Master thesis investigates reproductive barriers in diploid-polyploid complex of Vicia cracca. Complex with basic chromosome number x=7 consists of diploid (2x=14), tetraploid (4x=28) and rare triploid (3x=21) cytotype. I studied prereproducitve barriers between diploid and tetraploid cytotype: phenology of flowering, pollinators' behavior (preference of species of pollinators to cytotypes, sequence of visited cytotypes), variables, that could explain pollinators' behavior (amount of nectar as the main reward, size and amount of pollen grains as a potentional reward). To find out how strong the triploid block is I analyzed ploidy of seeds and seedlings from mixed-ploidy population. The habitat isolation showed up to be the strongest reproductive barrier. Pollinator's behavior meaningfully contributes to isolation, phenology of flowering contributes only minimally. Index expressing rate of prereproctive barriers is 0,956. Pollinator Bombus pascuorum visited on one locality preferably tetraploid plants and Andrena sp. preferred diploid plants. Even though tetraploid plants produce more nectar, no other analysis showed pollinators' preference to tetraploid plants. I prepared one squash of diploid V. cracca using method of in situ hybridization. This method needs to be optimilized for studied taxon.

Master thesis investigates reproductive barriers in diploid-polyploid complex of Vicia cracca. Complex with basic chromosome number x=7 consists of diploid (2x=14), tetraploid (4x=28) and rare triploid (3x=21) cytotype. I studied prezygotic barriers between diploid and tetraploid cytotype: phenology of flowering, pollinators' behavior (preference of spieces of pollinators to cytotypes, sequence of visited cytotypes), variables, that could explain pollinators' behavior (amount of nectar as the main reward, size and amount of pollen grains as a potentional reward). To find out how strong the triploid block is, I analyzed ploidy of seeds and seedlings from mixed-ploidy population. The habitat isolation showed up to be the strongest reproductive barrier. Pollinator's behavior meaningfully contributes to isolation, phenology of flowering contributes only minimally. Index expressing rate of prereproductive barriers is 0,990. Pollinator Bombus pascuorum visited on one locality preferably tetraploid plants and Andrena sp. preferred diploid plants. Even though tetraploid plants produce more nectar, no other analysis showed pollinators' preference to tetraploid plants.

