To investigate potential sources of novel grain protectors, this study evaluated, firstly, the bioactivity of ethanolic extracts (66) prepared from 29 species belonging to 11 different genera of Neotropical Annonaceae against the maize weevil Sitophilus zeamais (Coleoptera: Curculionidae). A screening assay demonstrated that the most pronounced effects (acute and chronic) on S. zeamais were caused by extracts from the Annona montana, A. mucosa, A. muricata and A. sylvatica seeds, and, to a lesser extent, by extracts prepared from leaves of A. montana, A. mucosa, A. muricata, and Duguetia lanceolata. However, the most active extracts (from seeds) did not affect fungal growth and aflatoxin production of Aspergillus flavus (Ascomycota: Trichocomaceae). Using the maize weevil as bioindicator, bioguided fractionations were then conducted in order to isolate, purify and characterize the possible active compound(s) from the most interesting extracts. By means of different chromatographic procedures, nine compounds (five acetogenins, three steroids, and one aromatic compound) were isolated. The acetogenins rolliniastatin-1 and ACG4 (structural determination in progress) and the aromatic compound 2,4,5- trimethoxystyrene as well as the steroids campesterol, stigmasterol, and sitosterol [tested in mixture (8.44 + 12.37 + 79.19%, respectively)] showed promising grain protective properties. Furthermore, the obtained results indicate that compounds from different chemical natures have a synergistic effect on the overall biological activity of the crude extracts. In a second study, the acute and chronic toxicity of selected ethanolic seed extracts from Annona species (A. montana, A. mucosa, A. muricata, and A. sylvatica) and an acetogenin-based commercial bioinsecticide (Anosom® 1EC) were investigated against the cabbage looper Trichoplusia ni (Lepidoptera: Noctuidae) and the green peach aphid Myzus persicae (Hemiptera: Aphididae). In the laboratory, extracts of A. mucosa and A. sylvatica as well as Anosom® were especially active through oral and topical administration. A greenhouse trial showed that a formulated A. mucosa extract and Anosom® were highly effective (>98% mortality) against third instar of T. ni larvae, and comparable to a pyrethrin-based commercial insecticide (Insect Spray®) used as a positive control. Similar to results with T. ni, A. mucosa extract showed the greatest aphicidal either in laboratory or greenhouse bioassays. In a third study, the acaricidal activity [against the citrus red mite Panonychus citri (Acari: Tetranychidae)] of the ethanolic extract from A. mucosa seeds (most active) was investigated. In laboratory tests, it exhibited levels of activity superior to commercial acaricides/insecticides of natural origin [Anosom® 1EC (annonin), Derisom® 2EC (karanjin), and Azamax® 1.2EC (azadirachtin + 3- tigloylazadirachtol)] and similar to a synthetic acaricide [Envidor® 24 SC (spirodiclofen)]. Finally, the compatibility of A. mucosa seed extract with three entomopathogenic fungi species (Beauveria bassiana, Isaria fumosorosea and Metarhizium anisopliae) was assessed. In overall, it was compatible with the three entomopathogenic fungi species when tested at recommended concentrations for target pest species control. Therefore, this study argues for the use of derivatives from Neotropical Annonaceae as a useful component in the framework of integrated pest management (IPM) programs. / Visando investigar potenciais fontes de novos protetores de grãos, este estudo avaliou, primeiramente, a bioatividade de extratos etanólicos (66) obtidos de 29 espécies pertencentes a 11 diferentes gêneros de anonáceas neotropicais sobre o gorgulho-do-milho Sitophilus zeamais (Coleoptera: Curculionidae). A triagem inicial demonstrou que os efeitos (agudos e crônicos) mais pronunciados sobre S. zeamais foram causados pelos extratos de sementes de Annona montana, de A. mucosa, de A. muricata e de A. sylvatica, seguidos pelos extratos de folhas de A. montana, de A. mucosa, de A. muricata e de Duguetia lanceolata. No entanto, os extratos mais ativos (sementes) não afetaram o crescimento vegetativo e a produção de aflatoxinas de um isolado de Aspergillus flavus (Ascomycota: Trichocomaceae). Fracionamentos biomonitorados foram então realizados a fim de isolar, purificar e caracterizar o(s) composto(s) ativo(s) majoritário(s) dos extratos mais promissores, utilizando-se, para isso, o gorgulho-do-milho como bioindicador. Por meio de diferentes procedimentos cromatográficos, foram isolados nove compostos: cinco acetogeninas, três esteroides e um composto aromático. As acetogeninas roliniastatina-1 e ACG4 (determinação estrutural em andamento), o composto aromático 2,4,5-trimetoxiestireno e os esteroides campesterol, estigmasterol e sitosterol [testados em mistura (8,44 + 12,37 + 79,19%, respectivamente)] mostraram promissoras propriedades protetoras de grãos. Em geral, os resultados obtidos indicaram que compostos de diferentes naturezas químicas têm efeito sinérgico sobre a atividade biológica dos extratos brutos. No segundo estudo, foi avaliada a toxicidade aguda e crônica dos extratos selecionados de sementes de Annona (A. montana, A. mucosa, A. muricata e A. sylvatica) e de um bioinseticida comercial à base de acetogeninas (Anosom® 1EC) sobre a lagarta-mede-palmo Trichoplusia ni (Lepidoptera: Noctuidae) e sobre o pulgão-verde Myzus persicae (Hemiptera: Aphididae). Em laboratório, os extratos de A. mucosa e de A. sylvatica e o bioinseticida Anosom® foram especialmente ativos através da administração oral e tópica. Em casa de vegetação, um extrato formulado de A. mucosa e Anosom® foram altamente eficazes contra larvas de terceiro ínstar de T. ni, com eficácia comparável ao de um inseticida comercial à base de piretrinas (Insect Spray®) utilizado como controle positivo. Similar aos resultados com T. ni, o extrato de A. mucosa apresentou a maior atividade aficida, tanto em bioensaios em laboratório