As relações entre flores e abelhas durante a evolução tiveram um importante papel na adaptação desses insetos, uma vez que eles se utilizam dos recursos florais (néctar e pólen) para sua alimentação. A adaptação a essa estratégia alimentar fica evidente pela presença de estruturas morfológicas nessas abelhas relacionadas à esse comportamento, como a corbícula. Apesar da utilização de recursos florais estar presente em praticamente todas as espécies de abelhas, algumas espécies de abelhas sem ferrão (Meliponini) apresentam um tipo de comportamento alimentar que difere bastante de todos os outros encontrados em abelhas sociais. No comportamento conhecido como cleptoparasita, as abelhas não visitam flores e conseguem seu alimento através do saque a outras colônias. Este comportamento está presente como estratégia exclusiva nos gêneros Lestrimelitta e Cleptotrigona. Durante o saque são levados da colônia pilhada, mel, pólen, cerúmen e, principalmente, alimento larval. Apesar de alguns estudos já terem sido realizados, muitas questões importantes relacionadas à ecologia química e comportamental do cleptoparasitismo ainda permaneciam sem respostas. Sendo assim, o presente estudo objetivou investigar esses dois aspectos do cleptoparasistismo em L. limao. Dentro do aspecto comportamental as análises se concentraram na investigação da escolha das colônias hospedeiras. As análises ecoquimícas, por sua vez, focaram na identificação dos compostos presentes nas glândulas mandibular e salivar cefálica e na identificação do papel ecológico desses compostos durante os saques. Os resultados obtidos trouxeram novas e importantes informações sobre o comportamento cleptoparasita de L. limao. Foi possível observar uma menor frequência dos ataques à espécie que apresenta respostas agressivas ao saque, em comparação às que agem passivamente. Além disso, os dados obtidos nas análises químicas trouxeram novidades importantes, relacionadas à proporção dos isômeros do citral presente na glândula mandibular das operárias, à presença de ésteres na glândula salivar cefálica e ao papel ecológico do citral. Ainda, a evolução do cleptoparasitismo em abelhas sem ferrão é amplamente discutida e, por fim, é explicitada a necessidade de uma nova classificação para os diferentes tipos de parasitismo presentes no grupo das abelhas. / The interactions between flowers and bees played an important role in the behavioral evolution of these insects since they feed on floral resources. Nevertheless, some species of stingless bees have a different feeding strategy, which is known as cleptoparasitism. These species obtain their food exclusively through pillaging of other beehives. The exclusivity of this stealing behavior can be found in two different groups of stingless bees, Lestrimelitta and Cleptotrigona. Although some studies concerning this interesting behaviour have already been made, there are many questions related to its chemical and behavioral ecology that remain virtually unknown. Thus, the aim of this study was to investigate these aspects of the cleptoparasitism in Lestrimelitta limao. The behavioral analyses focused on the investigation of host choice by the cleptoparasitic bees while eco-chemical analyses investigated the chemical profile of the cephalic glands (mandibular and cephalic salivary) and their ecological role during the pillage. The results brought new important information concerning the cleptoparasitic behavior of L. limao. The host choice analysis indicates a lower frequency of attacks in the aggressive hosts. Chemical investigations showed different proportions of the citrals isomers which are present in the mandibular gland, the chemical profile of cephalic salivary glands and the ecological role of citral. Furthermore, the evolution of cleptoparasitic behaviour is widely discussed and the necessity of a new classification for the parasitism among bees is presented.

The palm is an important crop, primarily for its forage potential, being an alternative to feeding livestock is scarce native grasslands, especially during periods of prolonged drought. One of the main problems in the cultivation of cactus pear is the infestation of pests such as mealybug in scale, Diaspis echinocacti, when uncontrolled can cause severe yield losses, reaching 100%. The Integrated Management is an alternative to control this pest and maintenance of production out of economic damage. Therefore, this study aimed to evaluate the interaction between the spineless cactus, cochineal scale and two of its natural enemies, Zagreus bimaculosus and Chilocorus nigrita. Volatile organic compounds (VOCs) released by ten varieties of cactus, healthy and infested by D. echinocacti, as well as the volatiles released by the two natural enemies studied were collected by aeration and extracted with n-hexane HPLC. The extracts were subjected to behavioral bioassays in olfatometre Y to identify their attractive activity for natural enemies, Z. bimaculosus and C. nigrita, and then subjected to chemical analysis by gas chromatography coupled to mass spectrometry (GC-MS) for identification of VOCs. Electrophysiologic testing with gas chromatography coupled to electroantennograph (GC-EAG) of the extracts that showed activity in behavioral bioassays were conducted. The results showed that the species of ladybugs are attracted by the extracts studied, mostly after the process of infestation with D. echinocacti, and that the profiles of the emission of VOCs of ten varieties studied differed. Among the VOCs identified in extracts of Z. bimaculosus and C. nigrita, are the 1-ethyl-hexanol, 3,7-dimethyloctanol, 1-heptacosanol, 1-decanol, 2-propyl-1-heptanol and farnesol, which were identified as components of the aggregation pheromone of beetles and beetle cuticular constituents. It also observe that new compounds were found in the extracts of ten varieties of cactus infested with cochineal scale of D. echinocacti, among these compounds are the n-octanal, geranyl linalool, linalool, n-nonanal, n-decanal and squalene, which are associated with plant defense, including the attraction of natural enemies. These results demonstrate the importance of VOCs produced by plants for their defense, after a process of herbivory, including with regard to the attraction of natural enemies. VOCs released by two species of ladybugs can be studied components of the aggregation pheromone, since attractiveness was demonstrated in behavioral bioassays. