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A obtenção de genótipos superiores no melhoramento de plantas depende da existência de variabilidade genética. A existência de coleções de germoplasma representativas e a utilização de um tamanho adequado de amostra são fundamentais para a preservação das frequências alélicas e genotípicas, diminuindo a perda de variabilidade genética e postergando o aparecimento dos efeitos da deriva genética. Assim, teve-se como objetivo avaliar os efeitos da deriva genética em caracteres quantitativos em subpopulações de milho. Este estudo foi realizado a partir das populações originais BR-105 e BR-106, das quais 10 subpopulações foram obtidas em cada um dos cinco ciclos sucessivos de amostragem com tamanho efetivo reduzido, totalizando 50 subpopulações para cada população original, as quais foram posteriormente autofecundadas, gerando um nível a mais de endogamia. Os tratamentos foram constituídos de 10 amostras da população original sem autofecundação, 10 amostras com autofecundação, 50 subpopulações obtidas da população original e 50 subpopulações autofecundadas, totalizando 120 tratamentos para cada população, avaliados separadamente. Utilizou-se o delineamento em blocos casualizados no esquema de parcelas subdivididas em faixas hierárquico, em quatro ambientes com duas repetições por ambiente. Os caracteres avaliados foram produção de grãos (PG), prolificidade (PROL), comprimento e diâmetro de espigas (CE e DE), número de fileiras por espiga (NFE), número de grãos por fileira (NGF), altura de planta e espiga (AP e AE), florescimento masculino e feminino (FM e FF) e número de ramificações do pendão (NRP). Foram estimados os efeitos da deriva genética entre as médias das subpopulações nos dois níveis de endogamia e os efeitos da depressão por endogamia nas subpopulações dentro dos ciclos. Posteriormente, realizaram-se análises de regressão linear para as subpopulações nos dois níveis de endogamia, separadamente, e em conjunto. Foi verificada uma grande variação nas médias das subpopulações ao longo dos ciclos, indicando que a deriva genética causou diferenciação entre as mesmas e que estas se diferenciaram das populações originais. Detectaram-se efeitos significativos da deriva genética nas populações não autofecundadas para todos os caracteres avaliados, em maior número para PG, já que este caráter é mais sensível à deriva genética por possuir maior grau de dominância que os demais. Houve diminuição no número de estimativas de deriva significativas para as populações autofecundadas, incluindo mudanças na magnitude e no sinal das mesmas em relação às populações não autofecundadas. Para as estimativas de depressão por endogamia, os caracteres PG, NGF, FM e FF apresentaram maior quantidade de estimativas significativas que os demais. Para a maioria dos caracteres, a regressão linear explicou a maior parte da variação encontrada com o aumento dos coeficientes de endogamia. As populações BR-105 e BR-106, por terem estruturas genéticas distintas, apresentaram performances diferentes quanto aos efeitos da deriva genética. Enfim, como a deriva genética interfere na integridade genética das populações, torna-se importante considerar seus efeitos na coleta e manutenção dos bancos de germoplasma e nas populações utilizadas no melhoramento genético de plantas. / Obtaining superior genotypes in plant breeding depends on the existence of genetic variability. The existence of representative germplasm collections and the use of appropriate sample size are essential for preserving allelic and genotypic frequencies, reducing loss of genetic variability and delaying genetic drift effects. This study aimed to evaluate the effects of genetic drift in quantitative traits in subpopulations of maize. The original populations used were BR-105 and BR-106, of which 10 subpopulations were obtained in each five successive sample cycles with reduced effective size, accounting 50 subpopulations for each original population that were subsequently selfed to generate an additional level of inbreeding. The treatments consisted in 10 samples of the original population, 10 samples of the selfed original population, 50 non selfed subpopulations obtained from the original population and 50 selfed subpopulations, accounting 120 treatments for each population evaluated separately. It was used the randomized block strip-plot design, in four environments with two replications. The traits assessed were grain yield (GY), prolificacy (PROL), ear length and ear diameter (EL and ED), number of rows per ear (NRE), kernel-row number (KRN), plant and ear height (PH and EH), days to anthesis and silking (DA and DS), and number of tassel branches (NTB). It was estimated the effects of genetic drift between subpopulations means at both inbreeding levels, and the effect of the inbreeding depression in subpopulations within cycles. It was also performed linear regression analysis for subpopulations at both levels of inbreeding separately and together. A large variation was observed in the subpopulations means over cycles, indicating that genetic drift caused differentiation between them, and that they differed from the original populations. The effects of genetic drift were significant for all traits in the non selfed subpopulations, especially for GY, which is more sensitive to genetic drift effects by having a greater degree of dominance than the other traits. There was a decrease in the number of significant genetic drift estimates for selfed populations, including changes in magnitude and signs, compared to the non selfed populations. GY, KRN, DA and DS had higher number of significant inbreeding depression estimates than the other traits. Linear regression analysis explained most of the variation found with increasing homozygosity. As BR-105 and BR-106 populations have distinct genetic structures, they showed different performances regarding the effects of genetic drift. Therefore, genetic drift interferes in the genetic integrity of populations and it is important to consider its effect on the collection and maintenance of germplasm banks and populations used in plant breeding.