A perda de polinizadores no mundo todo, especialmente abelhas, tem sido atribuída, entre outros fatores, à intensificação do uso de inseticidas em agro ecossistemas. A nova classe química de inseticidas, os neonicotinóides, tem sido o principal alvo de investigações, sobretudo em decorrência de sua ação sistêmica que transloca a substância ativa pela planta e atinge o pólen e néctar como resíduos. Dessa forma, o presente estudo teve como objetivo verificar os efeitos do neonicotinóide tiametoxam em abelhas sociais e avaliar a possibilidade da espécie Scaptotrigona aff. depilis ser utilizada como bioindicadora de contaminação ambiental por inseticidas. Para tanto, (i) operárias forrageiras de Bombus terrestris audax, Apis mellifera e Scaptotrigona bipunctata foram submetidas à exposição contínua de alimento contaminado; (ii) operárias de Scaptotrigona aff. depilis foram expostas ao inseticida, durante o estágio larval, através do alimento e; (iii) operárias forrageiras de Scaptotrigona aff. depilis foram coletadas no meliponário da Universidade de São Paulo e arredores, e submetidas a análises químicas, para a checagem de contaminação pelos neonicotinóides tiametoxam e/ ou imidacloprido. (i) Dentre as três espécies de abelhas sociais adultas, B. terrestris audax mostrou ser a mais sensível à ação de tiametoxam, seguida de S. bipunctata e A. mellifera, verificados através do decréscimo no consumo de alimento e da longevidade. Nesse sentido, os efeitos subletais não foram influenciados pelo tamanho do corpo dos indivíduos. (ii) As operárias de Scaptotrigona aff. depilis, quando expostas a doses do inseticida em fase larval, apresentaram desenvolvimento tardio e originaram indivíduos menores e assimétricos. Tanto em abelhas adultas como em imaturas, os efeitos subletais foram evidenciados nos indivíduos expostos às maiores doses de tiametoxam nos experimentos (39.37 e 98.43µg de substância ativa/kg de xarope em adultos, ou 0.044 e 4.375ng de substância ativa/larva). (iii) A detecção de tiametoxam e imidacloprido em operárias forrageiras de Scaptotrigona aff. depilis permitiu considerar essa espécie como bioindicadora de contaminação. Nesse estudo, a mortalidade não foi parâmetro conclusivo quanto aos potenciais efeitos adversos de tiametoxam para as abelhas. O potencial de Scaptotrigona aff. depilis como espécie bioindicadora é uma importante etapa para avaliar a contaminação ambiental por neonicotinóides em abelhas sem ferrão, podendo ser utilizada em políticas públicas. / Pollinator losses all over the world, especially bees disappearing, have been attributed, among others, to the intensification of insecticide use on crops. The new chemical class of insecticides, the neonicotinoids, has been the main target of investigation, mainly due to its systemic action, which translocates the active substance for the plant and to reach the pollen and nectar as residues. Thus, this study evaluated the effects of thiamethoxam neonicotinoid on bees and the possibility of the use of Scaptotrigona aff. depilis species as bioindicator of environmental contamination by insecticides. For this, (i) Bombus terrestris audax, Apis mellifera and Scaptotrigona bipunctata forager workers were induced to continue exposure of contaminated food; (ii) Scaptotrigona aff. depilis workers were exposed to the insecticide, during its larval phase, through the larval food and; (iii) Scaptotrigona aff. depilis forager workers were collected from University of São Paulo meliponary and surroundings and analyzed, in order to check for contamination by thiamethoxam and/ or imidacloprid neonicotinoids. (i) Among the three social bee species, B. terrestris audax showed to be more sensible to the thiamethoxam action, followed by S. bipunctata and A. mellifera, verified through decreasing food consumption and longevity. Thus, the sub lethal effects were not influenced by individual body size. (ii) Scaptotrigona aff. depilis workers, when exposed to insecticide doses on larval food, caused individuals with developmental delay and were smaller and asymmetric. Both adult bees and in immature, the sub lethal effects of thiamethoxam were evidenced on individuals exposed to the highest doses in the experiments. (iii) The detection of thiamethoxam and imidacloprid in Scaptotrigona aff. depilis forager workers allowed to consider this species as bioindicator of environmental contamination. In this study, the mortality was not a conclusive parameter as to the potential adverse effects of thiamethoxam for the bees. The potential of Scaptotrigona aff. depilis species as bioindicator is an important step to assess environmental contamination by neonicotinoids on stingless bees, may be used for public policy.

A abelha Bombus terrestris é um eficiente polinizador, prestando importantes serviços ecossistêmicos na Europa e adjacências, onde é nativa. Suas colônias têm sido criadas em larga escala para polinização agrícola, as quais são comercializadas internacionalmente, inclusive em países fora de sua área de ocorrência nativa. Deliberada ou acidentalmente a espécie tem sido introduzida em ambientes alóctones, em muitos casos tornando-se invasora. Quando invasora, a espécie é um potencial vetor de doenças e um competidor com outras abelhas; vários impactos têm sido relatados em áreas invadidas ao redor do mundo. Na América do Sul, a espécie foi inicialmente introduzida em ambientes naturais no Chile, mas rapidamente a invasão se espalhou; atualmente é encontrada ocupando ambientes naturais na Argentina. A elevada capacidade invasiva da espécie e a alta velocidade de sua expansão, conforme tem sido relatada por pesquisadores, levantou a possibilidade de a espécie alcançar o Brasil por meio de corredores ambientais favoráveis que se conectam com áreas já invadidas, gerando preocupações sobre potenciais impactos aos sistemas naturais e agrícolas. Esta pesquisa empregou uma abordagem interdisciplinar, integrando uma variedade de métodos analíticos oriundos de diferentes áreas da ecologia e os mais avançados recursos de sistemas de informações geográficas para detectar globalmente as áreas susceptíveis à invasão por Bombus terrestris, considerando-se também as mudanças climáticas. Para o sul da América do Sul foram identificados os corredores de invasão que poderão permitir a espécie se espalhar e alcançar Brasil a partir de locais invadidos. Para o Brasil, foram identificados os municípios mais vulneráveis à entrada da espécie, e também aqueles que estão na rota de expansão da invasão Brasil adentro. Para os municípios brasileiros, por sua vez, foram verificadas as culturas agrícolas e as espécies de Bombus nativas que a invasora poderá interagir ao longo da rota de invasão potencial; com estas informações foram apontadas áreas prioritárias, subsidiando o planejamento de monitoramento e ações de controle do processo de invasão, mas também medidas preventivas e mitigadoras de impactos ambientais e econômicos após a invasão, caso venha a ocorrer. / The bee Bombus terrestris is an efficient pollinator, providing important ecosystem services in Europe and surrounding areas, where it is a native species. Their colonies have been reared in large-scale for agricultural pollination, which are internationally traded, including for countries outside its native range. The species has been deliberately or accidentally introduced into non-native environments, becoming invasive in many cases. When invasive, the species is a potential vector of diseases and competes with other bees for resources; many impacts have been reported in invaded countries around the world. In South America, the species was introduced in natural environments of Chile at first, but the invasion was quickly spread; nowadays it is found living in natural environments of Argentina. The high invasiveness of the species and the high speed of its invasive expansion, as has been reported by researchers, raised the possibility of the species reach Brazil through suitable environmental corridors that are connected with areas already invaded, raising the concerns about potential impacts to natural and agricultural systems. An interdisciplinary approach was used in this research, composing a variety of analytical methods from different areas of ecology and applying the most advanced resources of geographic information systems to detect areas susceptible to invasion by Bombus terrestris at global scale, considering also climate change. For the southern South America, the corridors of invasion that could allow the spreading of the species and that potentially reach Brazil were identified. For Brazil, the most susceptible municipalities at the entrance of the species have been identified, and also those that are on the route of expansion inside the country. For Brazilian municipalities, crops and native species of Bombus that the invasive species can interact with along the potential invasion route were verified; with this information, priority areas were identified, supporting the planning of monitoring and control actions of the invasion process, but also preventive and mitigating measures of environmental and economic impacts after the invasion, if it eventually occurs.

