FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / The research was carried out between July and December 2009 using tropical soya bean (Glycine max (L.) Merril) cultivars irrigated by center pivots in the irrigation district of Jaguaribe-Apodi, belonging to the agribusiness Faedo Sementes situated at the Apodi plateau, Limoeiro do Norte, CearÃ, Brazil. The study aimed to investigate the use of Apis mellifera L. for pollination and productivity increment in the soya bean cultivars Monsoy 9144RR, BRS-CarnaÃba, BRS-SambaÃba, adapted for the tropical conditions of N and NE Brazil. The following aspects were investigated: floral biology, pollination requirements, pollination efficiency of A. mellifera ; behavior and pattern of foraging; use of A. mellifera to augment productivity per Ãrea, per plant, in oil and protein content; and potential for honey production in soya bean crops and risks of pesticide contamination of the honey. Results showed that the cultivars are attractive to honey bees and they concentrate their foraging for nectar collection in the morning shift. The varieties were capable of autopollination, however there were significant (p<0.05) increments of productivity when flowers were visited by biotic agents. A single honey bee visit raised productivity to equivalent (p>0.05) levels of those flowers open for pollination. Regarding absolute production, it differed significantly (p<0.05) from the areas where honey bees were introduced in the center or along the sideline of the plantation to that where flowers did not receive flower visits. The mean number of pods per plant was 49.64  2.64, 57.16  2.41 and 59.60  2.71; the mean number of seeds per plant was 104.68  5.13, 121.46  3.98 and 127.42  5.95; and the percentage of pods bearing three seeds was 18.65%, 20.43% and 23.72% for the caged, open and open + honey bees treatments, respectively. Oil and protein contents of seeds did not differ (p>0.05) among treatments. All colonies produced honey and by the end of the blooming period (30 days) total production reached 81.7 kg, with a mean production of 10.1  0.86 kg per hive and no pesticide residue was found. It was concluded that the soya bean cultivars adapted to N and NE Brazil can be used for honey production and the foraging behavior of A. mellifera visiting flowers only in the morning allows to recommend spraying the crop only in the late afternoon and early evening when bees are no longer in the crop to produce residue-free honey and prevent poisioning risks to the bees; also, introducing A. mellifera for pollination purpose can increase production over 25% especially due to augment in the percentage of three-seeded pods. / A pesquisa foi conduzida entre julho e dezembro de 2009 usando cultivares tropicais de soja (Glycine max (L.) Merril) irrigadas por pivà central no distrito de irrigaÃÃo Jaguaribe-Apodi, sendo o plantio pertencente à agroempresa Faedo Sementes situada na Chapada do Apodi, Limoeiro do Norte, CearÃ. O estudo teve por objetivo investigar o uso de Apis mellifera L. para polinizaÃÃo e incrementos de produtividade nas cultivares de soja Monsoy 9144RR, BRS-CarnaÃba, BRS-SambaÃba, adaptadas Ãs condiÃÃes tropicais do Norte e Nordeste brasileiro. Os seguintes aspectos foram investigados: biologia floral, requerimentos de polinizaÃÃo, eficiÃncia de polinizaÃÃo de A. mellifera; comportamento e padrÃo de forrageamento; uso de A. mellifera para elevar a produtividade por Ãrea, por planta, em conteÃdo de Ãleo e proteÃna dos grÃos; potencial para a produÃÃo de mel e os riscos de contaminaÃÃo deste mel. Os resultados mostraram que as cultivares estudadas sÃo atrativas para as abelhas melÃferas e elas concentram seu forrageio na coleta de nÃctar no turno da manhÃ. As variedades sÃo capazes de se autopolinizarem, contudo houve incrementos de produtividade significativos (p<0,05) quando as flores foram visitadas por agentes biÃticos. Uma Ãnica visita da abelha melÃfera elevou a produtividade para nÃveis equivalentes (p>0.05) ao das flores abertas para polinizaÃÃo. No que se refere à produÃÃo total, observou-se diferenÃas significativas (p<0,05) das Ãreas onde as abelhas foram introduzidas no centro ou ao longo da linha lateral do plantio para a Ãrea cujas flores nÃo receberam visitas. O nÃmero mÃdio de vagens por planta foi 49,64  2,64, 57,16  2,41 e 59,60  2,71; o nÃmero mÃdio de grÃos por planta foi 104,68  5,13, 121,46  3,98 e 127,42  5,95; e o percentual de vagens com trÃs grÃos foi 18,65%, 20,43% e 23,72% para os tratamentos