Diss. (sammanfattning) Umeå : Sveriges lantbruksuniv. / Härtill 4 uppsatser.

Submitted by JÚLIO HEBER SILVA (julioheber@yahoo.com.br) on 2016-11-29T18:59:55Z No. of bitstreams: 2 Dissertação - Aurilene Santos Oliveira 2013.pdf: 2646271 bytes, checksum: aa9174ef69bab8b407a57a5e249619fc (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Jaqueline Silva (jtas29@gmail.com) on 2016-11-30T15:36:35Z (GMT) No. of bitstreams: 2 Dissertação - Aurilene Santos Oliveira 2013.pdf: 2646271 bytes, checksum: aa9174ef69bab8b407a57a5e249619fc (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-11-30T15:36:35Z (GMT). No. of bitstreams: 2 Dissertação - Aurilene Santos Oliveira 2013.pdf: 2646271 bytes, checksum: aa9174ef69bab8b407a57a5e249619fc (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2013-08-23 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / With the possibility of planting corn in two seasons (normal and late crops) the Brazil has increased the total corn production, but it is necessary to develop new hybrids and varieties aiming to increase productivity in both planting dates, to attend increasing demand for this commodity. The objectives of the present work were directed for the study the genetic variability, evaluate the yield potential and estimate inbreeding depression in three semiexotic populations of maize, for purposes of recurrent selection. The field evaluation was in two experiments: Jataí (GO) and Anhembi (SP). The first experiment was represented by half-sib families, which were evaluated in randomized complete blocks with three replications with plots of 4m (0.90 m spacing) with 20 plants. The second experiment included half-sib and S1 families in two replications with plots of 3m (0,90 m spacing) with 15 plants. The following primary characters were evaluated: AP - plant height (cm), AE - ear height (cm), DE – ear diameter (cm), EC - ear length (cm), NP - number of plants, NE – ear of number, PE - ear weight (g parcela-1 ), PG – total grain weight (g parcela-1 ), PE4 - four ear weight, PG4 - four ear grain weight , NR -tassel branch number, CP - tassel length (cm), AD - evaluation of foliar disease and ACE - evaluation of corn stunt complex. The semiexótic populations CRE had an excellent pattern of genetic variability and a good productive potential, presenting an average yield of 70% compared to checks. Within the three populations it was observed differences of families in relation to resistance to corn stunt, indicating that selection for this trait can fairly effective. / RESUMO: Com a possibilidade do plantio de milho em duas épocas (safra e safrinha) o Brasil tem elevado a produção total de milho, porém é necessário desenvolver novos híbridos e variedades visando o aumento da produtividade em ambas as épocas de plantio, para atender a crescente demanda pelo grão de milho. Os objetivos do presente trabalho se dirigem ao estudo da variabilidade genética, avaliar o potencial produtivo, estimar a depressão por endogamia e avaliar o comportamento quanto a resistência ao complexo de enfezamento em três populações semiexóticas de milho, para fins de seleção recorrente. Foram instalados dois experimentos: Jataí (GO) e Anhembi (SP). No primeiro experimento foram utilizadas famílias de meios-irmãos, que foram avaliadas em experimentos delineados em blocos casualizados com três repetições de parcelas de 4m (espaçamento de 0,90m) com 20 plantas. No segundo experimento foram utilizadas famílias de meios-irmãos e famílias S1 com duas repetições de parcelas de 3m (espaçamento de 0,90m) com 15 plantas. Foram avaliados os seguintes caracteres: AP – altura de planta (cm), AE – altura de espiga (cm), DE – diâmetro de espiga (cm), CE – comprimento de espiga (cm), NP – número de plantas, NE – número de espiga, PE – peso de espiga (g.parcela-1 ), PG – peso de grãos (g.parcela-1 ), PE4 – peso de quatro espiga, PG4 – peso de grãos de quatro espiga, NR – número de ramificações do pendão, CP – comprimento de pendão (cm), AD – avaliação de doença foliar e ACE – avaliação do complexo de enfezamento. As populações semiexóticas apresentaram um excelente padrão de variabilidade genética e um bom potencial produtivo, apresentando em média produtividade de 70% em relação à testemunha. Dentro das três populações observou-se um comportamento diferente das famílias em relação à resistência ao complexo de enfezamento, indicando que seleções para este caráter nas populações pode trazer resultados satisfatórios.