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A palma é uma importante cultura, principalmente por seu potencial forrageiro, sendo uma alternativa para alimentação de rebanhos quando há escassez de pastagens nativas, inclusive em períodos de seca prolongada. Um dos principais problemas no cultivo da palma forrageira é a infestação de pragas, como a cochonilha em escama, Diaspis echinocacti, que quando não controlada pode causar perdas severas na produtividade, podendo chegar a 100%. O Manejo Integrado é uma alternativa para o controle desta praga e manutenção da produção fora de um dano econômico. Assim sendo, este estudo objetivou avaliar a interação entre a palma forrageira, a cochonilha de escama e dois de seus inimigos naturais, Zagreus bimaculosus e Chilocorus nigrita. Os compostos orgânicos voláteis (COVs) liberados por dez variedades de palma forrageira, sadias e infestadas por D. echinocacti, assim como os voláteis liberados pelos dois inimigos naturais estudados foram coletados por aeração e extraídos com hexano grau HPLC. Os extratos foram submetidos a bioensaios comportamentais em olfatômetrro Y, para identificação de sua atividade atraente para os inimigos naturais, Z. bimaculosus e C. nigrita, e em seguida submetidos a análises químicas por Cromatografia Gasosa acoplada a Espectrometria de Massas (CG-EM) para identificação dos COVs. Foram realizados testes eletrofisiológicos com Cromatografia Gasosa acoplada ao Eletroantenógrafo (CG-EAG) dos extratos que exibiram atividade nos bioensaios comportamentais. Os resultados obtidos demonstraram que as espécies de joaninhas estudadas são atraídas pelos extratos, em sua maioria, após o processo de infestação com D. echinocacti, e que os perfis de emissão dos COVs das dez variedades estudadas diferem entre si. Dentre os COVs identificados nos extratos de Z. bimaculosus e C. nigrita, encontram-se o 1-etil-hexanol, 3,7-dimetil-octanol, 1-heptacosanol, 1-decanol, 2-propil-1-heptanol e o farnesol, os quais foram identificados como componentes do feromônio de agregação de coleópteros e como constituintes cuticulares de coleópteros. Observa-se ainda que novos compostos foram encontrados nos extratos das dez variedades de palma forrageira infestadas com a cochonilha de escama, D. echinocacti, dentre estes compostos estão o n-octanal, geranil linalol, linalol, n-nonanal, n-decanal e esqualeno, os quais estão associados a defesa de plantas, inclusive na atração de inimigos naturais. Esses resultados comprovam a importância dos COVs produzidos por plantas para sua defesa, após um processo de herbivoria, inclusive no que diz respeito à atração de inimigos naturais. Os COVs liberados pelas duas espécies de joaninhas estudadas podem ser componentes do feromônio de agregação, uma vez que houve atratividade demonstrada em bioensaios comportamentais.

A idade das fêmeas colonizadoras de Hypothenemus hampei (Ferrari) durante o abandonamento do hospedeiro, assim como sua resposta aos voláteis de flores de café, Coffea arabica L. var. Tupi, foram investigadas neste trabalho. Um dispositivo experimental que simulasse as condições de dentro do fruto de café e permitisse a observação da saída dos insetos foi utilizado para a determinação da idade das fêmeas colonizadoras. Em média, os besouros apresentaram 15 dias de idade no momento em que abandonaram o dispositivo experimental. Nesta idade as fêmeas estavam acasaladas, possuiam o tegumento totalmente melanizado, foram capazes de voar e produzir ovos viáveis. Os voláteis de flores de café, foram coletados por aeração, analisados por GC-EAD e GC-MS, e testados em bioensaios olfatométricos. As fêmeas colonizadoras de H. hampei foram atraídas pelos voláteis de flores de café. Dos 50 compostos encontrados nos extratos naturais, sete foram eletrofisiologicamente ativos aos insetos. Destes sete, somente metil salicilato, neral e geranial puderam ser identificados, e a mistura de seus padrões sintéticos foi a mais atrativa nos bioensaios olfatométricos. Tomados juntos, estes resultados trazem novas informações sobre a bioecologia e ecologia química da broca-do-café, as quais poderão ser usadas futuramente em pesquisas de base e naquelas focadas no desenvolvimento de novas estratégias de manejo para H. hampei em cafezais. / The age of Hypothenemus hampei (Ferrari) females during the host abandonment, as well as their response to coffee flower volatiles were determinated under laboratory conditions. A experimental device that simulated the conditions inside a coffee berry, and that allowed the observation the insects coming out was used to determinate the age of colonizing females. On average, the beetles were 15 d-old at the moment they abandoned the experimental device. In this age, the females were mated, had a fully melanized tegument, were able to fly and to produce viable eggs. The volatiles of coffee, Coffea arabica L. var. Tupi, flowers were colected by aeration, analyzed in GC-EAD and GC-MS, and tested in olfactometer bioassays. The colonizing H. hampei females were attracted by the coffee flower volatiles. Of 50 compounds found in the natural extracts, seven were electrophysiologically actives to the insects. Of these seven, only methyl salicylate, neral and geranial could be identified, and the blend with their syntetic standards was the most atractive in the olfactometer bioassays. Taken together, these results bring new information on the bioecology and chemical ecology of the coffee berry borer, which may be used in the future by basic researchs or by those focused on the development of new strategies for the management of H. hampei in coffee farms.

