Marinus L. de Jager

Do pollinators influence the assembly of flower colours within plant communities

The co-occurrence of plant species within a community is influenced by local deterministic or neutral processes as well as historical regional processes. Floral trait distributions of co-flowering species that share pollinators may reflect the impact of pollinator preference and constancy on their assembly within local communities. While pollinator sharing may lead to increased visitation rates for species with similar flowers, the receipt of foreign pollen via interspecific pollinator movements can decrease seed set. We investigated the pattern of community flower colour assembly as perceived by native honeybee pollinators within 24 local assemblages of co-flowering Oxalis species within the Greater Cape Floristic Region, South Africa. To explore the influence of pollinators on trait assembly, we assessed the impact of colour similarity on pollinator choices and the cost of heterospecific pollen receipt. We show that flower colour is significantly clustered within Oxalis communities and that this is not due to historical constraint, as flower colour is evolutionarily labile within Oxalis and communities are randomly structured with respect to phylogeny. Pollinator observations reveal that the likelihood of pollinators switching between co-flowering species is low and increases with flower colour similarity. Interspecific hand pollination significantly reduced seed set in the four Oxalis species we investigated, and all were dependant on pollinators for reproduction. Together these results imply that flower colour similarity carries a potential fitness cost. However, pollinators were highly flower constant, and remained so despite the extreme similarity of flower colour as perceived by honeybees. This suggests that other floral traits facilitate discrimination between similarly coloured species, thereby likely resulting in a low incidence of interspecific pollen transfer (IPT). If colour similarity promotes pollinator attraction at the community level, the observed clustering of flower colour within communities might result from indirect facilitative interactions.

Floral polymorphism and the fitness implications of attracting pollinating and florivorous insects.

BACKGROUND AND AIMS Floral polymorphism is frequently attributed to pollinator-mediated selection. Multiple studies, however, have revealed the importance of non-pollinating visitors in floral evolution. Using the polymorphic annual daisy Ursinia calenduliflora, this study investigated the importance of different insect visitors, and their effects on fitness, in the maintenance of floral polymorphism. METHODS The spatial structure of a discrete floral polymorphism was characterized based on the presence/absence of anthocyanin floret spots in U. calenduliflora. A 3-year observational study was then conducted in polymorphic populations to investigate differences in visitation rates of dominant visitors to floral morphs. Experiments were performed to explore the floral preference of male and female Megapalpus capensis (the dominant insect visitor) and their effectiveness as pollinators. Next, floral damage by antagonistic florivores and the reproductive success of the two floral morphs were surveyed in multiple populations and years. KEY RESULTS Floral polymorphism in U. calenduliflora was structured spatially, as were insect visitation patterns. Megapalpus capensis males were the dominant visitors and exhibited strong preference for the spotted morph in natural and experimental observations. While this may indicate potential fitness benefits for the spotted morph, female fitness did not differ between floral morphs. However, as M. capensis males are very efficient at exporting U. calenduliflora pollen, their preference may likely increase the reproductive fitness of the spotted morph through male fitness components. The spotted morph, however, also suffered significantly greater costs due to ovule predation by florivores than the spotless morph. CONCLUSIONS The results suggest that pollinators and florivores may potentially exert antagonistic selection that could contribute to the maintenance of floral polymorphism across the range of U. calenduliflora. The relative strength of selection imposed by each agent is potentially determined by insect community composition and abundance at each site, highlighting the importance of community context in the evolution of floral phenotypes.

Does morphology matter? An explicit assessment of floral morphology in sexual deception

Summary Sexually deceptive orchids are renowned for their olfactory mimicry of female insect sex pheromones to lure male pollinators, but the role of floral morphology remains unknown. Here, we reveal compelling new experimental evidence that morphology also matters in sexual deception. Our study exploited two morphologically distinct Chiloglottis orchids that both employ the same semiochemical (chiloglottone 1) to attract their respective primary pollinator. In these experiments, we monitored attempted copulation of pollinators with orchid labella as this likely impacts plant fitness. Reciprocal pollinator choice tests revealed significant reductions in the frequency and duration of attempted copulation when pollinators were presented with alternate orchid species that differ in floral morphology, but nevertheless exhibit identical semiochemicals. Experimentally shortening the labellum also reduced the duration of attempted copulation in one of the species. Pollinators exhibited contrasting orientations during attempted copulation and pollination, which seem to be correlated with fundamental differences in the morphological adaptations for both mechanical fit and female mimicry in these orchids. Our findings confirm the overlooked importance of floral morphology for sexually deceptive orchid pollination and indicate that pollinator behaviour could impose strong selection on specific floral traits.