onde as plantas foram engaioladas para nÃo receberem visitantes florais, deixadas abertas para receberem visitantes florais e aberta + a introduÃÃo de colÃnias de abelhas melÃferas, respectivamente. Os conteÃdos de Ãleo e proteÃna nÃo diferiram (p>0,05) entre os tratamentos. Todas as colÃnias produziram mel e ao final do florescimento (30 dias) a produÃÃo total alcanÃou 81,7 kg, com uma produÃÃo mÃdia por colmÃia de 10,1  0,86 kg e nenhum resÃduo de pesticida foi identificado no mel. Concluiu-se que as cultivares de soja adaptadas ao Norte e Nordeste do Brasil podem ser usadas para a produÃÃo de mel e o comportamento de forrageio de A. mellifera visitando flores somente no perÃodo da manhà permite recomendar a aplicaÃÃo de defensivos apenas no final da tarde ou cedo da noite, quando as abelhas nÃo se encontram no plantio para produzir mel livre de resÃduos de pesticidas e prevenir riscos de envenenamento das abelhas; tambÃm, a introduÃÃo de A. mellifera para polinizaÃÃo pode elevar a produtividade em mais de 25%, principalmente devido ao aumento no percentual de vagens com trÃs grÃos.

Translocation involves the movement of organisms, by human intervention, from one area to other suitable (receptor) habitats. In a conservation context, translocation can be employed to support species preservation, population restoration and/or for ecological research. Despite decades of internationally published research, translocation remains a controversial endeavour. However, due to continual degradation and fragmentation of natural habitats in the face of human development, translocation is becoming a vital component of conservation efforts. Prior to the development of an Exxaro coal mine in Mpumalanga, a population of an endangered Highveld succulent species, Frithia humilis Burgoyne (Aizoaceae/Mesembryanthemaceae), was saved from extirpation by means of translocation. Three receptor habitats were identified within the distribution range of the species. The largest part of the donor population was transplanted to sandstone outcrops of the Ecca Group (Karoo Supergroup), resulting in four subpopulations residing on geological substrates typical of the species’ habitat. The remaining portion of the donor population was experimentally translocated to two habitats containing non-native geologies, namely sedimentary outcrops of the Wilge River Formation (Waterberg Group) and (igneous) felsite oucrops of the Rooiberg Group (Transvaal Supergroup). A control population was identified, occupying Ecca and Dwyka Group (Karoo Supergroup) sediments, as a measure to compare the response of translocated populations. A monitoring programme, utilising a plant age classification system, was initiated in February of 2010 to elucidate demographic trends and to gauge the response of translocated populations to novel environments. Plant survival, plant growth, flowering, fruiting (representing reproductive response) and seedling emergence were chosen as indicators to measure translocation success over the short term. Furthermore, quantitative and qualitative entomological investigations into the identity of possible F. humilis pollinators, as well as the presence of pollinator species at receptor habitats, were made. A repeatable methodology for post-translocation monitoring and scientifically sound baseline data for future comparative purposes were successfully established. Initial results showed that F. humilis subpopulations replanted on Ecca standstones had positive responses to translocation: Subpopulations survived and all but one increased in size. Individual plant growth increased, higher reproductive output was evident and seedling emergence was pervasive. Positive responses indicated that F. humilis populations translocated onto typical geologies had the potential to establish and persist over three years. Knowledge of this early success is of immense value to the conservation of the species, as a limited number of known natural populations remain. Coal mining, targeting coal seams underlying typical F. humilis habitats, is also likely to remain a threat. The viability of translocating F. humilis populations to non-typical geological substrates has shown limited efficacy. Poor survival along with inferior reproductive response confirmed Wilge River Formation outcrops as poor receptor sites for translocated F. humilis populations. Rooiberg felsite outcrops also proved to be dubious receptor sites, primarily since there was a downward trend in seedling emergence over