Faculdade De Ciências Agronômicas Câmpus De Botucatu

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Faculdade De Ciências Agronômicas Câmpus De Botucatu UNIVERSIDADE ESTADUAL PAULISTA “JÚLIO DE MESQUITA FILHO” FACULDADE DE CIÊNCIAS AGRONÔMICAS CÂMPUS DE BOTUCATU “CARACTERIZAÇÃO DE Amphobotrys ricini, AGENTE CAUSAL DO MOFO-CINZENTO DA MAMONEIRA (Ricinus communis L.), AVALIAÇÃO DE GENÓTIPOS, CONTROLE QUÍMICO E ALTERNATIVO.” HAROLDO ANTUNES CHAGAS Tese apresentada à Faculdade de Ciências Agronômicas da Unesp - Câmpus de Botucatu, para obtenção do título de Doutor em Agronomia (Agricultura). BOTUCATU - SP Outubro – 2012 UNIVERSIDADE ESTADUAL PAULISTA “JÚLIO DE MESQUITA FILHO” FACULDADE DE CIÊNCIAS AGRONÔMICAS CÂMPUS DE BOTUCATU “CARACTERIZAÇÃO DE Amphobotrys ricini, AGENTE CAUSAL DO MOFO-CINZENTO DA MAMONEIRA (Ricinus communis L.), AVALIAÇÃO DE GENÓTIPOS, CONTROLE QUÍMICO E ALTERNATIVO.” HAROLDO ANTUNES CHAGAS Orientador: Prof. Dr. Maurício Dutra Zanotto Co-orientador: Prof. Dr. Edson Luiz Furtado Tese apresentada à Faculdade de Ciências Agronômicas da Unesp - Câmpus de Botucatu, para obtenção do título de Doutor em Agronomia (Agricultura). BOTUCATU – SP Outubro – 2012 III Ao meu pai Veridiano Antunes Chagas, por sempre ter me ensinado que na vida, temos sempre que acreditar em nossos sonhos; À minha mãe Elvira Aparecida Chagas, por seu amor e carinho; sem eles nada disso seria possível; Ao meu querido irmão Rafael Antunes Chagas, meu melhor amigo, faltam palavras para expressar o quanto você é importante na minha vida; À minha querida sobrinha Helena, você é um presente de Deus, que chegou para alegrar ainda mais a nossa família. Obrigado, por vocês existirem e estarem sempre ao meu lado, nos bons e maus momentos. Dedico. IV AGRADECIMENTOS Em primeiro lugar agradeço a Deus, pela chance de poder realizar meus sonhos; À Faculdade de Ciências Agronômicas de Botucatu, pelo aprendizado; Aos Professores Maurício Dutra Zanotto e Edson Luiz Furtado, pela amizade, paciência, ensinamentos e pela orientação durante o período de doutorado; Aos amigos Willian Bucker Moraes, Renato Boreli, Ana Carolina Firmino, Cristiane Di Pieri, Diego Zied, Marilia Pizzeta, Caroline Geraldi Pierozzi, Martha Maria Passador, Juliana Sodário, João Zago, Junia, Tatiana Mituti, Mônica, Tadeu e Juan pela amizade, convivência, pela força e estímulo que me deram durante essa fase da minha vida; Às funcionárias da Seção de Pós-Graduação pela paciência e auxílio; Ao senhor Milton Pereira Marques, pela ajuda na instalação dos experimentos no campo, amizade, e convivência; Aos professores e funcionários do Departamento Produção Vegetal (Agricultura e Melhoramento Vegetal e Defesa Fitossanitária) por todos esses anos de ensinamentos e colaboração; A CAPES pela concessão da bolsa de estudos; Aos pesquisadores Neslon Dias Suassuna, Angelo Aparecido Barbosa Sussel da Embrapa Algodão e Embrapa Cerrados e Rafael Galbieri do Instituto Matogrossense do Algoão (IMA-MT), por cederem os isolados de Amphobotrys ricini; A todos aqueles que direta ou indiretamente contribuíram para a realização deste trabalho. V SUMÁRIO Página 1 RESUMO .................................................................................................................................... 1 2 SUMMARY ................................................................................................................................ 3 3. INTRODUÇÃO .......................................................................................................................... 5 4. REVISÃO BIBLIOGRÁFICA.................................................................................................. 8 4.1 Mamoneira (Ricinus communis L.)..................................................................................... 8 4.2 Mofo-cinzento (Amphobotrys ricini) e (Botrytis cinerea) .............................................. 11 4.3 Caracterização morfológica, cultural e molecular de fungos .......................................... 12 4.4 Controle químico ............................................................................................................... 14 4.5 Extratos vegetais ................................................................................................................ 16 4.6 Resistência de plantas a doenças fúngicas ....................................................................... 18 5. MATERIAL E MÉTODOS .................................................................................................... 21 5.1 Obtenção e manutenção de isolados de Amphobotrys ricini e Botrytis cinerea. ............ 21 5.2 Patogenicidade dos isolados ............................................................................................... 22 5.3 Microscopia eletrônica de varredura ................................................................................. 