O estudo foi dividido em capítulos que incluíram diversos aspectos da ecologia química de insetos e espécies de Piperaceae como se seguem: O Capítulo 1 descreve as informações taxonômicas, a história natural dos insetos, as observações de campo e a organização das espécies hospedeiras de Piperaceae segundo as preferências alimentares dos insetos. Observou-se forte especificidade química de espécies de Coleoptera (Naupactus bipes) e Homoptera (Membracis foliata, Callocanophora sp., e Aethalium reticulatum) por Piper aduncum, P. gaudichaudianum, P. arboreum e P. hispidum que possuem ácidos benzóicos prenilados como produtos predominantes. As Lepidoptera (Quadrus u-lucida, Heraclydes brasiliensis e H. helicorides) constituíram-se num segundo grupo de insetos para o qual constatou-se forte especificidade. Nesse caso as espécies de Piperaceae, P. regnellii e P. solmsianum, são nitidamente acumuladores de neolignanas e/ou lignanas. Os Capítulos 2 e 3 (coleópteros - besouros e lepidópteros - borboletas, respectivamente) descrevem os resultados relacionados aos estudos das reações que ocorreram durante o processo de digestão de folhas de espécies de Piper, seqüestros de metabólitos secundários pelos insetos. O estudo de possíveis fatores atrativos presentes em óleos essenciais foram investigados mediante o uso de ensaios eletrofisiológicos. Foram observadas reações de desmetilações em lignanas tetraidrofurânicas, esterificação de ácidos benzóicos, hidrólise de amidas, reação do tipo ozonólise de lignanas, neolignanas e fenilpropanóides durante o processo digestivo das borboletas (larvas) e dos besouros adultos. Foram observados seqüestros de neolignanas das folhas e raízes de P. regnellii pelas larvas da borboleta Heraclides hectorides e ácidos benzóicos prenilados das raízes de P. gaudichaudianum pelas larvas do besouro Naupactus bipes. Os experimentos de EAG com antenas do besouro N. bipes indicou que os óleos de P. gaudichaudianum, P. regnellii e P. hispidum foram ativos, e tal comportamento foi confirmado por observações em campo, sendo que as respostas mais intensas foram observadas para as fêmeas em relação às respostas para os machos. A análise de GC/EM-EAD permitiu a determinação dos compostos bioativos como os monoterpenos α- pineno, ß-pineno e ß-mirceno. O Capítulo 4 descreve as espécies de homópteros que têm especificidade por P. gaudichaudianum, P. arboreum e P. aduncum que são acumuladoras de ácidos benzóicos 7 prenilados. Outros possíveis fatores determinantes para tal especificidade foram atribuídos ao baixo teor de metabólitos secundários nas seivas, alto teor de micronutrientes, baixo teor de macronutrientes, ausência de lignanas ou neolignanas nas seivas, lignificação com predominância de resíduo siringila (S). O Capítulo 5 descreve a determinação e caracterização estrutural dos 49 metabólitos secundários envolvidos neste estudo. O Capítulo 6 descreve o estudo da estabilidade de lignanas tetraidrofurânicas realizado em função da rara ocorrência na natureza de isômeros com a configuração toda cis. O estudo da estabilidade para os dez possíveis estereoisômeros para a lignana tetraidrofurânica através do cálculo do funcional de densidade, B3LYP, com a base 6- 316(dp) mostrou que a configuração toda cis é de fato menos estável do que a toda trans. / The study was presented in six chapters that included several aspects of chemical ecology of insects and Piperaceae species as following: The Chapter 1 describes taxonomical aspects, natural history of the insects and field observation and the organization of hosts Piperaceae according to the observed feeding preferences of the associated insects. It was observed strong chemical specificity of Coleoptera and Homoptera species by Piper aduncum, P. gaudichaudianum, P. arboreum and P. hispidum which contain prenylated benzoic acids. The Lepidoptera showed preference for species containing neolignanas and/or lignanas such P. regnellii and P. solmsianum. The Chapters 2 and 3 (coleopterous - beetles and lepdopterous-butterflies, respectively), were addressed for the studies of biotransformation reactions, sequestration and attractive assays. The study of possible factors present attractions in essential oils was investigated by the use of electrophysiological essays. It were observed several reactions as esterification, hydrolyses and ozonolyse-type for lignans, neolignans, phenylpropanoids and amides occurring in from species of Piperaceae during the digestive process of the butterflies and of beetles. It was observed sequestration of neolignans of P. regnellii by larvae of Heraclides hectorides and prenylated of benzoic acids from roots of P. gaudichaudianum by the larva of the beetle Naupactus bipes. The experiments of EAG using antennas of the beetle N. bipes showed that the oils of P. gaudichaudianum, P. regnellii and P. hispidum were active and such results were in agreement with field observations and the most intense response was observed for the females in relation to the response for the males. The analyses of GC/EM-EAD allowed the determination of the bio-actives compounds as the monoterpenes α-pinene, β-pinene and β-mircene. The Chapter 4 describes the homopteros species that have specificity for the P. gaudichaudianum, P. arboreum and P. aduncum accumulative of prenylated benzoic acids. Further determinant factors for specificity included: low percentage of secondary metabolites in saps, high percentage of micro-nutrients and low percentage of macro-nutrients, absence of lignans or neolignans in the saps, lignifications with predominance of siringyl residues (S) which confer a pattern related to Angiosperms. The Chapter 5 describes the determination and structural characterization of the 49 secondary metabólitos involved in this study. The Chapter 6 shows the study of stability versus natural occurrence of tetrahydrofuran lignans was carried out due to the rare occurrence of all-cis configuration. The study of the stability for the ten possible stereoisomers for the tetrahydrofuran lignans was through the calculation of the functional of density, B3LYP, with the base 6-316(dp), it showed that the all-cis configuration is less stable than the all-trans tetrahydrofuran lignans.