time, suggesting inferior germination conditions. Nevertheless, translocation to non-native geological substrates did not have disastrous short-term consequences for these populations, since flowering, fruit production and seedling emergence continued, albeit at reduced (or continually declining) rates. Potential pollinator species of F. humilis were not revealed through quantitative surveys of insect diversity. Qualitative surveys proved more efficient and accurate at pinpointing insect pollinator species. This study provided the first evidence of Apidae, Megachilidae (Hymenoptera) and Bombyliidae (Diptera) insect species pollinating F. humilis. The generalist nature of the plant-pollinator relationship, as well as the presence of generalist pollinator species at some receptor habitats, probably contributed to the initial positive response of F. humilis flowering and fruiting after translocation. Results from this study, however promising, should be viewed as initial indications of translocation success. The literature review revealed a plethora of literature recommending post-translocation monitoring programmes for five years to several decades. This study confirmed that successful establishment of F. humilis can be determined after three years, but that long-term monitoring is required to evaluate persistence. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

Translocation involves the movement of organisms, by human intervention, from one area to other suitable (receptor) habitats. In a conservation context, translocation can be employed to support species preservation, population restoration and/or for ecological research. Despite decades of internationally published research, translocation remains a controversial endeavour. However, due to continual degradation and fragmentation of natural habitats in the face of human development, translocation is becoming a vital component of conservation efforts. Prior to the development of an Exxaro coal mine in Mpumalanga, a population of an endangered Highveld succulent species, Frithia humilis Burgoyne (Aizoaceae/Mesembryanthemaceae), was saved from extirpation by means of translocation. Three receptor habitats were identified within the distribution range of the species. The largest part of the donor population was transplanted to sandstone outcrops of the Ecca Group (Karoo Supergroup), resulting in four subpopulations residing on geological substrates typical of the species’ habitat. The remaining portion of the donor population was experimentally translocated to two habitats containing non-native geologies, namely sedimentary outcrops of the Wilge River Formation (Waterberg Group) and (igneous) felsite oucrops of the Rooiberg Group (Transvaal Supergroup). A control population was identified, occupying Ecca and Dwyka Group (Karoo Supergroup) sediments, as a measure to compare the response of translocated populations. A monitoring programme, utilising a plant age classification system, was initiated in February of 2010 to elucidate demographic trends and to gauge the response of translocated populations to novel environments. Plant survival, plant growth, flowering, fruiting (representing reproductive response) and seedling emergence were chosen as indicators to measure translocation success over the short term. Furthermore, quantitative and qualitative entomological investigations into the identity of possible F. humilis pollinators, as well as the presence of pollinator species at receptor habitats, were made. A repeatable methodology for post-translocation monitoring and scientifically sound baseline data for future comparative purposes were successfully established. Initial results showed that F. humilis subpopulations replanted on Ecca standstones had positive responses to translocation: Subpopulations survived and all but one increased in size. Individual plant growth increased, higher reproductive output was evident and seedling emergence was pervasive. Positive responses indicated that F. humilis populations translocated onto typical geologies had the potential to establish and persist over three years. Knowledge of this early success is of immense value to the conservation of the species, as a limited number of known natural populations remain. Coal mining, targeting coal seams underlying typical F. humilis habitats, is also likely to remain a threat. The viability of translocating F. humilis populations to non-typical geological substrates has shown limited efficacy. Poor survival along with