24 5.4 Caracterização cultural ....................................................................................................... 25 5.4.1 Avaliação do crescimento micelial dos isolados monospóricos de Amphobotrys ricini e de Botrytis cinerea, em diferentes temperaturas ....................................... 25 5.4.2 Aspecto de colônia e formação de escleródios ....................................................... 25 5.5 Caracterização morfológica................................................................................................ 26 5.5.1 Tamanho de conídios ................................................................................................ 26 5.5.2 Comprimento de conidióforos .................................................................................. 26 VI 5.6 Caracterização enzimática .................................................................................................. 27 5.6.1 Atividade celulolítica ................................................................................................ 27 5.6.2 Atividade lipolítica .................................................................................................... 27 5.6.3 Atividade proteolítica ................................................................................................ 28 5.6.4 Atividade pectinolítica .............................................................................................. 28 5.6.5 Análise de dados ........................................................................................................ 29 5.7 Caracterização molecular dos isolados .............................................................................. 29 5.8 Controle in vitro e in vivo do crescimento micelial de Amphobotrys ricini, utilizando diferentes fungicidas e extratos vegetais ........................................................................... 30 5.8.1 Coleta do material vegetal ........................................................................................ 30 5.8.2 Preparo dos extratos .................................................................................................. 31 5.9 Avaliação de genótipos de mamoneira visando a resistência ao mofo – cinzento em duas localidades diferentes em condições de campo ........................................................ 35 6. RESULTADOS E DISCUSSÂO ............................................................................................ 36 6.1 Patogenicidade dos isolados .......................................................................................... 36 6.2 Microscopia eletrônica de varredura ................................................................................. 40 6.3 Caracterização cultural ....................................................................................................... 44 6.3.1 Avaliação do crescimento micelial dos isolados monospóricos de Amphobotrys ricini e de Botrytis cinerea, em diferentes temperaturas ........................................ 44 6.3.2 Aspecto da colônia e formação de escleródios ........................................................ 47 6.4 Caracterização morfológica................................................................................................. 52 6.4.1 Tamanho de conídios e conidióforos de Amphobotrys ricini e de Botrytis cinerea52 6.4.2 Comprimento de conidióforos ................................................................................... 55 6.5 Caracterização enzimática .................................................................................................... 58 6.6 Caracterização molecular dos isolados ................................................................................ 62 6.7 Controle in vitro do crescimento micelial de Amphobotrys ricini utilizando diferentes VII fungicidas e controles alternativos ....................................................................................... 67 6.7.1 Controle in vivo do crescimento micelial de Amphobotrys ricini utilizando diferentes fungicidas e controles alternativos ........................................................... 74 6.8 Avaliação de genótipos de mamoneira visando a resistência ao mofo – cinzento em duas localidades diferentes ................................................................................................ 75 7. CONCLUSÃO.......................................................................................................................... 80 8. REFERÊNCIAS ........................................................................................................................ 82 VIII LISTA DE TABELAS Página 1.Relação de isolados de Amphobotrys ricini e Botrytis cinerea utilizados nos ensaios de laboratório..............................................................................................................................
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