... 42 4.1.1 – Estatística univariada .................................................................................... 42 4.1.2 – Herdabilidade ................................................................................................ 47 4.1.3 – Correlações fenotípicas ................................................................................ 48 4.2 – Análise multivariada ......................................................................................... 52 4.2.1 – Seleção de descritores .................................................................................. 52 4.2.2 – Divergência fenotípica ................................................................................... 57 4.2.3 – Análise de agrupamento ................................................................................ 58 4.2.3.1 Método de Otimização de Tocher ................................................................. 58 4.2.3.2 – Método UPGMA ......................................................................................... 60 4.2.3.3 – Variáveis canônicas .................................................................................... 61 4.3 – Análise molecular ............................................................................................. 62 4.3.1 – Ensaio de RAPD ........................................................................................... 62 4.3.1.1 – BOOTSTRAP ............................................................................................
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