Thesis (PhD)--Stellenbosch University, 2013. / Pollinators play a fundamental role in floral evolution. They can exert selection on the flowers they visit in a plethora of different ways, ranging from innate floral preferences to differences in body size and shape and behavioural elements such as flower constancy and learning capacity. Since different pollinators exhibit differences in these characters, shifts between pollinating species are often considered the most likely drivers of floral diversification. While many lines of evidence support this claim, numerous angiosperms pollinated by a single species also exhibit floral variation. Throughout my thesis, I explore and investigate floral diversification in such species in the absence of pollinator shifts. In Chapter 2, I investigate variation in the preference of conspecific male and female pollinators for the floral traits of a sexually deceptive daisy that comprises distinct floral forms. I show that its pollinator exhibits gender-specific variation in floral preferences, and that some floral forms have specialized on the male pollinator. This chapter thus illustrates the importance of intraspecific variation in pollinator preference for floral diversification, an underappreciated mechanism in this field of research. The innate preferences of pollinators are likely to have a genetic basis, especially innate preferences that govern mate choice. Genetic structure within the pollinators of sexually deceptive plants, which mimic female insects to achieve pollination, may thus provide an important source of selection on the plants they pollinate. This depends on an association between genetic divergence and divergent mate preferences, and I explore this intriguing idea in Chapter 3. While pollinators associated with sexually deceptive floral forms did exhibit significant genetic structuring, male pollinators from different phylogeographic clades all exhibited preference for the same sexually deceptive floral form, thus rejecting this hypothesis. Another behavioural attribute of pollinators that may affect floral evolution, particularly in deceptive plant species, is learning ability. Studies on sexually deceptive orchids often report that male pollinators tend to avoid sexually deceptive flowers with experience. In Chapter 4, I systematically investigate learning abilities within male pollinators and the costs they suffer on sexually deceptive floral forms that vary in deceptiveness. Results reveal a positive relationship between the level of floral deceptiveness and the 4 associated mating costs that deceived males suffer. Pollinator learning, however, appears to occur only on the most deceptive floral forms, suggesting a link between the costs suffered to the occurrence of learning. In Chapter 4, I systematically investigate learning abilities within male pollinators and the costs they suffer on sexually deceptive floral forms that vary in deceptiveness. Results reveal a positive relationship between the level of floral deceptiveness and the associated mating costs that deceived males suffer. Pollinator learning, however, appears to occur only on the most deceptive floral forms, suggesting a link between the costs suffered to the occurrence of learning. In Chapter 5, I explore the importance of florivory damage in a polymorphic daisy. Studies on floral evolution often overlook the significance of florivorous visits and focus only on pollinator-mediated selection. I show that floral polymorphism is maintained by antagonistic selection exerted by pollinators and florivores on the same floral traits. Lastly, I focus on evolutionary history to explore similarity in the patterns of South African angiosperm evolution and the pollinator species used throughout my thesis. Molecular dating shows this pollinator exhibits broadly congruent evolutionary patterns to these angiosperms, indicative of a shared biogeography. Taken together, my thesis demonstrates the vast impact of floral visitors, in particular pollinating insects, on the evolution of floral form. / My research was funded by the National Research Foundation of South Africa (NRF) and personal funding was provided by a NRF Innovation scholarship and merit bursaries from the Botany and Zoology department at Stellenbosch University. A WhiteSci Travel Grant and financial support from Prof. Erik Svensson at Lund University also allowed me to present parts of my research at international conference.

Understanding the functional relationship between characters and components of fitness is a central goal of evolutionary biology. The studies in this thesis examined the ecological causes and evolutionary consequences underlying differences in fitness among individuals of Lobelia cardinalis. Flowering plants experience selection from many sources, which may enhance or oppose selection by pollinators. In the second chapter of this thesis, the role of pollinators and herbivores in shaping selection on floral characters was investigated. Floral traits experienced pollinator-mediated selection and weak selection by weevil larvae and slugs. Because pollinators also forage according to local density of flowers, in the fourth chapter I explored how local density of individual plants and flowers influences fitness of individual plants. Plants at dense sites produced more seeds, consistent with pollinator preference for denser patches. Individual female-phase flowers produced more seeds as the density of surrounding male-phase flowers increased and female-phase flowers decreased. This study highlights how plant phenotype and local density influence pollination and subsequent plant fitness. In L. cardinalis rosette formation (a life-history character) partly shapes the distribution of plants, and may influence plant survival and fitness. In the fifth chapter, I explored how variation in allocation to clonal reproduction among plants (ramets) and genets influenced survival and fitness. Plants that produced more and larger rosettes realized higher survival independent of the phenotype of the parental. Plants that produced one rosette in 2009 produced more seeds in 2010 than plants that produced more than one rosette. This pattern was reversed in the following time period; plants that produced more rosettes in 2010 produced more seeds in 2011. The relative importance of pollinators versus other selective agents in shaping floral traits, as well as the intensity of competition among individual plants and flowers likely depend on the extent to which reproduction is pollen limited. In the third chapter, I explored how pollen limitation affected selection on floral traits via female fitness and found a weak relationship. Although this seems to contradict intuition, several reasons may limit the influence of pollen limitation on selection.

