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GABRYELE SILVA RAMOS CAPACIDADE DE DISPERSÃO E PARÂMETROS BIOLÓGICOS INERENTES À CRIAÇÃO MASSAL DE Telenomus podisi Ashmead (Hymenoptera: Platygastridae) PARA O CONTROLE DE PENTATOMÍDEOS-PRAGA EM SOJA Botucatu 2020 GABRYELE SILVA RAMOS CAPACIDADE DE DISPERSÃO E PARÂMETROS BIOLÓGICOS INERENTES À CRIAÇÃO MASSAL DE Telenomus podisi Ashmead (Hymenoptera: Platygastridae) PARA O CONTROLE DE PENTATOMÍDEOS-PRAGA EM SOJA Dissertação apresentada à Faculdade de Ciências Agronômicas da Unesp Câmpus de Botucatu, para obtenção do título de Mestre em Agronomia: Proteção de Plantas. Orientadora: Profª. Drª. Regiane Cristina Oliveira de Freitas Bueno Botucatu Ramos, Gabryele Silva R175c Capacidade de dispersão e parâmetros biológicos inerentes à criação massal de Telenomus podisi ashmead (hymenoptera: platygastridae) para o controle de pentatomídeos-praga em soja / Gabryele Silva Ramos. -- Botucatu, 2020 122 p. : il., tabs., fotos Tese (doutorado) - Universidade Estadual Paulista (Unesp), Faculdade de Ciências Agronômicas, Botucatu Orientadora: Regiane Cristina Oliveira de Freitas Bueno 1. Entomologia agrícola. 2. Manejo integrado de pragas. 3. Controle biológico. 4. Parasitoide de ovos. 5. Controle de qualidade. I. Título. Sistema de geração automática de fichas catalográficas da Unesp. Biblioteca da Faculdade de Ciências Agronômicas, Botucatu. Dados fornecidos pelo autor(a). Essa ficha não pode ser modificada. Aos meus amados pais, Júnia e Jorge, dedico AGRADECIMENTOS A Deus. Aos meus amados avós, Nilza e Perpedigno, aos meus pais, Júnia e Jorge, e irmão Gabryel pelo amor e apoio incondicionais. À Faculdade de Ciências Agronômicas da Universidade Estadual Paulista “Júlio de Mesquita Filho” – UNESP, Campus de Botucatu. À Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), pelo apoio financeiro, concedido por meio de bolsa de mestrado (Processo nº 180017). À Profª. Drª. Regiane Bueno, pela orientação, ensinamentos, paciência e inspiração profissional. Aos amigos do Grupo de Pesquisa em Manejo Integrado de Pragas na Agricultura (AGRIMIP), em especial, João Pedro, Nadja, Carol, Fábio e Pedro por todo o apoio, auxílio na execução dos experimentos e principalmente, pela amizade construída. Aos motoristas durante a bateria de ensaios a campo, Daniel, Thaís e Simone, e demais colegas que auxiliaram em toda a coleta de dados, pela colaboração e companheirismo. Aos professores do Departamento de Proteção Vegetal – FCA/UNESP pelos valiosos ensinamentos. Ao Prof. Dr. Carlos Frederico Wilcken e colegas do LCBPF pelo suporte e disponibilização do laboratório para a execução das análises morfométricas. Ao Prof. Dr. Luis Gustavo Frediani Lessa, Dr. Donizete e Amanda, pela disposição em ajudar e disponibilização do Laboratório de Geoprocessamento para a realização das análises geoestatísticas. À Vanessa Carvalho pelos importantes ensinamentos durante a nossa convivência e por toda a ajuda nas análises moleculares presentes neste trabalho e à Profª Drª Sílvia Renata pela concessão de uso do Laboratório de Biologia Molecular. Ao time da Agroterenas S/A (Unidade de Santa Cruz do Rio Pardo, SP), em especial ao Leonardo e Joselito, aos profissionais da Fazenda Santa Fé, João e Beatriz Burckas, e à BioPartner por toda a colaboração, auxílio, disponibilização de insumos e concessão de áreas necessárias à condução dos experimentos de campo. Aos amigos que fiz durante o Mestrado e aos que me acompanharam e apoiaram, mesmo que distantes, durante essa trajetória... Muito obrigada! “A ciência nunca resolve um problema sem criar pelo menos outros dez”. George Bernard Shaw RESUMO O sucesso de programas de controle biológico de insetos-praga é dependente do entendimento detalhado de parâmetros biológicos, comportamentais e da capacidade de dispersão dos agentes de controle nos agroecossistemas. Após a saída da especificação de referência para registro e comercialização, é esperada a otimização de pacotes tecnológicos para garantir a eficiência e eficácia de um agente macrobiológico. É o caso do parasitoide de ovos de pentatomídeos-praga, Telenomus podisi Ashmead (Hymenoptera: Platygastridae). Com as limitações do uso recorrente de inseticidas no controle de percevejos fitófagos em grandes culturas de elevada importância econômica como a soja, a liberação de T. podisi em áreas extensas vem sendo vista como uma estratégia de controle promissora e pertinente. Neste contexto, parâmetros biológicos inerentes ao desenvolvimento de um controle de qualidade para a criação massal e liberação a campo de T. podisi foram explorados no presente estudo. Bioensaios de preferência por ovos de Euschistus heros (Fabricius) (Hemiptera: Pentatomidae) frescos ou criopreservados, estudo morfométrico e identificação de endossimbiontes foram conduzidos em laboratório, bem