Orientador: Anita Jocelyne Marsaioli / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-21T20:30:57Z (GMT). No. of bitstreams: 1 Pianaro_Adriana_D.pdf: 8531703 bytes, checksum: 86c8399c69a5d82f0e333fed92903d03 (MD5) Previous issue date: 2012 / Resumo: Este trabalho teve como objetivo a comunicação química em insetos sociais e em particular de abelhas sem ferrão e formigas lava-pés brasileiras. Portanto, estes dois tópicos foram abordados em dois capítulos distintos e revelaram que: (1) As rainhas virgens de M. quadrifasciata com abdomes inflados liberam uma mistura de nerol/geraniol na razão 1:2, respectivamente, pelas glândulas tergais. Estes monoterpenos indicam o nível de maturidade destas rainhas e podem estar envolvidos na seleção de uma nova rainha. Rainhas fisogástricas de S. aff. depilis atraem machos e a partir da descoberta deste fenômeno iniciou¿se a procura dos compostos responsáveis pela atração sexual. As análises dos extratos epicuticulares, extratos abdominais, glândulas de Dufour e glândulas tergais das rainhas fisogástricas e virgens de S. aff. depilis revelaram ésteres no extrato das rainhas fisogástricas. Os testes de campo com os extratos, as frações dos compostos isolados de rainhas fisogástricas de S. aff. depilis e padrões sintéticos revelaram que os extratos epicuticulares das rainhas fisogástricas atraem e estimulam a cópula dos machos quando adsorvidos sobre feltro preto. As frações e os padrões não produziram nenhuma resposta. A presença de 2-alcanóis nos extratos cefálicos de rainhas de M. quadrifasciata e S. aff. depilis e machos de N. testaceicornis estimulou a realização de testes de campo com 2-alcanóis, os quais demonstraram que o (S)-(+)-2-heptanol e a mistura (S)-(+)-2-heptanol/ (S)-(+)-2-nonanol (1:1) atraem operárias de P. droryana, N. testaceicornis e F. silvestrii, sugerindo ser um feromônio de recrutamento de operárias e envolvidos na formação de leques de machos. (2) A análise dos alcalóides dos venenos de operárias e rainhas de S. saevissima revelou diferenças na composição química intra e inter-nidal de duas regiões diferentes do Brasil (São Paulo e Rio de Janeiro). Uma metodologia para acesso rápido das configurações relativas e absolutas dos alcalóides piperidínicos do veneno das formigas lava-pés foi elaborada empregando CG-EM e CG-DIC com coluna capilar quiral Chrompack CP-chirasil-Dex CB e derivados trifluoroacetamidas e acetamidas dos alcalóides piperidínicos sintéticos e naturais, onde se verificou pela primeira vez a presença dos quatro estereoisômeros da 2-metil-6-undecilpiperidina em venenos de formigas lava-pés. / Abstract: The aim of this thesis is the chemical communication of social insects and in particular of Brazilian stingless bees and fire ants. Therefore, the results obtained in these two topics are presented in two chapters as follows: (1) Virgin queens of M. quadrifasciata (stingless bee) with inflated abdomens release a mixture of nerol/ geraniol in 1:2 ratio, respectively, by tergal glands. These monoterpenes are related to the development of these queens and may be involved in the new queen selection. S. aff. depilis physogastric queens attract drones and this observation oriented the research to sexual pheromones. The analysis of the epicuticular, abdominal, Dufour¿s gland, and tergal gland extracts of S.aff. depilis from physogastric and virgin queens revealed the outstanding presence of esters in the epicuticular extracts of physogastric queens. Field tests with extracts, isolated compound fractions of S. aff. depilis physogastric queens and synthetic standards revealed that physogastric queen epicuticular extracts attract and stimulate mating of drones when adsorbed on black felt. The fractions and standards produced no response. The presence of 2-alkanols in cephalic extracts of M. quadrifasciata and S. aff. depilis queens and N. testaceicornis drones stimulated field tests with 2-alkanols, which showed that the (S)-(+)-2-heptanol and the mixture of (S)-(+)-2-heptanol/ (S)-(+)-2-nonanol (1:1) attract workers of P. droryana, N. testaceicornis and F. silvestrii. This biological response is consistent with worker recruit pheromones, involved in the formation of drone swarms. (2) Analysis of the venom alkaloids of S. saevissima workers and queens revealed differences in the intra and internidal chemical composition of fire ants from two different regions in Brazil (Sao Paulo and Rio de Janeiro). A methodology to rapidly access the relative and absolute configurations of the piperidinic alkaloids from the fire ant venom was developed using GC-MS and GC-FID with a chiral capillary column (Chrompack CP-chirasil-Dex CB) and trifluoroacetamides and acetamides of synthetic and natural piperidinic alkaloids. This methodology was applied to the analysis of 2-methyl-6-undecylpiperidine of S. saevissima gynes and worker venoms from Ubatuba-SP and Pedro do Rio-RJ revealing for the first time the presence of four stereoisomers of 2-methyl-6-undecylpiperidine in the fire ant venoms. / Doutorado / Quimica Organica / Doutora em Ciências