quanto em casa de vegetação. No terceiro estudo, a atividade acaricida [sobre o ácaro-purpúreo-dos-citros Panonychus citri (Acari: Tetranychidae)] do extrato de sementes de A. mucosa (mais ativo) foi avaliada em bioensaios laboratoriais. O extrato de A. mucosa apresentou eficácia superior aos acaricidas/inseticidas comerciais de origem natural [Anosom® 1EC (anonina), Derisom® 2EC (karanjina) e Azamax® 1.2EC (azadiractina + 3-tigloilazadiractol)] e similar a um acaricida sintético [Envidor® 24SC (espirodiclofeno)]. Finalmente, foi avaliada a compatibilidade do extrato de sementes de A. mucosa com três espécies de fungos entomopatogênicos (Beauveria bassiana, Isaria fumosorosea e Metarhizium anisopliae). De modo geral, o extrato de A. mucosa foi compatível com as três espécies quando testado nas concentrações preconizadas para o controle das espécies-praga alvo. Assim, este estudo fornece importantes subsídios para o uso de derivados de anonáceas neotropicais como um componente útil para os programas de manejo integrado de pragas (MIP).

Visando detectar alternativas de manejo para a lagarta-do-cartucho-do-milho, Spodoptera frugiperda (Lepidoptera: Noctuidae), realizou-se, primeiramente, uma triagem em extratos etanólicos preparados das estruturas (folhas, ramos e sementes) de diferentes espécies de Annonaceae (Annona cacans, A. montana, A. mucosa, A. reticulata, A. sylvatica e Duguetia lanceolata). Com base nessa etapa inicial, constatou-se que o extrato etanólico das sementes de A. mucosa (ESAM) foi o mais promissor, causando significativa toxicidade aguda (CL50 e CL90 de 842,97 e 1.882,00 mg kg-1, respectivamente) e pronunciada inibição do desenvolvimento larval (toxicidade crônica), após sete dias de exposição a dietas tratadas. Na CL50, o ESAM reduziu as viabilidades larval e pupal e o peso de pupas com 24 horas, e aumentou a duração da fase larval. No entanto, o ESAM não ocasionou efeito fagodeterrente para larvas de quarto ínstar de S. frugiperda, embora tenha reduzido o consumo ao longo do tempo. Feito isso, a eficácia de ESAM foi comparada com inseticidas comerciais de origem natural (Azamax® 1,2 EC) e sintética (Premio® SC). Nessa comparação, o ESAM (na CL90) apresentou eficácia similar (equitóxico) aos dois produtos comerciais (utilizados na dose registrada para o controle do inseto-praga). A seguir, foi realizado o fracionamento biomonitorado através de diferentes técnicas cromatográficas que conduziu ao isolamento da acetogenina roliniastatina-1, identificada (com base em técnicas espectroscópicas) como componente majoritário da fração mais ativa do extrato etanólico de A. mucosa. Roliniastatina-1 foi então novamente ensaiada frente a larvas de S. frugiperda e ocasionou significativos efeitos agudos (mortalidade larval) e crônicos (redução do desevolvimento larval). Por fim, investigou-se a bioatividade de uma formulação à base de acetogeninas (Anosom® 1 EC) recentemente registrada, tanto isoladamente quanto em mistura com uma formulação à base de limonoides (Azamax® 1,2 EC). Anosom® 1 EC, testado na CL90 estimada (2.959,00 mg kg-1) e Azamax® 1,2 EC, testado na concentração registrada para o controle do inseto-praga (4.000,00 mg kg-1), causaram significativa mortalidade larval, sem ocorrer diferença entre os tratamentos tanto quando testados isoladamente como em mistura binária. Anosom® 1 EC (na CL50) provocou ainda aumento significativo das mortalidades larval e pupal e da duração da fase larval, bem como redução do peso pupal do inseto. Dessa forma, derivados de Annonaceae podem constituir um componente útil para o manejo integrado de S. frugiperda em condições de campo. / A screening with ethanolic extracts prepared from structures (leaves, branches and seeds) from different species of Annonaceae (Annona cacans, A. montana, A. mucosa, A. reticulate, A. sylvatica and Duguetia lanceolata) aiming to detect management alternatives for Spodoptera frugiperda (Lepidoptera: Noctuidae) was done. Based on this initial phase, it was verified that the ethanolic extract from seeds of A. mucosa (AMSE) was the most promising one, causing significant acute toxicity (LC50 and LC90 of 842.97 and 1,822.00 mg kg-1, respectively) and pronounced inhibition of larval development (chronic toxicity) after a 7-day exposition to treated diets. At LC50, AMSE decreased larval and pupal viabilities and also the pupal weight at 24 hours, and increased the larval phase duration. However, AMSE didn\'t cause antifeedant effect for 4-instar S. frugiperda larvae, though it decreased consumption throughout the time. After that, AMSE efficacy was compared to commercial pesticides from natural (Azamax® 1.2 EC) and synthetic (Premio® SC) origin. In this confrontation, AMSE (at LC90) showed efficacy similar to that of commercial products (used at registered dose for S. frugiperda control). Then, the bioguided fractionation using different chromatographic techniques which led to isolation of acetogenin rolliniastatin-1 identified as the majoritary compound of the most active ethanolic extract from A. mucosa. After that, rolliniastatin-1 was again tested against S. frugiperda larvae and caused significant acute (larval mortality) and chronic (decrease of larval development) effects. Finally, the bioactivity of a newly-registered acetogenin-based (Anosom® 1 EC) formulation was investigated, both separately and mixed with a limonoid-based formulation (Azamax® 1.2 EC). Anosom® 1 EC, tested at estimated LC90 (2,959.00 mg kg-1) and Azamax® 1.2 EC, tested at registered dose for S. frugiperda (4,000.00 mg kg-1) led to significant larval mortality, without happening any differences between treatments either tested separately or in a binary mixture. Anosom® 1 EC (at LC50) also led to significant increase of larval and pupal mortality and larval phase, as well as insect pupal weight decrease. Thus, Annonaceae derivates can be a useful component for S. frugiperda integrated management under field conditions.