inferior reproductive response confirmed Wilge River Formation outcrops as poor receptor sites for translocated F. humilis populations. Rooiberg felsite outcrops also proved to be dubious receptor sites, primarily since there was a downward trend in seedling emergence over time, suggesting inferior germination conditions. Nevertheless, translocation to non-native geological substrates did not have disastrous short-term consequences for these populations, since flowering, fruit production and seedling emergence continued, albeit at reduced (or continually declining) rates. Potential pollinator species of F. humilis were not revealed through quantitative surveys of insect diversity. Qualitative surveys proved more efficient and accurate at pinpointing insect pollinator species. This study provided the first evidence of Apidae, Megachilidae (Hymenoptera) and Bombyliidae (Diptera) insect species pollinating F. humilis. The generalist nature of the plant-pollinator relationship, as well as the presence of generalist pollinator species at some receptor habitats, probably contributed to the initial positive response of F. humilis flowering and fruiting after translocation. Results from this study, however promising, should be viewed as initial indications of translocation success. The literature review revealed a plethora of literature recommending post-translocation monitoring programmes for five years to several decades. This study confirmed that successful establishment of F. humilis can be determined after three years, but that long-term monitoring is required to evaluate persistence. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

Guayule (Parthenium argentatum Gray) is one of two major plants in the world grown for natural rubber and therefore, is of potential importance to the U.S. One area of research interest relates to methods of enhancing production of viable seeds. Therefore, studies were conducted in 1984, 1985, and 1986, in Tucson, Arizona to determine the effect of honey bee pollination on: seed set; seed weight; total seed yield; percentage of seed germination; yield and percentage of rubber and resin content. This experiment involved four guayule cultivars and four pollination treatments: plants caged with bees; plants caged without bees; plants open-pollinated; and plants individually covered with Delnet bags. All four cultivars responded positively to honey bee pollination. Plots with bees produced at least 195% more seeds than plots from which bees were excluded. However, there were no qualitative differences in the seed weights between the treatments. The percentage of seed germination from plots serviced by bees was significantly greater (65%) than from plots without bees (50%). Highest seed germination rates were obtained when seeds were collected in May (80%) and September (76%). June, July, and August seed collections resulted in lower seed germination rates (40%, 26%, and 63%, respectively). The plots in which bees were present gave a higher rubber yield (323 kg/ha) than plots without bees (255 kg/ha). However, there were no quantitative differences in the percentage of resin content between the treatments. These studies demonstrated that (1) honey bees can increase seed yield, seed germination, and rubber content in guayule, and (2) seeds produced during summer months had poorer germination rates and lower yields.

Leafflower plant/leafflower moth brood pollination mutualisms are widespread in the Paleotropics. Leafflower moths pollinate leafflower plants, but their larvae consume a subset of the hosts' seeds. These interactions are highly phylogenetically constrained: six clades of leafflower plants are each associated with a unique clade of leafflower moths (Epicephala). Here, we report a previously unrecognized basal seventh pollinating Epicephala lineageassociated with the highly derived leafflower clade Glochidionin Asia. Epicephala lanceolaria is a pollinator and seed predator of Glochidion lanceolarium. Phylogenetic inference indicates that the ancestor of E. lanceolaria most likely shifted onto the ancestor of G. lanceolarium and displaced the ancestral allospecific Epicephala pollinator in at least some host populations. The unusual and apparently coadapted aspects of the G. lanceolarium/E. lanceolaria reproductive cycles suggest that plant-pollinator coevolution may have played a role in this displacement and provide insights into the dynamics of host shifts and trait coevolution in this specialized mutualism.