Studies assessing the evolution of plant traits frequently focus on pollinators as the primary drivers of floral trait evolution. However, herbivores can also play a role, and, under some circumstances, may even impose stronger selection on floral display than pollinators. This is especially true when the traits under selection are linked to anti-herbivore defense. Here I describe a study in which I quantified the relative role of herbivores and pollinators in selection for floral traits in the North American invasive plant, purple loosestrife (Lythrum salicaria). Because L. salicaria responds to leaf-chewing herbivores by producing compensatory tissue growth, and this in turn alters the architecture of the floral display, I further tested whether herbivores can indirectly modify pollinator-mediated selection through this pathway. Using a split-plot design, I measured pollen limitation and reproductive output in experimentally manipulated plants in the presence and absence of simulated herbivore damage in order to quantify the effects of damage and pollination on natural selection for floral display. My results showed that damage significantly increased direct selection (beta-i) for earlier flowering time and decreased selection on the number of inflorescences, even more than pollinators did. Because damage did not modify pollinator-mediated selection for floral display traits, the selection imposed by herbivores is likely only having direct effects. My findings demonstrate the importance of considering multiple agents of selection and their potential interactions when quantifying natural selection in a study system. In particular, it is important to consider that the agent most frequently studied may not be imposing the brunt of selection.

<p> Many floral characters have can affect plant fitness by altering the frequency or efficiency of pollinator visits. More frequent pollinator visitation may lead to larger pollen loads and greater outcrossing rates, which in tern may lead to greater numbers of offspring, with higher genetic variability and less inbreeding depression. Mating systems may moderate the strength of natural selection on traits that attract pollinators through its effects on inbreeding depression, and the ability to self-pollinate autogamously. Natural selection is expected to be stronger in largely outcrossing systems where inbreeding depression is high, but reduced in largely selfing systems where deleterious alleles have been purged.</p> <p> Here I investigate the role of floral display height in attracting pollinators, and its consequences for plant fitness in two closely related monocarpic herbs (Hesperis matronalis and Alliaria petiolata) with different mating systems. Though others have looked at height as a potential trait attracting pollinators, no one has yet clearly separated the effects of floral display height from that of plant size. I experimentally altered plant height independently of plant size by placing potted plants into either elevated or non-elevated treatments and observed pollinator foraging behaviour and reproduction.</p> <p> Pollinators preferred taller floral displays in the predominant outcrosser Hesperis matronalis. Elevated plants produced more seeds than control plants although their seeds were smaller, reflecting a strong seed size-number trade-off. Path analysis showed seed number was affected directly by plant height, presumably though greater pollinator visitation, and indirectly by plant mass through increased fruit number.</p> <p> In Alliaria petiolata, a predominant self-pollinating species, pollinators visited taller displays in field observations and experimentally elevated plants had significantly greater seed per fruit. Experimentally outcrossed and self-pollinated plants showed no difference in seed production, but offspring quality was significantly higher in outcrossed plants for early traits. Measures of maternal fitness increased significantly with both maternal and seed mass. For individual seeds, seed mass was the most important factor in seed fitness, where larger seeds germinated sooner, more frequently, and had higher survivorship than smaller seeds, regardless of pollination treatment. Competition strongly affected all seedling traits, with increased density having a significant negative effect on seedling mass, total leaf area, and maximum leaf length; and, a positive effect on hypocotyl length and petiole length. Seedling mass at harvest was not significantly affected by elevation treatment, seed mass, or maternal mass, however, this may have been due to lower competition brought about by low germination and survival; thus, competition did not accentuate the effect of maternal size or elevation treatment on seedling. Taken together, the results of these two studies suggest the importance of height, in attracting pollinators, independent of size in both species. The consequences of pollinator attraction were highly significant in the predominant outcrosser Hesperis matronalis, while maternal size and seed size played more important roles in the fitness of the predominant selfer Alliaria petiolata.</p> / Thesis / Master of Science (MSc)

It is well known that animals can exert strong selective pressures on plant traits. However, studies on the evolutionary consequences of plant–animal interactions have mainly focused on understanding how these interactions shape trait means, while overlooking its potential direct effect on the variability among structures within a plant (e.g. flowers and fruits). The degree of within-plant variability can have strong fitness effects but few studies have evaluated its role as a potential target of selection. Here we reanalysed data on Ipomoea wolcottiana stigma–anther distance to test alternate mechanisms driving selection on the mean as well as on intra-individual variance in 2 years. We found strong negative selection acting on intra-individual variation but not on mean stigma–anther distance, suggesting independent direct selection on the latter. Our result suggests that intra-individual variance has the potential to be an important target of selection in nature, and that ignoring it could lead to the wrong characterisation of the selection regime. We highlight the need for future studies to consider patterns of selection on the mean as well as on intra-individual variance if we want to understand the full extent of plant–animal interactions as an evolutionary force in nature.