Le palmier à huile africain Elaeis guineensis Jacqu. (Arecales : Arecaceae) est une plante tropicale d’intérêt économique et la première source mondiale d’huile végétale. La production d’huile dépend de la pollinisation par des charançons du genre Elaeidobius (Coleoptera : Curculionidae). Ces insectes sont impliqués dans une relation mutualiste spécialisée avec leur plante-hôte : ils se développent au dépend des inflorescences mâles qu’ils détectent par le biais des odeurs émises durant l’anthèse, tandis que les inflorescences femelles sont pollinisées en mimant les odeurs mâles. Une maladie mortelle sévissant en Amérique du sud et qui touche l’E. guineensis a contribué au développement de variétés commerciales d’hybrides interspécifiques, résultat du croisement entre E. guineensis et le palmier à huile sud-américain Elaeis oleifera (Knuth) Cortés. Mais ces hybrides s’avèrent naturellement mal pollinisées et nécessitent l’intervention d’une pollinisation assistée couteuse et contraignante. Cette thèse part de l’hypothèse que la communication chimique qui permet la rencontre entre les deux espèces de palmiers et leurs pollinisateurs respectifs est déficiente chez les hybrides.L’objectif principal de cette thèse a été d’étudier le fonctionnement du système de pollinisation des deux espèces de palmiers afin de comprendre pourquoi les hybrides sont mal pollinisés. L’étude a été conduite dans une plantation commerciale en Equateur, dans laquelle coexistent les deux espèces E. guineensis et E. oleifera avec leur insecte pollinisateur respectif, le charançon africain Elaeidobius kamerunicus Faust. et le charançon sud-américain Grasidius hybridus O’Brien & Beserra (Coleoptera : Curculionidae) et des palmiers hybrides interspécifiques. En utilisant des techniques de piégeages et l’échantillonnages des odeurs florales, nos résultats ont montré le système de pollinisation des deux espèces de palmier est basé sur l’émission d’un signal chimique spécifique et une duperie olfactive, renforcée par un phénomène de thermogénèse. Les deux insectes sont attirés en faible proportion sur les hybrides, qui possèdent une composition chimique intermédiaire à celle des espèces parentales. Des tests physiologiques et comportementaux ont permis d’identifier les composés organiques volatils clés responsables de l’attraction des insectes pollinisateurs.Ce travail devrait contribuer à apporter des connaissances théoriques sur le système de pollinisation du palmier à huile, et permettre d’élaborer des méthodes pratiques de gestion des pollinisateurs afin de réduire les coûts en pollinisation assistée. / The African oil palm Elaeis guineensis Jacqu. (Arecales: Arecaceae) is a tropical plant of economic interest and the world's leading source of vegetable oil. Oil production depends on pollination by weevils of the genus Elaeidobius (Coleoptera: Curculionidae). These insects are involved in a specialized mutualist relationship with the host plant: they develop at the expense of the male inflorescences they detect through the odors emitted during the anthesis, while the female inflorescences are pollinated by mimicking the male odors. E. guineensis is affected by a lethal disease in South America that leads to the development of commercial varieties of interspecific hybrids, resulting from artificial crossing between E. guineensis and the South American oil palm Elaeis oleifera (Knuth) Cortés. However, the natural pollination of these hybrids is inadequate and require the intervention of a costly assisted pollination. This thesis hypothesizes that the chemical communication underlying the successful encountering between each oil palm species and their respective pollinators is deficient in hybrid palms.The main objective of this work was to study the functioning of the pollination system for both oil palm species to understand why natural pollination in hybrids is inefficient. The study was conducted in a commercial plantation in Ecuador, including the two species E. guineensis and E. oleifera present with their respective pollinating insects, the African weevil Elaeidobius kamerunicus Faust. and the South American weevil Grasidius hybridus O'Brien & Beserra (Coleoptera: Curculionidae), as well as interspecific hybrid palms. Using techniques of trapping and sampling of floral odors, our results showed the pollination system of both species of palm is based on the emission of a specific chemical signal and an olfactory deception, reinforced by a phenomenon of thermogenesis. The two insects are attracted in a small proportion to hybrid palms, which have an intermediate chemical composition compare to parent species. Physiological and behavioral tests permit the identification of the key volatile organic compounds responsible for pollinator attraction.This work should contribute to theoretical knowledge about the oil palm pollination system and the development of practical methods for pollinator management to reduce costs in assisted pollination.