As angiospermas são um dos grupos mais diversos de organismos, e grande parte dessa diversidade deve-se às muitas formas florais existentes. Devido a isso, determinar os padrões de variação floral e os fatores históricos e ecológicos que levaram à evolução desta alta diversidade floral é essencial para entender os processos que dirigem a diversificação das angiospermas. A tribo Bignonieae (Bignoniaceae) representa um excelente modelo para o estudo de diversificação floral na região Neotropical, por ser o clado mais diverso de lianas neotropicais e apresentar uma altíssima diversidade floral. Nesta tese, utilizei a tribo Bignonieae para caracterizar: (i) o padrão de evolução de caracteres florais discretos e morfologias florais, assim como possíveis associações entre as morfologias florais e grupos de polinizadores, (ii) o sinal filogenético e as taxas de evolução de caracteres florais contínuos, (iii) o padrão e a magnitude de integração fenotípica entre caracteres florais contínuos ao longo da filogenia do grupo e (iv) a influência da filogenia, da morfologia floral e de fatores abióticos para a co-ocorrência de espécies e a estrutura de comunidades de Bignonieae. Os resultados indicam grande labilidade na evolução de caracteres discretos e morfologias florais, contrapondo o sinal filogenético encontrado para os 16 caracteres contínuos avaliados. O sinal filogenético, entretanto, difere entre caracteres de cálice-corola e estame-estigma. As taxas de evolução também variam entre caracteres, indicando a ação de diferentes pressões seletivas ou resposta diferencial à seleção em diferentes partes da flor. Os padrões de integração fenotípica se mantêm constantes ao longo da história evolutiva de Bignonieae, apesar da evolução homoplástica da magnitude das correlações entre caracteres. Essa aparente complexidade evolutiva, sugerida pela presença de padrões diferentes em diferentes tipos de caracteres, não se repete na estrutura ecológica das comunidades, já que nem a morfologia floral e nem a filogenia influenciam a co-ocorrência entre espécies. Por outro lado, há evidente especialização das espécies a fatores abióticos, sugerindo um papel crítico de filtro ambiental na estrutura das comunidades de Bignonieae. Esse resultado contraria a hipótese de que saturação causada por competição por polinizadores seria o fator determinante da estrutura interna de comunidades de Bignonieae. / Flowering plants represent one of the most diverse groups of organisms in the Planet. A large portion of this diversity results from the multitude of floral forms encountered nowadays. To understand the patterns of floral variation and of the historical and ecological factors that led to the evolution of such diversity in floral forms are critical for a better understanding of the processes that led to the diversification of angiosperms. The tribe Bignonieae (Bignoniaceae) is the most diverse clade of Neotropical lianas and represents an excellent model for the study of floral evolution due to the high diversity of floral forms of this group. In this Ph.D. thesis, I aimed to characterize: (i) the pattern of evolution of discrete floral traits and floral morphologies in Bignonieae, as well the potential associations between floral morphologies and pollinators; (ii) the phylogenetic signal and the rates of evolution of continuous floral traits; (iii) the pattern and the magnitude of phenotypic integration among floral traits across the phylogenetic history of the group; and, (iv) the influence of phylogeny, floral morphology and abiotic factors for the patterns of species co-occurrence and the structure of communities of Bignonieae in the Neotropics. The results indicate high lability in the evolution of discrete floral traits and floral forms, contrasting the significant phylogenetic signal encountered in all 16 continuous floral traits examined. However, the phylogenetic signal differs between traits of different floral whorls. The rates of evolution also varied among different characters, suggesting the action of different selective pressures or differential responses to selection in different floral parts. Overall, the patterns of phenotypic integration were constant during the history of Bignonieae, despite the homoplastic evolution of the magnitude of correlation among characters. This apparent evolutionary complexity, leading to different patterns in different traits, is not reflected in the ecological structure of communities, given that nor floral morphology or phylogeny influence species co-occurrence. On the other hand, species specialization to abiotic factors was encountered, suggesting that environmental filtering played a key role in the structure of communities of Bignonieae. The results reject the hypothesis that saturation caused by competition for pollinators would be the main factor determining the intra-community structure of Bignonieae.