Made available in DSpace on 2017-07-10T19:23:44Z (GMT). No. of bitstreams: 1 Marcia Mauli.pdf: 2078988 bytes, checksum: f7acd59f9f9540c3713e7315d3a18a92 (MD5) Previous issue date: 2013-11-12 / This trial aimed at isolating and identifying secondary compounds of winter cover crops as black oats, turnip and hairy vetch in order to meet the growing need for healthy products that do not harm the environment. It also enlarges the management knowledge of such crops to demonstrate their advantages on weed control and stimulate the use of a correct no-tillage system, since there is some evidence of their allelopathic potential. Parameters that help on elucidating decomposition of plant residues and releasing allelochemicals to inhibit Bidens pilosa have been evaluated as inhibition percentage according to the amount of mass, which used the amounts of 10, 20, 30, 40 and 50 g, equivalent to 7.5, 15, 22.5, 30 and 37.5 t ha -1, and as well as the control treatment (no mass cover). It was also evaluated influence of field capacity on the wastes decomposition, with 50 and 70% of available water plus the control (100%). There was some releasing of allelochemicals by permanence of plant material on the soil in order to evaluate their respective periods of vegetable permanence: 0, 2, 4, 6 and 8 weeks and the application of soil solution where cover crops were cultivated, which were tested in a concentrated way and at the following dilutions 100, 200, 300 and 400 mL L- 1, as well as the control. In all tests, emergence (inhibition emergence percentage, emergence speed index - ESI and emergence speed - ES) during 10 days and initial development (fresh and dry weight of shoot and root) during 30 days were evaluated. Five seeds or seedlings have been used in pot and 20 of them in gerbox, with three replications. The secondary compounds were obtained by exhaustive extraction during seven days, isolated by column chromatography by wet (CCW) and identified by nuclear magnetic resonance (NMR). The main chemical groups in plants were also identified through techniques such as fingerprint and phytochemical screen. The experimental design was completely randomized. Quantitative data were submitted to variance analysis to check significance and were submitted to regression analysis. The qualitative data were submitted to average comparison by Tukey test at 5 % probability. Generally, the greater residue amount on soil, the greater is B. pilosa inhibition. The best regarding water content in soil is that the result must range between 70 and 50%, which is when the greatest inhibitions occurred. During the evaluation of decomposition periods, there was greater interference on sterilized soil and the residue management than the period itself. Thus, the most significant results were found in no sterilized soil and residue incorporation. The soil solution showed considerable changes in B. pilosa emergence only during the last sampling collections, especially in oilseed radish and hairy vetch. The results, in this case, were more significant when germitest paper was used than sand as substrate. In relation to the compounds, it was detected the presence of tannins, steroids and triterpenoids, derived from coumarin, spumidic saponin and alkaloids. So, based on the observed positive changes, it is recommended to use these cover crops in crop rotation with soybeans to reduce B. pilosa weed / Este trabalho teve por objetivo isolar e identificar os compostos secundários das espécies de cobertura de inverno tais como aveia preta, nabo forrageiro e ervilhaca peluda a fim de suprir a crescente necessidade de busca por produtos saudáveis e que não agridam o meio ambiente. Além de ampliar os conhecimentos sobre o manejo dessas plantas, para demonstrar suas vantagens no controle de plantas invasoras e incentivar o uso do sistema plantio direto de forma adequada, já que as mesmas possuem evidências de potencial alelopático. Parâmetros que auxiliassem na elucidação da decomposição dos resíduos vegetais e liberação de aleloquímicos na inibição de picão preto foram avaliados, como a porcentagem de inibição em função da quantidade de massa, usando as quantidades de 10, 20, 30, 40 e 50 g, equivalentes a 7,5; 15; 22,5; 30 e 37,5 t ha-1, além da testemunha (sem cobertura). Também foi avaliada a influência da capacidade de campo na decomposição dos resíduos, com 70 e 50% de água disponível, mais a testemunha (100%). Houve a liberação de aleloquímicos pela permanência do material vegetal sobre o solo, em que foram avaliados os respectivos períodos de permanência vegetal: 0; 2; 4; 6 e 8 semanas e aplicação de solução do solo onde foram cultivadas as espécies de cobertura, as quais foram testadas de forma concentrada e nas diluições 100; 200; 300 e 400 mL L-1, mais a testemunha. Em todos os testes foram avaliados: emergência (porcentagem de inibição de emergência, índice de velocidade de emergência IVE e velocidade de emergência, VE) durante 10 dias e desenvolvimento inicial (massa fresca e seca de parte aérea e da raiz) durante 30 dias. Cinco sementes ou plântulas em vaso e 20 delas em gerbox foram utilizadas, com três repetições. Os compostos secundários