A obtenção de genótipos superiores no melhoramento de plantas depende da existência de variabilidade genética. A existência de coleções de germoplasma representativas e a utilização de um tamanho adequado de amostra são fundamentais para a preservação das frequências alélicas e genotípicas, diminuindo a perda de variabilidade genética e postergando o aparecimento dos efeitos da deriva genética. Assim, teve-se como objetivo avaliar os efeitos da deriva genética em caracteres quantitativos em subpopulações de milho. Este estudo foi realizado a partir das populações originais BR-105 e BR-106, das quais 10 subpopulações foram obtidas em cada um dos cinco ciclos sucessivos de amostragem com tamanho efetivo reduzido, totalizando 50 subpopulações para cada população original, as quais foram posteriormente autofecundadas, gerando um nível a mais de endogamia. Os tratamentos foram constituídos de 10 amostras da população original sem autofecundação, 10 amostras com autofecundação, 50 subpopulações obtidas da população original e 50 subpopulações autofecundadas, totalizando 120 tratamentos para cada população, avaliados separadamente. Utilizou-se o delineamento em blocos casualizados no esquema de parcelas subdivididas em faixas hierárquico, em quatro ambientes com duas repetições por ambiente. Os caracteres avaliados foram produção de grãos (PG), prolificidade (PROL), comprimento e diâmetro de espigas (CE e DE), número de fileiras por espiga (NFE), número de grãos por fileira (NGF), altura de planta e espiga (AP e AE), florescimento masculino e feminino (FM e FF) e número de ramificações do pendão (NRP). Foram estimados os efeitos da deriva genética entre as médias das subpopulações nos dois níveis de endogamia e os efeitos da depressão por endogamia nas subpopulações dentro dos ciclos. Posteriormente, realizaram-se análises de regressão linear para as subpopulações nos dois níveis de endogamia, separadamente, e em conjunto. Foi verificada uma grande variação nas médias das subpopulações ao longo dos ciclos, indicando que a deriva genética causou diferenciação entre as mesmas e que estas se diferenciaram das populações originais. Detectaram-se efeitos significativos da deriva genética nas populações não autofecundadas para todos os caracteres avaliados, em maior número para PG, já que este caráter é mais sensível à deriva genética por possuir maior grau de dominância que os demais. Houve diminuição no número de estimativas de deriva significativas para as populações autofecundadas, incluindo mudanças na magnitude e no sinal das mesmas em relação às populações não autofecundadas. Para as estimativas de depressão por endogamia, os caracteres PG, NGF, FM e FF apresentaram maior quantidade de estimativas significativas que os demais. Para a maioria dos caracteres, a regressão linear explicou a maior parte da variação encontrada com o aumento dos coeficientes de endogamia. As populações BR-105 e BR-106, por terem estruturas genéticas distintas, apresentaram performances diferentes quanto aos efeitos da deriva genética. Enfim, como a deriva genética interfere na integridade genética das populações, torna-se importante considerar seus efeitos na coleta e manutenção dos bancos de germoplasma e nas populações utilizadas no melhoramento genético de plantas. / Obtaining superior genotypes in plant breeding depends on the existence of genetic variability. The existence of representative germplasm collections and the use of appropriate sample size are essential for preserving allelic and genotypic frequencies, reducing loss of genetic variability and delaying genetic drift effects. This study aimed to evaluate the effects of genetic drift in quantitative traits in subpopulations of maize. The original populations used were BR-105 and BR-106, of which 10 subpopulations were obtained in each five successive sample cycles with reduced effective size, accounting 50 subpopulations for each original population that were subsequently selfed to generate an additional level of inbreeding. The treatments consisted in 10 samples of the original population, 10 samples of the selfed original population, 50 non selfed subpopulations obtained from the original population and 50 selfed subpopulations, accounting 