Floral scent evolution in the section Pseudophrys: pollinator-mediated selection or phylogenetic constraints?

Sexually deceptive orchid species from the Mediterranean genus Ophrys usually interact with one or a few pollinator species by means of specific floral scents. In this study, we investigated the respective role of pollinator-mediated selection and phylogenetic constraints in the evolution of floral scents in the section Pseudophrys. We built a phylogenetic tree of 19 Pseudophrys species based on three nuclear loci; we gathered a dataset on their pollination interactions from the literature and from our own field data; and we extracted and analysed their floral scents using solid phase microextraction and gas chromatography-mass spectrometry. We then quantified the phylogenetic signal carried by floral scents and investigated the link between plant-pollinator interactions and floral scent composition using phylogenetic comparative methods. We confirmed the monophyly of the section Pseudophrys and demonstrated the existence of three main clades within this section. We found that floral scent composition is affected by both phylogenetic relationships among Ophrys species and pollination interactions, with some compounds (especially fatty acid esters) carrying a significant phylogenetic signal and some (especially alkenes and alkadienes) generating dissimilarities between closely related Pseudophrys pollinated by different insects. Our results show that in the section Pseudophrys, floral scents are shaped both by pollinator-mediated selection and by phylogenetic constraints, but that the relative importance of these two evolutionary forces differ among compound classes, probably reflecting distinct selective pressures imposed upon behaviourally active and non-active compounds.