The ant Pachycondyla analis (formerly Megaponera foetens, commonly known as the Matabele ant) is a widespread ponerine in sub-Saharan Africa. It feeds solely on termites of economic importance belonging to the sub-family Macrotermitinae. These termites are captured during organised raids on their nests and galleries. Previous studies mostly concentrated on certain aspects of the raiding behaviour and trail laying pheromones in this species. Thus the detailed raiding behaviour and chemically-mediated communication between P. analis and its prey are virtually unknown. The aim of this study was to undertake detailed behavioural studies on termite raiding behaviour of P. analis, and to investigate whether P. analis uses olfactory cues for intra-specific communication during termite raids, and for detecting its prey. Termite raiding behaviour of P. analis was monitored at Mpala, a Kenyan savannah for six months (April to September, 2007). During this period, raids were found to occur mainly in the mornings and evenings, with late night raids occurring during dry periods. P. analis at Mpala mainly nests under rocks and in deserted termite mounds. Microtermes and Odontotermes were the main preyed termite genera, and ant raiding behaviour was synchronised with termite prey behaviour, and was influenced by foraging costs, prey defences and rewards. Olfactometric assays showed that P. analis workers used olfactory cues in their intra-specific chemical communication, with workers responding more to volatiles of individuals of the same size class (major to major and minor to minor) than between groups. Major workers discriminated more between the volatiles of the two groups than minor workers. GC-MS analysis of volatiles from major and minor workers revealed a cocktail of 48 compounds, majority of which were hydrocarbons. Volatile compounds were colony specific and quantitative analysis showed that major and minor workers alone released 2.5 fold more volatiles than the mixed stages. This suggests that ants have the innate ability to regulate the levels of the colony odour which they make up for with higher release levels when separated from each other. Using a Mandible Opening Response (MOR) bioassay, ants were able to distinguish between nestmates and non-nestmates based on cuticular hydrocarbon (CHC) profiles. This suggests that P. analis uses CHCs as short range contact recognition cues within the nest in traditional nest protection and during raids on termite species. GC-MS analyses revealed hydrocarbons of chain lengths in the range C8-C31 in the CHC profiles, comprising mainly alkanes, alkenes and methyl-branched alkanes. The CHCs were colony and individual worker specific. Nestmate recognition in P. analis may be encoded in the alkenes and methyl-branched alkanes. Dual choice olfactometric assays revealed that P. analis uses olfactory cues in locating potential termite sources with an average of 65% of workers choosing odours against the blank (clean air). When termite odours were offered to both major and minor workers, their choices were biased towards the termite odours, with minor workers attracted more to the odours than were major workers. Although ants responded to odours from the soil obtained for the termite gallery, overall, odours from termites inside their galleries were the most attractive to ants. These results suggest that the combined odours from both the termites and gallery components (in particular soil), serves as an effective nest location cue for the ants. Comparative GC-MS analyses showed that the composition of the volatiles from the gallery soil was richer than that released by the termites. Consistent with previous studies, the volatiles of the gallery soil were found to contain hydrocarbons, naphthalene and derivatives of this compound. In conclusion, these studies have revealed the rich diversity of chemical communication cues used by this ant species for nestmate recognition and for prey location during raids in search for its food source. / Thesis (PhD)--University of Pretoria, 2010. / Zoology and Entomology / unrestricted

Les interactions allélopathiques sont définies comme la libération par un organisme de composés chimiques inhibant les compétiteurs. Dans les milieux aquatiques, elles sont reconnues comme un facteur de la dynamique des communautés de phytoplancton. Malgré les conditions favorables créées par l'organisation des microorganismes en biofilms, l'allélopathie a été beaucoup moins étudiée dans les communautés de microorganismes phototrophes benthiques. L'objectif de cette thèse était d'étudier des interactions allélopathiques, au travers d'un cas particulier, entre les microorganismes dans les biofilms phototrophes d'eau douce. Ainsi le travail s'est focalisé sur les effets allélopathiques d'une algue verte filamenteuse, Uronema confervicolum Lagerheim, sur des diatomées benthiques. L'hypothèse principale est que, dans certaines conditions, U. confervicolum libère suffisamment de composés allélopathiques pour inhiber significativement le développement des diatomées épiphytes. Cette hypothèse soulève trois questions (i) quels sont les composés allélopathiques, (ii) leur production dépend-elle des conditions environnementales et (iii) quel est l'effet des composés allélopathiques produits sur les diatomées ? Cette interaction a été étudiée au travers de multiples approches. L'activité d'extraits de biomasse algale et de filtrat de culture a été testée sur la croissance, la photosynthèse, l'adhésion et la motilité des