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Ricardo De Nardi Fonoff 1 Universidade de São Paulo Escola Superior de Agricultura “Luiz de Queiroz” Sclerotinia sclerotiorum: características morfológicas, agressividade, sensibilidade “in vitro” a fungicidas e resistência de isolados a tiofanato metílico Patrícia Fabretti Kreyci Tese apresentada para obtenção do título de Doutora em Ciências. Área de concentração: Fitopatologia Piracicaba 2016 2 Patricia Fabretti Kreyci Engenheira Agrônoma Sclerotinia sclerotiorum: características morfológicas, agressividade, sensibilidade “in vitro” a fungicidas e resistência de isolados a tiofanato metílico versão revisada de acordo com a resolução CoPGr 6018 de 2011 Orientador: Prof. Dr. JOSÉ OTAVIO MACHADO MENTEN Tese apresentada para obtenção do título de Doutora em Ciências. Área de concentração: Fitopatologia Piracicaba 2016 Dados Internacionais de Catalogação na Publicação DIVISÃO DE BIBLIOTECA - DIBD/ESALQ/USP Kreyci, Patrícia Fabretti Sclerotinia sclerotiorum: características morfológicas, agressividade, sensibilidade “in vitro” a fungicidas e resistência de isolados a tiofanato metílico / Patrícia Fabretti Kreyci. - - versão revisada de acordo com a resolução CoPGr 6018 de 2011. - - Piracicaba, 2016. 146 p. : il. Tese (Doutorado) - - Escola Superior de Agricultura “Luiz de Queiroz”. 1. Sclerotinia sclerotiorum 2. Mofo-branco 3. Caracterização morfológica 4. Sensibilidade a fungicidas 5. Resistência 6. Tiofanato metílico I. Título CDD 632.43 K92s “Permitida a cópia total ou parcial deste documento, desde que citada a fonte – O autor” 3 Aos meus pais José e Magali, Que me ensinaram o valor da luta, E a felicidade da conquista, OFEREÇO À minha irmã Ana Paula, Meu exemplo de superação E meu sobrinho Mario, Por todo amor e afeto, DEDICO 4 5 AGRADECIMENTOS À Escola Superior de Agricultura “Luiz de Queiroz” – Universidade de São Paulo, pela formação nos cursos de Graduação e Pós Graduação. À Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) e ao Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) pela bolsa concedida. Ao Professor José Otávio Machado Menten pela oportunidade de desenvolver esta pesquisa. Ao Dr. Luiz Carlos Bhering Nasser e Dra. Silvana Petrofezza pelo fornecimento de grande parte dos isolados. Sem a generosidade de vocês esse projeto não seria possível. Aos demais colegas que colaboraram com isolados: Ériston, Marcos Freitas, Miller, Jair, Thomas, Ricardo, Meyriele, Gilson, Valter, Willian, Taurino, Basf, Ceres, Instituto Biológico, Fundação ABC, Vitabrás. À BASF, principalmente Kelly Simões, Eliara Xavier e Ronaldo Rodrigues pelo excelente treinamento em BPL – Boas Práticas Laboratoriais e no uso correto e seguro dos fungicidas. Ao Dr. Vicente Savino e Dr. Antônio Coelho pelo acolhimento e hospitalidade. Aos colegas de departamento: Miwa, Tirolez, Lico, Nuir, Sidney, Adilson (in memorian) pela agradável convivência. À todos os amigos de pós graduação e todos aqueles que de forma direta ou indireta contribuíram para realização desse trabalho. MUITO OBRIGADA! 6 7 “É melhor tentar e falhar, que preocupar-se e ver a vida passar. É melhor tentar, ainda que em vão que sentar-se, fazendo nada até o final. Eu prefiro na chuva caminhar, que em dias frios em casa me esconder. Prefiro ser feliz embora louco, que em conformidade viver.” Martin Luther King 8 9 SUMÁRIO RESUMO .............................................................................................................. 11 ABSTRACT .......................................................................................................... 13 LISTA DE FIGURAS ............................................................................................ 15 LISTA DE TABELAS ............................................................................................ 17 LISTA DE GRÁFICOS .......................................................................................... 19 1 INTRODUÇÃO .................................................................................................. 21 2 REVISÃO BIBLIOGRAFICA .............................................................................. 25 2.1 Histórico, hospedeiros e ocorrência de mofo branco ..................................... 25 2.2 Taxonomia e etiologia .................................................................................... 27 2.3 Ciclo de vida, epidemiologia e sintomatologia ............................................... 33 2.4 Manejo integrado ........................................................................................... 37 2.4.1 Controle cultural .......................................................................................... 38 2.4.2 Controle genético ........................................................................................ 