como a determinação da proporção ideal de fêmeas de T.podisi/ovo hospedeiro a fim de se diminuir a competição intraespecífica a aumentar as taxas de parasitismo em condições laboratoriais de multiplicação do parasitoide. Além disso, foi realizado o estudo da capacidade de dispersão de T. podisi a partir de matrizes de pontos equidistantes demarcadas e liberação central do número recomendado de fêmeas, segundo a especificação de referência de uso, em plantações de soja. As bactérias endossimbiontes Wolbachia pipientis e Serratia grimesii foram detectadas em T. podisi. Parasitoides multiplicados em ovos frescos de E. heros têm seu período de desenvolvimento ovo-adulto prolongado e tíbias mais compridas. A quantidade de fêmeas/ovo que permite uma maior taxa de parasitismo e de 8,87 fêmeas e a partir 9,28 ovos/fêmea há um estímulo de parasitismo de T. podisi. O emprego da proporção ideal de fêmeas de T. podisi/ovo hospedeiro e do número mínimo de ovos hospedeiros/fêmea em criações massais a fim de se diminuir a competição intraespecífica pode levar ao aumento das taxas de parasitismo em condições laboratoriais de multiplicação do parasitoide. A distância máxima de forrageamento atingida por T. podisi foi de 39 metros durante o estágio fenológico da soja Vn-R1 e de 30,90 metros no estágio R5-R6. Fêmeas oriundas de ovos de P. guildinii possuem asas mais estreitas e menor comprimento de corpo. Entre todas as espécies comparadas, os machos de T. podisi possuem asas mais largas que fêmeas. A presença de endossimbiontes foi bastante variável entre os ovos hospedeiros e os parasitoides oriundos destes ovos. Wolbachia pipientis foi detectada apenas em T. podisi oriundo de ovos de T. perditor. A espécie de bactéria endossimbionte Serratia grimesii também foi detectada entre as amostras. Os resultados obtidos fornecem informações que auxiliam o entendimento sobre a dinâmica ecológica de parasitismo a campo e adaptação hospedeira de T. podisi e norteiam a elaboração de programas de controle de qualidade da produção em massa e posterior liberação de T. podisi para o manejo de pentatomídeos-praga na cultura da soja. Palavras-chave: Controle biológico. Pentatomidae. Parasitoide de ovos. Dispersão. ABSTRACT The success of biological control programs for insect pests is dependent on a detailed understanding of biological, behavioral parameters and the dispersion capacity of control agents in agroecosystems. After the reference specification approval for registration and commercialization, the optimization of technological packages is expected to guarantee the efficiency and suitability of a macrobiological agent. This is the case of the pentatomid egg parasitoid, Telenomus podisi Ashmead (Hymenoptera: Platygastridae). With the limitations of the recurrent use of insecticides in the control of phytophagous stink bugs in large crops of high economic importance such as soy, the release of T. podisi in large areas has been seen as a promising and relevant control strategy. In this context, biological parameters inherent to the development of quality control for mass farming and field release of T. podisi were explored in the present study. Bioassays of preference for fresh or eggs cryopreserved Euschistus heros (Fabricius) (Hemiptera: Pentatomidae), morphometric study and identification of endosymbionts were carried out in the laboratory, as well as the determination of the ideal proportion of T. podisi females/ host egg in order to decrease the intraspecific competition to increase parasitism rates in laboratory conditions of parasitoid multiplication. In addition, the study of the dispersion capacity of T. podisi was carried out from matrices of demarcated equidistant points and central release of the recommended number of females, according to the reference specification of use, in soybean plantations. The endosymbiont bacteria Wolbachia pipientis and Serratia grimesii were detected in T. podisi. Parasitoids multiplied in fresh E. heros eggs have a prolonged egg-adult development period and longer tibiae. The number of females/ egg that allows a higher rate of parasitism and 8.87 females and from 9.28 eggs/ female there is a parasitism stimulus of T. podisi. The use of the ideal proportion of T. podisi females/ host egg and the minimum number of host eggs/ female in mass rearing in order to decrease intraspecific competition may lead to increased rates of parasitism in laboratory conditions of parasitoid multiplication. The maximum foraging distance reached by T. podisi was 39 meters during the phenological stage of soybean Vn-R1 and 30.90 meters in stage R5-R6. Females from P. guildinii eggs have narrower wings and shorter body length. Among all the species compared, males of T. podisi have wider wings than females. The presence
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