A perda de polinizadores no mundo todo, especialmente abelhas, tem sido atribuída, entre outros fatores, à intensificação do uso de inseticidas em agro ecossistemas. A nova classe química de inseticidas, os neonicotinóides, tem sido o principal alvo de investigações, sobretudo em decorrência de sua ação sistêmica que transloca a substância ativa pela planta e atinge o pólen e néctar como resíduos. Dessa forma, o presente estudo teve como objetivo verificar os efeitos do neonicotinóide tiametoxam em abelhas sociais e avaliar a possibilidade da espécie Scaptotrigona aff. depilis ser utilizada como bioindicadora de contaminação ambiental por inseticidas. Para tanto, (i) operárias forrageiras de Bombus terrestris audax, Apis mellifera e Scaptotrigona bipunctata foram submetidas à exposição contínua de alimento contaminado; (ii) operárias de Scaptotrigona aff. depilis foram expostas ao inseticida, durante o estágio larval, através do alimento e; (iii) operárias forrageiras de Scaptotrigona aff. depilis foram coletadas no meliponário da Universidade de São Paulo e arredores, e submetidas a análises químicas, para a checagem de contaminação pelos neonicotinóides tiametoxam e/ ou imidacloprido. (i) Dentre as três espécies de abelhas sociais adultas, B. terrestris audax mostrou ser a mais sensível à ação de tiametoxam, seguida de S. bipunctata e A. mellifera, verificados através do decréscimo no consumo de alimento e da longevidade. Nesse sentido, os efeitos subletais não foram influenciados pelo tamanho do corpo dos indivíduos. (ii) As operárias de Scaptotrigona aff. depilis, quando expostas a doses do inseticida em fase larval, apresentaram desenvolvimento tardio e originaram indivíduos menores e assimétricos. Tanto em abelhas adultas como em imaturas, os efeitos subletais foram evidenciados nos indivíduos expostos às maiores doses de tiametoxam nos experimentos (39.37 e 98.43µg de substância ativa/kg de xarope em adultos, ou 0.044 e 4.375ng de substância ativa/larva). (iii) A detecção de tiametoxam e imidacloprido em operárias forrageiras de Scaptotrigona aff. depilis permitiu considerar essa espécie como bioindicadora de contaminação. Nesse estudo, a mortalidade não foi parâmetro conclusivo quanto aos potenciais efeitos adversos de tiametoxam para as abelhas. O potencial de Scaptotrigona aff. depilis como espécie bioindicadora é uma importante etapa para avaliar a contaminação ambiental por neonicotinóides em abelhas sem ferrão, podendo ser utilizada em políticas públicas. / Pollinator losses all over the world, especially bees disappearing, have been attributed, among others, to the intensification of insecticide use on crops. The new chemical class of insecticides, the neonicotinoids, has been the main target of investigation, mainly due to its systemic action, which translocates the active substance for the plant and to reach the pollen and nectar as residues. Thus, this study evaluated the effects of thiamethoxam neonicotinoid on bees and the possibility of the use of Scaptotrigona aff. depilis species as bioindicator of environmental contamination by insecticides. For this, (i) Bombus terrestris audax, Apis mellifera and Scaptotrigona bipunctata forager workers were induced to continue exposure of contaminated food; (ii) Scaptotrigona aff. depilis workers were exposed to the insecticide, during its larval phase, through the larval food and; (iii) Scaptotrigona aff. depilis forager workers were collected from University of São Paulo meliponary and surroundings and analyzed, in order to check for contamination by thiamethoxam and/ or imidacloprid neonicotinoids. (i) Among the three social bee species, B. terrestris audax showed to be more sensible to the thiamethoxam action, followed by S. bipunctata and A. mellifera, verified through decreasing food consumption and longevity. Thus, the sub lethal effects were not influenced by individual body size. (ii) Scaptotrigona aff. depilis workers, when exposed to insecticide doses on larval food, caused individuals with developmental delay and were smaller and asymmetric. Both adult bees and in immature, the sub lethal effects of thiamethoxam were evidenced on individuals exposed to the highest doses in the experiments. (iii) The detection of thiamethoxam and imidacloprid in Scaptotrigona aff. depilis forager workers allowed to consider this species as bioindicator of environmental contamination. In this study, the mortality was not a conclusive parameter as to the potential adverse effects of thiamethoxam for the bees. The potential of Scaptotrigona aff. depilis species as bioindicator is an important step to assess environmental contamination by neonicotinoids on stingless bees, may be used for public policy.