foram extraídos por extração exaustiva durante sete dias; isolados por coluna cromatográfica por via úmida (CCVU) e identificados por ressonância magnética nuclear (RMN). Os principais grupos químicos presentes nas plantas também foram identificados por meio das técnicas fingerprint e screen fitoquímico. O delineamento experimental foi inteiramente casualizado (DIC). Os dados quantitativos foram submetidos à análise de variância para verificação de significância e submetidos à análise de regressão. Os qualitativos foram submetidos à comparação de médias pelo teste de Tukey a 5% de probabilidade. De modo geral, quanto maior a quantidade de resíduo sobre o solo, maior a inibição de picão preto. O ideal é que, no que se refere à quantidade de água no solo, o resultado fique entre 70 e 50%, quando ocorreram as maiores inibições. Na avaliação dos períodos de decomposição, houve maior interferência do solo esterilizado e do manejo do resíduo do que do próprio tempo. Todavia, os resultados mais expressivos foram encontrados em solo não esterilizado e com incorporação do resíduo. A solução do solo apresentou alterações consideráveis na emergência de picão preto apenas nas últimas coletas, principalmente em nabo forrageiro e ervilhaca peluda. Os resultados, neste caso, foram mais expressivos quando se utilizou papel germiteste do que areia como substrato. Com relação aos compostos presentes nas plantas, detectou-se a presença de taninos, esteroides e triterpenóides, derivados de cumarina, saponina espumídica e alcaloides. Em vista dos resultados observados, recomenda-se o uso dessas plantas de cobertura na rotação de culturas com a soja, visando à diminuição da planta invasora picão preto

Insect herbivores often secrete glucose oxidase (GOX) onto plants to counteract plant defenses and potential pathogens. Whether generalist herbivores always have significantly higher GOX activities than their specialist counterparts at any comparable stage or conditions and how this is realized remain unknown. To address these two general questions, we subjected larvae of a pair of sister species differed mainly in host range, the generalist Helicoverpa armigera and its specialist counterpart Helicoverpa assulta, to the same sets of stage, protein to digestible carbohydrate (P:C) ratio, allelochemical or host plant treatments for simultaneous analyses of GOX transcripts and activities in their labial glands. GOX activity and transcripts are upregulated concurrently with food ingestion and body growth, downregulated with stopping ingestion and wandering for pupation in both species. The three tested host plants upregulated GOX transcripts, and to a lesser extent, GOX activity in both species. There were significant differences in both GOX transcripts and activity elicited by allelochemicals, but only in GOX transcripts by P:C ratios in both species. GOX activities were higher in H. armigera than H. assulta in all the comparable treatments, but GOX transcripts were significantly higher either in generalists or in specialists, depending on the developmental stages, host plants, P:C ratio and allelochemicals they encounter. These data indicate that the greater GOX activity in generalist herbivores is not achieved by greater transcription rate, but by greater transcript stability, greater translation rate, better enzyme stability and/or their combination.

Visando contribuir para o manejo de moscas-das-frutas em parreirais de uva, inicialmente determinou-se, neste estudo, a concentração limiar dos limonoides azadiractina e 3-tigloilazadiractol (AzaMax®) capaz de inibir completamente a oviposição de Ceratitis capitata em frutos de uva. Em seguida, foram averiguados os estímulos envolvidos na inibição da oviposição da praga pelos referidos limonoides. Posteriormente, avaliou-se a proteção do parreiral de uva pelos limonoides contra C. capitata no polo de produção de Petrolina/PE, Brasil. Finalmente, foi avaliada a atividade biológica residual dos limonoides sobre a praga em parreiral de uva no polo de produção de Petrolina. A concentração limiar para a completa inibição da oviposição de C. capitata se dá com 100 ppm de azadiractina (+ 28,5 ppm de 3-tigloilazadiractol) na superfície de frutos de uva. A inibição da oviposição é drástica a partir de 50 ppm de azadiractina (+ 14,3 ppm de 3-tigloilazadiractol) em situação de chance de escolha do hospedeiro. A seleção hospedeira de C. capitata não é afetada por concentração igual ou inferior a 7,5 ppm de azadiractina (+ 2,14 ppm de 3-tigloilazadiractol) na superfície de frutos de uva. A concentração limiar de 100 ppm de azadiractina (+ 28,5 ppm de 3-tigloilazadiractol) impede que o hospedeiro seja reconhecido pela praga em função dos estímulos que promovem a inquietação (irritação), locomoção e consequente dispersão da praga. Assim, os limonoides (AzaMax®) podem ser qualificados como repelentes com baixa capacidade de difusão. Os limonoides em alta concentração ocasionam mortalidade sobre C. capitata, porém não evitam que os cachos ou frutos de uva sejam injuriados e ou infestados por C. capitata numa condição de baixa população natural ou artificial em campo. As conclusões