diatomées. Les techniques de fractionnement guidé par bioessais et de profilage métabolomique ont été utilisées pour identifier les composés allélopathiques produits. Les effets des facteurs environnementaux (limitations en nutriment, apports de lumière et de gaz) et de la phase de croissance sur la production et la libération de différents composés allélopathiques ont également été étudiés. Une étude approfondie de l'effet sur les diatomées des composés allélopathiques anti-adhésion a été réalisée en combinant le profilage métabolomique, la transcriptomique et la microscopie électronique à balayage. Les extraits d'U. confervicolum ont montré un effet négatif sur la croissance, la photosynthèse, l'adhésion et la motilité des diatomées. Deux acides gras polyinsaturés (acides linoléique et a-linolénique) ont été identifiés comme des composés allélopathiques inhibant la croissance et la photosynthèse des diatomées. L'inhibition de l'adhésion des diatomées est, par contre, causée par d'autres composés allélopathiques non identifiés. L'augmentation de l'intensité lumineuse a accru la production de composés allélopathiques, à l'instar des composés défensifs des plantes. Cependant, aucune des théories classiques sur les défenses chimiques des plantes n'a permis d'expliquer l'ensemble des résultats obtenus sur les questions de régulation. En plus de l'inhibition de l'adhésion, le filtrat d'U. confervicolum inhibe la formation de la matrice constituée de substances polymériques extracellulaires, impacte le métabolisme énergétique et induit une modification globale du transcriptome et du métabolome des diatomées. Une partie des gènes et métabolites les plus fortement impliqués dans la réponse des diatomées à l'inhibition de l'adhésion ont été identifiés. Ce travail a permis de mieux évaluer l'activité allélopathique de l'algue verte filamenteuse U. confervicolum et son impact potentiel sur la structure et le fonctionnement des biofilms phototrophes. D'autres approches seront nécessaires, à l'avenir, pour mieux comprendre le rôle global de l'allélopathie dans des biofilms phototrophes complexes. / Allelopathic interactions are defined as the release, by an organism, of chemicals that inhibit competitors. In aquatic environments, they are recognised as a driver of phytoplankton community dynamics. Despite the favourable conditions created by the organisation of microorganisms in biofilms, allelopathy has been much less studied in benthic phototrophic microorganism communities. The objective of this thesis was to increase our comprehension of allelopathic interactions amongst phototrophic microorganisms in biofilms by analysing several aspects of a specific allelopathic interaction. The model chosen was the allelopathic effect of Uronema confervicolum Lagerheim, a filamentous green alga isolated from river biofilms, on diatoms from the same environment. The main hypothesis was that, at least in specific environmental conditions, U. confervicolum produce and release sufficiently large amounts of allelopathic compounds to inhibit significantly the development of epiphytic diatoms. Three questions arised from this hypothesis: (i) what are the allelopathic compounds implied, (ii) do their production and release depend on environmental factors and (iii) what is the effect of these compounds on epiphytic diatoms? This interaction has been studied using multiple approaches. Activity of algal biomass extracts and culture filtrates have been tested on diatom growth, photosynthesis, adhesion and motility. Bioassay-guided fractionation and metabolomic profiling have been used to identify the allelopathic compounds produced. The effect of environmental factors (nutrient limitation, light and gas supply) and growth phase on the production and release of the different allelopathic compounds have also been studied. In-depth study of the effect of anti-adhesion allelochemicals on diatom by combining metabolomic profiling, transcriptomics and scanning electron microscopy have been carried out. The extracts of U. confervicolum exhibited negative effects on diatoms growth, photosynthesis, adhesion and motility in bioassays. Two polyunsaturated fatty acids (linoleic and a-linolenic acids) have been identified as allelopathic compounds affecting growth and photosynthesis but not adhesion ability of diatoms. The inhibition of diatom adhesion was caused by other allelopathic compounds, unidentified so far. Increasing light intensity enhanced the production of U. confervicolum allelopathic compounds as predicted by plant defence theories, but the regulation of allelopathic activity was not completely explainable by any classic plant defence theory. Finally, beside an inhibition of diatom adhesion U. confervicolum filtrate was also found to inhibit the formation of diatom extracellular polymeric substances matrix, affect energy metabolism and induce global modification of diatom transcriptome and metabolome. Some of the genes and metabolites implied in the response of diatoms to adhesion inhibition have been identified. These results indicate the strong potential of the filamentous green alga U. confervicolum to impact biofilm community by its allelopathic activity. Results on the allelopathic activity of this alga and the effect on diatoms in laboratory experiments provide the basis for further experiments to access the role of allelopathy in complex biofilms. These findings will encourage researchers working on biofilms to consider allelopathy when phototrophic biofilm ecology is studied.