Interações ecológicas interespecíficas que resultam em benefício para todos os organismos participantes são conhecidas como mutualismo. No entanto, a troca de benefícios apresenta custos para as espécies participantes. Este fato abre espaço para o surgimento de espécies que adotam estratégias oportunistas, denominadas parasitas de mutualismos. As plantas do gênero Ficus são adequadas para estudos dessas interações, pois apresentam relação mutualística com vespas polinizadoras da família Agaonidae e são exploradas por vespas não-polinizadoras (parasitas do mutualismo). O objetivo desse estudo foi elucidar a biologia larval de vespas não-polinizadoras do gênero Idarnes pertencentes aos grupos carme e flavicollis associadas a Ficus citrifolia no campus da USP em Ribeirão Preto. Foi possível elucidar que a espécie de Idarnes do grupo flavicollis é galhadora, ao passo que a espécie do grupo carme é incapaz de induzir galhas, sendo provavelmente inquilinas específicas da espécie polinizadora. Vespas flavicollis ovipositam nos sicônios antes da polinização e evitam a retaliação pela planta, adotando mecanismo de oviposição semelhante ao das vespas polinizadoras. Vespas carme, por sua vez, depositam seus ovos diretamente dentro do saco embrionário de flores já galhadas, onde se localiza a larva da polinizadora. Contrariando especulações prévias, os resultados mostram que mesmo em espécies próximas filogeneticamente, e.g. grupos flavicollis e carme do gênero Idarnes, podem ocorrer variações acentuadas na biologia e na estratégia de exploração de recursos. / Interspecific ecological interactions which benefit all participants are known as mutualisms. However, the benefit exchange implies in costs for the involved species. This fact enables the appearance of species adopting opportunist strategies, known as parasites of mutualism. Plants belonging to the genus Ficus are suitable for studying these ecological interactions since they show a mutualistic interaction with pollinating wasps belonging to Agaonidae family and are exploited by non-pollinating wasps (parasites of the mutualism). The aim of this study was to elucidate the larval biology of non-pollinating fig wasps, genus Idarnes, belonging to flavicollis and carme species groups, associated with Ficus citrifolia at USP campus in Ribeirão Preto. We showed that species of flavicollis group are gallers, while species of carme group were not able to induce galls, probably being specific inquilines of the pollinating species. flavicollis wasps oviposit in syconia before receptivity and avoid retaliation by the plant, adopting a similar mechanism of oviposition to the one of the pollinators. carme wasps, on the other hand, deposit their eggs directly inside the embrionary sac of already galled flowers, where the pollinator larva is located. In disagreement with previous speculations, the results show that even in phylogenetically close species, such as flavicollis and carme species groups, there may be considerable variation in biology and resource exploitation strategies.

Orientador: Elza Maria Guimarâes Santos / Coorientador: Silvia Rodrigues Machado / Banca: Juliana Hanna leite El Ottra / Banca: Tatiane Maria Rodrigues / Resumo: A família Bignoniaceae é um bom modelo dos tipos de diversificação evolutiva que deram origem à diversidade de comunidades de plantas tropicais, sendo sua interação com os polinizadores um fator determinante para tal. Zeyheria tuberculosa é uma espécie ameaçada de extinção e apresenta características relacionadas à síndrome de melitofilia. Informações sobre a reprodução desta espécie poderão auxiliar na compreensão dos processos ecológicos que podem ter levado esta espécie a ser incluída na lista vermelha das espécies ameaçadas de extinção. Assim, este estudo teve como objetivo central investigar a biologia floral de Z. tuberculosa, enfocando o padrão de secreção de néctar e as características funcionais e estruturais do nectário floral, responsável pela produção do recurso essencial à manutenção da interação com os visitantes florais. Os resultados obtidos foram divididos em dois capítulos que abordam questões específicas e complementares. O primeiro contempla informações sobre os eventos ocorridos durante a antese floral, a ecologia da polinização e o padrão de secreção de néctar em plantas de Z. tuberculosa situadas em fragmentos de floresta estacional semidecídua, imersos em uma paisagem alterada. O segundo aborda a estrutura e o funcionamento do nectário floral desta espécie, com enfoque nas características relacionadas ao tipo de secreção. Para o Capítulo 1, monitoramos flores durante todo o período de antese; fizemos observações focais dos visitantes e os capturamos para examinar a área de deposição de pólen; registramos a porcentagem de grãos de