Floral trait differences between related species may play a key role in reproductive isolation imposed by pollinators. Volatile emissions can influence pollinator choice, but how they act in combination with traits such as flower orientation is rarely studied. We compared flower-opening patterns, morphology, colour, orientation and volatile emissions for two closely related species of Zaluzianskya and their natural hybrids. Hawkmoth pollinators were tested for preference between flowers of the two species, and between flowers with manipulations of volatiles or orientation. Flowers of Z. natalensis and Z. microsiphon open at night and day, respectively, but they overlap during early evening, when hawkmoths showed a strong preference for Z. natalensis. The species have similar flower size and colour, but Z. natalensis emits more floral volatiles in the evening and presents flowers vertically face-up as opposed to horizontally in Z. microsiphon, whereas natural hybrids are intermediate. Adding methyl benzoate and linalool to flowers of Z. microsiphon did not increase hawkmoth attraction, but re-orientation of flowers to face vertically increased attraction when scent cues were present, whereas re-orientation of Z. natalensis flowers to face horizontally decreased attraction. This study highlights the importance of flower orientation in imposing reproductive isolation.

Background and aims

Large clades of angiosperms are often characterized by diverse interactions with pollinators, but how these pollination systems are structured phylogenetically and biogeographically is still uncertain for most families. Apocynaceae is a clade of >5300 species with a worldwide distribution. A database representing >10 % of species in the family was used to explore the diversity of pollinators and evolutionary shifts in pollination systems across major clades and regions.

Methods

The database was compiled from published and unpublished reports. Plants were categorized into broad pollination systems and then subdivided to include bimodal systems. These were mapped against the five major divisions of the family, and against the smaller clades. Finally, pollination systems were mapped onto a phylogenetic reconstruction that included those species for which sequence data are available, and transition rates between pollination systems were calculated.

Key results

Most Apocynaceae are insect pollinated with few records of bird pollination. Almost three-quarters of species are pollinated by a single higher taxon (e.g. flies or moths); 7 % have bimodal pollination systems, whilst the remaining approx. 20 % are insect generalists. The less phenotypically specialized flowers of the Rauvolfioids are pollinated by a more restricted set of pollinators than are more complex flowers within the Apocynoids + Periplocoideae + Secamonoideae + Asclepiadoideae (APSA) clade. Certain combinations of bimodal pollination systems are more common than others. Some pollination systems are missing from particular regions, whilst others are over-represented.

Conclusions

Within Apocynaceae, interactions with pollinators are highly structured both phylogenetically and biogeographically. Variation in transition rates between pollination systems suggest constraints on their evolution, whereas regional differences point to environmental effects such as filtering of certain pollinators from habitats. This is the most extensive analysis of its type so far attempted and gives important insights into the diversity and evolution of pollination systems in large clades.