registradas corroboram a hipótese de que são necessárias mais investigações pormenorizadas para avaliação do efeito de limonoides do nim sobre C. capitata em campo. A perspectiva de controle comportamental de moscas-das-frutas utilizando limonoides do nim foi discutida, com especial referência para C. capitata em parreirais de uva. / Aiming to contribute to the management of fruit flies in the vineyards, this research initially determined the concentration threshold of limonoids azadirachtin and 3-tigloylazadirachtol (AzaMax®) able to completely inhibit oviposition of Ceratitis capitata in grape fruits. Then the stimuli involved in the inhibition of oviposition of C. capitata by these limonoids were investigated. Subsequently, we evaluated the vineyard protection by limonoids against fruit flies in the production pole of Petrolina - PE, Brazil. Finally, we evaluated the residual biological activity of limonoids on C. capitata in vineyard in the production pole of Petrolina. The threshold concentration for complete inhibition of oviposition of C. capitata is 100 ppm of azadirachtin (+ 28.5 ppm of 3-tigloylazadirachtol) on the surface of grape fruits. The inhibition of oviposition is drastic from 50 ppm of azadirachtin (+ 14.3 ppm of 3-tigloylazadirachtol) with free choice of the host. The host selection by C. capitata is not affected by concentration equal to or less than 7.5 ppm of azadirachtin (2.14 ppm of 3-tigloylazadirachtol) on the fruits surface. The threshold concentration of 100 ppm of azadirachtin (+ 28.5 ppm of 3-tigloylazadirachtol) prevents the host to be recognized by C. capitata depending of the stimuli that promote caring (irritation), locomotion and consequent dispersion of insect. Thus, the limonoids (AzaMax®) can be qualified as repellents with low diffusion capacity. The limonoids in high concentration (AzaMax®) cause mortality of C. capitata, but do not prevent the injury and or infestation on grape clusters by C. capitata in a condition of low natural or artificial population in the field. The findings reported support the hypothesis that more detailed assessment for the effect of neem limonoids on C. capitata in field investigations are necessary. The perspective of behavioral control of fruit flies using neem limonoids was discussed, with special reference to C. capitata in vineyards.

Avaliou-se a bioatividade de derivados do nim (Azadirachta indica A. Juss.) como componente de isca para adultos e como regulador de crescimento de Ceratitis capitata (Wied.) e Anastrepha fraterculus (Wied.), determinando-se, ainda, o efeito de concentrações subletais de extratos de nim e o efeito deterrente de oviposição do extrato metanólico de folhas sobre C. capitata. Utilizando o extrato aquoso de amêndoas de nim, estimou-se a CL50 para adultos e imaturos, cujos valores foram 7.522 e 1.368 ppm para os adultos e 13.028 e 9.390 ppm para os imaturos de A. fraterculus e C. capitata, respectivamente. Subsequentemente, o óleo de amêndoas, os extratos de folhas, de ramos e de amêndoas de nim em diferentes solventes (água, metanol, diclorometano e hexano) foram comparados com base nas CL50 obtidas para adultos e para imaturos. Posteriormente, foram estimadas concentrações subletais (CL15 = 39 ppm, CL30 = 225 ppm e CL45 = 888 ppm) do extrato aquoso de amêndoas para adultos de C. capitata e avaliouse nessas concentrações o efeito do extrato aquoso de amêndoas, de folhas em metanol e de ramos em diclorometano sobre a reprodução e a longevidade dessa espécie. A deterrência de oviposição do extrato metanólico de folhas de nim em diferentes concentrações (10.000, 18.000, 32.000 e 56.000 ppm) para C. capitata foi avaliada tratando bagas de uva, que foram expostas às fêmeas com chance e sem chance de escolha. Verificou-se que os derivados do nim na concentração de até 7.522 e 1.368 ppm não afetaram os adultos de A. fraterculus e C. capitata, respectivamente. Já para a fase imatura, os extratos de amêndoas e de ramos em diclorometano na concentração de 13.028 ppm causaram mortalidade de A. fraterculus e na concentração de 9.390 ppm o extrato de amêndoas em metanol e em diclorometano causaram mortalidade de C. capitata. Houve redução da fecundidade total e da fertilidade no 8º dia quando os insetos foram alimentados com extrato de ramos em diclorometano a 888 ppm. A longevidade de machos e fêmeas e o período de pré-oviposição não foram afetados pelos tratamentos. O extrato de folhas em metanol provocou deterrência de oviposição para C. capitata em situação de chance de escolha, porém, em confinamento, o referido extrato não afetou o comportamento dessa espécie. A experiência recente de C. capitata não alterou a sua resposta em relação a bagas de uva tratadas. A atividade biológica dos derivados do nim em relação às moscas-das-frutas é discutida. / The bioactivity of neem (Azadirachta indica A. Juss.) as adult bait and as larval growth regulator for Ceratitis capitata (Wied.) and Anastrepha fraterculus (Wied.) was evaluated. Besides, the effect of sublethal concentrations of neem extracts and the oviposition deterrency of the methanolic extract from leaves on C. capitata was evaluated. The estimated LC50 