pólen viáveis e verificamos o período de receptividade do estigma; obtivemos dados relativos à morfometria das flores e das abelhas polinizadoras; fizemos testes de autoincompatibilidade para avaliar a dependência de vetores de pólen para a fecundação cruzada; avaliamos a dinâmica de secreção de néctar através dos... / Abstract: The Bignoniaceae family is a good model for studies about evolutionary diversification that gave rise to the diversity of tropical plant communities, and the interaction with pollinators is a determining factor. Zeyheria tuberculosa is an endangered species and has characteristics related to melittophily syndrome. Information on the reproduction of this species may help to understand the ecological processes that may have led this species to be included in the Red List of endangered species. This goal of this study was to investigate the floral biology of Z. tuberculosa, focusing on the pattern of nectar secretion and the functional and structural characteristics of the floral nectary, which is the responsible for producing the essential resource for maintaining the interaction with pollinators. The results were divided into two sections that deal with specific and complementary issues. The first includes information about the events of floral anthesis, the pollination ecology and the pattern of nectar secretion in plants of Z. tuberculosa located in semi-deciduous forest fragments, immersed in a modified landscape. The second deals with the structure and functioning of the floral nectary of this species, focusing on the characteristics related to the type of secretion. For Chapter 1, we monitored flowers throughout anthesis; we performed focal observations of floral visitors and captured them to examine the pollen deposition area; recorded the percentage of viable pollen grains and the period of stigma receptivity; obtained data on the morphometry of flowers and pollinator bees; made self-incompatibility tests to assess the dependence of pollen vectors for cross-fertilization; evaluated the dynamics of nectar secretion through the volume, concentration and total milligrams of sugars parameters, using the latter parameter to calculate the caloric supply available to pollinators by flower and plant, and we carried out analyzes on the ... / Mestre

I investigated the pollination ecology ofHesperidanthus suffrutescens, an endangered Utah shrub, as well as the effects that road dust and local plant density may have on successful reproduction. The breeding system study included four pollination treatments to determine the degree of self-compatibility. Flower visitors were collected and probable pollinators were identified. To determine the effects of road dust on reproduction, I measured dust deposition, plant size, and reproduction in plots of increasing distance from an unpaved road. I also measured dust effects on pollination success and stomatal conductance. I determined the number of individuals per plot and the relative isolation of individuals, and measured their reproductive success to determine any significant effects. Reproduction was measured as total fruit set per plant as well as estimates of total plant seeds and plant seed weight (mg). The results of the pollination treatments and the identity of pollinators are described in Chapter 2. Both autogamy and geitonogamy (selfing treatments) produced substantially and significantly fewer fruits, seeds, and seeds per fruit than xenogamy (outcrossing treatment). Additionally, outcrossed flowers produced significantly more fruits, seeds, and seeds per fruit than did open control flowers with no difference in mean seed weight. In total, 77 flower visitors were collected over the course of the study. Probable pollinators include severalAndrena,Halictus, andDialictusbees. The effects of road dust on the reproductive success ofH. suffrutescensare discussed in Chapter 3. Dust deposition decreased with increased distance from the road and was significantly correlated with decreased fruit set for plants of a given size. Other reproductive metrics showed the same negative patterns although not significantly. The results suggest that dust may disrupt pollination and affect the physiology of plants, resulting in decreased reproduction. Dust deposition did not decrease pollination success of dusted flowers but did negatively impact stomatal conductance of leaves. The main effects of patch size and relative isolation on reproductive success are discussed in Chapter 4. Both patch size and relative isolation had no significant impacts on reproductive success. Increased patch size did indicate negative patterns on reproductive metrics, although not significantly. Similarly, increased isolation indicated increased reproductive success, although not significantly.