120 treatments for each population evaluated separately. It was used the randomized block strip-plot design, in four environments with two replications. The traits assessed were grain yield (GY), prolificacy (PROL), ear length and ear diameter (EL and ED), number of rows per ear (NRE), kernel-row number (KRN), plant and ear height (PH and EH), days to anthesis and silking (DA and DS), and number of tassel branches (NTB). It was estimated the effects of genetic drift between subpopulations means at both inbreeding levels, and the effect of the inbreeding depression in subpopulations within cycles. It was also performed linear regression analysis for subpopulations at both levels of inbreeding separately and together. A large variation was observed in the subpopulations means over cycles, indicating that genetic drift caused differentiation between them, and that they differed from the original populations. The effects of genetic drift were significant for all traits in the non selfed subpopulations, especially for GY, which is more sensitive to genetic drift effects by having a greater degree of dominance than the other traits. There was a decrease in the number of significant genetic drift estimates for selfed populations, including changes in magnitude and signs, compared to the non selfed populations. GY, KRN, DA and DS had higher number of significant inbreeding depression estimates than the other traits. Linear regression analysis explained most of the variation found with increasing homozygosity. As BR-105 and BR-106 populations have distinct genetic structures, they showed different performances regarding the effects of genetic drift. Therefore, genetic drift interferes in the genetic integrity of populations and it is important to consider its effect on the collection and maintenance of germplasm banks and populations used in plant breeding.

Abstract Arabidopsis lyrata is a close outcrossing relative of Arabidopsis thaliana, the model organism of plant physiology and molecular biology. I studied variation in flowering time and the factors shaping the variation within and between A. lyrata populations in different environments. The role of the two important proximate factors determining flowering time, day length and temperature, were studied in climate chambers. The southern A. lyrata populations were found to flower in high frequency and quicker than northern A. lyrata populations in all studied environments, but the reaction of northern populations on long day length was found to be stronger than that of southern populations. Differences in vernalization requirement between A. lyrata populations were found in outdoor common garden, but in the climate chambers the results of vernalization experiments were not consistent. Strength and direction of selection on flowering time and other life history traits were studied in alpine and lowland A. lyrata populations in Scandinavia. Differences in selection were found both between populations and between years. Grazing sheep caused high levels of damage in inflorescences in the alpine population. In the lowland population there was less herbivory, caused by insects and hares. The difference in selection on flowering traits in the two study populations might be partly caused by selective grazing. Completely outcrossing mating system in A. lyrata is due to well developed self-incompatibility system. However, biparental inbreeding is likely to exist in natural populations and it may lead to spatial structuring of genetic variation within populations. I studied the effects of biparental inbreeding on components of fitness in A. lyrata in three different environments. I found inbreeding depression after sib-mating to be substantial. Stressful environment reduced the overall performance of the plants, but had no effect on the magnitude of inbreeding depression. A literature survey indicates that the observed levels of inbreeding depression in self-incompatible A. lyrata were higher than those of self-compatible species. This suggests that self-compatible species have purged some of their genetic load. The genetic basis of flowering time variation in A. lyrata can be further studied by using A. thaliana molecular tools.