<p>Natural selection mediated by pollinators has influenced the evolution of floral diversity of the flowering plants (angiosperms). The scope of this thesis was to study: 1) phenotypic selection, 2) mating systems, and 3) floral shifts involved in plant speciation. Model plant species were <i>Platanthera bifolia</i> and <i>P. chlorantha</i> (Orchidaceae). These orchids are moth-pollinated, strictly co-sexual (bisexual flowers), and produce a spike that displays 10-20 white flowers.</p><p>I explored the influence of characters on plant fitness by using multiple linear regressions. Pollen removal (male fitness) and fruit set (female fitness) increased with more flowers per plant in three <i>P. bifolia</i> populations. There was selection towards longer spurs in a dry year when average spur length was shorter than in normal-wet years. Female function was sensitive to drought, which enabled an application of the male function hypothesis of floral evolution (Bateman's principle). The results show that selection may vary between populations, years, and sex-functions.</p><p>I examined inbreeding by estimating levels of geitonogamy (self-pollination between flowers of an individual) with an emasculation method in two <i>P. bifolia</i> populations. Geitonogamy did not vary with inflorescence size. Levels of geitonogamy was 20-40% in the smaller, but non-significant in the larger population. This may relate to lower number of possible mates and pollinator activity in the smaller population.</p><p><i>Platanthera bifolia</i> exhibits the ancestral character state of tongue-attachment of pollinia on the pollinator. Its close relative <i>P. chlorantha</i> attaches its pollinia onto the pollinator's eyes. To explore the mechanism of a floral shift, pollination efficiency and speed was compared between the two species. The results showed no differences in pollination efficiency, but <i>P. chlorantha</i> had faster pollen export and import. Efficiency of pollination in terms of speed may cause floral shifts, and thus speciation.</p>