Floral trait variation and integration as a function of sexual deception in Gorteria diffusa

Phenotypic integration, the coordinated covariance of suites of morphological traits, is critical for proper functioning of organisms. Angiosperm flowers are complex structures comprising suites of traits that function together to achieve effective pollen transfer. Floral integration could reflect shared genetic and developmental control of these traits, or could arise through pollinator-imposed stabilizing correlational selection on traits. We sought to expose mechanisms underlying floral trait integration in the sexually deceptive daisy, Gorteria diffusa, by testing the hypothesis that stabilizing selection imposed by male pollinators on floral traits involved in mimicry has resulted in tighter integration. To do this, we quantified patterns of floral trait variance and covariance in morphologically divergent G. diffusa floral forms representing a continuum in the levels of sexual deception. We show that integration of traits functioning in visual attraction of male pollinators increases with pollinator deception, and is stronger than integration of non-mimicry trait modules. Consistent patterns of within-population trait variance and covariance across floral forms suggest that integration has not been built by stabilizing correlational selection on genetically independent traits. Instead pollinator specialization has selected for tightened integration within modules of linked traits. Despite potentially strong constraint on morphological evolution imposed by developmental genetic linkages between traits, we demonstrate substantial divergence in traits across G. diffusa floral forms and show that divergence has often occurred without altering within-population patterns of trait correlations.

Costs of deception and learned resistance in deceptive interactions

The costs that species suffer when deceived are expected to drive learned resistance, although this relationship has seldom been studied experimentally. Flowers that elicit mating behaviour from male insects by mimicking conspecific females provide an ideal system for such investigation. Here, we explore interactions between a sexually deceptive daisy with multiple floral forms that vary in deceptiveness, and the male flies that pollinate it. We show that male pollinators are negatively impacted by the interaction, suffering potential mating costs in terms of their ability and time taken to locate genuine females within deceptive inflorescences. The severity of these costs is determined by the amount of mating behaviour elicited by deceptive inflorescences. However, inexperienced male flies exhibit the ability to learn to discriminate the most deceptive inflorescences as female mimics and subsequently reduce the amount of mating behaviour they exhibit on them with increased exposure. Experienced males, which interact with sexually deceptive forms naturally, exhibit similar patterns of reduced mating behaviour on deceptive inflorescences in multiple populations, indicating that pollinator learning is widespread. As sexually deceptive plants are typically dependent on the elicitation of mating behaviour from male pollinators for pollination, this may result in antagonistic coevolution within these systems.

The impact of floral spot and ring markings on pollinator foraging dynamics

Linear areas of contrasting floral colour, or “nectar guides”, are recognized to enhance pollinator attraction. Few studies, however, have investigated the role of other types of floral marking in pollinator behaviour. In this study, we explore the impact of petal spots and petal rings on bumblebee foraging dynamics. Using model flowers with discrete spot or bullseye ring patterns we investigate the responses of Bombus terrestris foragers towards rewarding and unrewarding flowers. We find that the presence of petal spots and rings reduce the search time of pollinators to locate rewarding model flowers. Only the rewarding petal ring phenotype, however, is readily learned and significantly increases the foraging efficiency of experienced bees over four foraging bouts. Although the rewarding spot phenotype induces random foraging over this time frame, employing differential conditioning with a strong aversive stimulus over ten foraging bouts reveal that bees have the capacity to correctly identify rewarding spot phenotypes with more training. Once a strong association between petal marking and reward is formed, bees continue to respond to marked phenotypes even when unrewarding, suggesting rewardless plants that exhibit petal marking could potentially exploit their pollinators. We conclude that petal marking, whether organised discretely in a spot or in a continuous ring around the centre of a flower, have a significant and complex influence on pollinator foraging dynamics.