Insular fauna face disproportionate risks of extinction owing to direct human perturbation and intrinsic factors that are enhanced at small population sizes. Currently, our understanding of the processes that promote long-term persistence of naturally small populations and the cryptic processes that may contribute to accelerating their decline is limited by lack of empirical investigations across the range of natural conditions. Implementing effective protections for rare and understudied taxa requires the identification and examination of factors that limit recruitment at critical life stages. Predicting population health outcomes of future perturbations further necessitates an understanding a taxon’s behavioral ecology. Finally, cryptic threats to viability, such as inbreeding depression, must be investigated with an appreciation for taxon-specific life history, as these attributes can alter the context in which severe fitness reductions are expressed. In this project I enlist integrative and cross-disciplinary approaches to study the behavioral ecology and conservation genetics of a critically endangered West Indian Rock Iguana, Cyclura nubila caymanensis, on Little Cayman Island. I demonstrate how coastal communal nesting areas, a critical limiting resource on the island, serve a diverse population demographic and contribute to significantly enhanced nesting outcomes. These data emphasize the importance of expanding protections for major sites, as aggregative nesting appears to be perpetuated by both habitat suitability and adaptive fitness benefits. I next evaluate the possibility of evolved inbreeding avoidance strategies, including natal dispersal, non-assortative mate choice, and genetic bet-hedging. I conclude that the contribution of pre-reproductive dispersal to inbreeding avoidance likely outweighs that of active mate choice. Importantly, observed patterns of siring success imply constrained female choice and sexual conflict over genetic mating outcomes – a pattern that may extend to many territorial, male-driven mating systems and therefore should be an important consideration in genetic management. Finally, I investigate age-dependent inbreeding effects and the degree to which inbreeding depression may limit recruitment to the breeding population. I fail to reveal significant correlations of multi-locus heterozygosity with hatchling fitness; however, negative effects of parental inbreeding on fecundity and hatching success imply fitness consequences of inbreeding depression could be felt at other life stages.

Inbreeding occurs when relatives mate and have offspring. Inbreeding depression is hypothesized to have influenced the evolution of mating systems and behavioural mechanisms of inbreeding avoidance in the animal kingdom. Inbreeding in the wild is difficult to measure, as in order to build a pedigree allowing us to identify matings between relatives, the identity of as many as possible members of a population needs to be known. For a long time, the main source of knowledge about inbreeding depression was based on laboratory and agricultural studies, which did not reflect the array of environmental pressures wild populations have to cope with. In consequence, the deleterious consequences of inbreeding have often been underestimated. This is problematic because accurate estimates of the effect size of inbreeding depression are needed to study the strength of selection on inbreeding avoidance mechanisms, and are also of importance to conservation genetics. The aim of this thesis was to use pedigree data to infer the occurrence and effects of inbreeding using over forty years of breeding events of the great tit Parus major from Wytham Woods, Oxfordshire. The effects of inbreeding on fitness were investigated across a life-history continuum, and across environments. I found that close inbreeding (f=0.25) resulted in pronounced inbreeding depression, which acted independently on hatching success, fledging success, and recruitment success, and reduced the number of fledged grand-offspring by 55%. My results therefore suggest that estimates of fitness costs of inbreeding must focus on the entire life cycle. I also show that the variation in the strength of inbreeding depression varies across environments, particularly so the more the environmental variable considered is linked to fitness. These results emphasise the need of using relevant environmental contrasts when investigating inbreeding by environment interactions. I further asked whether individuals involved in matings with relatives differed relative to individuals mating with unrelated partners. I did not find any evidence for clear predictors of inbreeding, and I show that inbreeding depression in our population is entirely independent of any tendency for low quality parental genotypes, or phenotypes, to inbreed. Neither did I find any evidence for active inbreeding avoidance: great tits did not mate less often with kin than expected based on several scenarios of random mating, nor did I find increased rates of extra-pair paternity among birds breeding with relatives. In fact, I observed quite the contrary, as birds mating with kin exhibited a higher than average rate of close inbreeding relative to all scenarios of random mating investigated, showed lower rates of extra-pair paternity and divorce than birds mated to unrelated partners. I hypothesise that cases of occasional inbreeding in this population may result from mis-imprinting or a related process whereby some birds develop particularly strong bonds that are at odds with all predictions of avoiding inbreeding. Finally, I asked to what extent natal dispersal, a behaviour that is often hypothesized to play an important role in avoiding inbreeding, indeed reduces the likelihood of inbreeding. I found that male and female individuals breeding with a relative dispersed over several-fold shorter distances than those outbreeding. This led to a 3.4 fold increase (2.3-5, 95% CI) in the likelihood of close inbreeding relative to the population average when individuals dispersed less than 200m. This thesis demonstrates that inbreeding has deleterious effects on a wild population of birds, occurring throughout an individual’s life, and is of varying strength across environments. My findings strongly support the theory that natal dispersal should be considered as a mechanism of prime importance for inbreeding avoidance.