Natural selection mediated by pollinators has influenced the evolution of floral diversity of the flowering plants (angiosperms). The scope of this thesis was to study: 1) phenotypic selection, 2) mating systems, and 3) floral shifts involved in plant speciation. Model plant species were Platanthera bifolia and P. chlorantha (Orchidaceae). These orchids are moth-pollinated, strictly co-sexual (bisexual flowers), and produce a spike that displays 10-20 white flowers. I explored the influence of characters on plant fitness by using multiple linear regressions. Pollen removal (male fitness) and fruit set (female fitness) increased with more flowers per plant in three P. bifolia populations. There was selection towards longer spurs in a dry year when average spur length was shorter than in normal-wet years. Female function was sensitive to drought, which enabled an application of the male function hypothesis of floral evolution (Bateman's principle). The results show that selection may vary between populations, years, and sex-functions. I examined inbreeding by estimating levels of geitonogamy (self-pollination between flowers of an individual) with an emasculation method in two P. bifolia populations. Geitonogamy did not vary with inflorescence size. Levels of geitonogamy was 20-40% in the smaller, but non-significant in the larger population. This may relate to lower number of possible mates and pollinator activity in the smaller population. Platanthera bifolia exhibits the ancestral character state of tongue-attachment of pollinia on the pollinator. Its close relative P. chlorantha attaches its pollinia onto the pollinator's eyes. To explore the mechanism of a floral shift, pollination efficiency and speed was compared between the two species. The results showed no differences in pollination efficiency, but P. chlorantha had faster pollen export and import. Efficiency of pollination in terms of speed may cause floral shifts, and thus speciation.

Bees provide vital ecosystem services for cropping systems as well as natural landscapes. Declines in both both native bee and managed honey bee (Apis mellifera L.) populations has brought attention to the significance of their role as pollinators in managed and native ecosystems. As a result, conservation efforts have been undertaken to preserve them. While considerable attention has been given to honey bees, relatively little is known about many native bee species. Of particular interest is the family Halictidae, which can comprise the majority of observed individuals in many habitats. These, often small, bees are difficult to identify, and, as a result, relatively little is known about their preferred floral hosts. Because bee species assemblages vary significantly from region to region, it is important to have an understanding of local populations and their floral hosts. It is also important to understand factors which affect the quantity and quality of floral resources, such as anthropogenic disturbance. The Pacific Northwest is a diverse landscape, with rich agricultural and wildland environments that require pollinators in order to continue to thrive. Two studies examine the interface between these two systems, the first explores how roadside disturbance, which is prevalent across the world, impacts native pollinators across habitat types, and the second explores the diet of common native species, and how that diet changes across habitat types. In these studies, it was found that roadside disturbance was associated with reduced native bee diversity and abundance in the seasonally wet Willamette Valley of Oregon, but not in the more xeric Central Oregon. Bee abundance was positively correlated with temperature. Bee diet was more diverse in areas of scarce floral resources. In both regions, exotic plants were important floral hosts, representing nearly half of observed floral visitations. This thesis presents results of species analysis, floral richness and density correlations, and comparisons of floral resources used by different bee species. Implications and recommendations for land management are discussed. / Graduation date: 2013

The objective of this study was to characterize the wild bee fauna of a managed mixed-grass prairie in southwestern Manitoba. Weekly sampling using two methods, sweep-netting and bee bowls, was conducted over two years (2005-2006) at three sites within the Yellow Quill Mixed-grass Prairie Preserve. Spatial and seasonal patterns in diversity indices, taxonomic composition and ecological composition of the bee fauna were identified and investigated in relation to sampling method, environmental conditions, and floral resource availability. A total of 7014 individual bees representing five families and 100 species were collected. Numerically, social nesters from the genera Lasioglossum and Bombus dominated, while mining species of Andrena represented the greatest species richness. Observed spatial and seasonal patterns in the abundance, diversity and composition of the bee community were strongly modified by sampling method, resource limitation in the second year and the presence of the exotic invasive plant leafy spurge (Euphorbia esula). / October 2007

The objective of this study was to characterize the wild bee fauna of a managed mixed-grass prairie in southwestern Manitoba. Weekly sampling using two methods, sweep-netting and bee bowls, was conducted over two years (2005-2006) at three sites within the Yellow Quill Mixed-grass Prairie Preserve. Spatial and seasonal patterns in diversity indices, taxonomic composition and ecological composition of the bee fauna were identified and investigated in relation to sampling method, environmental conditions, and floral resource availability. A total of 7014 individual bees representing five families and 100 species were collected. Numerically, social nesters from the genera Lasioglossum and Bombus dominated, while mining species of Andrena represented the greatest species richness. Observed spatial and seasonal patterns in the abundance, diversity and composition of the bee community were strongly modified by sampling method, resource limitation in the second year and the presence of the exotic invasive plant leafy spurge (Euphorbia esula).