of aqueous extract from seed kernels were 7,522 and 1,368 ppm for adults and 13,028 and 9,390 ppm for immatures of A. fraterculus and C. capitata, respectively. In addition, the oil from seed kernels and the extracts from leaves, branches and seed kernels in different solvents (water, methanol, dichloromethane and hexane) were compared based on the LC50 obtained for adults and immatures. After that, sublethal concentrations from aqueous extract from neem seed kernels for C. capitata adults were estimated (LC15 = 39 ppm, LC30 = 225 ppm and LC45 = 888 ppm) and the effects of such concentrations on the reproduction and logenvity of this species were also accessed. Oviposition deterrency of C. capitata to grapes treated with leaf methanolic extract at different concentrations (10,000; 18,000; 32,000 and 56,000 ppm) was evaluated in free-choice or no-choice test. It was verified that the neem extracts on the concentrations up to 7,522 and to 1,368 ppm did not affect the adults of A. fraterculus and C. capitata, respectively. On the other hand, the extract from seed kernels and branches in dichloromethane at 13,028 ppm provoked mortality in A. fraterculus larvae and the seed kernel extract in methanol and dichloromethane at 9,390 ppm provoked high mortality in C. capitata larvae. The total fecundity was reduced significantly and the fertility decreased on the 8th day when the insects were fed on branch extract in dichloromethane at 888 ppm. The longevity of males and females and the pre-oviposition period were not affected by treatments. The leaf extract in methanol provoked oviposition deterrency for C. capitata in free-choice test, but did not affect its behavior in no-choice test. The C. capitatas early experience did not affect its answer to the grapes treated in free-choice and no-choice situations. The biological activity of the neem extracts to fruit flies is discussed.

L’objectif de cette thèse est d’étudier les interactions allélopathiques dans la forêt de chêne pubescente en région méditerranéenne et d’envisager comment ces interactions peuvent être affectées par le changement climatique. Nous avons montré que l'arbre dominant Q. pubescens et ses espèces compagnes A. monspessulanum et C. coggygria ont des potentialités allélopathiques différentes en fonction du stade phénologique des feuilles ce qui pourrait affecter la germination et la croissance des plantes herbacées de sous-étage. Ces différences d'effet allélopathique peuvent résulter non seulement de différences dans la quantité des composés allélopathiques produits, mais aussi de différences qualitatives observées en fonction du stade phénologique. Cela suggère que l’effet allélopathique puisse varier dans le temps de manière à correspondre au stade de développement des plantes herbacées cibles. Sur l’une des plantes cibles utilisées dans les bioessais, les effets allélopathiques ont d’autre part été accentués en changeant les conditions environnementales (hydriques et/ou thermiques) ce qui suggère des modifications éventuelles des potentiels allélopathiques des plantes à attendre avec le changement climatique. Nos résultats en pépinière ont montré le potentiel allélopathique de cotinus et pinus sur la croissance des plantules de chêne, quelques soient les conditions d’arrosage (stressé ou non stressé. En affectant notamment la biomasse du système racinaire dont la croissance est un élément clé des espèces méditerranéennes pour résister à la sécheresse estivale prolongée, ces interactions pourraient jouer un rôle important dans les processus de régénération du chêne pubescent. / The aim of this thesis is to study allelopathic interactions in the downy oak forest in the mediterranean region and to consider how these interactions can be affected by climate change. We have shown that the dominant tree Q. pubescens and its companion species A. monspessulanum and C. coggygria have different allelopathic potentialities, evaluated in vitro on herbaceous target plants, depending on the phenological stage of the leaves, which could in natura affect the germination and growth of understory herbaceous plants. These differences in allelopathic effect may result not only from differences in the quantity of allelopathic compounds produced, but also from qualitative differences observed as a function of the phenological stage. This suggests that the allelopathic effect may vary over time to match the stage of development of the target herbaceous plants. On one of the target plants used in the bioassays, the allelopathic effects were further accentuated by changing the water and / or thermal conditions which suggests possible modifications of the allelopathic potential of the plants to be expected with climate change.Our results in the plant nursery showed the allelopathic potential of cotinus and pinus on the growth of oak seedlings, whatever the watering conditions (stressed or unstressed). By affecting in particular the biomass of the root system, the growth of which is a key element of the mediterranean species to resist the prolonged summer drought, these interactions could play an important role in the regeneration processes of downy oak.