The evolution of extra-group paternity (EGP) is a contentious issue in evolutionary biology. This thesis examines the factors and adaptive benefits driving EGP in a high-density, group-living population of European badgers (Meles meles). To improve power to assign parentage, I isolated and characterised 21 new polymorphic microsatellite markers. I genotyped 83% of 1410 badger trapped 1987‒2010 using 35 autosomal microsatellite markers. Maternity and paternity were assigned at 80% confidence ca. 82% of individuals. 48% of paternities were extra-group, where 85% were attributable to neighbouring-group males and EGP was detected in 47% of litters; thus badger social group do not correspond with a breeding unit. I tested whether indirect genetic benefits explain these high EGP rates. (1) ‘Good-gene-as-heterozygosity Hypothesis’: Paternal heterozygosity, but not maternal or an individual’s own heterozygosity, associated positively with first-year survival probability. Under benign environmental conditions, cubs fathered by more heterozygous males had a higher first year survival probability. Despite this correlation, the EGP rate per litter correlated with neither average nor maximum within-group heterozygosity of candidate fathers. (2) Fitness benefit Hypothesis: Extra-group offspring (EGO) had lower first-year survival probability and lived 1.3 years less than within-group offspring (WGO). Female WGO produced more litters and offspring over their lifetime than female EGO, whereas male EGO produced more offspring than male WGO. (3) Inbreeding avoidance hypothesis: The EGP rate within a litter increased with greater average pair-wise relatedness between mothers and within-group candidate fathers. No inbreeding depression on first-year survival probability was detected, but small sample sizes limited statistical power. Socio-ecologically, at the litter level, EGP correlated negatively with the number of within-group candidate fathers, and positively with neighbouring-group candidate fathers. In conclusion, EGP in badgers may reduce inbreeding and be maintained in the population through a sex-specific antagonistic selection and indirect genetic benefits may occur when the total fitness benefits of producing extra-group sons outweigh the costs of producing extra-group daughters. These indirect genetic benefits only partially explain the evolution of promiscuity in European badgers, highlighting that evolutionary factors underlying promiscuity remain unclear.

Doctor of Philosophy (PhD) / A renewed focus on generalised pollinator systems has inspired a conceptual framework which highlights that spatial and temporal interactions among plants and their assemblage of pollinators can vary across the individual, population, regional and species levels. Pollination is clearly a dynamic interaction, varying in the number and interdependence of participants and the strength of the outcome of the interaction. Therefore, the role of variation in pollination is fundamental for understanding ecological dynamics of plant populations and is a major factor in the evolution and maintenance of generalised and specialised pollination systems. My study centred on these basic concepts by addressing the following questions: (1) How variable are pollinators in a generalised pollination system? To what degree do insect visitation rates and assemblage composition vary spatially among populations and temporally among flowering seasons? (2) How does variation in pollinators affect plant reproductive success? I chose to do this using a model system, Trachymene incisa subsp. incisa (Apiaceae), which is a widespread Australian herbaceous species with simple white flowers grouped into umbels that attract a high diversity of insect visitors. The Apiaceae are considered to be highly generalist in terms of pollination, due to their simple and uniform floral display and easily accessible floral rewards. Three populations of T. incisa located between 70 km and 210 km apart were studied over 2-3 years. The few studies investigating spatial and temporal variation simultaneously over geographic and yearly/seasonal scales indicate that there is a trend for more spatial than temporal variation in pollinators of generalist-pollinated plants. My study showed both spatial and temporal variation in assemblage composition among all populations and variation in insect visitation rates, in the form of a significant population by year interaction. However, removing ants from the analyses to restrict the assemblage to flying insects and the most likely pollinators, resulted in a significant difference in overall visitation rate between years but no difference in assemblage composition between the Myall Lakes and Tomago populations. These results indicate more temporal than spatial variation in the flying insect visitor assemblage of T. incisa. Foraging behaviour provides another source of variation in plant-pollinator interactions. Trachymene incisa exhibits umbels that function as either male or female at any one time