Diacetin, a reliable cue and private communication channel in a specialized pollination system.

The interaction between floral oil secreting plants and oil-collecting bees is one of the most specialized of all pollination mutualisms. Yet, the specific stimuli used by the bees to locate their host flowers have remained elusive. This study identifies diacetin, a volatile acetylated glycerol, as a floral signal compound shared by unrelated oil plants from around the globe. Electrophysiological measurements of antennae and behavioural assays identified diacetin as the key volatile used by oil-collecting bees to locate their host flowers. Furthermore, electrophysiological measurements indicate that only oil-collecting bees are capable of detecting diacetin. The structural and obvious biosynthetic similarity between diacetin and associated floral oils make it a reliable cue for oil-collecting bees. It is easily perceived by oil bees, but can't be detected by other potential pollinators. Therefore, diacetin represents the first demonstrated private communication channel in a pollination system.

Strong phylogenetic effects on floral scent variation of oil-secreting orchids in South Africa.

PREMISE OF THE STUDY: Evolution involves the interplay between natural selection and phylogenetic constraint. This is particularly evident among the flowering plants where form and diversity of flowers attest to the importance of both pollinator-mediated selection and phylogenetic constraint. Although this has been studied mostly using visible floral characters, invisible volatile chemicals emitted by the flowers should be subject to these same evolutionary forces. Unfortunately, most analyses of floral volatiles have over-emphasized the importance of natural selection and underplayed phylogenetic constraint without quantifying their respective roles in the evolution and composition of floral scents. METHODS: We used multivariate analyses to test the relative importance of pollinators vs. phylogeny in determining the composition of floral scents among oil-secreting orchids in southern Africa. Floral scents of 42 oil-secreting taxa/ecotypes distributed among 12 subclades in the tribe Diseae were sampled using headspace adsorption and gas chromatography-mass spectroscopy. KEY RESULTS: We identified 257 scent compounds distributed over nine different compound classes, with the majority of scents dominated by aliphatic or benzenoid compounds. The only significant predictor of floral scent among these orchids above the species level was phylogeny. Nevertheless, in two of the clades there were differences in scent profiles at the species and ecotype level that corresponded to different pollinators and were thus suggestive of pollinator-mediated selection. CONCLUSIONS: Scent variation was greater than expected and phylogeny was more important than pollinator-mediated selection in predicting the composition of floral scents of oil-secreting orchids, despite the specialized nature of the pollinator reward system.

Twin oil sacs facilitate the evolution of a novel type of pollination unit (meranthium) in a South African orchid.

The unique floral morphology of the South African orchid H. pulchra, with its twin meranthia, is best explained as an adaptation to pollination by oil-collecting bees. Flowers consisting of meranthia (floral parts that function as single pollination units; commonly observed in garden Iris) are extremely rare among the angiosperms and their significance poorly understood. Unlike all other known examples of meranthia, the novel type described for H. pulchra is not bilabiate. All Huttonaea species are unique in having twin petal sacs with glandular verrucae that secrete oil and are pollinated by Rediviva (Melittidae) oil-collecting bees. But only Huttonaea pulchra has long and widely divergent petal claws that place the oil sacs well beyond the reach of a centrally positioned bee. The wide separation of these sacs forces the pollinator, R. colorata, to visit each side of the flower independently and effectively divides the flower into two meranthia. Molecular data indicate that the evolution of the Huttonaea-type meranthium was dependent on the prior evolution of the oil flower/oil bee relationship. Meranthium evolution was also facilitated by the presence of oil in two separate structures (petal sacs) that were not physically constrained to remain in close proximity.

OIL FLOWERS AND OIL BEES: FURTHER EVIDENCE FOR POLLINATOR ADAPTATION.

We examined foreleg length and body size variation in two species of oil-collecting bees (Rediviva; Melittidae) in southern Africa. Oil-collecting bees harvest oil from host flowers by rubbing their forelegs against oil-secreting trichomes. Significant differences in foreleg length occur among populations of both species. Rediviva "pallidula" populations vary significantly in mean foreleg length (11.34 ± 0.42 mm to 12.67 ± 0.36 mm), but not in body length (10.59 ± 0.74 to 10.80 ± 0.64), and foreleg length and body size are not significantly correlated. Instead, foreleg variation appears to be a function of host plant spur length. Ninety-two percent of the variance in foreleg length of R. "pallidula" is explained by mean Diascia spur length. Rediviva rufocincta populations vary significantly in mean foreleg length (10.12 ± 0.70 mm to 12.34 ± 0.68 mm) and in body length (9.03 ± 0.26 mm to 10.56 ± 0.24 mm). Foreleg length scales allometrically with body size in this species as 90.5% of the variance in foreleg length can be explained as a function of body length. Body size appears to be constrained by the morphology of the oil-secreting host plant. Both bees collect floral oil with specially modified setae on the tarsi of their forelegs. The length of the disti- + mediotarsus (refered to here as "tarsus") in relation to the entire foreleg is shorter in R. rufocincta and does not increase as rapidly with increasing foreleg length as for R. "pallidula." These differences in variation can be attributed to differences in position of oil within the flowers of the respective host plants. Rediviva "pallidula" collects oil from Diascia species that have the oil deeply situated in narrow floral spurs of varying length, while R. rufocincta collects oil from the broadly saccate flowers of Bowkeria verticillata and B. citrina.