Pollinators can prefer rewarding models to mimics: consequences for the assumptions of Batesian floral mimicry

Many members of the Orchidaceae offer no rewards to pollinators and attract them through deception. One common approach is to mimic the specific floral signals of co-flowering species that are rewarding (Batesian floral mimicry) to exploit their pollinators. We investigated two putative Batesian floral mimics, Disa gladioliflora subsp. capricornis and Disa ferruginea, within a single community to determine whether their pollinators are able to discriminate them from their models. Despite mimics being indistinguishable from models in terms of floral colour as perceived by pollinators, strong preference for model flowers was observed in both systems. We nevertheless recorded frequent pollen transfer in D. gladioliflora subsp. capricornis, suggesting pollination still occurs in the face of pollinator discrimination. Although the inability of pollinators to discriminate between models and mimics is often cited as a requirement for Batesian floral mimicry, we suggest that this need not be the case. Pollination by discriminating visitors may still lead to fruit set, with the most accurate mimics expected to have the highest fitness. Batesian floral mimicry may thus be best indicated when mimic fitness is positively correlated with its resemblance to model flowers.

Experimental examination of pollinator-mediated selection in a sexually deceptive orchid

Background and Aims Selection exerted by pollinators on flowers is predicted to occur along two distinct axes. While pollinator attraction to flowers is governed by pollinator preferences, pollen transfer efficiency is mediated by the mechanical fit of pollinators to flower morphology. Although pollinator attraction in sexually deceptive orchids is typically underpinned by floral odour, morphological traits are expected to play a vital role in mechanical fit during floral contact with pollinators. Methods Here we utilize a comprehensive and novel procedure to test for pollinator-mediated selection through mechanical fit with the flower labellum in the orchid Chiloglottis trapeziformis. This approach combines detailed pollinator observations related to plant reproductive fitness with complementary experimental manipulation and phenotypic selection analysis. Key Results Experiments with virgin flowers revealed that pollen removal occurs only during vigorous pseudocopulation. This behaviour involves male wasps that grasp the insectiform callus structure on the labellum while probing the labellum tip in a forward orientation. Both orientation and duration of pseudocopulation were significant predictors of pollen removal, confirming a direct relationship between pollinator behaviour and plant fitness. Controlled floral manipulation that either shortened or elongated the distance between the callus and the labellum tip detected no change in pollinator attraction. The duration of pseudocopulation, however, was significantly reduced on flowers with shortened or elongated callus-tip distances, consistent with stabilizing selection. Phenotypic selection analysis confirmed this prediction in natural populations by uncovering evidence for stabilizing selection on the distance between the callus and the labellum tip. Conclusions Our experimental manipulations and selection analysis in natural populations thus demonstrate stabilizing selection on the distance from the callus to the labellum tip, and illustrate the utility of employing multiple approaches to confirm selection exerted by pollinators on floral form.

Evolutionary history of a keystone pollinator parallels the biome occupancy of angiosperms in the Greater Cape Floristic Region

The Greater Cape Floristic Region (GCFR) in South Africa has been extensively investigated for its phenomenal angiosperm diversity. A key emergent pattern is the occurrence of older plant lineages in the southern Fynbos biome and younger lineages in the northern Succulent Karoo biome. We know practically nothing, however, about the evolutionary history of the animals that pollinate this often highly-specialized flora. In this study, we explore the evolutionary history of an important GCFR fly pollinator, Megapalpus capensis, and ask whether it exhibits broadly congruent genetic structuring and timing of diversification to flowering plants within these biomes. We find that the oldest M. capensis lineages originated in Fynbos during the Miocene, while younger Succulent Karoo lineages diverged in the Pliocene and correspond to the proposed age of this recent biome. A strong signature of population expansion is also recovered for flies in this arid biome, consistent with recent colonization. Our first investigation into the evolutionary history of GCFR pollinators thus supports a recent origin of the SK biome, as inferred from angiosperm phylogenies, and suggests that plants and pollinators may have co-diverged within this remarkable area.