This thesis describes the synthetic preparation of somecompounds, which can serve as chemical signals for use in thedevelopment of control methods for pest insects. The compoundssynthesised are of the isoprenoid type and of two kinds:carvone derivatives and germacranes. The derivatives of carvoneare based on modifications of this compound, by reactions ofeither its endocyclic or its exocyclic double bond. One type ofmodifications was accomplished by chemoselective additions ofthiophenol. The latter ones imply additions to the exocyclicdouble bond and seem to constitute general, previously rarelystudied reactions. In other modifications of its exocyclic side chain, carvoneafforded some sesqui- and diterpeniod natural products. Thefollowing compounds were synthesised in an enantioselectiveway: (-)-epi-delobanone, (-)-delobanone,(-)-7-hydroxy-3,10-prenylbisaboladien- 2-one (an insecticidalconstituent of Croton linearis) as well as its diastereomer andsome other compounds with similar structures. All of thesecompounds weretested for their antifeedant/feeding deterrentcapability against gnawing of the pine weevil, Hylobiusabietis. The germacranes prepared by means of enantioselective totalsyntheses are: (–)- 1(10),5-germacradien-4-ol and(–)-germacrene D. The former is a constituent of thedefence secretion (an allomone) from the larvae of the pinesawfly, and the needles of Scots pine. (–)-Germacrene D isa ubiquitous compound in nature. For example, it occurs in thepeels of apples and acts as one component of a lure (akairomone) to the apples, which attracts the codling moth,Cydia pomonella. The main problem in the total syntheses of the germacraneswas the formation of the unsaturated monocyclic 10-memberedring. This was achieved by intramolecular alkylation with asuitably functionalised/protected cyanohydrin derivative,which, after further elaboration, afforded a monocyclic10-membered enone, that was used in the syntheses of the twogermacranes mentioned above. In the initial steps in thesynthetic sequence the stereochemistry was established byalkylation of an amide enolate attached to a chiral auxiliary.This approach could most likely also readily furnish the(+)-enantiomers of these germacrenes (of the germacraneterpenoid class) using the opposite enantiomer of the chiralauxiliary in the initial steps. <b>Keywords</b>: isoprenoids, natural product synthesis,allelochemicals, kairomones, allomones, bisabolane terpenoids,Hylobius abietis, germacrane terpenoids, Neodiprion sertifer,stereoselective synthesis.

<p>This thesis describes the synthetic preparation of somecompounds, which can serve as chemical signals for use in thedevelopment of control methods for pest insects. The compoundssynthesised are of the isoprenoid type and of two kinds:carvone derivatives and germacranes. The derivatives of carvoneare based on modifications of this compound, by reactions ofeither its endocyclic or its exocyclic double bond. One type ofmodifications was accomplished by chemoselective additions ofthiophenol. The latter ones imply additions to the exocyclicdouble bond and seem to constitute general, previously rarelystudied reactions.</p><p>In other modifications of its exocyclic side chain, carvoneafforded some sesqui- and diterpeniod natural products. Thefollowing compounds were synthesised in an enantioselectiveway: (-)-epi-delobanone, (-)-delobanone,(-)-7-hydroxy-3,10-prenylbisaboladien- 2-one (an insecticidalconstituent of Croton linearis) as well as its diastereomer andsome other compounds with similar structures. All of thesecompounds weretested for their antifeedant/feeding deterrentcapability against gnawing of the pine weevil, Hylobiusabietis.</p><p>The germacranes prepared by means of enantioselective totalsyntheses are: (–)- 1(10),5-germacradien-4-ol and(–)-germacrene D. The former is a constituent of thedefence secretion (an allomone) from the larvae of the pinesawfly, and the needles of Scots pine. (–)-Germacrene D isa ubiquitous compound in nature. For example, it occurs in thepeels of apples and acts as one component of a lure (akairomone) to the apples, which attracts the codling moth,Cydia pomonella.</p><p>The main problem in the total syntheses of the germacraneswas the formation of the unsaturated monocyclic 10-memberedring. This was achieved by intramolecular alkylation with asuitably functionalised/protected cyanohydrin derivative,which, after further elaboration, afforded a monocyclic10-membered enone, that was used in the syntheses of the twogermacranes mentioned above. In the initial steps in thesynthetic sequence the stereochemistry was established byalkylation of an amide enolate attached to a chiral auxiliary.This approach could most likely also readily furnish the(+)-enantiomers of these germacrenes (of the germacraneterpenoid class) using the opposite enantiomer of the chiralauxiliary in the initial steps.</p><p><b>Keywords</b>: isoprenoids, natural product synthesis,allelochemicals, kairomones, allomones, bisabolane terpenoids,Hylobius abietis, germacrane terpenoids, Neodiprion sertifer,stereoselective synthesis.</p>

Submitted by Izabel Franco (izabel-franco@ufscar.br) on 2016-10-03T14:33:55Z No. of bitstreams: 1 TeseLPC.pdf: 5483250 bytes, checksum: fa7b00ca116fc99170337aefc31b986d (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-10-20T18:29:53Z (GMT) No. of bitstreams: 1 TeseLPC.pdf: 5483250 bytes, checksum: fa7b00ca116fc99170337aefc31b986d (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-10-20T18:30:01Z (GMT) No. of bitstreams: 1 TeseLPC.pdf: 5483250 bytes, checksum: fa7b00ca116fc99170337aefc31b986d (MD5) / Made available in DSpace on 2016-10-20T18:30:08Z (GMT). No. of bitstreams: 1 TeseLPC.pdf: 5483250 bytes, checksum: fa7b00ca116fc99170337aefc31b986d (MD5) Previous issue date: 2016-03-16 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Brazil by presenting incomparable biodiversity, distributed in mosaic wonderfully distinct biomes becomes a promising the field of study in seeks of new