Parallel chemical switches underlying pollinator isolation in A sian M itella

Floral scents are among the key signals used by pollinators to navigate to specific flowers. Thus, evolutionary changes in scents should have strong impacts on plant diversification, although scent-mediated plant speciation through pollinator shifts has rarely been demonstrated, despite being likely. To examine whether and how scent-mediated plant speciation may have occurred, we investigated the Asimitellaria plant lineage using multidisciplinary approaches including pollinator observations, chemical analyses of the floral scents, electroantennographic analyses and behavioural bioassays with the pollinators. We also performed phylogenetically independent contrast analyses of the pollinator/floral scent associations. First, we confirmed that the pairs of the sympatric, cross-fertile Asimitellaria species in three study sites consistently attract different pollinators, namely long-tongued and short-tongued fungus gnats. We also found that a stereoisomeric set of floral volatiles, the lilac aldehydes, could be responsible for the pollinator specificity. This is because the compounds consistently elicited responses in the antennae of the long-tongued fungus gnats and had contrasting effects on the two pollinators, that is triggering the nectaring behaviour of long-tongued fungus gnats while repelling short-tongued fungus gnats in a laboratory experiment. Moreover, we discovered that volatile composition repeatedly switched in Asimitellaria between species adapted to long-tongued and short-tongued fungus gnats. Collectively, our results support the idea that recurrent scent-mediated speciation has taken place in the Asimitellaria-fungus gnat system.