Orientador: Anita Jocelyne Marsaioli / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-22T10:44:31Z (GMT). No. of bitstreams: 1 Fonseca_FrancineSouzaAlvesda_D.pdf: 8642062 bytes, checksum: fa3ebef40c6304d2e6d2b02ba701f86e (MD5) Previous issue date: 2013 / Resumo: Capítulo I. Este trabalho revelou aspectos inéditos da ecologia química de Phloeophana longirostris e Phloea subquadrata, duas espécies de percevejo da família Phloeidae. Esta família, representada por apenas três gêneros e quatro espécies, é caracterizada por apresentar cuidado maternal com ovos e ninfas e camuflagem sobre o tronco das árvores hospedeiras. O objetivo deste trabalho foi localizar, identificar e caracterizar os principais compostos voláteis emitidos por percevejos e verificar a relação destes com as árvores hospedeiras. Os resultados revelaram que os insetos, após estímulo físico, liberam aldeídos e hidrocarbonetos insaturados, oxoaldeídos e ésteres como compostos majoritários. Além disso, foi possível discriminar compostos característicos de cada grupo estudado. Capítulo II. Na segunda parte deste trabalho, foi explorada a atividade enzimática do micro-organismo Bacillus safensis isolado do petróleo brasileiro. Esse micro-organismo apresenta um arsenal enzimático capaz de degradar compostos orgânicos aromáticos. O objetivo deste trabalho foi purificar, isolar e identificar oxirredutases presentes na cepa (CFA06) de Bacillus safensis. Através dessa bactéria Gram-positiva, altamente resistente à radiação ultravioleta, foi possível identificar duas oxirredutases, sendo uma delas inédita. A oxirredutase inédita, aqui denominada de BsP315, apresenta 21 kDa e ferro e molibdênio ligados a essa proteína. Além dessa oxirredutase, uma catalase, aqui denominada de BsCat, também foi identificada / Abstract: Chapter I. This study revealed novel aspects of the chemical ecology of Phloeophana longirostris and Phloea subquadrata from Phloeidae family. This family, has only three genders and four species with characteristic maternal care of eggs and nymphs and camouflage on the trunk of the host tree. The aim of this study was to the identification of major volatile compounds produced by the stink bugs and their relationship with the host trees. The results revealed that after physical stimulation, the insects release aldehydes and unsaturated hydrocarbons, esters and oxoaldehyde as major compounds. These volatile compounds were important in the discrimination of male, female and nymphs of each species. Chapter II. In the second part of this research, we investigated the enzymatic activity of the micro-organism Bacillus safensis isolated from Brazilian oil. This micro-organism has an enzymatic arsenal capable of degrading aromatic organic compounds. The aim of this study was to purify, isolate and identify oxidoreductases present in strain (CFA06) Bacillus safensis. Through this Gram-positive bacterium, highly resistant to ultraviolet radiation, it was possible to identify two oxidoreductases. The novel oxidoreductase, here named BsP315 has 21 kDa and iron and molybdenum bound to its catalytic site, additionally we have isolated a catalase here named BsCat / Doutorado / Quimica Organica / Doutora em Ciências

Orientador: José Roberto Trigo / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-26T19:23:05Z (GMT). No. of bitstreams: 1 Massuda_KamilaFerreira_D.pdf: 3476211 bytes, checksum: a20550d22526cbcb5a72c290cf3e8ff1 (MD5) Previous issue date: 2015 / Resumo: As interações entre os insetos herbívoros e as plantas foram moldadas ao longo de um extenso processo coevolutivo, que gerou adaptações das plantas para evitarem a herbivoria e contra-adaptações dos herbívoros para superarem as defesas impostas pelas plantas. As folhas de Ipomoea carnea subsp. fistulosa (Convolvulaceae) são atacadas pelos besouros Cassidinae (Chrysomelidae), Chelymorpha reimoseri, Chelymorpha cribraria e Charidotella rubicunda. Esses besouros depositam seus ovos nas folhas e se alimentam das mesmas nos estágios de larva e adulto. Nesse trabalho, demonstramos a eficácia de dois mecanismos de defesa de I. carnea contra a deposição de ovos por C. reimoseri: (1) o descolamento de posturas, devido a formação de neoplasmas abaixo da postura e (2) a atração de parasitoides de ovos, por voláteis liberados de plantas induzidas pela própria postura ou por um indivíduo realmente capaz de ovipor, uma fêmea que copulou. Além disso, demonstramos duas estratégias de defesa para as três espécies cassidíneas supramencionadas contra predadores: (1) a impalatabilidade, devido ao alcaloide swainsonina, sequestrado da planta hospedeira e cuja atividade deterrente foi demonstrada para Gallus gallus e (2) a camuflagem química das larvas de C. reimoseri nas folhas da planta hospedeira, devido à alta similaridade química entre seus compostos cuticulares e os da planta hospedeira larval, que dificulta o reconhecimento por predadores quimicamente orientados / Abstract: The interactions between herbivorous insects and plants have been shaped over a co-evolutionary process that spawned adaptations of plants to avoid herbivory and counter adaptation of herbivores to overcome the defenses imposed by plants. The plant Ipomoea carnea subsp. fistulosa (Convolvulaceae) has as herbivores, beetles of Cassidinae subfamily (Chelymorpha reimoseri, Chelymorpha cribraria, and Charidotella rubicunda), which use the leaves to feed larvae and adults, as well as places for depositing their eggs. This study demonstrated the efficiency of two I. carnea defensive mechanisms against oviposition by C. reimoseri: (1) the detachment of eggs neoplasm formation, below the eggs, and (2) the attraction of eggs parasitoids by volatile released from posture-induced plants and plants induced by an individual capable of laying eggs, a mated female. Furthermore, we demonstrated the effectiveness of two defensive strategies in the three above mentioned cassidine species; (1) the unpalatability, due to swainsonine, an akaloid provided by the host plant, and its deterrent effect against predator, the chicken Gallus gallus; (2) the chemical camouflage of C. reimoseri larvae on its host plant, due to high chemical similarity among their cuticular compounds that turns difficult recognition by a chemical oriented predator / Doutorado / Doutor em Ecologia