and offer different floral rewards in each phase. For successful pollination, pollinators must visit both male and female umbels during a foraging trip. Insects showed both preferences and non-preferences for umbel phases in natural patches where the gender ratio was male biased. In contrast, insects showed no bias in visitation during a foraging trip or in time spent foraging on male and female umbels in experimental arrays where the gender ratio was equal. Pollinator assemblages consisting of a mixture of different pollinator types coupled with temporal variation in the assemblages of populations among years maintains generalisation at the population/local level. In addition, spatial variation in assemblages among populations maintains generalisation at the species level. Fire alters pollination in T. incisa by shifting the flowering season and reducing the abundance of flying insects. Therefore, fire plays an important role in maintaining spatial and temporal variation in this fire-prone system. Although insect pollinators are important in determining the mating opportunities of 90% of flowering plant species worldwide, few studies have looked at the effects of variation in pollinator assemblages on plant reproductive success and mating. In T. incisa, high insect visitation rates do not guarantee high plant reproductive success, indicating that the quality of visit is more important than the rate of visitation. This is shown by comparing the Agnes Banks and Myall Lakes populations in 2003: Agnes Banks received the highest visitation rate from an assemblage dominated by ants but produced the lowest reproductive output, and Myall Lakes received the lowest visitation rate by an assemblage dominated by a native bee and produced the highest seedling emergence. Interestingly, populations with different assemblage composition can produce similar percentage seed set per umbel. However, similar percentage seed set did not result in similar percentage seedling emergence. Differences among years in reproductive output (total seed production) were due to differences in umbel production (reproductive effort) and proportion of umbels with seeds, and not seed set per umbel. Trachymene incisa is self-compatible and suffers weak to intermediate levels of inbreeding depression through early stages of the life cycle when seeds are self-pollinated and biparentally inbred. Floral phenology, in the form of synchronous protandry, plays an important role in avoiding self-pollination within umbels and reducing the chance of geitonogamous pollination between umbels on the same plant. Although pollinators can increase the rate of inbreeding in T. incisa by foraging on both male and female phase umbels on the same plant or closely related plants, most consecutive insect movements were between plants not located adjacent to each other. This indicates that inbreeding is mostly avoided and that T. incisa is a predominantly outcrossing species, although further genetic analyses are required to confirm this hypothesis. A new conceptual understanding has emerged from the key empirical results in the study of this model generalised pollination system. The large differences among populations and between years indicate that populations are not equally serviced by pollinators and are not equally generalist. Insect visitation rates varied significantly throughout the day, highlighting that sampling of pollinators at one time will result in an inaccurate estimate and usually underestimate the degree of generalisation. The visitor assemblage is not equivalent to the pollinator assemblage, although non-pollinating floral visitors are likely to influence the overall effectiveness of the pollinator assemblage. Given the high degree of variation in both the number of pollinator species and number of pollinator types, I have constructed a model which includes the degree of ecological and functional specialisation of a plant species on pollinators and the variation encountered across different levels of plant organisation. This model describes the ecological or current state of plant species and their pollinators, as well as presenting the patterns of generalisation across a range of populations, which is critical for understanding the evolution and maintenance of the system. In-depth examination of pollination systems is required in order to understand the range of strategies utilised by plants and their pollinators, and I advocate a complete floral visitor assemblage approach to future studies in pollination ecology. In particular, future studies should focus on the role of introduced pollinators in altering generalised plant-pollinator systems and the contribution of non-pollinating floral visitors to pollinator assemblage effectiveness. Comparative studies involving plants with highly conserved floral displays, such as those in the genus Trachymene and in the Apiaceae, will be useful for investigating the dynamics of generalised pollination systems across a range of widespread and restricted species.