phytochemicals extracted from native plants with biological activity on weeds and insects of medical importance. Species of Lauraceae and Dilleniaceae family have an immense potential in the production of secondary compounds with herbicide and insecticide properties. Thus, this study objective was to clarify the following questions: leaf extracts the stem and root Ocotea pulchella and Davilla elliptica could interfere with the growth and development of Triticum aestivum, Euphorbia heterophylla and Panicum maximum and thus make a new alternative for the control of weeds? Amongst the donor species and crude extracts which more promising to start a bioprospecting, allowing isolate and identify the present compounds? The extracts show insecticidal activity against eggs and larvae of Aedes aegypti? Coleoptile bioassays were performed using hexane extracts, dichloromethanic, acetoetilicos, methanolic and aqueous leaf, stem and root of O. pulchella and D. elliptica 0.2, 0.5 and 0.8 mg.mL-1. Most active extracts have been evaluated on the germination and early seedling growth of E. heterophylla and P.maximum using doses of 0.25, 0.5, 1 and 2 mg.mL-1. The larvicidal activity was evaluated at concentrations of 0.25, 0.5, 1, 2 and 3 mg.mL-1 and ovicidal effect with the LC50 and LC90 lethal concentrations estimated in test A. aegypti larvae. The leaf extracts O. pulchella inhibit coleoptile growth of wheat and seed germination and seedling growth of weeds and standard species. In general, the ethyl acetate extract O. pulchella showed higher phytotoxic activity. Eleven compounds were isolated and characterized. Compounds 1, 2, 4 and 6 have not been identified before in O. pulchella and Ocoteol was first described in the literature. The compounds 5, 10 and 11 were most active with superior inhibition values of 90% in the standard species. P. maximum has been the target species most affected by O. pulchella and D.elliptica leaf extracts in all evaluated parameters. The reduction in root growth of wild poinsettia seedlings can be related to the decrease in the elongation of metaxylem cells submitted at the ethyl acetate extract (O. pulchella) and methanol (D.elliptica). The extracts from root hexane and dichloromethane O. pulchella at a concentration 3 mg.mL-1 have produced bigger effect larvicide. Lower values of CL50 and CL90 were estimated for larvae subjected to the root extract from O. pulchella dichloromethane. Hatching rate, TME and VME have been affected by DCM and hexane root extracts from O. pulchella. Specie O. pulchella showed higher potential herbicide on the germination and the growth pattern and weed species and insecticide on larvae and eggs of A. aegypti. / O Brasil, por apresentar uma grande biodiversidade, torna-se um campo de estudo promissor na busca de novos fitoquímicos extraídos de plantas nativas, com atividades biológicas sobre plantas infestantes e insetos de importância médica. As espécies da família Lauraceae e Dillineaceae possuem potencial na produção de compostos secundários com propriedades herbicida e inseticida. Sendo assim, o presente estudo objetivou esclarecer os seguintes questionamentos: os extratos de folha, caule e raiz de Ocotea pulchella e Davilla elliptica interferirem no crescimento e desenvolvimento de Triticum aestivum, Euphorbia heterophylla e Panicum maximum possibilitando, assim, uma nova alternativa no controle de plantas infestantes? Entre as espécies doadoras , qual o mais promissor para se iniciar uma bioprospecção, permitindo isolar e identificar os compostos presentes? Os extratos brutos de ambas as espécies apresentam atividade inseticida sobre ovos e larvas de Aedes aegypti? Para tanto, foram realizados bioensaios de coleóptilos de trigo utilizando os extratos hexânicos, diclorometânicos, acetoetílicos, acetônicos, metanólicos e aquosos de folha, caule e raiz de O. pulchella e D. elliptica a 0,2, 0,4 e 0,8 mg.mL-1. Os extratos mais ativos foram avaliados sobre a germinação e o crescimento inicial de plântulas de E. heterophylla e P. maximum utilizando as doses de 0,25, 0,5, 1 e 2 mg.mL-1. Para os compostos puros foi utilizado as doses de 10-3, 3x10-4, 10-4, 3x10-5 e 10-5. A atividade larvicida foi avaliada nas concentrações de 0,25, 0,5, 1, 2 e 3 mg.mL-1 e o efeito ovicida por meio das concentrações letais CL50 e CL90, estimadas no teste com larvas de A. aegypti. Os extratos foliares de O. puchella inibiram o crescimento de coleóptilos de trigo, a germinação e o crescimento das plântulas das espécies daninhas e das espécies padrão. Em geral, o extrato de acetato de etila extraído de folha de O. pulchella apresentou maior atividade fitotóxica. Onze compostos do extrato de acetato de etila foram isolados e caracterizados. Os compostos dihidroxi-eudesma, hidroxi-eudesma, loliolide e ácido dimorfecolico não foram descritos antes em O. pulchella e o Ocoteol foi descrito pela primeira vez na literatura. Os compostos espatulenol, benzil salicilate e benzil benzoate foram os mais ativos, com valores de inibição superior a 90% nas espécies padrão. P. maximum foi a espécie alvo mais afetada pelos extratos de folha de O. pulchella e D.elliptica em todos os parâmetros avaliados. A redução no crescimento radicular das plântulas de amendoim-bravo pode estar relacionada com a diminuição no alongamento das células do metaxilema submetidas ao extrato de acetato de etila (O. pulchella) e metanólico (D.elliptica). Os extratos hexânico e diclorometânico de raiz de O. pulchella, na concentração de 3 mg.mL-1 produziram maior efeito larvicida. Os menores valores das CL50 e CL90 foram estimados para larvas submetidas ao extrato de raiz de O. pulchella extraído com diclorometano. A taxa de eclosão, tempo médio e velocidade média de eclosão foram afetados pelos extratos diclorometano e hexano de raiz de O. pulchella. A espécie O. pulchella apresentou maior potencial como herbicida sobre a germinação e o crescimento de espécies padrão e infestantes e atividade inseticida sobre larvas e ovos de A. aegypti.