Cotton is a high-value commercial crop in Australia. Although cotton is largely self-pollinating, previous researchers have reported that honeybees, Apis mellifera, can assist in cross-pollination and contribute to improved yield. Until recently, use of bees in cotton had, however, been greatly limited by excessive use of pesticides to control arthropod pests. With the widespread use of transgenic (Bt) cotton varieties and the associated reduction in pesticide use, I decided to investigate the role and importance of honeybees in Bt cotton, under Australian conditions. I conducted two major field trials at Narrabri, in the centre of one of Australia’s major cotton-growing areas, in the 2005-6 and 2006-7 seasons. In the first trial, I particularly assessed methods of manipulating honeybee colonies by feeding pollen supplements of pollen/soybean patties, and by restricting pollen influx by the fitting of 30% efficient pollen traps. I aimed to test whether either of these strategies increased honeybee flight activity and, thus, increased foraging on cotton flowers. My results showed that although supplementary feeding increased bee flight activity and brood production, it did not increase pollen collection on cotton. Pollen traps initially reduced flight activity. They also reduced the amount of pollen stored in colonies, slowed down brood rearing activity, and honey production. However, they did not contribute to increased pollen collection in cotton. In the second trial, I spent more time investigating honeybee behaviour in cotton as well as assessing the effect of providing flowering cotton plants with access to honeybees for different time periods (e.g. 25 d, 15 d, 0 d). In this year, I used double the hive stocking rate of (16 colonies / ha) than in the previous year, because in 2005-6 I observed few bees in cotton flowers. I also conducted a preliminary investigation to assess whether there was any gene flow over a 16 m distance from Bt cotton to conventional cotton, in the presence of a relatively high honeybee population. Both of my field experiments showed that honeybees significantly increased cotton yield via increased boll set, mean weight of bolls, number of seeds / boll, and weight of lint / boll. It was obvious that cotton flowers, and particularly cotton pollen, were not attractive to honeybees, and this was also reflected in the low proportion (5.3% w/w) of pollen from cotton collected in the pollen traps. However, flower visitation rate was generally above the 0.5% level regarded as optimal for cross-pollination in cotton, and this was reflected in increased yield parameters. I recorded a gene flow of 1.7 % from Bollgard®II cotton to conventional cotton, over a distance of 16 m. This is much higher than had previously been reported for Australia, and may have been a result of high honeybee numbers in the vicinity, associated with my managed hives. In an attempt to attract more honeybees to cotton flowers, I conducted an investigation where I applied synthetic Queen Mandibular Pheromone (QMP) (Fruit Boost®) at two rates, 50 QEQ and 500 QEQ / ha, and for two applications, 2 d apart. Neither rate of QMP increased the level of bee visitation to flowers, either on the day of application or the subsequent day. There was also no increase in boll set or yield in plants treated with QMP. My observations of honeybee behaviour in cotton brought some interesting findings. First, honeybees totally ignored extra floral nectaries. Second, most flower-visiting honeybees collected nectar, but the overwhelming majority of them (84%) collected floral nectar from outside flowers: this meant these bees did not contribute to pollination. Those nectar gatherers which entered flowers did contribute to pollination. However, they were observed to exhibit rejection of cotton pollen by scraping pollen grains from their body and discarding them, prior to returning to their hives. Pollen gatherers collected only small, loose pellets from cotton. SEM studies showed that cotton pollen grains were the largest of all pollen commonly collected by bees in my investigations, and that they also had large spines. It is likely that these characteristics make cotton pollen unattractive to honeybees. Another possible reason for the unattractiveness of cotton flowers was the presence of pollen beetles, Carpophilus aterrimus, in them. I conducted a series of studies to determine the role of pollen beetles in pollination of cotton. I found that they did not contribute to pollination at low levels; at high populations they damaged flowers (with ≥ 10 beetles / flower, no flowers set bolls); and that honeybees, when given the choice, avoid flowers with pollen beetles. Because the insecticide fipronil was commonly used in Australian cotton at flowering time, and because I had some experience of its toxic effects against honeybees in my field investigations, I conducted a series of laboratory and potted plant bioassays, using young worker bees. The studies confirmed its highly toxic nature. I recorded an acute dermal LD50 of 1.9 ng / bee, and an acute oral LC50 of 0.62 ppm. Fipronil’s residual toxicity also remained high for an extended period in both laboratory and potted plant trials. For example, when applied to cotton leaves in weather-exposed potted cotton plants, it took 25 d and 20 d for full and half recommended rates of fipronil, respectively, to become non- toxic to honeybees. I had previously investigated whether a shorter period of exposure of cotton plants to honeybees would contribute adequately to increased yield, and concluded that a 10 d window within a 25 d flowering period would contribute 55% of the increase in total weight of bolls contributable to honeybee pollination, but only 36% of the increase in weight of lint. Given the highly residual activity of fipronil I recorded, the only opportunity for an insecticide-free period during flowering would be at its commencement. I concluded that, while there is evidence that honeybees can contribute to increased cotton yield in Bt cotton in Australia, this is unlikely with the continued use of fipronil at flowering. / Doctor of Philosophy (PhD)