40 2.4.3 Controle biológico ....................................................................................... 41 2.4.4 Controle químico ......................................................................................... 43 2.4.4.1 Tratamento de sementes ......................................................................... 45 2.4.4.2 Fungigação .............................................................................................. 46 2.4.4.3 Herbicidas ................................................................................................ 48 3 CARACTERIZAÇÃO MORFOLOGICA DE COLÔNIA EM CULTURA PURA E AGRESSIVIDADE DE ISOLADOS DE S. SCLEROTIORUM............................... 49 3.1 Introdução ...................................................................................................... 49 3.2 Materiais e métodos ....................................................................................... 52 3.2.1 Obtenção dos Isolados ............................................................................... 52 3.2.2 Desenvolvimento e morfologia de isolados de Sclerotinia Sclerotiorum em cultura pura .......................................................................................................... 55 3.2.2.1 Índice de Velocidade de Crescimento Micelial (IVCM) ............................ 56 3.2.2.2 Caracterização micelial das colônias ....................................................... 56 3.2.2.3 Características dos escleródios produzidos ............................................. 57 3.2.3 Agressividade de isolados de S. sclerotiorum em trifólios de soja e feijão...................................................................................................................... 57 3.3 Discussão de Resultados .............................................................................. 58 3.3.1 Índice de velocidade de crescimento micelial (IVCM) ................................. 58 10 3.3.2 Caracterização micelial das colônias .......................................................... 61 3.3.3 Características dos escleródios produzidos ................................................ 62 3.3.4 Agressividade de isolados de S. sclerotiorum em trifólios de soja e feijão.. 65 4 SENSIBILIDADE DE ISOLADOS DE SCLEROTINIA SCLEROTIORUM A FUNGICIDAS “IN VITRO” .................................................................................... 71 4.1 Introdução ...................................................................................................... 71 4.2 Materiais e métodos ....................................................................................... 73 4.2.1 Descrição dos Produtos Utilizados ............................................................. 73 4.2.1.1 Fluazinam ................................................................................................. 73 4.2.1.2 Procimidona ............................................................................................. 74 4.2.1.3 Tiofanato Metílico ..................................................................................... 75 4.2.2 Sensibilidade “in vitro” de isolados de S. sclerotiorum a fungicidas .......... 76 4.2.3 Avaliação da sensibilidade fungicida “in vitro” utilizando doses discriminatórias .................................................................................................... 78 4.3 Discussão de resultados ................................................................................ 79 4.3.1 Sensibilidade “in vitro” de isolados de S. sclerotiorum a fungicidas ........... 79 4.3.2 Avaliação da sensibilidade fungicida “in vitro” utilizando doses discriminatórias .................................................................................................... 81 5 RESISTÊNCIA DE ISOLADOS DE SCLEROTINIA SCLEROTIORUM A TIOFANATO METÍLICO ....................................................................................... 85 5.1 Introdução ...................................................................................................... 85 5.2 Materiais e métodos ....................................................................................... 88 5.3 Discussão De Resultados .............................................................................. 90 6 CONSIDERAÇÕES FINAIS ............................................................................. 95 REFERÊNCIAS ................................................................................................... 97 ANEXOS .............................................................................................................. 117 11 RESUMO Sclerotinia sclerotiorum: características morfológicas, agressividade, sensibilidade “in vitro” a fungicidas e resistência de isolados a tiofanato metílico
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